TRPV4 ANTAGONISTS

FIELD OF THE INVENTION

The present invention relates to diazabicyclo[2.2.1 ]hept-2-yl analogs,

pharmaceutical compositions containing them and their use as TRPV4 antagonists.

BACKGROUND OF THE INVENTION

TRPV4 is a member of the Transient Receptor Potential (TRP) superfamily of cation channels and is activated by heat, demonstrating spontaneous activity at physiological temperatures (Guler et al. 2002. J Neurosci 22: 6408-6414). Consistent with its polymodal activation property TRPV4 is also activated by hypotonicity and physical cell stress/pressure (Strotmann et al. 2000. Nat Cell Biol 2: 695-702), through a mechanism involving phospholipase A2 activation, arachidonic acid and epoxyeicosatrienoic acid generation (Vriens et al. 2004. Proc Natl Acad Sci U S A 101 : 396-401 ), In addition, amongst other mechanisms proposed, tyrosine kinase activity may also regulate TRPV4 (Wegierski et al. 2009. J Biol Chem. 284: 2923-33.

Heart failure results in the decreased ability of the left ventricle to pump blood into the peripheral circulation as indicated by a reduced ejection fraction and/or left ventricular dialation. This increases the left ventricular end diastolic pressure resulting in

enhancedpulmonary blood pressures. This places the septal barrier, which separates the circulatory aqueous environment and the alveolar airspaces of the lung, at risk. Increased pulmonary pressure results in the flow of fluid from the pulmonary circulation into the alveolar space resulting in lung edema/congestion, as is observed in patients with congestive heart failure.

TRPV4 is expressed in the lung (Delany et al. 2001. Physiol. Genomics 4: 165- 174) and has been shown to mediate Ca ²⁺ entry in isolated endothelial cells and in intact lungs (Jian et al. 2009 Am J Respir Cell Mol Biol 38: 386-92). Endothelial cells are responsible for forming the capillary vessels that mediate oxygen/carbon dioxide exchange and contribute to the septal barrier in the lung. Activation of TRPV4 channels results in contraction of endothelial cells in culture and cardiovascular collapse in vivo (Willette et al., 2008 J Pharmacol Exp Ther 325: 466-74), at least partially due to the enhanced filtration at the septal barrier evoking lung edema and hemorrage (Alvarez et al. 2006. Circ Res 99: 988-95). Indeed filtration at the septal barrier is increased in response to increased vascular and/or airway pressures and this response is dependent on the activity of TRPV4 channels (Jian et al. 2008 Am J Respir Cell Mol Biol 38: 386-92). Overall this suggests a clinical benefit of inhibiting TRPV4 function in the treatment of heart failure associated lung congestion.

Additional benefit is suggested in inhibiting TRPV4 function in pulmonary-based pathologies presenting with symptoms including lung edema/congestion, infection, inflammation, pulmonary remodeling and/or altered airway reactivity. A genetic link between TRPV4 and chronic obstructive pulmonary disorder (COPD) has recently been identified (Zhu et al., 2009. Hum Mol Genetics, 18: 2053-62) suggesting potential efficacy for TRPV4 modulation in treatment of COPD with or without coincident emphysema.

Enhanced TRPV4 activity is also a key driver in ventilator-induced lung injury (Hamanaka et al., 2007. Am J Physiol 293: L923-32) and it is suggested that TRPV4 activation may underlie pathologies involved in acute respiratory distress syndrome (ARDS), pulmonary fibrosis and asthma (Liedtke & Simon, 2004. Am J Physiol 287: 269-71 ) A potential clinical benefit for TRPV4 blockers in the treatment of sinusitis, as well as allergic and non-allergic rhinitis is also supported (Bhargave et al., 2008. Am J Rhinol 22:7-12).

In addition, TRPV4 channels have recently been implicated in urinary bladder function (Thorneloe et al., 2008. J Pharmacol Exp Ther 326 : 432-42) and are likely to provide therapeutic benefit for conditions of bladder overactivity, characterized by an increased urge to urinate and an enhancement of micturition frequency. These data suggest a clinically beneficial effect of inhibiting TRPV4, located on multiple cell types, on urinary bladder function that is likely to be effective in bladder disorders such as overactive bladder, interstitial cystitis and painful bladder syndrome.

Furthermore TRPV4 has in recent years been implicated in a number of other physiological/pathophysiological processes in which TRPV4 antagonists are likely to provide significant clinical benefit. These include various aspects of pain (Todaka et al. 2004. J Biol Chem 279: 35133-35138; Grant et al. 2007. J Physiol 578: 715-733;

Alessandri-Haber et al. 2006. J Neurosci 26: 3864-3874), genetic motor neuron disorders (Auer-Grumbach et al. 2009. Nat Genet. PMID: 20037588; Deng et al. 2009. Nat Genet PMID: 20037587; Landoure et al. 2009. Nat Genet PMID: 20037586), cardiovascular disease (Earley et al. 2005. Circ Res 97: 1270-9; Yang et al. 2006. Am. J Physiol.

290:L1267-L1276), and bone related disorders; including osteoarthritis (Muramatsu et al. 2007. J. Biol. Chem. 282: 32158-67) genetic gain-of function mutations (Krakow et al., 2009. Am J Hum Genet 84: 307-15; Rock et al., 2008 Nat Genet 40: 999-1003) and osteoclast differentiation (Masuyama et al. 2008. Cell Metab 8: 257-65). SUMMARY OF THE INVENTION

In one aspect this invention provides for diazabicyclo[2.2.1]hept-2-yl analogs, pharmaceutically acceptable salts thereof, and pharmaceutical compositions containing them.

In a second aspect, this invention provides for the use of the compounds of Formula (I) as TRPV4 antagonists.

In another aspect, this invention provides for the use of the compounds of Formula (I) for treating and preventing conditions associated with TRPV4 imbalance.

In yet another aspect, this invention provides for the use of the compounds of Formula (I) for the treatment or prevention of atherosclerosis, disorders related to intestinal edema, post-surgical abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone related dysfunctions and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator induced lung injury, high altitude induced pulmonary edema, acute respiratory distress syndrome, pulmonary fibrosis, sinusitis/rhinitis, asthma, overactive bladder, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction and osteoarthritis.

The TRPV4 antagonist may be administered alone or in conjunction with one or more other therapeutic agents, eg. agents being selected from the group consisting of endothelin receptor antagonists, angiotensin converting enzyme (ACE) inhibitors, angiotension II receptor antagonists, vasopeptidase inhibitors, vasopressin receptor modulators, diuretics, digoxin, beta blocker, aldosterone antagonists, iontropes, NSAIDS, nitric oxide donors, calcium channel modulators, muscarinic antagonists, steroidal antiinflammatory drugs, bronchodilators, anti-histamines, leukotriene antagonist, HMG-CoA reductase inhibitors, dual non-selective β-adrenoceptor and a-| -adrenoceptor antagonists, type-5 phosphodiesterase inhibitors, and renin inhibitors.

Other aspects and advantages of the present invention are described further in the following detailed description of the preferred embodiments thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides for compounds of Formula (I):

wherein: R-ι is C _1-3 alkyl, C _1-3 alkoxy, CF ₃ , halo, S0 ₂ C _1-3 alkyl, N(R ₄ ) ₂ , OCF ₃ , or CN;

R ₂ is C _1-4 alkyl, -CH ₂ C _3-6 cycloalkyl, or -CH ₂ -phenyl;

R ₃ is

all of which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OCi _-5 alkyl, Ci _-5 alkyl, OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C(0)NR ₄ C _1-5 alkyl, C(0)NHphenyl, C(0)R ₆ , C0 ₂ C _1-3 alkyl, S0 ₂ C _1-3 alkyl, S0 ₂ phenyl, S0 ₂ NR ₄ C _1-5 alkyl, -O-phenyl, phenyl, morpholinyl, pyrimidinyl, tetrahydropyranyl, pyridizinyl, oxazolyl, pyrazinyl, pyrrolyl, tetrazolyl, oxadiazolyl, triazolyl, dihydropyranyl, C _3-6 cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl;

wherein the OCi _-5 alkyl, d _-5 alkyl, C(0)NR ₄ Ci _-5 alkyl, S0 ₂ NR ₄ Ci _-5 alkyl, phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, tetrahydropyranyl, pyridizinyl, pyrazinyl, oxazolyl, pyrollyl, C _{3- 6} cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , morpholinyl, piperidinyl, pyrollidinyl, piperazinyl, tetrazolyl, C0 ₂ Ci _-4 alkyl, S0 ₂ NHCi _-3 alkyl, C(0)N(R ₄ ) ₂ , NHS0 ₂ Ci _-3 alkyl and S0 ₂ Ci _-3 alkyl;

G is *

S,S

which is substituted by one or two substituients selected from the group consisting of: Ci _-2 alkyl, OH, =0, or -CH ₂ OH;

X is a bond or CH ₂ ; Y is NR ₄ or S;

R ₄ is independently H or Cr ₃ alkyl;

R ₅ is hydrogen or d- ₅ alkyl;

R is

i is 0, 1 , 2, or 3;

or a pharmaceutically acceptable salt thereof.

"Alkyl" refers to a monovalent saturated hydrocarbon chain having the specified number of member atoms. For example, C _1-4 alkyl refers to an alkyl group having from 1 to 4 member atoms. Alkyl groups may be straight or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), and butyl (n-butyl, isobutyl, and t-butyl).

"Cycloalkyl" refers to a monovalent saturated or unsaturated hydrocarbon ring having the specified number of member atoms. For example, C ₃ - ₆ cycloalkyl refers to a cycloalkyl group having from 3 to 6 member atoms. Unsaturated cycloalkyl groups have one or more carbon-carbon double bonds within the ring. Cycloalkyl groups are not aromatic. Cycloalkyl includes cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, and cyclohexenyl.

"Alkoxy" as used herein refers to an -0-Ci _-3 alkyl group wherein Ci _-3 alkyl is as defined herein. Examples of such groups include methoxy, ethoxy, propoxy, and the like.

When used herein, the terms 'halogen' and 'halo' mean fluorine, chlorine, bromine and iodine, and fluoro, chloro, bromo, and iodo, respectively.

