WO2011025799A1 - Cathepsin c inhibitors - Google Patents

Cathepsin c inhibitors Download PDF

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Publication number
WO2011025799A1
WO2011025799A1 PCT/US2010/046566 US2010046566W WO2011025799A1 WO 2011025799 A1 WO2011025799 A1 WO 2011025799A1 US 2010046566 W US2010046566 W US 2010046566W WO 2011025799 A1 WO2011025799 A1 WO 2011025799A1
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Prior art keywords
pyrrolidinyl
cyano
alkyl
mmol
amino
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PCT/US2010/046566
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French (fr)
Inventor
Ann Marie Bullion
Jakob Busch-Petersen
Brian Evans
Christopher E. Neipp
Brent W. Mccleland
Neysa Nevins
Michael D. Wall
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Glaxo Group Limited
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Publication of WO2011025799A1 publication Critical patent/WO2011025799A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/14—Nitrogen atoms not forming part of a nitro radical
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/10—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to certain 3-aminopyrrolidines that are cathepsin C inhibitors, pharmaceutical compositions containing these compounds, and their use in the treatment of diseases mediated by the cathepsin C enzyme such as chronic obstructive pulmonary disease.
  • Cathepsins are a family of enzymes included in the papain superfamily of cysteine proteases. Cathepsins B, C, F, H, K, L, S, V, and X have been described in the scientific literature. Cathepsin C is also known in the literature as Dipeptidyl Peptidase I or "DPPI.” A number of recently published studies have begun to describe the role cathepsin C plays in certain inflammatory processes. See e.g. Adkison et al., The Journal of Clinical Investigation 109:363-371 (2002); Tran et al., Archives of Biochemistry and Biophysics 403:160-170 (2002); Thiele et al., The Journal of Immunology 158: 5200-5210 (1997);
  • proteases Once activated, these proteases have a number of functions including degradation of various extracellular matrix components, which together can propagate tissue damage and chronic inflammation.
  • COPD Chronic Obstructive Pulmonary Disease
  • COPD chronic Obstructive Pulmonary Disease
  • chronic bronchitis and emphysema usually occur together in COPD patients.
  • Chronic bronchitis is generally characterized by a chronic productive cough, whereas emphysema is generally
  • Cigarette smoking is a significant risk factor for developing COPD. Exposure to cigarette smoke and other noxious particles and gases may result in chronic inflammation of the lung. In response to such exposure, inflammatory cells such as CD8+ Tcells, macrophages, and neutrophils are recruited to the area. These recruited inflammatory cells release proteases, which are believed to play a major role in the disease etiology by a number of mechanisms. Proteases believed to be involved in this process include the serine proteases neutrophil elastase ("NE"), cathepsin G, and proteinase 3, all released from neutrophils; granzymes A and B, released from cytotoxic T cells or natural killer cells; and chymases, released from mast cells. Cathepsin C appears to be involved in activating all of these enzymes.
  • NE neutrophil elastase
  • RA Rheumatoid arthritis
  • Cathepsin C may play a role.
  • Neutrophils are recruited to the site of joint inflammation and release cathepsin G, NE, and proteinase 3, which are believed to be responsible in part for cartilage destruction associated with RA (Hu, Y. and Pham, C. T. (2005) Arthritis Rheum 52: 2553-2558).
  • cathepsin C may play a role
  • Other conditions where cathepsin C may play a role include osteoarthritis, asthma, and Multiple Sclerosis. See e.g. Matsui, K.; Yuyama, N.; Akaiwa, M.; Yoshida, N. L.; Maeda, M.; Sugita, Y.; Izuhara, K., Identification of an alternative splicing variant of cathepsin C/dipeptidyl-peptidase I, Gene. 293(1-2): 1-7, 2002 Jun 26; Wolters, P. J.; Laig- Webster, M.; Caughey, G. H., Dipeptidyl peptidase I cleaves matrix-associated proteins and is expressed mainly by mast cells in normal dog airways, American Journal of Respiratory Cell & Molecular Biology. 22(2): 183-90, 2000.
  • One approach to treating these conditions is to inhibit the activity of the serine proteases involved in the inflammatory process, especially NE activity. See e.g.,
  • cathepsin C There are additional activities of cathepsin C that may also be related to disease etiology. Cathepsin C has been demonstrated to have a role in neutrophil migration in the development of aortic aneurysms by a mechanism which has not been clearly elucidated (Pagano, M. B. et al. (2007) PNAS 104: 2855-2860). Thus, disease processes that involve neutrophil migration, as well as proteolytic enzyme release can be modulated by cathepsin C inhibition. Also, cathepsin C is highly expressed in the lung epithelium where it may play a role in the processing of other enzymes not yet identified.
  • Cathepsin C has also been reported to cleave kallikrein-4, which is believed to play a role in dental enamel maturation (Tye, C. E. et al. (2009) J. Dental Res. 88: 323-327). Finally, cathepsin C is itself released from cells and may play a direct role in the degradation of matrix proteins.
  • the present invention involves novel compounds according to Formula (I) or a pharmaceutically acceptable salt thereof:
  • A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , (C 3 -Ce)cycloalkyl, heterocycloalkyl, hydroxyl, (Ci-C 4 )alkoxy, aryl, aryloxy, heteroaryl, cyano, -CO 2 (C i-C 4 )alkyl, -CONH(C i-C 4 )alkyl, -CON(Ci-C 4 )alkyl(Ci-C 4 )alkyl,
  • A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 5- or 6-membered heterocycloalkyl group;
  • R 1 is hydrogen, (Ci-C 4 )alkyl, (C 2 -C 5 )alkenyl, (C 2 -C 5 )alkynyl, (C 3 -C 5 )cycloalkyl,
  • heteroaryl(Ci-C 2 )alkyl methoxy(Ci-C 2 )alkyl, aryl(Ci-C 2 )alkyl, or heteroaryl(Ci-C 2 )alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C 2 )alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms; and
  • R 2 is hydrogen or methyl
  • the present invention is also directed to the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in the prevention, management or treatment of a respiratory or inflammatory disease, such as chronic obstructive pulmonary disease or rhinitis.
  • a respiratory or inflammatory disease such as chronic obstructive pulmonary disease or rhinitis.
  • this invention relates to a pharmaceutically acceptable formulation
  • a pharmaceutically acceptable formulation comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
  • alkyl refers to a straight- or branched-chain
  • hydrocarbon radical having the specified number of carbon atoms.
  • (Ci-C 4 )alkyl and “(Ci-C 8 )alkyl” refer to an alkyl group having at least 1 and up to 4 or 8 carbon atoms respectively. Examples of such branched or straight-chained alkyl groups useful in the present invention include, but are not limited to, methyl, ethyl, n -propyl, isopropyl, /?
  • alkenyl refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1 and up to 3 carbon- carbon double bonds. Examples include ethenyl and propenyl.
  • alkynyl refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1 and up to 3 carbon- carbon triple bonds. Examples include ethynyl and propynyl.
  • cycloalkyl refers to a non-aromatic, saturated, cyclic hydrocarbon ring containing the specified number of carbon atoms.
  • (C 3 -C 8 )cycloalkyl refers to a non-aromatic cyclic hydrocarbon ring having from three to eight ring carbon atoms.
  • Exemplary "(C 3 -C 8 )cycloalkyl” groups useful in the present invention include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • heterocycloalkyl refers to a group or moiety comprising a non-aromatic, monovalent monocyclic or bicyclic radical, which is saturated or partially unsaturated, containing 3 to 10 ring atoms, which includes 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, and which may be unsubstituted or substituted by one or more of the substituents defined herein.
  • heterocycloalkyls include, but are not limited to, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3- dithianyl, hexahydro-lH-l,4-diazepinyl, azabicylo[3.2.1]octyl, azabicylo[3.2.1]o
  • Alkoxy means an alkyl radical containing the specified number of carbon atoms attached through an oxygen linking atom.
  • the term "(Ci-C 4 )alkoxy” refers to a straight- or branched-chain hydrocarbon radical having at least 1 and up to 4 carbon atoms attached through an oxygen linking atom.
  • Exemplary "(Ci-C 4 )alkoxy” groups useful in the present invention include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n- butoxy, 5-butoxy, and t-butoxy.
  • Aryl refers to optionally substituted monocyclic or fused bicyclic groups having
  • aryl groups are phenyl, naphthyl, indenyl, dihydroindenyl, anthracenyl, phenanthrenyl, and the like.
  • aryl refers to optionally substituted phenyl.
  • Aryloxy means an aryl radical attached through an oxygen linking atom.
  • aryloxy groups useful in the present invention include, but are not limited to, phenyloxy (or phenoxy) and naphthyloxy.
  • Heteroaryl means an optionally substituted aromatic monocyclic ring or fused bicyclic ring system wherein at least one ring complies with H ⁇ ckel's Rule, has the specified number of ring atoms, and that ring contains at least one heteroatom selected from N, O, and/or S.
  • 5-membered “heteroaryl” groups include furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, and isothiazolyl.
  • heteroaryl groups include oxo-pyridyl, pyridinyl (or pyridyl), pyridazinyl, pyrazinyl, and pyrimidinyl.
  • 6,6-fused "heteroaryl” groups include quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1,6- naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, and pteridinyl.
  • 6,5- fused "heteroaryl” groups include benzofuranyl, benzothienyl, benzimidazolyl, benzthiazolyl, indolizinyl, indolyl, isoindolyl, and indazolyl.
  • bicyclic ring systems may be attached at any suitable position on either ring.
  • Optionally substituted indicates that a group, such as alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, aryl, aryloxy, or heteroaryl, may be unsubstituted, or the group may be substituted with one or more substituent(s) as defined. In the case where groups may be selected from a number of alternative groups the selected groups may be the same or different.
  • halogen or halo refers to F, Cl, Br, or I.
  • A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , (C 3 -Ce)cycloalkyl, heterocycloalkyl, hydroxyl, (Ci-C 4 )alkoxy, aryl, aryloxy, heteroaryl, cyano, -CO 2 (C i-C 4 )alkyl, -CONH(C i-C 4 )alkyl, -CON(Ci-C 4 )alkyl(Ci-C 4 )alkyl, -SO 2 (Ci-C 4 )alkyl, -SO 2 NH(C i-C 4 )alkyl, -SO 2 N(Ci-C 4 )alkyl(Ci-C 4 )alkyl, amino,
  • A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 5- or 6-membered heterocycloalkyl group.
  • A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, aryl, aryloxy, heteroaryl, cyano,
  • A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy.
  • A represents a 5-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxa
  • A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 10-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group.
  • A represents thienyl, 5-carboxyethylthienyl, or 3-chloroisothiazolyl.
  • A represents a 5-membered aromatic ring containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is fused to an aromatic or non-aromatic carbocyclic ring moiety to form an 8- to 10- membered bicyclic group, wherein said aromatic carbocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group.
  • A represents benzothienyl, 5,6-dihydro-4H-cyclopenta[ ⁇ ]thienyl, 4,5,6,7- tetrahydro- 1 -benzothienyl, 4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'-[l ,3]dioxolanyl, or 5,6,7,8-tetrahydro-4H-cyclohepta[ ⁇ ]thienyl.
  • A represents indolyl or JV-methylindolyl.
  • A represents
  • A represents a 6-membered aromatic ring optionally containing one or two nitrogen atoms, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1,4- dioxanyl, 1,3-oxathiolanyl, 1,3-
  • A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 11-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy.
  • A represents pyridinyl, pyrazinyl, or quinolinyl.
  • A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (d-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3- dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1, 3 -dithianyl,
  • A represents phenyl, 4-fluorophenyl, 4-bromophenyl, 4,5-difluorophenyl, 4- trifluoromethylphenyl, 5-trifluoromethylphenyl, 4-biphenyl, 5-biphenyl, 4'-fluoro-4- biphenyl, 3'-nitro-4'-methyl-4-biphenyl, 3'-acetyl-4-biphenyl, 3'-carboxyethyl-4-biphenyl, 4'-methoxy-4-biphenyl, 3 '-fluoro-4'-methoxy-4-biphenyl, 4-(l ,3-benzodioxol-5- yl)phenyl, 3'-amino-4-biphenyl, 4-pyridin-4-ylphenyl, 4-pyrimidin-2-ylphenyl, A- pyrimidin-5-ylphenyl, 4-furan-3-yl
  • A represents a thienyl, 4,5,6,7-tetrahydro-l- benzothienyl, isothiazolyl, indolyl, pyridinyl, pyrazinyl, quinolinyl, or phenyl ring optionally substituted one to two times, independently, by fluoro, chloro, bromo, methyl, trifluoromethyl, carboxyethyl, phenyl, 4-fluorophenyl, pyrimidinyl, furanyl, thienyl, thiazolyl, methoxy, phenyloxy, amino, dimethylamino, or nitro.