"Substituted" in reference to a group indicates that one or more hydrogen atom attached to a member atom within the group is replaced with a substituent selected from the group of defined substituents. It should be understood that the term "substituted" includes the implicit provision that such substitution be in accordance with the permitted valence of the substituted atom and the substituent and that the substitution results in a stable compound (i.e. one that does not spontaneously undergo transformation such as by rearrangement, cyclization, or elimination and that is sufficiently robust to survive isolation from a reaction mixture). When it is stated that a group may contain one or more substituents, one or more (as appropriate) member atoms within the group may be substituted. In addition, a single member atom within the group may be substituted with more than one substituent as long as such substitution is in accordance with the permitted valence of the atom. Suitable substituents are defined herein for each substituted or optionally substituted group. With regard to stereoisomers, the compounds of Formula (I) may have one or more asymmetric carbon atom and may occur as racemates, racemic mixtures and as individual enantiomers or diastereomers. All such isomeric forms are included within the present invention, including mixtures thereof.

As used herein, "pharmaceutically acceptable" refers to those compounds, materials, compositions, and dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The skilled artisan will appreciate that pharmaceutically acceptable salts of the compounds according to Formula (I) may be prepared. These pharmaceutically acceptable salts may be prepared in situ during the final isolation and purification of the compound, or by separately treating the purified compound in its free acid or free base form with a suitable base or acid, respectively.

In certain embodiments, compounds according to Formula (I) may contain an acidic functional group and are, therefore, capable of forming pharmaceutically acceptable base addition salts by treatment with a suitable base. Examples of such bases include a) hydroxides, carbonates, and bicarbonates of sodium, potassium, lithium, calcium, magnesium, aluminium, and zinc; and b) primary, secondary, and tertiary amines including aliphatic amines, aromatic amines, aliphatic diamines, and hydroxy alkylamines such as methylamine, ethylamine, 2-hydroxyethylamine, diethylamine, triethylamine, ethylenediamine, ethanolamine, diethanolamine, and cyclohexylamine.

In certain embodiments, compounds according to Formula (I) may contain a basic functional group and are therefore capable of forming pharmaceutically acceptable acid addition salts by treatment with a suitable acid. Suitable acids include pharmaceutically acceptable inorganic acids and organic acids. Representative pharmaceutically acceptable acids include hydrogen chloride, hydrogen bromide, nitric acid, sulfuric acid, sulfonic acid, phosphoric acid, acetic acid, hydroxyacetic acid, phenylacetic acid, propionic acid, butyric acid, valeric acid, maleic acid, acrylic acid, fumaric acid, malic acid, malonic acid, tartaric acid, citric acid, salicylic acid, benzoic acid, tannic acid, formic acid, stearic acid, lactic acid, ascorbic acid, methylsulfonic acid, p-toluenesulfonic acid, oleic acid, lauric acid, and the like.

As used herein, the term "a compound of Formula (I)" or "the compound of Formula (I)" refers to one or more compounds according to Formula (I). The compound of Formula (I) may exist in solid or liquid form. In the solid state, it may exist in crystalline or noncrystalline form, or as a mixture thereof. The skilled artisan will appreciate that pharmaceutically acceptable solvates may be formed for crystalline compounds wherein solvent molecules are incorporated into the crystalline lattice during crystallization.

Solvates may involve non-aqueous solvents such as, but not limited to, ethanol, isopropanol, DMSO, acetic acid, ethanolamine, or ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes all such solvates.

The skilled artisan will further appreciate that certain compounds of the invention that exist in crystalline form, including the various solvates thereof, may exhibit polymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs." The invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. The skilled artisan will appreciate that different polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.

The subject invention also includes isotopically-labelled compounds, which are identical to those recited in formula (I) and following, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention and pharmaceutically acceptable salts thereof include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulphur, fluorine, iodine, and chlorine, such as 2H, 3H, 1 1 C, 13C, 14C, 15N, 170, 180, 31 P, 32P, 35S, 18F, 36CI, 1231 and 1251.

Compounds of the present invention and pharmaceutically acceptable salts of said compounds that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as 3H, 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. 1 1 C and 18F isotopes are particularly useful in PET

(positron emission tomography), and 1251 isotopes are particularly useful in SPECT (single photon emission computerized tomography), all useful in brain imaging. Further, substitution with heavier isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labelled compounds of formula I and following of this invention can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labelled reagent for a non-isotopically labelled reagent.

Representative Embodiments

In one embodiment:

Ri is Ci-3 alkyl, d _-3 alkoxy, CF ₃ , halo, S0 ₂ Ci _-3 alkyl, N(R ₄ ) ₂ , OCF ₃ , or CN;

R ₂ is Ci _-4 alkyl, -CH ₂ C ₃ - ₆ cycloalkyl, or -CH ₂ -phenyl;

R ₃ is

all of which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OCi _-5 alkyl, Ci _-5 alkyl, OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C(0)NR ₄ Ci _-5 alkyl, C(0)NHphenyl, C(0)R ₆ , C0 ₂ Ci _-3 alkyl, S0 ₂ Ci _-3 alkyl, S0 ₂ phenyl, S0 ₂ NR ₄ Ci _-5 alkyl, -O-phenyl, phenyl, morpholinyl, pyrimidinyl, tetrahydropyranyl, pyridizinyl, oxazolyl, pyrazinyl, pyrrolyl, tetrazolyl, oxadiazolyl, triazolyl, dihydropyranyl, C _3-6 cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl;

wherein the OCi _-5 alkyl, Ci _-5 alkyl, C(0)NR ₄ Ci _-5 alkyl, S0 ₂ NR ₄ Ci _-5 alkyl, phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, tetrahydropyranyl, pyridizinyl, pyrazinyl, oxazolyl, pyrollyl, C _3-6 cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , morpholinyl, piperidinyl, pyrollidinyl, piperazinyl, tetrazolyl, C0 ₂ C _1-4 alkyl, S0 ₂ NHC _1-3 alkyl, C(0)N(R ₄ ) ₂ , NHS0 ₂ C _1-3 alkyl and

S0 ₂ C _1-3 alkyl;

G is *

S,S

which is substituted by one or two substituients selected from the group consisting of: Ci _-2 alkyl, OH, =0, or -CH ₂ OH;

X is a bond or CH ₂ ;

Y is NR ₄ or S;

R ₄ is independently H or C1-3 alkyl;

R ₅ is hydrogen or C1-5 alkyl;

R is independently

i is 0, 1 , 2, or 3.

In another embodiment:

R-i is C _1-3 alkyl, C _1-3 alkoxy, CF ₃ , halo, OCF ₃ , or CN;

R ₂ is C _1-4 alkyl, -CH ₂ C _3-6 cycloalkyl, or -CH ₂ -phenyl;

R is:

all of which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C0 ₂ Ci _-3 alkyl, S0 ₂ Ci _-3 alkyl, S0 ₂ phenyl, phenyl and pyridyl; wherein the phenyl or pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N (R ₄ ) ₂ , C0 ₂ C _1-3 alkyl, and S0 ₂ C _1-3 alkyl;

G is *

S,S

which is substituted by one or two substituents selected from the group consisting of: Ci _-2 alkyl, OH, =0, or -CH ₂ OH;

X is a bond or CH ₂ ;

Y is NR ₄ or S;

R ₄ is independently H or Ci- ₃ alkyl;

R ₅ is hydrogen;

R is independently

i is 0, 1 , 2, or 3.

In another embodiment:

R-i is CF ₃ , halo, OCF ₃ , or CN;

R ₂ is C _1-4 alkyl;

all of which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C0 ₂ Ci _-3 alkyl, S0 ₂ Ci _-3 alkyl, S0 ₂ phenyl, -O-phenyl, phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, C _3-6 cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl;

wherein the phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, C _3-6 cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C0 ₂ C _1-3 alkyl, and S0 ₂ C _1-3 alkyl;

G is substituted by C _1-2 alkyl,

X is a bond;

Y is NR ₄ or S;

R ₄ is independently C ₃ alkyl;

R ₅ is hydrogen;

R is independently

i is 0, 1 , or 2.

In another embodiment:

R-i is CF ₃ , halo, OCF ₃ , or CN;

R ₂ is C _1-4 alkyl;

R ₃ is:

all of which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C0 ₂ C _1-3 alkyl, S0 ₂ C _1-3 alkyl, S0 ₂ phenyl, -O-phenyl, phenyl, and pyridyl;

wherein the phenyl and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , C0 ₂ C _1-3 alkyl, and S0 ₂ C _1-3 alkyl;

G is substituted by methyl;

X is a bond;

Y is NR ₄ or S;

R ₄ is independently C ₃ alkyl;

R ₅ is hydrogen or Ci- ₃ alkyl;

R ₆ is independently

i is 0, 1 , or 2.

In another embodiment:

Ri is Ci-3 alkyl, d _-3 alkoxy, CF ₃ , halo, S0 ₂ Ci _-3 alkyl, N(R ₄ ) ₂ , OCF ₃ , or CN;

R ₂ is Ci _-4 alkyl, -CH ₂ C ₃ - ₆ cycloalkyl, or -CH ₂ -phenyl;

which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OCi _-5 alkyl, Ci _-5 alkyl, OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ ,

C(0)NR ₄ Ci _-5 alkyl, C(0)NHphenyl, C(0)R ₆ , C0 ₂ Ci _-3 alkyl, S0 ₂ Ci _-3 alkyl, S0 ₂ phenyl, S0 ₂ NR ₄ Ci _-5 alkyl, -O-phenyl, phenyl, morpholinyl, pyrimidinyl, tetrahydropyranyl, pyridizinyl, oxazolyl, pyrazinyl, pyrrolyl, tetrazolyl, oxadiazolyl, triazolyl, dihydropyranyl, C _{3- 6} cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl;

wherein the OCi _-5 alkyl, Ci _-5 alkyl, C(0)NR ₄ Ci _-5 alkyl, S0 ₂ NR ₄ Ci _-5 alkyl, phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, tetrahydropyranyl, pyridizinyl, pyrazinyl, oxazolyl, pyrollyl, C _{3- 6} cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , morpholinyl, piperidinyl, pyrollidinyl, piperazinyl, tetrazolyl, C0 ₂ Ci _-4 alkyl, S0 ₂ NHCi _-3 alkyl, C(0)N(R ₄ ) ₂ , NHS0 ₂ Ci _-3 alkyl and S0 ₂ Ci _-3 alkyl;

G is substituted by methyl;

X is a bond or CH ₂ ;

Y is NR ₄ or S;

R ₄ is independently H or Cr ₃ alkyl;

R ₅ is hydrogen or d- ₅ alkyl;

R is independently

i is 0, 1 , 2, or 3.