  • R 1 is hydrogen, (Ci-C 4 )alkyl, (C 2 -C 5 )alkenyl, (C 2 -C 5 )alkynyl,
  • heteroaryl(Ci-C 2 )alkyl hydroxy(Ci-C 2 )alkyl, methoxy(Ci-C 2 )alkyl, aryl(Ci-C 2 )alkyl, or heteroaryl(Ci-C 2 )alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C 2 )alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms.
  • R 1 is hydrogen, (Ci-C 4 )alkyl or heteroaryl(Ci-C 2 )alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C 2 )alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms.
  • R 1 is (Ci-C 4 )alkyl or thienyl(Ci-C 2 )alkyl.
  • R 1 is methyl, ethyl, n-propyl, isopropyl, or 2-thienylmethyl.
  • R 1 is methyl.
  • R 1 is ethyl.
  • R 1 is 2-thienylmethyl.
  • R 2 is hydrogen or methyl. In a selected embodiment, R 2 is hydrogen. In another embodiment, R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF 3 , cyano, (Ci-C 4 )alkyl, amino, (Ci-C 4 )alkylamino, ((Ci-C 4 )alkyl)((Ci-C 4 )alkyl)amino, hydroxyl, (Ci-C 4 )alkoxy, or (Ci-C 4 )alkylthio-; wherein said ring is optionally fused to a (C 3 -Cs)cycloalkyl ring.
  • R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF 3 , cyano, methyl, amino, hydroxyl, methoxy, or methylthio-; wherein said ring is optionally fused to a cyclopropyl ring.
  • R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF3, cyano, methyl, methoxy, or methylthio-.
  • R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F.
  • R 1 and R 2 taken together represent -CH 2 CH 2 -.
  • R 1 and R 2 taken together represent -CH 2 CH 2 CH 2 -.
  • R 1 and R 2 taken together represent -CH 2 CHFCH 2 -.
  • R 1 and R 2 taken together represent -CH 2 CF 2 CH 2 -.
  • R 1 and R 2 taken together represent -CH 2 CH(CH 3 )CH 2 -.
  • R 1 and R 2 taken together represent -CHOHCH 2 CH 2 -.
  • R 1 and R 2 taken together represent -CH 2 CH 2 CH 2 CH 2 -.
  • One particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
  • A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (d-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3- oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, pheny
  • R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF 3 , cyano, methyl, methoxy, or methylthio-.
  • Another particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
  • A represents a 5-membered aromatic ring containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is fused to an aromatic or non-aromatic carbocyclic ring moiety to form an 8- to 10-membered bicyclic group, wherein said aromatic carbocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF 3 , (Ci-C 4 )alkyl, hydroxyl, or (Ci-C 4 )alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group; and
  • R 1 and R 2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF 3 , cyano, methyl, methoxy, or methylthio-.
  • Another particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
  • A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (Ci-C 4 )alkyl, -CF 3 , hydroxyl, (Ci-C 4 )alkoxy, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, cyano, -CO 2 (C i-C 4 )alkyl, -CONH(C i-C 4 )alkyl,
  • R 1 is (Ci-C 4 )alkyl or thienyl(Ci-C 2 )alkyl
  • R 2 is hydrogen
  • the invention also includes various isomers of the compounds of Formula (I) and mixtures thereof.
  • “Isomer” refers to compounds that have the same composition and molecular weight but differ in physical and/or chemical properties. The structural difference may be in constitution (geometric isomers) or in the ability to rotate the plane of polarized light (stereoisomers).
  • the compounds according to Formula (I) contain two or more asymmetric centers, also referred to as chiral centers, and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomeric forms, or as mixtures thereof. All such isomeric forms are included within the present invention, including mixtures thereof.
  • Chiral centers may also be present in a substituent such as an alkyl group. Where the stereochemistry of a chiral center present in Formula (I), or in any chemical structure illustrated herein, is not specified the structure is intended to encompass any stereoisomer and all mixtures thereof. Thus, compounds according to Formula (I) containing two or more chiral centers may be used as racemic mixtures, enantiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers.
  • Individual stereoisomers of a compound according to Formula (I) which contain two or more asymmetric centers may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer-specif ⁇ c reagent, for example by enzymatic oxidation or reduction; or (3) by gas-liquid or liquid chromatography in a chiral environment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent.
  • stereoisomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired form.
  • specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation.
  • Enantiomerically enriched refers to products whose enantiomeric excess is greater than zero.
  • enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee, and greater than 90% ee.
  • Enantiomeric excess or "ee” is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%).
  • Enantiomerically pure means products whose enantiomeric excess is 99% ee or greater.
  • the invention also includes various deuterated forms of the compounds of Formula (I).
  • Each available hydrogen atom attached to a carbon atom may be independently replaced with a deuterium atom.
  • a person of ordinary skill in the art will know how to synthesize deuterated forms of the compounds of Formula (I).
  • ⁇ -deuterated ⁇ -amino acids are commercially available or may be prepared by conventional techniques (see for example: Elemes, Y. and Ragnarsson, U. J. Chem. Soc, Perkin Trans. 1, 1996, 6, 537-40).
  • Employing such compounds according to Scheme 1 below will allow for the preparation of compounds of Formula (I) in which the hydrogen atom at the chiral center baring R 1 is replaced with a deuterium atom.
  • ⁇ -amino acids in which deuterium atoms have been incorporated into the sidechains are commercially available or may be prepared by conventional techniques. Employing such compounds according to Scheme 1 below will allow for the preparation of compounds of Formula (I) in which deuterium atoms have been incorporated in R 1 .
  • solvate refers to a complex of variable stoichiometry formed by a solute and a solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute.
  • suitable solvents include, but are not limited to, water, methanol, ethanol and acetic acid.
  • the solvent used is a pharmaceutically acceptable solvent.
  • suitable pharmaceutically acceptable solvents include, without limitation, water, ethanol and acetic acid.
  • Solvates wherein water is the solvent molecule are typically referred to as "hydrates". Hydrates include compositions containing stoichiometric amounts of water, as well as compositions containing variable amounts of water. Solvates, particularly hydrates, of the compounds of Formula (I) and salts thereof, are within the scope of the invention.
  • the compound or salt including solvates (particularly, hydrates) thereof, may exist in crystalline forms, non-crystalline forms or a mixture thereof.
  • the compound or salt, or solvates (particularly, hydrates) thereof may also exhibit polymorphism (i.e. the capacity to occur in different crystalline forms). These different crystalline forms are typically known as "polymorphs.”
  • polymorphs typically known as “polymorphs.”
  • the disclosed compound, or solvates (particularly, hydrates) thereof also include all polymorphs thereof. 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. One of ordinary skill in the art will appreciate that different polymorphs may be produced, for example, by changing or adjusting the conditions used in crystallizing/recrystallizing the compound.
  • salts of the compounds of Formula (I) are preferably pharmaceutically acceptable.
  • Suitable pharmaceutically acceptable salts can include acid or base addition salts.
  • salts and solvates e.g. hydrates and hydrates of salts
  • the counterion or associated solvent is pharmaceutically acceptable.
  • salts and solvates having non-pharmaceutically acceptable counterions or associated solvents are within the scope of the present invention, for example, for use as
  • Compounds of Formula (I) have one or more nitrogen(s) basic enough to form pharmaceutically acceptable acid addition salts by treatment with a suitable acid.
  • Suitable acids include pharmaceutically acceptable inorganic acids and pharmaceutically acceptable organic acids.
  • Representative pharmaceutically acceptable acid addition salts include acetate, aspartate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, formate, fumarate, galacturonate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexanoate, hydrobromide, hydrochloride,
  • phosphate/diphosphate polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate, and tosylate salts.
  • salts include pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; carbonates and bicarbonates of a pharmaceutically acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; pharmaceutically acceptable organic 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, cyclohexylamine, triethanolamine, choline, arginine, lysine, and histidine.
  • pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts
  • carbonates and bicarbonates of a pharmaceutically acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc
  • non-pharmaceutically acceptable salts e.g. trifluoroacetate
  • Other non-pharmaceutically acceptable salts e.g. trifluoroacetate, may be used, for example in the isolation of compounds of the invention, and are included within the scope of this invention.
  • the invention includes within its scope all possible stoichiometric and non- stoichiometric forms of the salts of the compounds of Formula (I).
  • pro-drugs examples include Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31, pp 306 - 316 and in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as “pro-moieties”, for example as described by H. Bundgaard in “Design of Prodrugs” (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within compounds of the invention.
  • Preferred "pro- moieties" for compounds of the invention include: ester, carbonate ester, hemi-ester, phosphate ester, nitro ester, sulfate ester, sulfoxide, amide, carbamate, azo-, phosphamide, glycoside, ether, acetal, and ketal derivatives of the compounds of Formula (I).
  • the compounds of the invention inhibit the cathepsin C enzyme and can be useful in the treatment of conditions wherein the underlying pathology is (at least in part) attributable to cathepsin C involvement or in conditions wherein cathepsin C inhibition offers some clinical benefit even though the underlying pathology is not (even in part) attributable to cathepsin C involvement.
  • Examples of such conditions include COPD, rheumatoid arthritis, osteoarthritis, asthma, and multiple sclerosis. Accordingly, in another aspect the invention is directed to methods of treating such conditions.
  • the methods of treatment of the invention comprise administering an effective amount of a compound of the invention to a patient in need thereof.
  • treatment in reference to a condition means: (1) the amelioration or prevention of the condition being treated or one or more of the biological
  • prevention of a condition includes prevention of the condition.
  • prevention is not an absolute term.
  • 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.
  • An "effective amount” means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician.
  • therapeutically effective amount means any amount which, as compared to a
  • corresponding subject who has not received such amount results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder.
  • the term also includes within its scope amounts effective to enhance normal physiological function.
  • patient refers to a human or 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.
  • suitable dosing regimens including the amount administered and 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 particular route of administration chosen, 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 range from 1 mg to 1000 mg.
  • the invention includes the use of compounds of the invention for the preparation of a composition for treating or ameliorating diseases mediated by the cathepsin C enzyme in a subject in need thereof, wherein the composition comprises a mixture of one or more of the compounds of the invention and an optional pharmaceutically acceptable excipient.
  • the invention further includes the use of compounds of the invention as an active therapeutic substance, in particular in the treatment of diseases mediated by the cathepsin C enzyme.
  • the invention includes the use of compounds of the invention in the treatment of COPD, rheumatoid arthritis, osteoarthritis, asthma, and multiple sclerosis.
  • the invention includes the use of compounds of the invention in the manufacture of a medicament for use in the treatment of the above disorders.
  • the compounds of the invention will normally, but not necessarily, be formulated into a pharmaceutical composition prior to administration to a patient. Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of the invention and a pharmaceutically acceptable excipient.
  • compositions of the invention may be prepared and packaged in bulk form wherein an effective amount of a compound of the invention can be extracted and then given to the patient such as with powders, syrups, and solutions for injection.
  • the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains an effective amount of a compound of the invention.
  • 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. Conversely, the pharmaceutical compositions of the invention typically contain more than one pharmaceutically acceptable excipient. However, in certain embodiments, the pharmaceutical compositions of the invention contain one pharmaceutically acceptable excipient.
  • pharmaceutically acceptable excipient means a material, composition or vehicle involved in giving form or consistency to the composition and which is safe when administered to a patient.
  • 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.
  • each excipient must of course be of sufficiently high purity to render it pharmaceutically acceptable.
  • dosage forms include those adapted for (1) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, 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 aerosols and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.
  • oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets
  • parenteral administration such as sterile solutions, suspensions, and powders for reconstitution
  • transdermal administration such as transdermal patches
  • rectal administration such as supposi
  • Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen.
  • suitable pharmaceutically acceptable excipients may be chosen for a particular function that they may serve in the composition.
  • 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 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, sweeteners, flavoring agents, flavor masking agents, coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents.
  • 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.
  • 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).
  • 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
  • the invention is directed to a solid oral dosage form such as a tablet or capsule comprising an 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.
  • 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, magnesium stearate, calcium stearate, and talc.
  • the invention is directed to a dosage form adapted for administration to a patient by inhalation.
  • the compound of the invention may be inhaled into the lungs as a dry powder, an aerosol, a suspension, or a solution.
  • Dry powder compositions for delivery to the lung by inhalation typically comprise a compound of the invention as a finely divided powder together with one or more pharmaceutically acceptable excipients as finely divided powders.