In another embodiment:

Ri is Ci _-3 alkyl, Ci _-3 alkoxy, CF ₃ , halo, or CN;

R ₂ is Ci _-4 alkyl;

which may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OCi _-5 alkyl, Ci _-5 alkyl, OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ ,

C(0)NR ₄ Ci _-5 alkyl, C(0)NHphenyl, C(0)R ₆ , C0 ₂ Ci _-3 alkyl, S0 ₂ Ci _-3 alkyl, S0 ₂ phenyl, S0 ₂ NR ₄ Ci _-5 alkyl, -O-phenyl, phenyl, morpholinyl, pyrimidinyl, tetrahydropyranyl, pyridizinyl, oxazolyl, pyrazinyl, pyrrolyl, tetrazolyl, oxadiazolyl, triazolyl, dihydropyranyl, C _{3- 6} cycloalkyl, cyclohexenyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl;

wherein the phenyl, morpholinyl, pyrimidinyl, dihydropyranyl, tetrahydropyranyl,

C _3-6 cycloalkyl, piperazinyl, pyrrolidinyl, piperadinyl, and pyridyl may be unsubstituted or substituted with one, two, or three substituents selected from the group consisting of: OR ₄ , R ₄ , OCF ₃ , halo, CF ₃ , CN, N(R ₄ ) ₂ , morpholinyl, piperidinyl, pyrollidinyl, piperazinyl, tetrazolyl, C0 ₂ C _1-4 alkyl, S0 ₂ NHC _1-3 alkyl, C(0)N(R ₄ ) ₂ , NHS0 ₂ C _1-3 alkyl and S0 ₂ C _1-3 alkyl; G is substituted by methyl;

X is a bond;

Y is NR ₄ or S;

R ₄ is independently H or Ci- ₃ alkyl;

R ₅ is hydrogen or C1-5 alkyl;

R is independently

i is 0, 1 , 2, or 3.

It is to be understood that the present invention covers all combinations of particular groups described hereinabove.

Specific examples of compounds of the present invention include the following: A/-[(1 S)-2,2-dimethyl-1-({(3S)-3-methyl-5-[(5-phenyl-2-pyridinyl)c arbonyl]-2,5- diazabicyclo[2.2.1]hept-2-yl}carbonyl)propyl]-1 /-/-indole-2-carboxamide;

N-((S)-3,3-dimethyl-1 -((1 S,4S,7R)-7-methyl-5-(5-phenylpicolinoyl)-2,5- diazabicyclo[2.2.1]heptan-2-yl)-1 -oxobutan-2-yl)-1 H-indole-2-carboxamide; and

A/-[(1 S)-2,2-dimethyl-1-({(1 S,4S,6S)-6-methyl-5-[(5-phenyl-2-pyridinyl)carbonyl]-2,5- diazabicyclo[2.2.1]hept-2-yl}carbonyl)propyl]-1 /-/-indole-2-carboxamide;

or a pharmaceutically acceptable salt thereof. Compound Preparation

The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999). In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.

The synthesis of the compounds of the general formula (I) and pharmaceutically acceptable derivatives and salts thereof may be accomplished as outlined below in Schemes 1 - 4. In the following description, the groups are as defined above for compounds of formula (I) unless otherwise indicated. Starting materials are commercially available or are made from commercially available starting materials using methods known to those skilled in the art.

Scheme 1

The free secondary amine of appropriately substituted A/-Boc-(1 S,4S)-2,5- diazabicyclo[2.2.1]heptane can be coupled to an appropriate carboxylic acid 1 under conditions common to the art such as EDC in the presence of a base such as N- methylmorpholine or triethylamine, a coupling modifier such as HOBt to provide the amide intermediate 2. Subsequent CBz deprotection under standard conditions such as by treatment with Pd/C with ammonium formate provides the amine intermediate 3.

Treatment of intermediate 3 with an appropriate carboxylic acid under conditions common to the art such as EDC in the presence of a base such as /V-methylmorpholine and a coupling modifier such as HOBt provides the amide intermediate 4. Subsequent Boc deprotection can be accomplished under conditions common to the art such as treatment with an acid such as hydrochloric acid in 1 ,4-dioxane and methanol or TFA in

dichloromethane to provide intermediate 5. Treatment of intermediate 5 with an appropriate carboxylic acid under conditions common to the art such as EDC in the presence of a base and a coupling modifier provides the compound of Formula (I).

Scheme 2

(I)

The free amine of appropriately substituted /V-Boc-(1 S,4S)-2,5-diazabicyclo[2.2.1]heptane can be coupled to an appropriate carboxylic acid under conditions common to the art such as EDC in the presence of a base such as /V-methylmorpholine or triethylamine, a coupling modifier such as HOBt to provide the amide intermediate 6. Subsequent Boc deprotection can be accomplished under conditions common to the art such as treatment with an acid such as hydrochloric acid in 1 ,4-dioxane and methanol or TFA in

dichloromethane to provide intermediate 7. Treatment of intermediate 7 with an appropriate carboxylic acid under conditions common to the art such as EDC in the presence of a base such as /V-methylmorpholine and a coupling modifier such as HOBt provides the amide intermediate 8. Subsequent CBz deprotection under standard conditions such as by treatment with Pd/C with ammonium formate provides the amine intermediate 9. Treatment of intermediate 9 with an appropriate carboxylic acid under conditions common to the art such as EDC in the presence of a base and a coupling modifier provides the compound of Formula (I).

Scheme 3

The free alcohol of a hydroxy proline derivative can be protected as a silyl ether with chloro-ferf-butyldimethylsilane to yield intermediate 10. Subsequent hydrolysis of the methyl ester and acylation with 3,5-dimethyl-1 H-pyrazole yields amide 11. Reduction of intermediate 11 and subsequent Grignard reaction with methyl magnesium bromide yields intermediate 12. Deprotection with TBAF provides free alcohol which can then be mesylated with methanesulfonyl chloride to yield intermediate 13. Cyclization of intermediate 13 with benzylamine provides intermediate 14. Deprotection of the tosyl amine with HBr and subsequent protection as a tert-butyl carbamate yields intermediate 15. Finally, deprotection of intermediate 15 under hydrolysis conditions provides intermediate 16. Scheme 4

22

Trans-4-hydroxy-L-proline can be converted to the ethyl ester via an acid chloride intermediate and then can be protected as benzyl carbamate to yield intermediate 17. Subsequent protection of the alcohol of intermediate 17 as a mesylate, displacement of mesylate with phenylselane, and and subsequent oxidation/elimination provides dihydropyrrole intermediate 18. Epoxidation of intermediate 18 follows by methyl cuprate addition resulting in a mixture of diastereomers which can be deprotected under hydrogenation conditions to yield the amine intermediate 19. The secondary amine of the mixture of diastereomers in intermedidate 19 can be protected as a tosyl group. Methyl ester can then be reduced to alcohol using lithium aluminum hydride..Subsequent protection of both alcohols as a mesylate yields intermediate 20. Cyclization of intermediate 20 with benzylamine and subsequent ferf-butyl carbamate protection provides intermediate 21. Finally, deprotection of intermediate 21 under hydrogenation conditions yields intermediate 22.

Biological Activity

As stated above, the compounds according to Formula I are TRPV4 antagonists, and are useful in the treatment or prevention of atherosclerosis, disorders related to intestinal edema, post-surgical abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone related dysfunctions and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator induced lung injury, high altitude induced pulmonary edema, acute respiratory distress syndrome, pulmonary fibrosis, sinusitis/rhinitis, asthma, overactive bladder, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction and osteoarthritis.

The biological activity of the compounds according to Formula I can be determined using any suitable assay for determining the activity of a candidate compound as a TRPV4 antagonist, as well as tissue and in vivo models.

The biological activity of the compounds of Formula (I) are demonstrated by the following tests.

Liqand-qated assay:

TRP channel activation/opening results in an influx of divalent and monovalent cations including calcium. The resulting changes in intracellular calcium are monitored using a calcium selective fluorescent dye Fluo4 (MDS Analytical Technologies). Dye loaded cells are initially exposed to test compound to verify a lack of agonist activity. Cells are subsequently activated by addition of an agonist and inhibition of the agonist-induced activation is recorded. Human embryonic kidney 293 cells stably expressing the macrophage scavenger receptor class II (HEK-293-MSR-II) and transduced with 1 % BacMam (J. P. Condreay, S.M. Witherspoon, W.C. Clay and T.A. Kost, Proc Natl Acad Sci 96 (1999), pp. 127-132) virus expressing the human TRPV4 gene are plated at 15000 cells/well in a volume of 50 uL in a 384 well poly-D lysine coated plate. Cells are incubated for 24 hours at 37 degrees and 5% C0 ₂ . Media is then aspirated using a Tecan Plate- washer and replaced with 20 uL of dye loading buffer: HBSS, 500 uM Brilliant Black (MDS Analytical Technologies), 2 uM Fluo-4. Dye loaded plates are then incubated in the dark at room temperature for 1 -1 .5 hours. 10 uL of test compound diluted in HBSS + 0.01 % Chaps is added to the plate, incubated for 10 min at room temperature in the dark and then 10 uL of agonist is added at a final cone, equal to the agonist EC80. Calcium release is measured using the FLIPRtetra (MDS Analytical Technologies).

All examples described herein possessed TRPV4 biological activity with IC ₅₀ s ranges from .5 nM - 100 nM.