  • Pharmaceutically acceptable excipients particularly suited for use in dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides.
  • the dry powder may be administered to the patient via a reservoir dry powder inhaler (RDPI) having a reservoir suitable for storing multiple (un-metered doses) of medicament in dry powder form.
  • RDPIs typically include a means for metering each medicament dose from the reservoir to a delivery position.
  • the metering means may comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation.
  • the dry powder may be presented in capsules (e.g. gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPI).
  • MDPIs are inhalers wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple defined doses (or parts thereof) of medicament.
  • the dry powder is presented as a blister pack, it comprises multiple blisters for containment of the medicament in dry powder form.
  • the blisters are typically arranged in regular fashion for ease of release of the medicament therefrom.
  • the blisters may be arranged in a generally circular fashion on a disc-form blister pack, or the blisters may be elongate in form, for example comprising a strip or a tape.
  • Each capsule, cartridge, or blister may, for example, contain between 20 ⁇ g-10mg of the compound of the invention.
  • Aerosols may be formed by suspending or dissolving a compound of the invention in a liquified propellant.
  • Suitable propellants include halocarbons, hydrocarbons, and other liquified gases.
  • Representative propellants include: trichlorofluoromethane
  • Aerosols comprising a compound of the invention will typically be administered to a patient via a metered dose inhaler (MDI). Such devices are known to those skilled in the art.
  • the aerosol may contain additional pharmaceutically acceptable excipients typically used with multiple dose inhalers such as surfactants, lubricants, cosolvents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
  • additional pharmaceutically acceptable excipients typically used with multiple dose inhalers such as surfactants, lubricants, cosolvents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
  • Suspensions and solutions comprising a compound of the invention may also be administered to a patient via a nebulizer.
  • the solvent or suspension agent utilized for nebulization may be any pharmaceutically acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g., ethanol, isopropylalcohol, glycerol, propylene glycol,
  • Saline solutions utilize salts which display little or no pharmacological activity after administration.
  • organic salts such as alkali metal or ammonium halogen salts, e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc. may be used for this purpose.
  • compositions may be added to the suspension or solution.
  • the compound of the invention may be stabilized by the addition of an inorganic acid, e.g., hydrochloric acid, nitric acid, sulfuric acid and/or phosphoric acid; an organic acid, e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc., a complexing agent such as EDTA or citric acid and salts thereof; or an antioxidant such as antioxidant such as vitamin E or ascorbic acid.
  • Preservatives may be added such as benzalkonium chloride or benzoic acid and salts thereof.
  • Surfactant may be added particularly to improve the physical stability of suspensions. These include lecithin, disodium dioctylsulphosuccinate, oleic acid and sorbitan esters. Methods of Preparation.
  • the compounds of Formula (I) may be obtained by using synthetic procedures illustrated in the Schemes below or by drawing on the knowledge of a skilled organic chemist.
  • the synthesis provided in these Schemes are applicable for producing compounds of the invention having a variety of different R 1 and R 2 groups employing appropriate precursors, which are suitably protected if need be, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, where needs be, and then affords compounds of the nature generally disclosed. While the Schemes are shown with compounds only of Formula (I), they are illustrative of processes that may be used to make the compounds of the invention.
  • the compounds of Formula (I) can be prepared in a multi- step sequence starting from a protected 3-aminopyrrolidine, such as the commercially available 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate.
  • an appropriate base such as K2CO3 or Et 3 N
  • an appropriate solvent such as CH 3 CN, THF, DMF, or DMSO
  • an appropriate catalyst such as Pd 2 (dba) 3
  • an appropriate ligand such as BINAP
  • an appropriate base such as Cs 2 CO 3
  • Boc-protected amino acid such as N-(tert-butoxy- carbonyl)glycine, iV-(te/t-butoxycarbony l)-L-alanine, (2S)-2-( ⁇ [(1,1 -dimethylethyl)- oxy]carbonyl ⁇ -amino)butanoic acid, iV-(te/t-butoxycarbonyl)-L-valine, N-(tert-butoxy- carbonyl)-L-norvaline, iV-(tert-butoxycarbonyl)-L-leucine, iV-(te/t-butoxycarbonyl)-L- phenylalanine, iV-(te/t-butoxycarbonyl)-L-serine, ⁇ /- ⁇ [(1,1 -dimethylethyl)oxy]carbonyl ⁇ - 3-(l,3-thiazol-4-
  • Boc-protected amino acid such as N
  • the free base form of a compound of Formula (I) may be prepared by any suitable method known to the art, including treatment of the salt with an inorganic or organic base, suitably an inorganic or organic base having a higher pKa than the free base form of the compound.
  • Reagents and conditions a) K 2 CO 3 or Et 3 N, CH 3 CN, THF, DMF, or DMSO; or Pd 2 (dba) 3 , BINAP, Cs 2 CO 3 , toluene; b) HCl, 1,4-dioxane; c) Et 3 N, HATU, CH 2 Cl 2 or CH 2 Cl 2 and MeOH; or Et 3 N, 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphorinane 2,4,6- trioxide, CH 2 Cl 2 ; d) HCl, 1,4-dioxane; or TFA, CH 2 Cl 2 .
  • Quadrupole LC-MS API-150a or Waters ZQ instruments.
  • the compound is analyzed using a reverse phase column, e.g., Thermo Aquasil/Aquasil C18, Acquity UPLC C18, Thermo Hypersil Gold eluted using an CH 3 CN and water gradient with a low percentage of an acid modifier such as 0.02% TFA or 0.1% formic acid.
  • a reverse phase column e.g., Thermo Aquasil/Aquasil C18, Acquity UPLC C18, Thermo Hypersil Gold eluted using an CH 3 CN and water gradient with a low percentage of an acid modifier such as 0.02% TFA or 0.1% formic acid.
  • Celite ® is a filter aid composed of acid- washed diatomaceous silica, and is a registered trademark of Manville Corp., Denver, Colorado.
  • Isolute ® is a functionalized silica gel based sorbent, and is a registered trademark of Biotage AB Corp., Sweden.
  • Nuclear magnetic resonance spectra were recorded at 400 MHz using a Bruker AVANCE 400 or Brucker DPX400 spectrometer.
  • CDCl 3 is deuteriochloroform
  • DMSO-D 6 is hexadeuteriodimethylsulfoxide
  • MeOD is tetradeuteriomethanol.
  • Biotage Initiator ® microwave reactor typically employing the high absorbance setting.
  • Cartridges or columns containing polymer based functional groups can be used as part of compound workup.
  • the "amine” columns or cartridges are used to neutralize or basify acidic reaction mixtures or products. These include NH2 Aminopropyl SPE-ed SPE Cartridges available from Applied Separations and diethylamino SPE cartridges available from United Chemical Technologies, Inc.
  • the reaction mixture was concentrated under a stream of nitrogen at 50 0 C, and the residue was taken up in DMF (1.0 mL).
  • the reaction mixture was heated to 100 0 C for 24 h following by increasing to 150 0 C for a further 53 h.
  • a solution of 1 M aq. HCl (2 mL) and EtOAc (2 mL) were added.
  • the layers were separated, and the aqueous layer was washed with EtOAc (2 x 1 mL).
  • the aqueous layer was made basic with 6 M aq. NaOH (1 mL) and extracted with EtOAc (4 x 1 mL).
  • the reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0 C.
  • Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL).
  • EtOAc 3 x 1 mL
  • the combined organic layers were concentrated under a stream of nitrogen at 50 0 C onto Isolute ® .
  • the vial was capped and the reaction mixture was heated in a Biotage Initiator ® microwave reactor to 140 0 C for 10 min. Water (5 mL) and CH 2 Cl 2 (2 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with CH 2 Cl 2 (3 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0 C. The crude product was dissolved in DMSO (1 mL) and purified by reverse phase
  • 1,1-dimethylethyl [(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3- pyrrolidinyl] carbamate
  • the vial was capped and the reaction mixture was heated in a Biotage Initiator ® microwave reactor to 140 0 C for 10 min. Water (5 mL) and CH 2 Cl 2 (2 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with CH 2 Cl 2 (3 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0 C.
  • Pd 2 (dba) 3 (27 mg, 0.029 mmol), BINAP (55 mg, 0.088 mmol), Cs 2 CO 3 (96 mg, 0.294 mmol) and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (71 mg, 0.381 mmol) were added to a 2 dram vial containing a solution of 2-bromo-l-benzothiophene-3- carbonitrile (70 mg, 0.294 mmol) in degassed toluene (1.5 mL) (degassed by bubbling argon through it for 10 min) under argon.
  • Pd 2 (dba) 3 (182 mg, 0.199 mmol), BINAP (372 mg, 0.597 mmol), Cs 2 CO 3 (649 mg, 1.991 mmol), and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (482 mg, 2.59 mmol) were added to a vial containing a solution of 2-bromo-4-(l-pyrrolidinyl)benzonitrile (500 mg, 1.991 mmol) in degassed toluene (10 mL) (degassed by bubbling argon through it for 10 min) under argon. The vial was capped, and the reaction mixture was heated at 100 0 C (bath temp) for 16.5 h.
  • Pd 2 (dba) 3 (141 mg, 0.154 mmol), BINAP (288 mg, 0.462o mmol), Cs 2 CO 3 (502 mg, 1.541 mmol), and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (373 mg, 2.003 mmol) were added to a 20 mL vial containing a solution of 2-bromo-5-(l- pyrrolidinyl)benzonitrile (387 mg, 1.541 mmol) in degassed toluene (8 mL) (degassed by bubbling argon through it for 10 min) under argon.
  • 1,1 -dimethylethyl ⁇ (35)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3- pyrrolidinyl ⁇ carbamate (659 mg, 1.849 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (10 mL, 40.0 mmol). The reaction mixture was stirred at RT for 45 min and then Et 2 O (50 mL) was added. The cloudy mixture was filtered and washed with Et 2 O (2 x 10 mL). The solid was set aside, and the residual product in the funnel and reaction flash was transferred to a 2 dram vial with MeOH.

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Abstract

Disclosed are 3-aminopyrrolidines of Formula (I) having pharmacological activity, pharmaceutical compositions containing them, and methods for the treatment of diseases mediated by the cathepsin C enzyme such as chronic obstructive pulmonary disease.

Description

Cathepsin C Inhibitors
FIELD OF THE INVENTION
The present invention relates to certain 3-aminopyrrolidines that are cathepsin C inhibitors, pharmaceutical compositions containing these compounds, and their use in the treatment of diseases mediated by the cathepsin C enzyme such as chronic obstructive pulmonary disease.
BACKGROUND OF THE INVENTION
Cathepsins are a family of enzymes included in the papain superfamily of cysteine proteases. Cathepsins B, C, F, H, K, L, S, V, and X have been described in the scientific literature. Cathepsin C is also known in the literature as Dipeptidyl Peptidase I or "DPPI." A number of recently published studies have begun to describe the role cathepsin C plays in certain inflammatory processes. See e.g. Adkison et al., The Journal of Clinical Investigation 109:363-371 (2002); Tran et al., Archives of Biochemistry and Biophysics 403:160-170 (2002); Thiele et al., The Journal of Immunology 158: 5200-5210 (1997);
Bidere et al., The Journal of Biological Chemistry 277: 32339-32347 (2002); Mabee et al., The Journal of Immunology 160: 5880-5885; McGuire et al., The Journal of Biological Chemistry, 268: 2458-2467; and Paris et al., FEBS Letters 369: 326-330 (1995). From these studies, it appears that cathepsin C is co-expressed in granules with certain serine proteases and functions to process the pro-forms of these proteases to active forms, which are then released from the granules of inflammatory cells recruited to sites of
inflammation. Once activated, these proteases have a number of functions including degradation of various extracellular matrix components, which together can propagate tissue damage and chronic inflammation.
For example, Chronic Obstructive Pulmonary Disease ("COPD") is a chronic inflammatory disease where cathepsin C appears to play a role. Chronic bronchitis and emphysema usually occur together in COPD patients. Chronic bronchitis is generally characterized by a chronic productive cough, whereas emphysema is generally
characterized by permanent enlargement of the airspaces distal to the terminal bronchioles and airway wall destruction.
Cigarette smoking is a significant risk factor for developing COPD. Exposure to cigarette smoke and other noxious particles and gases may result in chronic inflammation of the lung. In response to such exposure, inflammatory cells such as CD8+ Tcells, macrophages, and neutrophils are recruited to the area. These recruited inflammatory cells release proteases, which are believed to play a major role in the disease etiology by a number of mechanisms. Proteases believed to be involved in this process include the serine proteases neutrophil elastase ("NE"), cathepsin G, and proteinase 3, all released from neutrophils; granzymes A and B, released from cytotoxic T cells or natural killer cells; and chymases, released from mast cells. Cathepsin C appears to be involved in activating all of these enzymes.