Hypotonicity assay (BHK cells):

BHK/AC9_DMEM/F12 conditioned (Baby Hamster Kidney) cells are transduced with 2% BacMam virus expressing the human TRPV4 gene and are plated at 10K cells per well in a volume of 50uL in 384 well poly-D-lysine coated plates. Cells are incubated for 18-24 hours at 37 degrees and 5% C0 _2. The following day, the media is aspirated using a Tecan Plate-washer and replaced with 20uL of dye loading buffer: HBSS buffer, 2.5mM Probenecid, 500 uM Brilliant Black, 2 uM Fluo-4. The dye loaded cells are incubated for 1-1 .5 hours at room temperature in the dark. 10 uL of test compound diluted in HBSS/H ₂ 0 (-1 :2.3) + 0.01 % Chaps is added to the plate, incubated for 10 min at room temperature in the dark, and then 10uL of hypotonic buffer (H ₂ 0 + 1 .5mM CaCI ₂ + ~68mM NaCI; 140mOsm stock/260mOsm FAC) is used to test the inhibition of the hypotonicity- induced activation. Reaction is measured on a heated stage (37 degrees) using the FLIPRtetra. (range 1 nM to 100 nM)

Methods of Use

The compounds of the invention are TRPV4 antagonists, and are useful in the treatment or prevention of atherosclerosis, disorders related to atherosclerosis, disorders related to intestinal edema, post-surgical abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone related dysfunctions and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator induced lung injury, high altitude induced pulmonary edema, acute respiratory distress syndrome, pulmonary fibrosis, sinusitis/rhinitis, asthma, overactive bladder, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction and osteoarthritis. Accordingly, in another aspect the invention is directed to methods of treating such conditions.

The methods of treatment of the invention comprise administering a safe and effective amount of a compound according to Formula I or a pharmaceutically-acceptable salt thereof to a patient in need thereof.

As used herein, "treat" in reference to a condition means: (1 ) to ameliorate or prevent the condition or one or more of the biological manifestations of the condition, (2) to interfere with (a) one or more points in the biological cascade that leads to or is responsible for the condition or (b) one or more of the biological manifestations of the condition, (3) to alleviate one or more of the symptoms or effects associated with the condition, or (4) to slow the progression of the condition or one or more of the biological manifestations of the condition.

As indicated above, "treatment" of a condition includes prevention of the condition. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.

As used herein, "safe and effective amount" in reference to a compound of the invention or other pharmaceutically-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. A safe and effective amount of a compound will vary with the particular compound chosen (e.g.

consider the potency, efficacy, and half-life of the compound); the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.

As used herein, "patient" refers to a human or other animal.

The compounds of the invention may be administered by any suitable route of administration, including both systemic administration and topical administration.

Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation.

Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. Topical administration includes application to the skin as well as intraocular, otic, intravaginal, and intranasal administration.

The compounds of the invention may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change.

Typical daily dosages may vary depending upon the particular route of

administration chosen. Typical dosages for oral administration range from 1 mg to 1000 mg per person per dose. Additionally, the compounds of the invention may be administered as prodrugs. As used herein, a "prodrug" of a compound of the invention is a functional derivative of the compound which, upon administration to a patient, eventually liberates the compound of the invention in vivo. Administration of a compound of the invention as a prodrug may enable the skilled artisan to do one or more of the following: (a) modify the onset of the compound in vivo; (b) modify the duration of action of the compound in vivo; (C) modify the transportation or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome or overcome a side effect or other difficulty encountered with the compound. Typical functional derivatives used to prepare prodrugs include modifications of the compound that are chemically or enzymatically cleaved in vivo. Such modifications, which include the preparation of phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art.

Compositions

The compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient. Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of the invention and one or more pharmaceutically-acceptable excipient.

The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be extracted and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a safe and effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from 1 mg to 1000 mg.

The pharmaceutical compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. For example, in certain embodiments the pharmaceutical compositions of the invention contain two compounds of the invention. In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional pharmaceutically active compounds.

As used herein, "pharmaceutically-acceptable excipient" means a

pharmaceutically acceptable material, composition or vehicle involved in giving form or consistency to the pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of the invention when administered to a patient and interactions which would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided. In addition, each excipient must of course be of sufficiently high purity to render it pharmaceutically-acceptable.

The compound of the invention and the pharmaceutically-acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixers, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as dry powders, aerosols, suspensions, and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

Suitable pharmaceutically-acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically-acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically- acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceutically-acceptable excipients may be chosen for their ability to enhance patient compliance.

Suitable pharmaceutically-acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweetners, flavoring agents, flavor masking agents, coloring agents, anticaking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutically-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically-acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically-acceptable excipients and may be useful in selecting suitable pharmaceutically-acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing

Company).

In one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesuim stearate, calcium stearate, and talc.

The compounds may be administered alone or in conjunction with one or more other therapeutic agents, said agents being selected from the group consisting of endothelin receptor antagonists, angiotensin converting enzyme (ACE) inhibitors, angiotension II receptor antagonists, vasopeptidase inhibitors, vasopressin receptor modulators, diuretics, digoxin, beta blocker, aldosterone antagonists, iontropes, NSAIDS, nitric oxide donors, calcium channel modulators, muscarinic antagonists, steroidal antiinflammatory drugs, bronchodilators, anti-histamines, leukotriene antagonist, HMG-CoA reductase inhibitors, dual non-selective β-adrenoceptor and a-| -adrenoceptor antagonists, type-5 phosphodiesterase inhibitors, and renin inhibitors.

EXAMPLES

The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention. While particular embodiments of the present invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention.

In the Examples:

Chemical shifts are expressed in parts per million (ppm) units. Coupling constants (J) are in units of hertz (Hz). Splitting patterns describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), dd (double doublet), dt (double triplet), m (multiplet), br (broad).

Flash column chromatography was performed on silica gel.

The naming program used is ACD Name Pro 6.02 or ChemDraw.

The following abbreviations and terms had the indicated meanings throughout:

AcOH acetic acid

Aq aqueous

Boc ₂ 0 di-tert-butyl dicarbonate

BnNH ₂ benzyl amine

Brine saturated aqueous sodium chloride

CBzCI benzyl chloroformate

CH ₂ CI ₂ methylene chloride

CHCI ₃ chloroform

DCC dicyclohexylcarbodiimide

DCM methylene chloride

DMF /V,/V-dimethylformamide

DMSO dimethylsulfoxide

EA ethyl acetate

EDC 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride

Et ethyl

EtOH ethanol

Et ₂ 0 diethyl ether

EtOAc ethyl acetate

h, hr hour

HBr hydrogen bromide

HCI hydrochloric acid

H ₂ 0 ₂ hydrogen peroxide

HOBt 1 -hydroxybenzotriazole

Imid imidazole

UAIH4 lithium aluminum hydride K ₂ C0 ₃ potassium carbonate

KOH potassium hydroxide

LCMS liquid chromatography-mass spectroscopy

m-CPBA mefa-chloroperoxybenzoic acid

Me methyl

MeMgBr methyl magnesium bromide

MeOH or CH30H methanol

MgS04 magnesium sulfate

Min minute

MsCI methanesulfonyl chloride

MS mass spectrum

NaHC03 sodium bicarbonate

NaOH sodium hydroxide

Na2S04 sodium sulfate

NH4CI ammonium chloride

NMM N-methylmorpholine

Pd/C palladium on carbon

Pd(dppf)CI2 1 ,1 '-bis(diphenylphosphino)ferrocene] dichloropalladi

PE petroleum ether

Ph phenyl

rt room temperature

satd saturated

SOCI2 thionyl chloride

TBAF tetrabutyl ammonium fluoride

TBDPSCI tert-butylchlorodimethylsilane

TEA triethylamine

TFA trifluoroacetic acid

THF tetrahydrofuran

TsCI tosyl chloride Example 1

N-\( 1 SV2.2-dimethyl-1 - (3SV3-methyl-5-r(5-Dhenyl-2-Dyridinvncarbonyll-2.5- diazabicvclo[2.2.1 lhept-2-yl}carbonyl)propyll-1 /-/-indole-2-carboxamide

TBDPSO

4-(te t-Butyl-diphenyl-silanyloxy)-1 -(toluene-4-sulfonyl)-pyrrolidine-2-carboxylic acid methyl ester

To a solution of 4-hydroxy-1 -(toluene-4-sulfonyl)-pyrrolidine-2-carboxylic acid methyl ester (69 g, 230 mmol) and imidazole (35.9 g, 529 mmol) in DMF (300 mL) was added

TBDPSCI (75.8 g, 277 mmol). The mixture was stirred at rt. for 18 h, followed by 500 mL of H ₂ 0 added to the mixture. The solution was extracted with EA (3 x 500 mL), washed with H ₂ 0 (3 x 500 mL), brine (500 mL), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product which was purified by silica gel column (PE:EA=8:1 ) to yield 124 g of product. LCMS (m/z): 460.1 (M+H).

TBDPS

4-(terf-Butyl-diphenyl-silanyloxy)-1 -(toluene-4-sulfonyl)-pyrrolidine-2-carboxylic acid

To an ice-cold solution of 4-(fe/t-butyl-diphenyl-silanyloxy)-1 -(toluene-4-sulfonyl) - pyrrolidine-2-carboxylic acid methyl ester (124 g, 231 mmol) in EtOH (600 mL) was added 50 mL of 2N aqueous KOH. The mixture was stirred at rt for 1 .5 h. The mixture was cooled in an ice bath and acidified with 2N aqueous HCI, (pH ~5). The mixture was concentrated to remove the organic solvent, extracted with EA (3 x 200 mL), washed with H ₂ 0 (3 x 500 mL), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product (121 g), which was directly used without further purification. LCMS (m/z): 446.1 (M+H).

r4-(te t-Butyl-diphenyl-silanyloxy)-1-(toluene-4-sulfonyl)-pyrrolid in-2-yll- pyrazol-1 -vD-methanone

To a solution of 4-(fe/t-butyl-diphenyl-silanyloxy)-1-(toluene-4-sulfonyl) -pyrrolidine-2- carboxylic acid (121 g, 231 mmol) and 3,5-dimethyl-1 H-pyrazole (22.2 g, 231 mmol) in CHCIs (600 mL) was added DCC (47.66 g, 231 mmol) at 10 °C. The mixture was stirred at rt for 18 h. The mixture was filtered and concentrated to give the crude product which was purified by silica gel column (PE: EA=10:1 ) to yield 87.7 g. LCMS (m/z): 602.0 (M+H).