Rheumatoid arthritis ("RA") is another chronic inflammatory disease where cathepsin C may play a role. Neutrophils are recruited to the site of joint inflammation and release cathepsin G, NE, and proteinase 3, which are believed to be responsible in part for cartilage destruction associated with RA (Hu, Y. and Pham, C. T. (2005) Arthritis Rheum 52: 2553-2558).
Other conditions where cathepsin C may play a role include osteoarthritis, asthma, and Multiple Sclerosis. See e.g. Matsui, K.; Yuyama, N.; Akaiwa, M.; Yoshida, N. L.; Maeda, M.; Sugita, Y.; Izuhara, K., Identification of an alternative splicing variant of cathepsin C/dipeptidyl-peptidase I, Gene. 293(1-2): 1-7, 2002 Jun 26; Wolters, P. J.; Laig- Webster, M.; Caughey, G. H., Dipeptidyl peptidase I cleaves matrix-associated proteins and is expressed mainly by mast cells in normal dog airways, American Journal of Respiratory Cell & Molecular Biology. 22(2): 183-90, 2000.
One approach to treating these conditions is to inhibit the activity of the serine proteases involved in the inflammatory process, especially NE activity. See e.g.,
Ohbayashi, "Neutrophil elastase inhibitors as treatment for COPD", Expert Opin. Investig. Drugs 11(7): 965-980 (2002); Shapiro, "Neutrophil Elastase: Path Clearer, Pathogen Killer, or Just Pathologic?", Am. J. Respir. Cell MoI. Biol. 26: 266-268 (2002). In light of the role cathepsin C plays in activating certain serine proteases, especially NE, it is desirable to prepare compounds that inhibit its activity, which thereby inhibit serine protease activity. Thus, there is a need to identify compounds that inhibit cathepsin C, which can be used in the treatment of a variety of conditions mediated by cathepsin C.
There are additional activities of cathepsin C that may also be related to disease etiology. Cathepsin C has been demonstrated to have a role in neutrophil migration in the development of aortic aneurysms by a mechanism which has not been clearly elucidated (Pagano, M. B. et al. (2007) PNAS 104: 2855-2860). Thus, disease processes that involve neutrophil migration, as well as proteolytic enzyme release can be modulated by cathepsin C inhibition. Also, cathepsin C is highly expressed in the lung epithelium where it may play a role in the processing of other enzymes not yet identified. Cathepsin C has also been reported to cleave kallikrein-4, which is believed to play a role in dental enamel maturation (Tye, C. E. et al. (2009) J. Dental Res. 88: 323-327). Finally, cathepsin C is itself released from cells and may play a direct role in the degradation of matrix proteins.
SUMMARY OF THE INVENTION
The present invention involves novel compounds according to Formula (I) or a pharmaceutically acceptable salt thereof:
Figure imgf000004_0001
wherein:
A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, (C3-Ce)cycloalkyl, heterocycloalkyl, hydroxyl, (Ci-C4)alkoxy, aryl, aryloxy, heteroaryl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl,
-SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said aryl, aryloxy, or heteroaryl is optionally substituted by -OCH2O- or is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, formyl, -CO(Ci-C4)alkyl, -CO2(C i-C4)alkyl, hydroxyl, (Ci-C4)alkoxy, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 5- or 6-membered heterocycloalkyl group;
R1 is hydrogen, (Ci-C4)alkyl, (C2-C5)alkenyl, (C2-C5)alkynyl, (C3-C5)cycloalkyl,
(C3-C4)cycloalkyl(Ci-C2)alkyl, cyano(Ci-C2)alkyl, hydroxy(Ci-C2)alkyl,
methoxy(Ci-C2)alkyl, aryl(Ci-C2)alkyl, or heteroaryl(Ci-C2)alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C2)alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms; and
R2 is hydrogen or methyl;
or R1 and R2 taken together with atoms through which they are connected form a
4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF3, cyano, (Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, hydroxyl, (Ci-C4)alkoxy, or (Ci-C4)alkylthio-; wherein said ring is optionally fused to a (C3-Cs)cycloalkyl ring.
The present invention is also directed to the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in the prevention, management or treatment of a respiratory or inflammatory disease, such as chronic obstructive pulmonary disease or rhinitis.
In a further aspect, this invention relates to a pharmaceutically acceptable formulation comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
DETAILED DESCRIPTION OF THE INVENTION
Terms and Definitions
As used herein, the term "alkyl" refers to a straight- or branched-chain
hydrocarbon radical having the specified number of carbon atoms. As used herein, the terms "(Ci-C4)alkyl" and "(Ci-C8)alkyl" refer to an alkyl group having at least 1 and up to 4 or 8 carbon atoms respectively. Examples of such branched or straight-chained alkyl groups useful in the present invention include, but are not limited to, methyl, ethyl, n -propyl, isopropyl, /? -butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, and branched analogs of the latter 3 normal alkanes. As used herein, the term "alkenyl" refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1 and up to 3 carbon- carbon double bonds. Examples include ethenyl and propenyl.
As used herein, the term "alkynyl" refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1 and up to 3 carbon- carbon triple bonds. Examples include ethynyl and propynyl.
As used herein, the term "cycloalkyl" refers to a non-aromatic, saturated, cyclic hydrocarbon ring containing the specified number of carbon atoms. The term
"(C3-C8)cycloalkyl" refers to a non-aromatic cyclic hydrocarbon ring having from three to eight ring carbon atoms. Exemplary "(C3-C8)cycloalkyl" groups useful in the present invention include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
As used herein, the term "heterocycloalkyl" refers to a group or moiety comprising a non-aromatic, monovalent monocyclic or bicyclic radical, which is saturated or partially unsaturated, containing 3 to 10 ring atoms, which includes 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, and which may be unsubstituted or substituted by one or more of the substituents defined herein. Illustrative examples of heterocycloalkyls include, but are not limited to, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3- dithianyl, hexahydro-lH-l,4-diazepinyl, azabicylo[3.2.1]octyl, azabicylo[3.3.1]nonyl, azabicylo[4.3.0]nonyl, oxabicylo[2.2.1]heptyl, and 1,5,9-triazacyclododecyl.
"Alkoxy" means an alkyl radical containing the specified number of carbon atoms attached through an oxygen linking atom. The term "(Ci-C4)alkoxy" refers to a straight- or branched-chain hydrocarbon radical having at least 1 and up to 4 carbon atoms attached through an oxygen linking atom. Exemplary "(Ci-C4)alkoxy" groups useful in the present invention include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n- butoxy, 5-butoxy, and t-butoxy.
"Aryl" refers to optionally substituted monocyclic or fused bicyclic groups having
6 to 14 carbon atoms and having at least one aromatic ring that complies with Ηϋckel's Rule. Examples of "aryl" groups are phenyl, naphthyl, indenyl, dihydroindenyl, anthracenyl, phenanthrenyl, and the like. Preferably aryl refers to optionally substituted phenyl.
"Aryloxy" means an aryl radical attached through an oxygen linking atom.
Exemplary "aryloxy" groups useful in the present invention include, but are not limited to, phenyloxy (or phenoxy) and naphthyloxy.
"Heteroaryl" means an optionally substituted aromatic monocyclic ring or fused bicyclic ring system wherein at least one ring complies with Hϋckel's Rule, has the specified number of ring atoms, and that ring contains at least one heteroatom selected from N, O, and/or S. Examples of 5-membered "heteroaryl" groups include furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, and isothiazolyl. Examples of 6-membered
"heteroaryl" groups include oxo-pyridyl, pyridinyl (or pyridyl), pyridazinyl, pyrazinyl, and pyrimidinyl. Examples of 6,6-fused "heteroaryl" groups include quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1,6- naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, and pteridinyl. Examples of 6,5- fused "heteroaryl" groups include benzofuranyl, benzothienyl, benzimidazolyl, benzthiazolyl, indolizinyl, indolyl, isoindolyl, and indazolyl.
For the avoidance of doubt, all bicyclic ring systems may be attached at any suitable position on either ring.
"Optionally substituted" indicates that a group, such as alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, aryl, aryloxy, or heteroaryl, may be unsubstituted, or the group may be substituted with one or more substituent(s) as defined. In the case where groups may be selected from a number of alternative groups the selected groups may be the same or different.
The term "independently" means that where more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.
As used herein, "halogen" or "halo" refers to F, Cl, Br, or I.
Suitably, A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, (C3-Ce)cycloalkyl, heterocycloalkyl, hydroxyl, (Ci-C4)alkoxy, aryl, aryloxy, heteroaryl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said aryl, aryloxy, or heteroaryl is optionally substituted by -OCH2O- or is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, formyl, -CO(Ci-C4)alkyl, -CO2(C i-C4)alkyl, hydroxyl, (Ci-C4)alkoxy, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 5- or 6-membered heterocycloalkyl group.
In another embodiment, A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, aryl, aryloxy, heteroaryl, cyano,
-CO2(Ci-C4)alkyl, -CONH(Ci-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(C i-C4)alkyl, -SO2NH(Ci-C4)alkyl, -SO2N(C i-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said aryl, aryloxy, or heteroaryl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy.
In another embodiment, A represents a 5-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3- oxathianyl, 1,3-dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1,6- naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano,
-CO2(Ci-C4)alkyl, -CONH(Ci-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(C i-C4)alkyl, -SO2NH(Ci-C4)alkyl, -SO2N(C i-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 10-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group. In selected embodiments, A represents thienyl, 5-carboxyethylthienyl, or 3-chloroisothiazolyl.
In another embodiment, A represents a 5-membered aromatic ring containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is fused to an aromatic or non-aromatic carbocyclic ring moiety to form an 8- to 10- membered bicyclic group, wherein said aromatic carbocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group. In selected
embodiments, A represents benzothienyl, 5,6-dihydro-4H-cyclopenta[δ]thienyl, 4,5,6,7- tetrahydro- 1 -benzothienyl, 4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'-[l ,3]dioxolanyl, or 5,6,7,8-tetrahydro-4H-cyclohepta[δ]thienyl. In further selected embodiments, A represents indolyl or JV-methylindolyl. In a specific embodiment, A represents
JV-methylindolyl.
In a further embodiment, A represents a 6-membered aromatic ring optionally containing one or two nitrogen atoms, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1,4- dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl,
-SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8- naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 11-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy. In selected embodiments, A represents pyridinyl, pyrazinyl, or quinolinyl.
In yet another embodiment, A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (d-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3- dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1, 3 -dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl,
-SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8- naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy. In selected embodiments, A represents phenyl, 4-fluorophenyl, 4-bromophenyl, 4,5-difluorophenyl, 4- trifluoromethylphenyl, 5-trifluoromethylphenyl, 4-biphenyl, 5-biphenyl, 4'-fluoro-4- biphenyl, 3'-nitro-4'-methyl-4-biphenyl, 3'-acetyl-4-biphenyl, 3'-carboxyethyl-4-biphenyl, 4'-methoxy-4-biphenyl, 3 '-fluoro-4'-methoxy-4-biphenyl, 4-(l ,3-benzodioxol-5- yl)phenyl, 3'-amino-4-biphenyl, 4-pyridin-4-ylphenyl, 4-pyrimidin-2-ylphenyl, A- pyrimidin-5-ylphenyl, 4-furan-3-ylphenyl, 4-thien-2-ylphenyl, 4-thien-3-ylphenyl, 4-(5- formyl-2-thienyl)phenyl, 4-thiazol-2-ylphenyl, 4-quinolin-3-ylphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 5-methoxyphenyl, 4-phenoxyphenyl, 4,5-dimethoxyphenyl, 3- aminophenyl, 5-dimethylaminophenyl, 4-pyrrolidinylphenyl, 5-pyrrolidinylphenyl, or 4-nitrophenyl.
In yet another embodiment, A represents a thienyl, 4,5,6,7-tetrahydro-l- benzothienyl, isothiazolyl, indolyl, pyridinyl, pyrazinyl, quinolinyl, or phenyl ring optionally substituted one to two times, independently, by fluoro, chloro, bromo, methyl, trifluoromethyl, carboxyethyl, phenyl, 4-fluorophenyl, pyrimidinyl, furanyl, thienyl, thiazolyl, methoxy, phenyloxy, amino, dimethylamino, or nitro.
Suitably, R1 is hydrogen, (Ci-C4)alkyl, (C2-C5)alkenyl, (C2-C5)alkynyl,
(C3-C5)cycloalkyl, (C3-C4)cycloalkyl(Ci-C2)alkyl, cyano(Ci-C2)alkyl,
hydroxy(Ci-C2)alkyl, methoxy(Ci-C2)alkyl, aryl(Ci-C2)alkyl, or heteroaryl(Ci-C2)alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C2)alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms.
In another embodiment, R1 is hydrogen, (Ci-C4)alkyl or heteroaryl(Ci-C2)alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C2)alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms. In a further embodiment, R1 is (Ci-C4)alkyl or thienyl(Ci-C2)alkyl. In selected embodiments, R1 is methyl, ethyl, n-propyl, isopropyl, or 2-thienylmethyl. In a selected embodiment, R1 is methyl. In another selected embodiment, R1 is ethyl. In yet another selected embodiment, R1 is 2-thienylmethyl.
Suitably, R2 is hydrogen or methyl. In a selected embodiment, R2 is hydrogen. In another embodiment, R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF3, cyano, (Ci-C4)alkyl, amino, (Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, hydroxyl, (Ci-C4)alkoxy, or (Ci-C4)alkylthio-; wherein said ring is optionally fused to a (C3-Cs)cycloalkyl ring. In a further embodiment, R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF3, cyano, methyl, amino, hydroxyl, methoxy, or methylthio-; wherein said ring is optionally fused to a cyclopropyl ring. In a further embodiment, R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF3, cyano, methyl, methoxy, or methylthio-. In a further embodiment, R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F. In a selected embodiment, R1 and R2 taken together represent -CH2CH2-. In another selected embodiment, R1 and R2 taken together represent -CH2CH2CH2-. In another selected embodiment, R1 and R2 taken together represent -CH2CHFCH2-. In another selected embodiment, R1 and R2 taken together represent -CH2CF2CH2-. In another selected embodiment, R1 and R2 taken together represent -CH2CH(CH3)CH2-. In another selected embodiment, R1 and R2 taken together represent -CHOHCH2CH2-. In another selected embodiment, R1 and R2 taken together represent -CH2CH2CH2CH2-. One particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (d-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3- oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1,6- naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano,
-CO2(Ci-C4)alkyl, -CONH(Ci-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(C i-C4)alkyl, -SO2NH(Ci-C4)alkyl, -SO2N(C i-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy; and
R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF3, cyano, methyl, methoxy, or methylthio-.
Another particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
A represents a 5-membered aromatic ring containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is fused to an aromatic or non-aromatic carbocyclic ring moiety to form an 8- to 10-membered bicyclic group, wherein said aromatic carbocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group; and
R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF3, cyano, methyl, methoxy, or methylthio-.
Another particular embodiment of the invention is a compound of Formula (I) or a pharmaceutically acceptable salt thereof wherein:
A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl,
-CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(Ci-C4)alkyl, -SO2NH(Ci-C4)alkyl,
-SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, or pyrimidinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
R1 is (Ci-C4)alkyl or thienyl(Ci-C2)alkyl; and
R2 is hydrogen.
Specific compounds exemplified herein are:
N1-[(35)-l-(3-chloro-4-cyano-5-isothiazolyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-alaninamide;
N1 - [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] glycinamide;
N1-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-L- alaninamide;
N1 - [(35)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-valinamide;
(25)-2-amino-JV-[(3iS)- 1 -(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]butanamide;
N1 - [(35)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-norvalinamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-leucinamide; N- [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-phenylalaninamide;
N1-[(35)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(4-cyano-3-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
ethyl 5-[(35)-3-(L-alanylamino)- 1 -pyrrolidinyl]-4-cyano-2-thiophenecarboxylate; N1- [(35)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-serinamide;
N1 -[(3S)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(1,3 -thiazol-4-yl)-L- alaninamide;
(25)-2-amino-3 -cyano-iV- [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl]- propanamide;
N1- [(3S)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(l,3-thiazol-2-yl)phenyl]-3-pyrrolidinyl}- butanamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3-pyrrolidinyl}- butanamide;
N1- [(3S)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -cyclopropyl-L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-4-pentynamide;
N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- butanamide;
N1-[(35)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1- [(3S)- l-(4-bromo-2-cyanophenyl)-3 -pyrrolidinyl] -L-alaninamide;
N1-[(35)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1 - [(3S)- 1 -(3 -cyano- 1 -methyl- lH-indol-2-yl)-3 -pyrrolidinyl] -L-alaninamide;
N1-[(35)-l-(3-cyano-l-methyl-lH-indol-2-yl)-3-pyrrolidinyl]-3-(2-thienyl)-L- alaninamide;
N1 - [(3S)- 1 -(2-cyano-4-fluorophenyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]butanamide;
N1- [(3S)- l-(2-cyano-4-fluorophenyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide;
N1 -[(3S)- 1 -(3 -cyano-4'-fluoro-4-biphenylyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L- alaninamide;
N1 - [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(3 -thienyl)-L-alaninamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-norleucinamide; (25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopropyl- ethanamide;
N1-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-3-(2- thienyl)-L-alaninamide;
(25)-2-amino-JV-[(35)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3- pyrrolidinyljbutanamide;
N1 - [(35)- 1 -(3 -cyano-2-pyrazinyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-Λ/-[(35)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]butanamide;
N1 - [(35)- 1 -(3 -cyano-2-pyrazinyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide; (25)-2-amino-JV-[(35)- 1 -(3-cyano-2-pyridinyl)-3-pyrrolidinyl]butanamide;
N1 - [(35)- 1 -(3 -cyano-2-pyridinyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide; N1-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]-L-alaninamide;
N1- [(35)- l-(2-cyano-3-thienyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide; (25)-2-amino-iV-[(35)- 1 -(3-cyano-4-biphenylyl)-3-pyrrolidinyl]butanamide;
N1- {(35)- 1 -[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide; (25)-2-amino-N-{(35)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- butanamide;
N1- {(35)- 1 -[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1 - [(35)- 1 -(2-cyanophenyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyanophenyl)-3-pyrrolidinyl]butanamide;
N1-[(35)-l-(2-cyanophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1 - [(35)- 1 -(2-cyano-4, 5 -difluorophenyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]butanamide; N1-[(35)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopentyl- ethanamide;
(25)-2-amino-N-[(35)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]butanamide; N1- [(3S)- 1 -[2-cyano-4-(phenyloxy)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1 - [(35)- 1 -(3 -cyano-4-pyridinyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide; N1 - [(3S)- 1 -(2-cyano-3 -pyridinyl)-3 -pyrrolidinyl] -L-alaninamide;
N1-[(3S)-l-(2-cyano-3-pyridinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1 - [(3S)- 1 -(3 -cyano-2-quinolinyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]butanamide;
N1-[(35)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide; N1-[(35)-l-(3-cyano-2-thienyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -N2-methyl-L- alaninamide;
N1- {(35)- 1 -[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(methyloxy)phenyl]-3- pyrrolidinyl}butanamide;
N1- [(3S)- 1 -[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- butenamide;
(25)-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-2-azetidine- carboxamide;
N- [(3S)- 1 -(3 -cyano-4'-fluoro-4-biphenylyl)-3 -pyrrolidinyl] -L-prolinamide;
N1-{(35)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-L-alaninamide-(ii; N1- [(3S)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide-^; N1- [(3S)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
N1- [(3S)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(4-pyridinyl)phenyl]-3- pyrrolidinyl}butanamide;
N1- [(3S)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide; N1- [(3S)- 1 -[2-cyano-4-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(2S)-N- [(3S)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -2- azetidinecarboxamide; (25)-N-[(35)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-2-azetidinecarboxamide; (2S)-N-[QS)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-2- azetidinecarboxamide;
N-[QS)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-L- prolinamide;
(2S)-N-[QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-2- azetidinecarboxamide;
N- [QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-L-prolinamide;
(2S)-N- [QS)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -2- azetidinecarboxamide;
N- [QS)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
(2S)-N-[QS)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-2- piperidinecarboxamide;
(2S)-N-[QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-2- piperidinecarboxamide;
(4S)-N-[QS)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-4- fluoro-L-prolinamide;
(4i?)-N-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-4- fluoro-L-prolinamide;
N-[(35)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-4,4- difluoro-L-prolinamide;
(3i?)-N-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-3- hydroxy-L-prolinamide;
N-[(3S)-l-(3-cyano-5,6-dihydro-4H-cyclopenta[ό]thien-2-yl)-3-pyrrolidinyl]-L- prolinamide;
N-[(35)-l-(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thien-2-yl)-3- pyrrolidinyl] -L-prolinamide;
(25)-N-[(35)-l-(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thien-2-yl)-3- pyrrolidinyl]-2-azetidinecarboxamide;
N- [QS)- 1 -(3-cyano-4'-methyl-3'-nitro-4-biphenylyl)-3-pyrrolidinyl]-L- prolinamide;
ethyl 3'-cyano-4'-[(35)-3-(L-prolylamino)- 1 -pyrrolidinyl]-3-biphenylcarboxylate; N- [(3S)- 1 -[4-(1 ,3-benzodioxol-5-yl)-2-cyanophenyl]-3-pyrrolidinyl} -L- prolinamide;
N- [(3S)- 1 -[2-cyano-4-(5-formyl-2-thienyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
Λ/-[(35)-l-(3'-acetyl-3-cyano-4-biphenylyl)-3-pyrrolidinyl]-L-prolinamide;
N- [(3S)- 1 -[2-cyano-4-(3-quinolinyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
N- [(3S)- 1 -[3-cyano-4'-(methyloxy)-4-biphenylyl]-3-pyrrolidinyl} -L-prolinamide;
N- [(3S)- 1 - [3 -cyano-3 '-fluoro-4'-(methyloxy)-4-biphenylyl] -3 -pyrrolidinyl} -L- prolinamide;
N- [(3 S)- 1 -(3 '-amino-3-cyano-4-biphenylyl)-3 -pyrrolidinyl] -L-prolinamide;
JV-[(3iS)- 1 -(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-L-prolinamide;
Λ/-[(35)- 1 -(3-cyano- lH-indol-2-yl)-3-pyrrolidinyl]-L-prolinamide;
N-[(35)- 1 -(3-cyano-4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'-[ 1 ,3]dioxolan]-2- yl)-3-pyrrolidinyl]-L-prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-4-methyl-L- prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide;
(4S)-N-[(3S)- 1 -(3 -cyano-4-biphenylyl)-3 -pyrrolidinyl] -4-fluoro-L-prolinamide;
(45)-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide;
(4S)-N- [(3S)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -4-fluoro-L- prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -4- methyl-L-prolinamide;
JV-[(3IS)- 1 -(3-cyano- 1 -benzothien-2-yl)-3-pyrrolidinyl]-L-prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -benzothien-2-yl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -4-methyl-L- prolinamide;
(4S)-N-[(3S)- 1 -(3 -cyano-4'-fluoro-4-biphenylyl)-3 -pyrrolidinyl] -4-methyl-L- prolinamide; (4S)-N- [(3S)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -4-methyl-L- prolinamide; and
(2S)-N- [(3S)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -2- piperidinecarboxamide.
The invention also includes various isomers of the compounds of Formula (I) and mixtures thereof. "Isomer" refers to compounds that have the same composition and molecular weight but differ in physical and/or chemical properties. The structural difference may be in constitution (geometric isomers) or in the ability to rotate the plane of polarized light (stereoisomers). The compounds according to Formula (I) contain two or more asymmetric centers, also referred to as chiral centers, and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomeric forms, or as mixtures thereof. All such isomeric forms are included within the present invention, including mixtures thereof.
Chiral centers may also be present in a substituent such as an alkyl group. Where the stereochemistry of a chiral center present in Formula (I), or in any chemical structure illustrated herein, is not specified the structure is intended to encompass any stereoisomer and all mixtures thereof. Thus, compounds according to Formula (I) containing two or more chiral centers may be used as racemic mixtures, enantiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers.
Individual stereoisomers of a compound according to Formula (I) which contain two or more asymmetric centers may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer-specifϊc reagent, for example by enzymatic oxidation or reduction; or (3) by gas-liquid or liquid chromatography in a chiral environment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent. The skilled artisan will appreciate that where the desired stereoisomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation. "Enantiomerically enriched" refers to products whose enantiomeric excess is greater than zero. For example, enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee, and greater than 90% ee.
"Enantiomeric excess" or "ee" is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%).
"Enantiomerically pure" means products whose enantiomeric excess is 99% ee or greater.