TBDPS

4-(terf-Butyl-diphenyl-silanyloxy)-1-(toluene-4-sulfonyl)-py rrolidine-2-carbaldehvde To a solution of LiAIH ₄ (8.32 g, 219 mmol) in THF (600 mL) was added dropwise [4-(ferf- butyl-diphenyl-silanyloxy)-1 -(toluene-4-sulfonyl)-pyrrolidin-2-yl]-(3,5-dimethyl-pyrazol -1-yl)- methanone (87.7 g, 146 mmol) in THF (200 mL) at -40°C. The mixture was stirred at -40°C for 0.5 h then 219 mL of 1 N aqueous HCI was added to the mixture between -40°C and -30°C and after 15 mins the insoluble material was filtered off. The filtrate was concentrated to dryness, 600 mL of EA was added to the residue, washed with H ₂ 0 (800 mL), dried over Na ₂ S0 ₄ and the mixture was filtered and concentrated to give the crude product which was purified by silica gel column (PE: EA=10:1 ) to yield 54 g. LCMS (m/z): 432.1 (M+H).

(SV1-((2S. 4RV4-(tert-butyldiphenylsilyloxyV1 -tosylpyrrolidin-2-vnethanol

To a solution of MeMgBr (54 mL) in THF (100 mL) was added dropwise 4-(ferf-butyl- diphenyl-silanyloxy)-1-(toluene-4-sulfonyl)-pyrrolidine-2-ca rbaldehyde (54.4 g, 107 mmol) in THF (400 mL) at -10°C. The mixture was stirred at 0°C for 15 mins and then satd.

NH ₄ CI was added to the mixture at 0°C slowly. The organic layer was then washed with H ₂ 0 (400 mL) and brine (400 mL), dried over Na ₂ S0 ₄ . The organic layer was filtered and concentrated to give the crude product (54 g), which was directly used without further purification. LCMS (m/z): 446.1 (M+H).

(3R, 5SV5-((S)-1 -hvdroxyethvn-1 -tosylDyrrolidin-3-ol

To a solution of (S)-1 -((2S, 4/?)-4-(fert-butyldiphenylsilyloxy)-1 -tosylpyrrolidin-2-yl) ethanol (50 g, 95.6 mmol) in THF (500 mL) was added TBAF (37.4 g, 143.4 mmol). The mixture was stirred at rt for 3 h and then 500 mL of ethyl acetate was added to the mixture. The organic layer was washed with H ₂ 0 (2 x 400 mL), extracted with EA (3 x 200 mL) and dried over Na ₂ S0 ₄ . The mixture was then filtered and concentrated to give the crude product which was purified by silica gel column (PE: EA=1 :1 ) to yield 22.8 g. LCMS (m/z): 286.0 (M+H).

Methanesulfonic acid 5-methanesulfonyloxymethyl-1 -(toluene-4-sulfonyl)-pyrrolidin-3-yl ester

To a solution of {3R, 5S)-5-((S)-1 -hydroxyethyl)-1 -tosylpyrrolidin-3-ol (22.8 g, 80 mmol) and TEA (36.4 g, 360 mmol) in DCM (300 mL) was added dropwise MsCI (27.49 g, 240 mmol) at 0 °C. The mixture was stirred at rt for 18h. The mixture was then quenched with H ₂ 0 (100 mL), extracted with DCM (2 x 200 mL), washed with NaHC0 ₃ (2 x 300 mL) and dried over Na ₂ S0 ₄ . The mixture was filtered and concentrated to give the crude product (36 g), which was directly used without further urification. LCMS (m/z): 442.0 (M+H).

(1 S, 3S, 4S)-2-benzyl-3-methyl-5-tosyl-2,5-diazabicvclor2.2.1 lheptane A mixture of 5-methanesulfonyloxymethyl-1 -(toluene-4-sulfonyl)-pyrrolidin-3-yl ester (33 g, 74.83 mmol) and benzylamine (24 g, 224.49 mmol) in toluene (300 mL) was heated to reflux for 4 days. The mixture was filtered and the filtrate was concentrated to give the crude product which was purified by silica gel column (PE: EA=8:1 ) to yield 1 1 g. LCMS (m/z): 357.1 (M+H).

(1 S,3S,4/?V2-benzyl-3-methyl-2,5-diazabicvclor2.2.1lheDtane hvdrobromide

A mixture of (1 S, 3S, 4S)-2-benzyl-3-methyl-5-tosyl-2, 5-diazabicyclo[2.2.1 ]heptane (1 1 g, 30.89 mmol) was added to 22 ml. of HBr in AcOH (200 ml_). The mixture was stirred at 80°C for 3 days. The mixture was concentrated to give the crude product (14.7 g), which was directly used without further urification. LCMS (m/z): 203.0 (M+H).

C\ S, 4S, 6SHerf-butyl 5-benzyl-6-methyl-2,5-diazabicvclor2.2.1 lheDtane-2-carboxylate To a solution of (1 S, 3S, 4R)-2-benzyl-3-methyl-2,5-diazabicyclo[2.2.1]heptane hydrobromide (1 1.2 g, 30.9 mmol) and TEA (15.6 g, 154.5 mmol) in DCM (150 ml.) was added dropwise Boc ₂ 0 (6.7 g, 30.9 mmol) at 0°C. The mixture was stirred at rt for 18h. 50 ml. of H ₂ 0 was added to the mixture, extracted with DCM (3 x 50 ml_), washed with NaHC0 ₃ (2 x 200 ml.) and dried over Na ₂ S0 ₄ . The mixture was filtered and concentrated to give the crude product (7.8 g), which was purified by silica gel column (PE: EA=10:1 ) to yield 6.9 g of material. A portion of the product (1 .5 g) was separated by SFC (Berger MultiGram; OJ column 250 mm x 20 mm, A = C0 ₂ : B = IPA; A:B = 65:35 at 80 mL/min; column temp 38°C) to yield a 1 g mixture of isomers. It was subjected to a second SFC separation (Berger MultiGram; AD column 250 mm x 30 mm, A = C0 ₂ : B = MeOH; A:B = 90:10 at 80 mL/min; column temp 38°C) to yield 800 mg of pure material. LCMS (m/z): 303.1 (M+H).

(1 S, 4S, 6SHerf-butyl 6-methyl-2,5-diazabicvclor2.2.1 lheptane-2-carboxylate

5-benzyl-6-methyl-2,5-diazabicyclo[2.2.1 ]heptane-2-carboxylate (100 mg) was dissolved in MeOH (5 mL) and 20 mg of Pd/C was added under N ₂ atmosphere. The reaction mixture was hydrolyzed under an average pressure of 50 psi with H ₂ for 18 h at 50 °C. The mixture was filtered and the filtrate was concentrated to give the product as a white solid (90 mg). LCMS (m/z): 213.1 (M+H).

2-Benzyloxycarbonylamino-3,3-dimethyl-butyric acid

To a solution of 2-amino-3, 3-dimethyl-butyric acid (67.5 g, 514 mmol) in H ₂ 0 (700 ml) was added CbzCI (96.5 g, 565 mmol) at 0 °C. The pH was kept at -8-9 by adding a solution of NaOH (41 .1 g, 1030 mmol) in H ₂ 0 (500 mL). After addition, the mixture was stirred at 0 °C for 30 mins and then stirred at rt for 16h. The mixture was extracted with Et ₂ 0 (3 x 500 mL), then pH was adjusted to -2-3 by adding 2N HCI, extracted with EA (4 x 500 mL), dried over Na ₂ S0 ₄ and concentrated to give the crude product (13.2 g) which was used without further purification. L MS (m/z): 266.1 (M+H).

5-(2-Benzyloxycarbonylamino-3,3-dimethyl-butyryl)-6-methyl-2 ,5-diaza- bicvclor2.2.1lheptane-2-carboxylic acid tert-butyl ester To a solution of (1 S, 4S, 6S)-fert-butyl 6-methyl-2,5-diazabicyclo[2.2.1] heptane-2- carboxylate (240 mg, 1 .132 mmol) in DCM (5 mL) was added 2-benzyloxycarbonylamino- 3,3-dimethyl-butyric acid (300 mg, 1 .132 mmol), EDC (260 mg, 1.36 mmol), HOBt (31 mg, 0.226 mmol) and NMM (343 mg, 3.396 mmol). The mixture was stirred at rt for 16h. The mixture was diluted with DCM (10 mL), washed with H ₂ 0 (2 x 20 mL), sat. NaHC0 ₃ (10 mL), 1 N HCI (10 mL), brine (10 mL), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product, which was purified by silica gel column (PE: EA=2:1 ) to yield 545 mg of pure product. LCMS (m/z): 404.1 (M+

5-(2-Amino-3,3-dimethyl-butyryl)-6-methyl-2,5-diaza-bicvc lo[2.2.1lheptane-2-carboxylic acid ferf-butyl ester

5-(2-Benzyloxycarbonylamino-3,3-dimethyl-butyryl)-6-methy l-2,5-diaza- bicyclo[2.2.1 ]heptane-2-carboxylic acid tert-butyl ester (545 mg, 1 .187 mmol) was dissolved in MeOH (10 mL), 100 mg of Pd/C was added under N ₂ atmosphere. The reaction mixture was hydrolyzed under an average pressure of 50 psi with H ₂ for 18 h. The mixture was filtered and the filtrate was concentrated to give the product as a white solid (315 mg). LCMS (m/z): 326.1 (M+H).

5-{2-r(1 H-lndole-2-carbonyl)-aminol-3,3-dimethyl-butyryl}-6-methyl-2 ,5-di

bicvclor2.2.1lheptane-2-carboxylic acid tert-butyl ester

To a solution of 5-(2-amino-3,3-dimethyl-butyryl)-6-methyl-2,5-diaza-bicyclo[ 2.2.1 ] heptane-2-carboxylic acid ferf-butyl ester (315 mg, 0.969 mmol) in DCM (5 mL) was added 1 H-indole-2-carboxylic acid (156 mg, 0.969 mmol), EDC (223 mg, 1.16 mmol), HOBt (26 mg, 0.19 mmol) and NMM (294 mg, 2.9 mmol). The mixture was stirred at rt for 2h. The mixture was diluted with DCM (10 mL), washed with H ₂ 0 (2 x 10 mL), sat.