The invention also includes various deuterated forms of the compounds of Formula (I). Each available hydrogen atom attached to a carbon atom may be independently replaced with a deuterium atom. A person of ordinary skill in the art will know how to synthesize deuterated forms of the compounds of Formula (I). For example, α-deuterated α-amino acids are commercially available or may be prepared by conventional techniques (see for example: Elemes, Y. and Ragnarsson, U. J. Chem. Soc, Perkin Trans. 1, 1996, 6, 537-40). Employing such compounds according to Scheme 1 below will allow for the preparation of compounds of Formula (I) in which the hydrogen atom at the chiral center baring R1 is replaced with a deuterium atom. Similarly, α-amino acids in which deuterium atoms have been incorporated into the sidechains are commercially available or may be prepared by conventional techniques. Employing such compounds according to Scheme 1 below will allow for the preparation of compounds of Formula (I) in which deuterium atoms have been incorporated in R1.
The term "solvate" refers to a complex of variable stoichiometry formed by a solute and a solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include, without limitation, water, ethanol and acetic acid. Solvates wherein water is the solvent molecule are typically referred to as "hydrates". Hydrates include compositions containing stoichiometric amounts of water, as well as compositions containing variable amounts of water. Solvates, particularly hydrates, of the compounds of Formula (I) and salts thereof, are within the scope of the invention.
When a disclosed compound or its salt is named or depicted by structure, it is to be understood that the compound or salt, including solvates (particularly, hydrates) thereof, may exist in crystalline forms, non-crystalline forms or a mixture thereof. The compound or salt, or solvates (particularly, hydrates) thereof, may also exhibit polymorphism (i.e. the capacity to occur in different crystalline forms). These different crystalline forms are typically known as "polymorphs." It is to be understood that when named or depicted by structure, the disclosed compound, or solvates (particularly, hydrates) thereof, also include all polymorphs thereof. 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. One of ordinary skill in the art will appreciate that different polymorphs may be produced, for example, by changing or adjusting the conditions used in crystallizing/recrystallizing the compound.
Because of their potential use in medicine, the salts of the compounds of Formula (I) are preferably pharmaceutically acceptable. Suitable pharmaceutically acceptable salts can include acid or base addition salts.
As used herein, the term "pharmaceutically acceptable" means a compound which is suitable for pharmaceutical use. Salts and solvates (e.g. hydrates and hydrates of salts) of the compounds of the invention which are suitable for use in medicine are those wherein the counterion or associated solvent is pharmaceutically acceptable. However, salts and solvates having non-pharmaceutically acceptable counterions or associated solvents are within the scope of the present invention, for example, for use as
intermediates in the preparation of other compounds of the invention and their salts and solvates.
Compounds of Formula (I) have one or more nitrogen(s) basic enough to form pharmaceutically acceptable acid addition salts by treatment with a suitable acid. Suitable acids include pharmaceutically acceptable inorganic acids and pharmaceutically acceptable organic acids. Representative pharmaceutically acceptable acid addition salts include acetate, aspartate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, formate, fumarate, galacturonate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexanoate, hydrobromide, hydrochloride,
hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate,
phosphate/diphosphate, polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate, and tosylate salts.
Other iterations of compounds of the invention have an acidic functional group, one acidic enough to form salts. Representative salts include pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; carbonates and bicarbonates of a pharmaceutically acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; pharmaceutically acceptable organic 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, cyclohexylamine, triethanolamine, choline, arginine, lysine, and histidine.
Other non-pharmaceutically acceptable salts, e.g. trifluoroacetate, may be used, for example in the isolation of compounds of the invention, and are included within the scope of this invention.
The invention includes within its scope all possible stoichiometric and non- stoichiometric forms of the salts of the compounds of Formula (I).
It will be appreciated by those skilled in the art that certain protected derivatives of compounds of Formula (I), which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolized in the body to form compounds of the invention which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". Further, certain compounds of the invention may act as prodrugs of other compounds of the invention. All protected derivatives and prodrugs of compounds of the invention are included within the scope of the invention. Examples of suitable pro-drugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31, pp 306 - 316 and in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as "pro-moieties", for example as described by H. Bundgaard in "Design of Prodrugs" (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within compounds of the invention. Preferred "pro- moieties" for compounds of the invention include: ester, carbonate ester, hemi-ester, phosphate ester, nitro ester, sulfate ester, sulfoxide, amide, carbamate, azo-, phosphamide, glycoside, ether, acetal, and ketal derivatives of the compounds of Formula (I).
The compounds of the invention inhibit the cathepsin C enzyme and can be useful in the treatment of conditions wherein the underlying pathology is (at least in part) attributable to cathepsin C involvement or in conditions wherein cathepsin C inhibition offers some clinical benefit even though the underlying pathology is not (even in part) attributable to cathepsin C involvement. Examples of such conditions include COPD, rheumatoid arthritis, osteoarthritis, asthma, and multiple sclerosis. Accordingly, in another aspect the invention is directed to methods of treating such conditions.
The methods of treatment of the invention comprise administering an effective amount of a compound of the invention to a patient in need thereof.
As used herein, "treatment" in reference to a condition means: (1) the amelioration or prevention of the condition being treated or one or more of the biological
manifestations of the condition being treated, (2) the interference with (a) one or more points in the biological cascade that leads to or is responsible for the condition being treated or (b) one or more of the biological manifestations of the condition being treated, or (3) the alleviation of one or more of the symptoms or effects associated with the condition being treated.
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. An "effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician. Furthermore, the term
"therapeutically effective amount" means any amount which, as compared to a
corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function.
As used herein, "patient" refers to a human or 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 amount administered and 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 particular route of administration chosen, 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 range from 1 mg to 1000 mg.
The invention includes the use of compounds of the invention for the preparation of a composition for treating or ameliorating diseases mediated by the cathepsin C enzyme in a subject in need thereof, wherein the composition comprises a mixture of one or more of the compounds of the invention and an optional pharmaceutically acceptable excipient.
The invention further includes the use of compounds of the invention as an active therapeutic substance, in particular in the treatment of diseases mediated by the cathepsin C enzyme. Specifically, the invention includes the use of compounds of the invention in the treatment of COPD, rheumatoid arthritis, osteoarthritis, asthma, and multiple sclerosis.
In another aspect, the invention includes the use of compounds of the invention in the manufacture of a medicament for use in the treatment of the above disorders.
Compositions
The compounds of the invention will normally, but not necessarily, be formulated into a pharmaceutical composition prior to administration to a patient. Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of the invention and a pharmaceutically acceptable excipient.
The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein an effective amount of a compound of the invention can be extracted and then given to the patient such as with powders, syrups, and solutions for injection. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains an 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. Conversely, the pharmaceutical compositions of the invention typically contain more than one pharmaceutically acceptable excipient. However, in certain embodiments, the pharmaceutical compositions of the invention contain one pharmaceutically acceptable excipient.
As used herein, "pharmaceutically acceptable excipient" means a material, composition or vehicle involved in giving form or consistency to the composition and which is safe when administered to a patient. 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 compounds 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, elixirs, 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 aerosols 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 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, sweeteners, flavoring agents, flavor masking agents, coloring agents, anti-caking agents, humectants, 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 an 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, magnesium stearate, calcium stearate, and talc.
In another aspect, the invention is directed to a dosage form adapted for administration to a patient by inhalation. For example, the compound of the invention may be inhaled into the lungs as a dry powder, an aerosol, a suspension, or a solution.
Dry powder compositions for delivery to the lung by inhalation typically comprise a compound of the invention as a finely divided powder together with one or more pharmaceutically acceptable excipients as finely divided powders. Pharmaceutically acceptable excipients particularly suited for use in dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides.
The dry powder may be administered to the patient via a reservoir dry powder inhaler (RDPI) having a reservoir suitable for storing multiple (un-metered doses) of medicament in dry powder form. RDPIs typically include a means for metering each medicament dose from the reservoir to a delivery position. For example, the metering means may comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation.
Alternatively, the dry powder may be presented in capsules (e.g. gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPI). MDPIs are inhalers wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple defined doses (or parts thereof) of medicament. When the dry powder is presented as a blister pack, it comprises multiple blisters for containment of the medicament in dry powder form. The blisters are typically arranged in regular fashion for ease of release of the medicament therefrom. For example, the blisters may be arranged in a generally circular fashion on a disc-form blister pack, or the blisters may be elongate in form, for example comprising a strip or a tape. Each capsule, cartridge, or blister may, for example, contain between 20μg-10mg of the compound of the invention.
Aerosols may be formed by suspending or dissolving a compound of the invention in a liquified propellant. Suitable propellants include halocarbons, hydrocarbons, and other liquified gases. Representative propellants include: trichlorofluoromethane
(propellant 11), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane
(propellant 114), tetrafluoroethane (HFA- 134a), 1,1-difluoroethane (HFA- 152a), difluoromethane (HFA-32), pentafluoroethane (HFA- 12), heptafluoropropane (HFA- 227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane, and pentane. Aerosols comprising a compound of the invention will typically be administered to a patient via a metered dose inhaler (MDI). Such devices are known to those skilled in the art.
The aerosol may contain additional pharmaceutically acceptable excipients typically used with multiple dose inhalers such as surfactants, lubricants, cosolvents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
Suspensions and solutions comprising a compound of the invention may also be administered to a patient via a nebulizer. The solvent or suspension agent utilized for nebulization may be any pharmaceutically acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g., ethanol, isopropylalcohol, glycerol, propylene glycol,
polyethylene glycol, etc. or mixtures thereof. Saline solutions utilize salts which display little or no pharmacological activity after administration. Both organic salts, such as alkali metal or ammonium halogen salts, e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc. may be used for this purpose.
Other pharmaceutically acceptable excipients may be added to the suspension or solution. The compound of the invention may be stabilized by the addition of an inorganic acid, e.g., hydrochloric acid, nitric acid, sulfuric acid and/or phosphoric acid; an organic acid, e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc., a complexing agent such as EDTA or citric acid and salts thereof; or an antioxidant such as antioxidant such as vitamin E or ascorbic acid. These may be used alone or together to stabilize the compound of the invention. Preservatives may be added such as benzalkonium chloride or benzoic acid and salts thereof. Surfactant may be added particularly to improve the physical stability of suspensions. These include lecithin, disodium dioctylsulphosuccinate, oleic acid and sorbitan esters. Methods of Preparation.
The compounds of Formula (I) may be obtained by using synthetic procedures illustrated in the Schemes below or by drawing on the knowledge of a skilled organic chemist. The synthesis provided in these Schemes are applicable for producing compounds of the invention having a variety of different R1 and R2 groups employing appropriate precursors, which are suitably protected if need be, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, where needs be, and then affords compounds of the nature generally disclosed. While the Schemes are shown with compounds only of Formula (I), they are illustrative of processes that may be used to make the compounds of the invention.
Compounds names were generated using the software naming program
ACD/Name Pro V6.02 available from Advanced Chemistry Development, Inc., 110 Yonge Street, 14th Floor, Toronto, Ontario, Canada, M5C 1T4 (http://www.acdlabs.com/).