NaHCOs (10 mL), 1 N HCI (10 mL), brine (10 mL), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product (480 mg), which was directly used without further purification. LCMS (m/z): 469.0 (M+H .

1 /-/-lndole-2-carboxylic acid Γ2, 2-dimethyl-1 -(3-methyl-2,5-diaza-bicvclor2.2.1 lheptane-2- carbonvD-propyll-amide

To a solution of 5-{2-[(1 /-/-indole-2-carbonyl)-amino]-3,3-dimethyl-butyryl}-6-methyl -2,5- diaza-bicyclo[2.2.1 ]heptane-2-carboxylic acid ferf-butyl ester (480 mg, 1 .03 mmol) in DCM (6 mL) was added TFA (1 .5 mL). The mixture was stirred at rt for 1 h. Then, satd. NaHC0 ₃ was added to the mixture, pH adjusted to -8-9, extracted with DCM (3 x 10 mL), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product (300 mg), which was directly used without further purification. LCMS m/z): 369.1 (M+H).

5-Phenyl-pyridine-2-carboxylic acid

A mixture of compound 5-bromo-pyridine-2-carboxylic acid (8.0 g, 39.6 mmol),

phenylboronic acid (6.28 g, 51 .5 mmol), K ₂ C0 ₃ (7.56 g, 71.3 mmol), Pd(dppf)CI ₂ (0.88g, 1.2 mmol) in dioxane/water (100 mL, v/v, 3/1 ) was stirred at 1 10 °C under N ₂ over 16 h. The reaction mixture was cooled to rt, charged with satd. NaHC0 ₃ solution to pH -9-10 and filtered. The aqueous layer was washed with Et ₂ 0 (2 x 50ml_). The separated aqueous layer was adjusted to pH -4-5 with 1 N HCI and extracted with EA (6 x 50 ml_). The combined organic layer was dried over Na ₂ S0 ₄ and concentrated to give the crude product which was recrystallized from EtOH/water (50ml_, v/v, 2/1 ) to give 5.6 g (71 % yield) as a pale brown solid. LCMS m/z): 200.1 (M+H).

A/-r(1 SV2.2-dimethyl-1- (3SV3-methyl-5-r(5-Dhenyl-2-Dyridinvncarbonyll-2.5- diazabicvclor2.2.1lhept-2-yl}carbonyl)propyll-1 /-/-indole-2-carboxamide To a solution of 1 H-indole-2-carboxylic acid [2, 2-dimethyl-1-(3-methyl-2, 5- diaza-bicyclo[2.2.1 ]heptane-2-carbonyl)-propyl]-amid (150 mg, 0.408 mmol) in DCM (5 ml.) was added 5-phenyl-pyridine-2-carboxylic acid (81 mg, 0.408 mmol), EDC (94 mg, 0.489 mmol), HOBt (1 1 mg, 0.082 mmol) and NMM (124 mg, 1 .22 mmol). The mixture was stirred at rt for 16 h. The mixture was then diluted with DCM (20 ml_), washed with H ₂ 0 (2 x 10 ml_), sat. NaHC0 ₃ (10 ml_), 1 N HCI (10 ml_), brine (10 ml_), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product. The material was purified by HPLC (DYO 250 ^* 150 mm) and re-crystallized from water and dried by lyophilization to afford 55 mg of the title compound: LCMS (m/z): 550.2 (M+H). ¹ H NMR (400 MHz, CDCI ₃ ) δ ppm 9.24-9.19 (m, 1 H), 8.78-8.75 (m, 1 H), 8.06-7.98 (m, 2H), 7.69-7.59 (m, 3H), 7.56- 7.43 (m, 4H), 7.36-7.33 (m, 1 H), 7.19-7.13 (m, 1 H), 6.97-6.80 (m, 2H), 5.91 (m, 0.5H), 5.22-5.17(m, 1 H), 4.92-4.82(m, 2H), 4.16-4.04 (m, 2H), 3.87-3.61 (m, 1 H), 2.16-2.09 (m, 1 H), 1 .98-1.83 (m, 1 H), 1 .37-1.36 (m, 1 H), 1.27-1 .25 (m, 1 H), 1.1 1 (s, 9H).

Example 2

N-((SV3.3-dimethyl-1 -((1 S.4S.7/?V7-methyl-5-(5-phenylpicolinovn-2.5- diazabicvclor2.2.1lheptan-2-yl)-1-oxobutan-2-yl)-1 /-/-indole-2-carboxamide H

(2S,4/?)-ethyl 4-hvdroxypyrrolidine-2-carboxylate hydrochloride

To a solution of (2S,4/?)-4-hydroxypyrrolidine-2-carboxylic acid (100 g, 762 mmol) in EtOH (1 L) was added dropwise SOCI ₂ (1 10 ml_, 1524 mmol) at rt. The mixture was stirred for 16 h at rt. Then the solvent was evaporated in vacuo to give the crude product (150 g, 100%) as a white solid.

(2S,4/?)-1 -benzyl 2-ethyl 4-hydroxypyrrolidine-1 ,2-dicarboxylate

To a mixture of (2S,4/?)-1 -benzyl 2-ethyl 4-hydroxypyrrolidine-1 ,2-dicarboxylate

(150 g, 762 mmol) and CbzCI (130 g, 762 mmol) in THF/H ₂ 0 (1 :1 , 2.6 L) was added dropwise TEA (266 ml_, 1905 mmol) at 0°C. The mixture was stirred for 16 h at rt. After which the mixture was extracted with EtOAc (2 x 800 ml_). The organic layers were washed with 1 N aq.HCI, satd. NaHC0 ₃ , dried over Na ₂ S0 ₄ and concentrated to give the crude product (220 g, 100%) as a brown oil which used without further purification. LCMS (m/z): 294.1 (M+H).

(2S,4/?)-1 -benzyl 2-ethyl 4-(methylsulfonyloxy)pyrrolidine-1 ,2-dicarboxylate

To a mixture of (2S,4/?)-1 -benzyl 2-ethyl 4-hydroxypyrrolidine-1 ,2-dicarboxylate

(220 g, 760 mmol) and TEA (212 ml_, 1520 mmol) in dry CH ₂ CL ₂ (2.2 L) was added dropwise MsCI (130 g, 1 140 mmol) at 0°C. The mixture was stirred for 16 h at room temperature. The mixture was washed with 1 N aq. HCI, satd. NaHC0 ₃ , dried over Na ₂ S0 ₄ and concentrated to give the product (280 g, 100%) as a brown oil. LCMS (m/z): 372.0 (M+H).

(2S,4S)-1 -benzyl 2-ethyl 4-(phenylselanyl)pyrrolidine-1 ,2-dicarboxylate NaBH ₄ (13.4 g, 350 mmol) was added in small portions to a solution of Ph ₂ Se ₂ (46 g, 148 mmol) in EtOH (1 .6 L) at 0°C. The mixture was stirred for 5 min until the bright yellow color disappeared. Then to the mixture was added dropwise (2S,4S)-1 -benzyl 2-ethyl 4- (phenylselanyl)pyrrolidine-1 ,2-dicarboxylate (100 g, 270 mmol) in 400 mL of EtOH. The mixture was refluxed for 5 h. The solvent was evaporated in vacuo to dryness. The residue was diluted with 2 L EtOAc, washed with water, dried over Na ₂ S0 ₄ and

concentrated to dryness. The residue was purified by silica column (PE: EA, 10:1-6:1 ) to give the product (50 g, 43 %) as a pale ellow oil. LCMS (m/z): 434.0 (M+H).

(SV1 -benzyl 2-ethyl 1 H-pyrrole-1 ,2(2H,5H)-dicarboxylate To a solution of (2S,4S)-1 -benzyl 2-ethyl 4-(phenylselanyl)pyrrolidine-1 ,2-dicarboxylate (50 g, 1 15.5 mmol) in CH ₂ CI ₂ (2 L) was added dropwise pyridine (13.7 g, 173 mmol) at 0°C. H ₂ 0 ₂ (64 g, 288 mmol) was gradually added over 5 min. The mixture was stirred for 1 h at 20°C and diluted with 500 mL of CH ₂ CI ₂ , washed with 0.1 M aq. HCI, satd. NaHC0 ₃ , brine, dried over Na ₂ S0 ₄ and evaporated to dryness. The residue was purified by silica gel column (PE: EA, 20:1-8:1 ) to give the product (28g, 88 %) as a pale yellow oil. LCMS (m/z): 276.0 (M+H).

(1 R2S,5S)-3-benzyl 2-ethyl 6-oxa-3-azabicvclor3.1 .01hexane-2,3-dicarboxylate and (1 S,2S,5ffl-3-benzyl 2-ethyl 6-oxa-3-azabicvclor3.1.01hexane-2,3-dicarboxylate

Under an atmosphere of N ₂ , a solution of m-CPBA (60 g, 356 mmol) in 300 mL of CHCI ₃ was added dropwise to a mixture of (S)-1 -benzyl 2-ethyl 1 H-pyrrole-1 ,2(2H,5/-/)- dicarboxylate (28 g, 102 mmol) and 2,6-di-tert-butyl-phenol (3.0 g, 15 mmol) in 200 mL of CHCI ₃ . Then the mixture was stirred for 6 h at 80°C, then cooled and filtered. The filtrate was washed with 10 % aq Na ₂ C0 ₃ (3 x 200 mL), dried over Na ₂ S0 ₄ and evaporated to dryness. The residue was purified by silica column (PE/EA, 10:1 ) to give 16.5 g of material as a pale yellow oil. L -43).