As shown in Scheme 1, the compounds of Formula (I) can be prepared in a multi- step sequence starting from a protected 3-aminopyrrolidine, such as the commercially available 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate. Coupling with an appropriate optionally substituted cyano-A-ring wherein X is a leaving group, such as a halogen, using an appropriate base, such as K2CO3 or Et3N, in an appropriate solvent, such as CH3CN, THF, DMF, or DMSO, or with an appropriate catalyst, such as Pd2(dba)3, with an appropriate ligand, such as BINAP, and an appropriate base, such as Cs2CO3, in an appropriate solvent, such as toluene, is followed by Boc deprotection with an appropriate reagent, such as HCl, in an appropriate solvent, such as 1,4-dioxane. Coupling of the liberated amine with an appropriate Boc-protected amino acid, such as N-(tert-butoxy- carbonyl)glycine, iV-(te/t-butoxycarbony l)-L-alanine, (2S)-2-( {[(1,1 -dimethylethyl)- oxy]carbonyl}-amino)butanoic acid, iV-(te/t-butoxycarbonyl)-L-valine, N-(tert-butoxy- carbonyl)-L-norvaline, iV-(tert-butoxycarbonyl)-L-leucine, iV-(te/t-butoxycarbonyl)-L- phenylalanine, iV-(te/t-butoxycarbonyl)-L-serine, Λ/-{[(1,1 -dimethylethyl)oxy]carbonyl} - 3-(l,3-thiazol-4-yl)-L-alanine, (25)-3-cyano-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)propanoic acid, Λ/-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)-L-alanine, N- {[(1,1 -dimethylethyl)oxy]carbonyl} -3-(3-thienyl)-L-alanine, 3-cyclopropyWV- {[(1 , 1 - dimethylethyl)oxy]-carbonyl} -L-alanine, (2S)-2-( {[(1 , 1 -dimethyl-ethyl)oxy]carbonyl} - amino)-4-pentynoic acid, iV-(te/t-butoxycarbonyl)-L-norleucine, (25)-cyclopropyl({[(l,l- dimethylethyl)oxy] -carbonyl} amino)ethanoic acid, (25)-cyclopentyl( {[(1,1- dimethylethyl)oxy]-carbonyl} amino)ethanoic acid, Λ/-(te/t-butoxycarbonyl)-JV-methyl-L- alanine, (2S)- 1 - { [( 1 , 1 -dimethylethyl)oxy]carbonyl} -2-azetidinecarboxylic acid,
Λ/-(tert-butoxycarbonyl)-L-proline, Λ/-(tert-butoxycarbonyl)-L-alanine-<ii , or
JV-(te/t-butoxycarbonyl)-L-alanine-<i4 with an appropriate coupling reagent, such as HATU or 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphorinane 2,4,6-trioxide, and an appropriate base, such as Et3N, in an appropriate solvent or solvents, such as CH2Cl2 or CH2Cl2 and MeOH is followed by Boc deprotection with an appropriate reagent, such as HCl or TFA, in an appropriate solvent, such as 1,4-dioxane or CH2Cl2, resulting in the formation of the desired compounds of Formula (I), which may be isolated as the corresponding salt form or converted to the free base. The free base form of a compound of Formula (I) may be prepared by any suitable method known to the art, including treatment of the salt with an inorganic or organic base, suitably an inorganic or organic base having a higher pKa than the free base form of the compound.
Scheme 1
Figure imgf000032_0001
Reagents and conditions: a) K2CO3 or Et3N, CH3CN, THF, DMF, or DMSO; or Pd2(dba)3, BINAP, Cs2CO3, toluene; b) HCl, 1,4-dioxane; c) Et3N, HATU, CH2Cl2 or CH2Cl2 and MeOH; or Et3N, 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphorinane 2,4,6- trioxide, CH2Cl2; d) HCl, 1,4-dioxane; or TFA, CH2Cl2. SYNTHETIC EXAMPLES
The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. All temperatures are given in degrees Celsius, all solvents are highest available purity and all reactions run under anhydrous conditions in an argon (Ar) or nitrogen (N2) atmosphere where necessary.
Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography. Both flash and gravity chromatography were carried out on E. Merck Kieselgel 60 (230-400 mesh) silica gel. The CombiFlash® system used for purification in this application was purchased from Isco, Inc. CombiFlash® purification was carried out using prepacked silica gel columns, a detector with UV wavelength at 254 nm and a variety of solvents or solvent combinations. Preparative HPLC was performed using a Gilson Preparative System with variable wavelength UV detection or an Agilent Mass Directed AutoPrep (MDAP) system with both mass and variable wavelength UV detection. A variety of reverse phase columns, e.g., Luna 5u
C 18(2) 10OA, SunFire C 18, XBridge C18 were used in the purification with the choice of column support dependent upon the conditions used in the purification. The compounds are eluted using a gradient of CH3CN and water. Neutral conditions used an CH3CN and water gradient with no additional modifier, acidic conditions used an acid modifier, usually 0.1% TFA (added to both the CH3CN and water) and basic conditions used a basic modifier, usually 0.1% NH4OH (added to the water). Analytical HPLC was run using an Agilent system with variable wavelength UV detection using reverse phase
chromatography with an CH3CN and water gradient with a 0.05 or 0.1% TFA modifier (added to each solvent). LC-MS was determined using either a PE Sciex Single
Quadrupole LC-MS API-150a, or Waters ZQ instruments. The compound is analyzed using a reverse phase column, e.g., Thermo Aquasil/Aquasil C18, Acquity UPLC C18, Thermo Hypersil Gold eluted using an CH3CN and water gradient with a low percentage of an acid modifier such as 0.02% TFA or 0.1% formic acid.
Celite® is a filter aid composed of acid- washed diatomaceous silica, and is a registered trademark of Manville Corp., Denver, Colorado. Isolute® is a functionalized silica gel based sorbent, and is a registered trademark of Biotage AB Corp., Sweden. Nuclear magnetic resonance spectra were recorded at 400 MHz using a Bruker AVANCE 400 or Brucker DPX400 spectrometer. CDCl3 is deuteriochloroform, DMSO-D6 is hexadeuteriodimethylsulfoxide, and MeOD is tetradeuteriomethanol.
Chemical shifts are reported in parts per million (δ) downfϊeld from the internal standard tetramethylsilane (TMS) or calibrated to the residual proton signal in the NMR solvent (e.g., CHCl3 in CDCI3). Abbreviations for NMR data are as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, dt = doublet of triplets, app = apparent, br = broad. J indicates the NMR coupling constant measured in Hertz.
Heating of reaction mixtures with microwave irradiations was carried out on a
Biotage Initiator® microwave reactor, typically employing the high absorbance setting.
Cartridges or columns containing polymer based functional groups (acid, base, metal chelators, etc) can be used as part of compound workup. The "amine" columns or cartridges are used to neutralize or basify acidic reaction mixtures or products. These include NH2 Aminopropyl SPE-ed SPE Cartridges available from Applied Separations and diethylamino SPE cartridges available from United Chemical Technologies, Inc.
Abbreviations are listed in the table below. All other abbreviations are as described in the ACS Style Guide (American Chemical Society, Washington, DC, 1986).
Table of Abbreviations
Figure imgf000035_0001
INTERMEDIATE COMPOUNDS
Intermediate 1
1,1-dimethylethyl [(3S)-l-(3-chloro-4-cyano-5-isothiazolyl)-3-pyrrolidinyl] carbamate
Figure imgf000036_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (46 mg, 0.247 mmol) in THF (1.5 mL) were added 3,5-dichloro-4-isothiazolecarbonitrile (44 mg, 0.246 mmol) and Et3N (0.034 mL, 0.246 mmol). After stirring for 1 h at RT, water (1 mL) and EtOAc (1 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 1 mL). The combined organic layers were washed with brine (1 mL), dried over Na2SO4, filtered, concentrated in vacuo, and dried under high vacuum to afford the title compound (79.3 mg, 98%). LC-MS m/z 329 (M+H)+, 1.03 min (ret time).
Intermediate 2
5-[(3S)-3-amino-l-pyrrolidinyl]-3-chloro-4-isothiazolecarbonitrile hydrochloride
Figure imgf000036_0002
A solution of 1,1-dimethylethyl [(35)-l-(3-chloro-4-cyano-5-isothiazolyl)-3- pyrrolidinyl] carbamate (55 mg, 0.167 mmol) in HCl (I M solution in 1,4-dioxane, 1 mL, 1.000 mmol) was stirred at RT for 1 h. Et2O (2 mL) was added, and the resultant white precipitate was filtered through a plug of cotton. The solid was dissolved in MeOH (3 mL) and concentrated under a stream of nitrogen at 50 0C to afford the title compound (37 mg, 83%). LC-MS m/z 229 (M+H)+, 0.58 min (ret time). Intermediate 3
l,l-dimethylethyl ((15)-2-{[(3S)-l-(3-chloro-4-cyano-5-isothiazolyl)-3-pyrrolidinyl]- amino}-l-methyl-2-oxoethyl)carbamate
Figure imgf000037_0001
To a solution of 5-[(35)-3-amino- 1 -pyrrolidinyl]-3-chloro-4-isothiazole- carbonitrile hydrochloride (37 mg, 0.140 mmol), JV-(te/t-butoxycarbonyl)-L-alanine (27 mg, 0.143 mmol), and HATU (58 mg, 0.153 mmol) in CH2Cl2 (1.0 mL) was added Et3N (0.058 mL, 0.419 mmol). The reaction mixture was stirred at RT for 30 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were washed with brine (1 mL), dried over Na2SO4, filtered, concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum to afford the title compound (60 mg, >100%, contained some residual solvent). LC-MS m/z 400 (M+H)+, 0.95 min (ret time).
Intermediate 4
1,1-dimethylethyl [(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl] carbamate
Figure imgf000037_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (3.36 g, 18.06 mmol) and 2-fluoro-5-nitrobenzonitrile (3 g, 18.06 mmol) in CH3CN (30 mL) was added K2CO3 (2.5 g, 18.09 mmol). After stirring for 1 h at RT, the reaction mixture was concentrated in vacuo, followed by the addition of water (40 mL) and EtOAc (30 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, concentrated in vacuo, and dried under high vacuum. The crude product was washed with Et2O (30 mL), filtered, and dried under high vacuum to afford the title compound (5.16 g, 86%). LC-MS m/z 333 (M+H)+, 1.06 min (ret time). Intermediate 5
2-[(3S)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride
Figure imgf000038_0001
A solution of 1,1-dimethylethyl [(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]- carbamate (5.16 g, 15.53 mmol) in HCl (4 M solution in 1,4-dioxane, 80 rnL, 320 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (300 mL) and the slurry was filtered through a medium porosity glass fritted filter. Washing the solid with Et2O (2 x 50 mL) afforded the title compound (4.04 g, 97%). LC-MS m/z 233 (M+H)+, 0.58 min (ret time). Intermediate 6
1,1-dimethylethyl ((lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000038_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (129 mg, 0.480 mmol), JV-(ter£-butoxycarbonyl)-L-alanine (91 mg, 0.480 mmol), and HATU (201 mg, 0.528 mmol) in CH2Cl2 (2.5 mL) was added Et3N (0.20 mL, 1.435 mmol). The reaction mixture was stirred at RT for 3 h 20 min. Water (4 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 4 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (50-70% EtOAc/hexanes) afforded the title compound (190 mg, 81%). LC-MS m/z 404 (M+H)+, 0.98 min (ret time).
Intermediate 7
1 , 1-dimethylethyl (2- { [(3S)- l-(2-cyano-4-nitr ophenyl)-3-pyrrolidinyl] amino}-2- oxoethyl)carbamate
Figure imgf000039_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride
(31 mg, 0.133 mmol), Λ/-(tøt-butoxycarbonyl)glycine (23.4 mg, 0.133 mmol), and HATU (55.8 mg, 0.147 mmol) in CH2Cl2 (1.0 mL) was added Et3N (0.056 mL, 0.400 mmol). The reaction mixture was stirred at RT for 18 h 30 min. Water (1 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (50-70% EtOAc/hexanes) afforded the title compound (42.4 mg, 82%). LC-MS m/z 390 (M+H)+, 0.96 min (ret time).
Intermediate 8
2-chloro-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile
Figure imgf000039_0002
To a slurry of CuCl2 (91 mg, 0.673 mmol) and tert-hutyl nitrite (0.100 mL, 0.841 mmol) in CH3CN (2.0 mL) at 0 0C was added 2-amino-4,5,6,7-tetrahydro-l- benzothiophene-3-carbonitrile (100 mg, 0.561 mmol) portionwise over 20 sec. The ice bath was removed, and the reaction was stirred at RT for 10 min. A solution of 9:1 saturated aq. NH4Cl/saturated aq. NH4OH (4 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were washed with brine (2 mL), dried over Na2SO4, filtered, and concentrated in vacuo onto Isolute®. Purification via flash column chromatography (0- 20% EtOAc/hexanes) afforded the title compound (25 mg, 23%). LC-MS m/z 198
(M+H) λ+ , 1.22 min (ret time). Intermediate 9
2-[(3S)-3-amino-l-pyrrolidinyl]-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile
Figure imgf000040_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (24 mg, 0.129 mmol) and 2-chloro-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile (25 mg, 0.126 mmol) in CH3CN (1 niL) was added K2CO3 (18 mg, 0.130 mmol). After stirring for 45 min at RT, the reaction was heated to 50 0C. After 1 h, 10 min, the reaction was heated to reflux. LC-MS of an aliquot of the reaction mixture taken after 17 h showed only starting materials. The reaction mixture was concentrated under a stream of nitrogen at 50 0C, and the residue was taken up in DMF (1.0 mL). The reaction mixture was heated to 100 0C for 24 h following by increasing to 150 0C for a further 53 h. Upon cooling, a solution of 1 M aq. HCl (2 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was washed with EtOAc (2 x 1 mL). The aqueous layer was made basic with 6 M aq. NaOH (1 mL) and extracted with EtOAc (4 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL), dried over Na2SO4, filtered, and concentrated under a stream of nitrogen at 50 0C to afford the title compound (5.1 mg, 16.3%). LC-MS m/z 248 (M+H)+, 0.86 min (ret time).