(2R3S,4/?)-1 -benzyl 2-ethyl 4-hydroxy-3-methylpyrrolidine-1 ,2-dicarboxylate and (2S,3S,4/?)-1 -benzyl 2-ethyl 4-hydroxy-3-methylpyrrolidine-1 ,2-dicarboxylate and (2S,3S,4/?)-1 -benzyl 2-ethyl 3-hydroxy-4-methylpyrrolidine-1 ,2-dicarboxylate

To a suspension of Cul (25.6 g, 134.8 mmol) in 350 mL of Et ₂ 0 at -10°C was carefully added MeLi (3 M in ether, 270 mmol), while keeping the temperature below -5°C. After stirring for 30 min at -10°C, a mixture of (1 R,2S,5S)-3-benzyl 2-ethyl 6-oxa-3- azabicyclo[3.1.0]hexane-2,3-dicarboxylate and (1 S,2S,5R)-3-benzyl 2-ethyl 6-oxa-3- azabicyclo[3.1.0]hexane-2,3-dicarboxylate (16.4 g, 56.2 mmol) in 150 mL of Et ₂ 0 was added dropwise below -5°C. The mixture was stirred for 1 h at 0°C, quenched with 200 ml of water and then diluted with 200 mL of CH ₂ CI ₂ . The mixture was filtered over a celite pad, the organic layer was decanted, dried over Na ₂ S0 ₄ and evaporated in vacuo to provide 15 g (87%) of an inseparable mixture of the product which was used without further purification. LC +H).

(2R3S,4/?)-ethyl 4-hydroxy-3-methylpyrrolidine-2-carboxylate and (2S,3S,4/?)-ethyl 4-hydroxy-3-methylpyrrolidine-2-carboxylate and (2S,3S,4/?)-ethyl 3-hydroxy-4-methylpyrrolidine-2-carboxylate

A mixture of (2R,3S,4R)-ethyl 4-hydroxy-3-methylpyrrolidine-2-carboxylate, (2S,3S,4R)- ethyl 4-hydroxy-3-methylpyrrolidine-2-carboxylate and (2S,3S,4/?)-ethyl 3-hydroxy-4- methylpyrrolidine-2-carboxylate (15 g) and 50% of Pd/C ( 4.0 g) in 300 mL of EtOH was hydrogenated for 4 h under 45 Psi at rt. The mixture was filtered off and the filtrate was concentrated to dryness. The residue was purified by silica column (PE: EA, 3:1 -1 :2) to give 6.59 g (78%) a mixture of the crude product as a pale yellow oil.

(2R3S,4/?)-ethyl 4-hvdroxy-3-methyl-1 -tosylpyrrolidine-2-carboxylate and (2S,3S,4/?)-ethyl 4-hvdroxy-3-methyl-1-tosylpyrrolidine-2-carboxylate and

(2S,3S,4/?)-ethyl 3-hvdroxy-4-methyl-1-tosylpyrrolidine-2-carboxylate To a solution of a mixture of (2/?,3S,4/?)-ethyl 4-hydroxy-3-methyl-1-tosylpyrrolidine-2- carboxylate, (2S,3S,4/?)-ethyl 4-hydroxy-3-methyl-1-tosylpyrrolidine-2-carboxylate and (2S,3S,4R)-ethyl 3-hydroxy-4-methyl-1 -tosylpyrrolidine-2-carboxylate (6.59 g, 38.1 mmol) in 100 mL of dry CH ₂ CI ₂ was added TsCI (7.63 g, 40 mmol) in one portion at 0°C, followed by TEA (6.36 mL, 45.7 mmol) slowly. Then the mixture was stirred for 3h at rt. The mixture was diluted with 200 mL of CH ₂ CI ₂ , washed with 0.5 N aq HCI, dried over Na ₂ S0 ₄ and concentrated to dryness. The residue was purified by silica column (PE: EA, 3:1 -1 :2) to give the crude product in 85 % yield as pale yellow oil. LCMS (m/z): 328.0 (M+H).

(3R4S,5S)-5-(hvdroxymethyl)-4-methyl-1 -tosylpyrrolidin-3-ol and (3R4S,5/?)-5-(hvdroxymethyl)-4-methyl-1 -tosylpyrrolidin-3-ol

UAIH4 (1.84 g, 48.4 mmol) was suspended in 100 mL of dry THF under N ₂ and cooled to 0°C with vigorous stirring. Then a solution of a mixture of (2/?,3S,4/?)-ethyl 4-hydroxy-3- methyl-1 -tosylpyrrolidine-2-carboxylate, (2S,3S,4/?)-ethyl 4-hydroxy-3-methyl-1 - tosylpyrrolidine-2-carboxylate and (2S,3S,4/?)-ethyl 3-hydroxy-4-methyl-1 -tosylpyrrolidine- 2-carboxylate (10.56 g, 32.3 mmol) in 50 mL of dry THF was added dropwise below 5°C. Then the mixture was stirred for 1 h at room temperature. The reaction was treated with Fieser work up (1 .84 mL of water, 1 .84 mL of 30% aq NaOH, 1 .84 mL of water and dried over MgS0 ₄ and finally filtered off). The filtrate was concentrated to give the crude product. The crude product was purified by prep HPLC (TFA, 45%~75%) to give 1.53 g of (3R,4S,5S)-5-(hydroxymethyl)-4-methyl-1-tosylpyrrolidin-3-ol and 5.3 g of (3R,4S,5R)- (hydroxymethyl)-4-methyl-1-tosylpyrrolidin-3-ol as white solid. LCMS (m/z): 286.0(M+H).

((2S.3S.4 ?V3-methyl-4-(methylsulfonyloxyV1-tosylpyrrolidin-2-vnmethyl

methanesulfonate

To a mixture of (3R,4S,5S)-5-(hydroxymethyl)-4-methyl-1-tosylpyrrolidin-3-ol (1 .2 g, 4.2 mmol) and TEA (2.05 mL, 14.7 mmol) in dry CH ₂ CI ₂ (8 mL) was added dropwise MsCI (0.8 mL, 10.5 mmol) at 0°C. Then the mixture was stirred for 2 h at rt. The mixture was diluted with 30 mL of CH ₂ CI ₂ , washed with 0.5 N aq HCI, dried over Na ₂ S0 ₄ and concentrated to give the crude product (1 .92 g, 100 % as a pale yellow oil. LCMS (m/z): 442.0(M+H).

( 1 S,4S,7/?V2-benzyl-7-methyl-5-tosyl-2,5-diazabicvclor2.2.1 lheptane A mixture of ((2S,3S,4/?)-3-methyl-4-(methylsulfonyloxy)-1 -tosylpyrrolidin-2-yl)methyl methanesulfonate (1 .92 g, 4.2 mmol) and phenylmethanamine (1 .5 mL, 12.6 mmol) in xylene (20 mL) was stirred for 8 h at 135°C. Then the solvent was evaporated in vacuo to dryness. The residue was purified by silica column (PE/EA, 20:1 -10:1 ) to give the product (1 .13 g, 75 %) as a pale yellow oil. LCMS (m/z): 357.0(M+H).

(1 S,4S,7S)-2-benzyl-7-methyl-2,5-diazabicvclo[2.2.1lheptane dihydrobromide salt A mixture of (1 S,4S,7 ?)-2-benzyl-7-methyl-5-tosyl-2,5-diazabicyclo[2.2.1]heptane( 1.4 g, 3.93 mmol) and HBr (33% in AcOH, 10 mL) in 10 ml. of AcOH was stirred for 18 h at 80°C. Then the solvent was evaporated to dryness to give (1 S,4S,7S)-2-benzyl-7-methyl- 2,5-diazabicyclo[2.2.1]heptane dihydrobromide salt (1 .1 g, 100%) as a brown solid. LCMS (m/z): 203.2(M+H).

(1 S,4S,7/?)-tert-butyl 5-benzyl-7-methyl-2.5-diazabicvclor2.2.1 lheptane-2-carboxylate To a mixture of (1 S,4S,7S)-2-benzyl-7-methyl-2,5-diazabicyclo[2.2.1 ]heptane

dihydrobromide salt (1.1 g, 3.9 mmol) and TEA (1.63 mL, 1 1 .7 mmol) in 10 mL of CH ₂ CI ₂ was added dropwise (Boc) ₂ 0 (893 mg, 4.1 mmol) at 0 °C . Then the mixture was stirred for 16 h at rt. The mixture was diluted with 20 mL of CH ₂ CI ₂ , washed with 0.5 N aq HCI, brine, dried over Na ₂ S0 ₄ and concentrated to dryness. The residue was purified by silica column (PE/EA, 20:1-10:1 ) to give the product (880 mg, 75%) as a white solid. LCMS (m/z): 303.1 (M+H).

(1 S,4S,7/?He/f-butyl 7-methyl-2,5-diazabicvclor2.2.1lheptane-2-carboxylate

A mixture of (1 S,4S,7R)-tert-butyl 5-benzyl-7-methyl-2,5-diazabicyclo[2.2.1]heptane-2- carboxylate (880 mg, 2.91 mmol) and Pd/C (300 mg) in 30 mL of MeOH was

hydrogenated for 16 h under 50 Psi at 50°C. The mixture was filtered off and the filtrate was evaporated to give the product 500 mg, 81 %) as a pale yellow oil.

(1 S,4S,7/?Herf-butyl 5-((S)-2-(benzyloxycarbonylamino)-3,3-dimethylbutanoyl)-7-me thyl-

2,5-diazabicyclo[2.2.1lheptane-2-carboxylate

To a mixture of (1 S,4S,7R)-ferf-butyl 7-methyl-2,5-diazabicyclo[2.2.1 ]heptane-2- carboxylate (500 mg, 2.36 mmol), (S)-2-(benzyloxycarbonylamino)-3,3-dimethylbutanoic acid (687 mg, 2.86 mmol), EDCI (676 mg, 3.54 mmol), HOBT (190 mg, 1.42 mmol) in CH ₂ CI ₂ (6 mL) was added NMM (1 mL). The mixture was stirred for 16 h at rt. Then water (20 mL) was added to the mixture, extracted with CH ₂ CI ₂ (2 x 10 mL), washed with satd. NaHC0 ₃ , brine, dried over Na ₂ S0 ₄ , filtered and concentrated to dryness. The residue was purified by silica column (PE/EA, 15:1-10:1 ) to give the product (400 mg, 37%) as a pale yellow oil. LCMS (m/z): 404.1 (M-55).

(1 S,4S,7 ?Hert-butyl 5-((S)-2-amino-3,3-dimethylbutanoyl)-7-methyl-2,5- diazabicyclo[2.2.1lheptane-2-carboxylate

A mixture of (1 S,4S,7R)-tert-butyl 5-((S)-2-(benzyloxycarbonylamino)-3,3- dimethylbutanoyl)-7-methyl-2,5-diazabicyclo[2.2.1]heptane-2- carboxylate (400 mg, 0.87 mmol) and Pd/C (200 mg) in 20 mL of MeOH was hydrogenated for 6 h under 45 Psi at rt. The mixture was filtered off and the filtrate was evaporated to give the product (240 mg, 85%) as a pale yellow oil.

(1 S,4S,7 ?Hert-butyl 5-((S)-2-(1 H-indole-2-carboxamido)-3,3-dimethylbutanoyl)-7-methyl-

2,5-diazabicvclor2.2.1lheptane-2-carboxylate

To a mixture of (1 S,4S,7R)-tert-butyl 5-((S)-2-amino-3,3-dimethylbutanoyl)-7-methyl-2,5- diazabicyclo[2.2.1]heptane-2-carboxylate (240 mg, 0.74 mmol), 1 H-indole-2-carboxylic acid (125 mg, 0.77 mmol), EDCI (212 mg, 1 .1 1 mmol), HOBT (60 mg, 0.44 mmol) in CH ₂ CI ₂ (5 mL) was added NMM (0.3 mL). The mixture was stirred for 16 h at rt. Then water (20 mL) was added to the mixture, extracted with CH ₂ CI ₂ (2 x 10 mL), washed with satd. NaHC0 ₃ , brine, dried over Na ₂ S0 ₄ , filtered and concentrated to dryness. The residue was purified by silica column (PE/EA, 15:1-10:1 ) to give the product (280 mg, 81 %) as a brown solid. LCMS (m/z : 413.1 (M-55).

A/-((SV3.3-dimethyl-1-((1 S.4S.7SV7-methyl-2.5-diazabicvclor2.2.1 lheDtan-2-vn-1 - oxobutan-2-yl)-1 /-/-indole-2-carboxamide To a solution of (1 S,4S,7R)-fert-butyl 5-((S)-2-(1 H-indole-2-carboxamido)-3,3- dimethylbutanoyl)-7-methyl-2,5-diazabicyclo[2.2.1]heptane-2- carboxylate (140 mg, 0.3 mmol) in CH ₂ CI ₂ (3 mL) was added TFA (0.6 mL). The resulting mixture was stirred for 2h at rt. The reaction mixture was quenched by the addition of satd. NaHC0 ₃ and extracted with CH ₂ CI ₂ (2 x 10 mL). The organic layer was separated, washed with brine, dried over MgS0 ₄ and concentrated to give the product (100 mg, 90%) as a brown solid. LCMS (m/z): 413.1 (M-55).

A/-((S)-3,3-dimethyl-1 -((1 S,4S,7/?)-7-methyl-5-(5-phenylpicolinoyl)-2,5- diazabicvclor2.2.1lheptan-2-yl)-1-oxobutan-2-yl)-1 /-/-indole-2-carboxamide

To a mixture of A/-((S)-3, 3-dimethyl-1-((1 S,4S,7S)-7-methyl-2,5- diazabicyclo[2.2.1]heptan-2-yl)-1 -oxobutan-2-yl)-1 H-indole-2-carboxamide (100 mg, 0.27 mmol), 5-phenylpicolinic acid (59 mg, 0.30 mmol), EDCI (77 mg, 0.40 mmol), HOBT (22 mg, 0.16 mmol) in CH ₂ CI ₂ (3 mL) was added NMM (0.1 1 mL). The mixture was stirred for 16 h at rt. Then water (15 mL) was added to the mixture, extracted with CH ₂ CI ₂ (10 mL), washed with brine, dried over Na ₂ S0 ₄ , filtered and concentrated to dryness. The residue was purified by HPLC (YMC C18 5.0 um 150 ^* 30 mm) and dried by lypophilization to afford 67 mg of the title compound: LCMS (m/z): 550.3 (M+H); ¹ H NMR (400 MHz, DMSO-d6) δ ppm 0.85-1.12(12 H, m), 2.22-2.42 (1 H, m), 3.51-4.05(4 H, m), 4.55-4.92 (3 H, m), 6.95- 7.62 (9 H, m), 7.76-8.28 (4 H, m), 8.92-9.02 (1 H, m), 1 1 .60-1 1 .78 (1 H, m).

Example 3

A/-r(1 S)-2,2-dimethyl-1 -({(1 S,4S,6S)-6-methyl-5-r(5-phenyl-2-pyridinyl)carbonyll-2,5- diazabicvclo[2.2.1lhept-2-yl}carbonyl)propyll-1 /-/-indole-2-carboxamide

6-Methyl-5-(5-phenyl-pyridine-2-carbo

acid ferf-butyl ester

To a solution of 6-methyl-2,5-diaza-bicyclo[2.2.1]heptane-2-carboxylic acid ferf-butyl ester (265 mg, 1.25 mmol) in DCM (5 ml.) was added 5-phenyl-pyridine-2-carboxylic acid (249 mg, 1 .25 mmol), EDC (287 mg, 1.5 mmol), HOBt (34 mg, 0.25 mmol) and NMM (379 mg, 3.75 mmol). The mixture was stirred at rt for 4h. The mixture was then diluted with DCM (10 ml_), washed with H ₂ 0 (2 x 20 ml_), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product (550 mg) which was used in the next step without further purification. LCMS (m/z): 394.0 (M+H).

(3-Methyl-2,5-diaza-bicvclor2.2.1lhept-2-yl)-(5-phenyl-pyrid in-2-yl)-methanone

To a solution of 6-methyl-5-(5-phenyl-pyridine-2-carbonyl)-2,5-diaza-bicyclo[ 2.2.1 ] heptane-2-carboxylic acid ferf-butyl ester (550 mg, 1 .4 mmol) in DCM (5 ml.) was added TFA (1.5 ml_). The mixture was stirred at rt for 2 h, then satd. NaHC0 ₃ was added to the mixture, pH adjusted to -8-9, extracted with DCM (3 x 10 ml_), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product (243 mg), which was directly used to the next step without further purification. LCMS (m/z): 294.1 (M+H).

{2,2-Dimethyl-1-[6-methyl-5-(5-phenyl-pyridine-2-carbonyl)-2 ,5-diaza- bicyclo[2.2.1lheptane-2-carbonyll-propyl}-carbamic acid benzyl ester To a solution of (3-methyl-2,5-diaza-bicyclo[2.2.1 ]hept-2-yl)-(5-phenyl-pyridin-2-yl)- methanone (243 mg, 0.829 mmol) in DCM (5 ml.) was added 2-benzyloxycarbonylamino- 3,3-dimethyl-butyric acid (220 mg, 0.829 mmol), EDC (191 mg, 0.995 mmol), HOBt (22 mg, 0.166 mmol), NMM (252 mg, 2.487 mmol). The mixture was stirred at rt for 16h. The mixture was then diluted with DCM (10 ml_), washed with H ₂ 0 (2 x 10 ml_), dried over Na ₂ S0 ₄ , filtered and concentrated to give the crude product, which was purified by silica gel column (PE: EA=1 :1 ) to yield 330 mg of the desired compound. LCMS (m/z): 541 .3 (M+H).

2-Amino-3,3-dimethyl-1-r6-methyl-5-(5-phenyl-pyridine-2-carb onyl)-2,5-^

bicvclor2.2.1 lhept-2-yll-butan-1-one

{2,2-Dimethyl-1-[6-methyl-5-(5-phenyl-pyridine-2-carbonyl )-2,5-diaza-bicycloP

heptane-2-carbonyl]-propyl}-carbamic acid benzyl ester (330 mg, 0.61 mmol) was dissolved in MeOH (8 mL), and 80 mg of Pd/C was added under N ₂ atmosphere. The reaction mixture was hydrolyzed under an average pressure of 50 psi with H ₂ for 12 h. The mixture was then filtered and the filtrate was concentrated to give the product (220 mg) as a white solid which was directly used to the next step without further purification. LCMS (m/z): 407.1 (M+H).

1 /-/-lndole-2-carboxylic acid {2,2-dimethyl-1 -[6-methyl-5-(5-phenyl-pyridine-2-carbonyl)- 2,5-diaza-bicvclor2.2.1 lheptane-2-carbonyll-propyl}-amide To a solution of 2-amino-3,3-dimethyl-1-[6-methyl-5-(5-phenyl-pyridine-2-carb onyl)-2,5- diaza-bicyclo[2.2.1]hept-2-yl]-butan-1 -one (220 mg, 0.542 mmol) in CH ₂ CI ₂ (5 mL) was added 1 H-indole-2-carboxylic acid (87 mg, 0.542 mmol), EDC (125 mg, 0.65 mmol), HOBt (15 mg, 0.1 1 mmol) and NMM (164 mg, 1.63 mmol). The reaction mixture was stirred at rt for 16 h. The reaction was then diluted with CH ₂ CI ₂ (5 mL) and washed with H ₂ 0 (2 x 10 mL). The organic layer was dried over Na ₂ S0 ₄ , filtered, and concentrated to yield the crude product. The material was purified by HPLC (DYO 250 ^* 150 mm) and then recrystallized from water, dried by lypophilization to afford 146 mg of the title compound: LCMS (m/z): 550.2 (M+H); ¹ H NMR (400 MHz, CDCI ₃ ) δ ppm 0.98 - 1.18 (9 H, m), 1.23 - 1.34 (1 H, m), 1.43-1 .54 (1 H, m), 1 .65 - 2.13 (4 H, m), 3.54-3.62 (1 H, m), 3.82-4.04 (1 H, m), 4.21 - 4.25 (1 H, m), 4.62-5.21 (4 H, m), 6.87-7.03 (1 H, m), 7.06-7.16 (1 H, m), 7.27- 7.33 (2 H, m), 7.35-7.52 (4 H, m), 7.56-7.68 (3 H, m), 7.94-8.07 (2 H, m), 8.86 (1 H, s), 9.24-9.27 (1 H, m).