Intermediate 10
1,1-dimethylethyl ((15)-2-{[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate
Figure imgf000040_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5,6,7-tetrahydro-l-benzo- thiophene-3-carbonitrile (5.1 mg, 0.021 mmol), JV-(tert-butoxycarbonyl)-L-alanine (3.9 mg, 0.021 mmol), and HATU (9.4 mg, 0.025 mmol) in CH2Cl2 (0.5 mL) was added Et3N (0.009 mL, 0.065 mmol). The reaction mixture was stirred at RT for 1 h 15 min. Water (2 mL) and EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 1 mL). The combined organic layers were washed with brine (1 mL), dried over Na2SO4, filtered, and concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-60% EtOAc/hexanes) afforded the title compound (7.5 mg, 87%). LC-MS m/z 419 (M+H)+, 1.43 min (ret time).
Intermediate 11
7V1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-7V2-{[(l,l-dimethylethyl)oxy]- carbonyl}-L-valinamide
Figure imgf000041_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), JV-(tert-butoxycarbonyl)-L-valine (70 mg, 0.322 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 30 min. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 5 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. Purification via flash column chromatography (30-100% EtOAc/hexanes) afforded the title compound (131 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 432 (M+H)+, 1.02 min (ret time).
Intermediate 12
1,1-dimethylethyl [(lS)-l-({[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000041_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride
(75 mg, 0.279 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}-amino)butanoic acid (62.4 mg, 0.307 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 30 min. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (2 x 3 mL). The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (10 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (20 mL), and water (20 mL). The solvent was removed and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (110 mg, 94%). LC-MS m/z 418 (M+H)+, 0.99 min (ret time).
Intermediate 13
7V1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-7V2-{[(l,l-dimethylethyl)oxy]- carbonyl}-L-norvalinamide
Figure imgf000042_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), N-(tert-butoxycarbonyl)-L-norvaline (66.7 mg, 0.307 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 30 min. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (2 x 3 mL). The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (10 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (20 mL), and water (20 mL). The solvent was removed and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (127 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 432 (M+H)+, 1.02 min (ret time).
Intermediate 14
7V1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-7V2-{[(l,l-dimethylethyl)oxy]- carbonyl}-L-leucinamide
Figure imgf000042_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), N-(tert-butoxycarbonyl)-L-leucine (64.6 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 rnL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (121.7 mg, 98%) as a yellow residue. LC-MS m/z 446 (M+H)+, 1.07 min (ret time).
Intermediate 15
7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-7V-{[(l,l-dimethylethyl)oxy]- carbonyl}-L-phenylalaninamide
Figure imgf000043_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), JV-(te/t-butoxycarbonyl)-L-phenylalanine (81 mg, 0.307 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (139.6 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 480 (M+H)+, 1.09 min (ret time).
Intermediate 16
4-fluoro-3-biphenylcarbonitrile
Figure imgf000043_0002
To a mixture of 5-bromo-2-fluorobenzonitrile (500 mg, 2.500 mmol), phenyl boronic acid (305 mg, 2.500 mmol), and K2CO3 (1.037 g, 7.50 mmol) in 1,4-dioxane (6.0 mL) and water (2.0 mL) was added PdCl2(dppf) (10 mg, 0.014 mmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 140 0C for 30 min. Water (5 rnL) and EtOAc (10 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (3 x 5 mL), dried over Na2SO4, filtered, concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum to afford the title compound (500 mg, 90%). LC-MS m/z 198 (M+H)+, 1.10 min (ret time).
Intermediate 17
1,1-dimethylethyl [(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl] carbamate
Figure imgf000044_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (47.2 mg, 0.254 mmol) and 4-fluoro-3-biphenylcarbonitrile (50 mg, 0.254 mmol) in DMF (1 mL) was added K2CO3 (35.0 mg, 0.254 mmol). The reaction vial was capped and heated in a Biotage Initiator® microwave at 150 0C for 30 min followed by conventional heating in an oil bath at 100 0C overnight. Upon cooling, water (3 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-50% EtOAc/hexanes) afforded the title compound (31 mg, 34%). LC-MS m/z 364 (M+H)+, 1.28 min (ret time).
Intermediate 18
4-[(3S)-3-amino-l-pyrrolidinyl]-3-biphenylcarbonitrile hydrochloride
Figure imgf000044_0002
HCl
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-4-biphenylyl)-3- pyrrolidinyl] carbamate (30 mg, 0.083 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. Washing the solid with Et2O (2 x 0.5 mL), followed by dissolving in 1 : 1 MeOH/CH2Cl2 and then concentrating under a stream of nitrogen at 50 0C afforded the title compound (22.9 mg, 93%). LC-MS m/z 264 (M+H)+, 0.86 min (ret time). Intermediate 19
l,l-dimethylethyl ((lS)-2-{[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000045_0001
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-3-biphenylcarbonitrile
hydrochloride (22.9 mg, 0.076 mmol), Λ/-(te/t-butoxycarbonyl)-L-alanine (14.5 mg, 0.077 mmol), and HATU (29.0 mg, 0.076 mmol) in CH2Cl2 (0.5 mL) was added Et3N (0.032 mL, 0.229 mmol). The reaction mixture was stirred at RT for 25 min and then
concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (10-70% EtOAc/hexanes) afforded the title compound (7.8 mg, 24%). LC-MS m/z 435 (M+H)+, 1.14 min (ret time). Intermediate 20
3-fluoro-4-biphenylcarbonitrile
Figure imgf000045_0002
To a mixture of 4-bromo-2-fluorobenzonitrile (500 mg, 2.500 mmol), phenyl boronic acid (305 mg, 2.500 mmol), and K2CO3 (691 mg, 5.00 mmol) in 1,4-dioxane (6.0 mL) and water (2.0 mL) was added PdCl2(dppf) (10 mg, 0.014 mmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 140 0C for 30 min. Water (5 mL) and EtOAc (10 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (3 x 5 mL), dried over Na2SO4, filtered, concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum to afford the title compound (490 mg, 88%). LC-MS m/z 198 (M+H)+, 1.10 min (ret time). Intermediate 21
1,1-dimethylethyl [(3S)-l-(4-cyano-3-biphenylyl)-3-pyrrolidinyl] carbamate
Figure imgf000046_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (47.2 mg, 0.254 mmol) and 3-fluoro-4-biphenylcarbonitrile (50 mg, 0.254 mmol) in DMF (1 mL) was added K2CO3 (35.0 mg, 0.254 mmol). The reaction vial was capped and heated to 100 0C for 5O h. Upon cooling, water (3 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (10-40% EtOAc/hexanes) afforded the title compound (60 mg, 65%). LC-MS m/z 364 (M+H)+, 1.28 min (ret time).
Intermediate 22
3-[(3S)-3-amino-l-pyrrolidinyl]-4-biphenylcarbonitrile hydrochloride
Figure imgf000046_0002
A solution of 1 , 1 -dimethylethyl [(3S)- 1 -(4-cyano-3-biphenylyl)-3- pyrrolidinyl] carbamate (60 mg, 0.165 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. Washing the solid with Et2O (2 x 1 mL), followed by dissolving in 1 : 1 MeOHZCH2Cl2, concentrating under a stream of nitrogen at 50 0C, and drying under high vacuum afforded the title compound (46.4 mg, 93%). LC-MS m/z 264 (M+H)+, 0.77 min (ret time). Intermediate 23
l,l-dimethylethyl ((lS)-2-{[(3S)-l-(4-cyano-3-biphenylyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000047_0001
To a solution of 3-[(35)-3-amino- 1 -pyrrolidinyl]-4-biphenylcarbonitrile hydrochloride (46 mg, 0.153 mmol), Λ/-(te/t-butoxycarbonyl)-L-alanine (29.0 mg, 0.153 mmol), and HATU (58.3 mg, 0.153 mmol) in CH2Cl2 (0.5 mL) was added Et3N (0.064 mL, 0.460 mmol). The reaction mixture was stirred at RT for 25 min and then
concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-60% EtOAc/hexanes) afforded the title compound (41.6 mg, 62%). LC-MS m/z 435 (M+H)+, 1.13 min (ret time). Intermediate 24
ethyl 4-cyano-5-[(3S)-3-({[(l,l-dimethylethyl)oxy]carbonyl}amino)-l-pyrrolidinyl]-2- thiophenecarboxylate
Figure imgf000047_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (16.4 mg, 0.088 mmol) and ethyl 5-chloro-4-cyano-2-thiophenecarboxylate (19 mg, 0.088 mmol) in DMF
(0.5 mL) was added K2CO3 (26 mg, 0.188 mmol). The reaction vial was capped and heated to 100 0C for 40 min. Upon cooling, water (3 mL) and EtOAc (2 mL) were added.
The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL).
The combined organic layers were washed with brine (3 x 1 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (10-40% EtOAc/hexanes) afforded the title compound (20.7 mg, 64%). LC-MS m/z 366 (M+H)+, 1.09 min (ret time).
Intermediate 25
ethyl 5-[(3S)-3-amino-l-pyrrolidinyl]-4-cyano-2-thiophenecarboxylate hydrochloride
Figure imgf000048_0001
A solution of ethyl 4-cyano-5-[(3S)-3-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)-l-pyrrolidinyl]-2-thiophenecarboxylate (20 mg, 0.055 mmol) in HCl (0.5 M solution in 1,4-dioxane, 0.5 mL, 0.250 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (1 mL), followed by dissolving in MeOH and concentrating under a stream of nitrogen at 50 0C afforded the title compound (16 mg, 97%). LC-MS m/z 266 (M+H)+, 0.63 min (ret time).
Intermediate 26
ethyl 4-cyano-5-{(3S)-3-[(7V-{[(l,l-dimethylethyl)oxy]carbonyl}-L-alanyl)amino]-l- pyrrolidinyl}-2-thiophenecarboxylate
Figure imgf000048_0002
To a solution of ethyl 5-[(35)-3-amino-l-pyrrolidinyl]-4-cyano-2- thiophenecarboxylate hydrochloride (16 mg, 0.053 mmol), 7V-(tert-butoxycarbonyl)-L- alanine (10 mg, 0.053 mmol), and HATU (20.2 mg, 0.053 mmol) in CH2Cl2 (0.5 mL) was added Et3N (0.022 mL, 0.159 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-60% EtOAc/hexanes) afforded the title compound (18.1 mg, 78%). LC-MS m/z 437 (M+H)+, 1.00 min (ret time). Intermediate 27
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-l-
(hydroxymethyl)-2-oxoethyl]carbamate
Figure imgf000049_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride
(75 mg, 0.279 mmol), Λ/-(te/t-butoxycarbonyl)-L-serine (57.3 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (70-100% EtOAc/hexanes) to afford the title compound (93.4 mg, 88%) as a yellow residue. LC-MS m/z 420 (M+H)+, 0.86 min (ret time). Intermediate 28
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-2- oxo-l-(l,3-thiazol-4-ylmethyl)ethyl]carbamate
Figure imgf000049_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), N-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(l,3-thiazol-4-yl)-L- alanine (76 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (105 mg, 77%) as a yellow solid. LC-MS m/z 487 (M+H)+, 0.95 min (ret time).
Intermediate 29
l,l-dimethylethyl ((lS)-l-(cyanomethyl)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-2-oxoethyl)carbamate
Figure imgf000050_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), (2S)-3-cyano-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- propanoic acid (59.8 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (134.7 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 429 (M+H)+, 0.96 min (ret time).
Intermediate 30
l,l-dimethylethyl [(15)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000050_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), Λ/-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)-L-alanine (76 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 rnL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (157 mg, >100%, contained some residual solvent) as a yellow residue. LC- MS m/z 486 (M+H)+, 1.08 min (ret time).
Intermediate 31
2-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzonitrile
Figure imgf000051_0001
To a mixture of 5-bromo-2-fluorobenzonitrile (500 mg, 2.500 mmol),
6ώ(pinacolato)diboron (1.9 g, 7.48 mmol), and KOAc (1.47 g, 14.98 mmol) in 1,4- dioxane (10.0 mL) that was degassed with argon for 10 min was added PdCl2(dppf) (10 mg, 0.014 mmol). The reaction mixture was heated to 100 0C overnight. The reaction mixture was filtered through Celite 521® and rinsed through with EtOAc (10 mL). The filtrate was concentrated in vacuo and treated with CH2Cl2 (10 mL) and water (10 mL). The layers were separated, and the aqueous layer was extracted with CH2Cl2 (5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated in vacuo onto Isolute®. Purification via flash column chromatography (0- 20% EtOAc/hexanes) afforded the title compound (1.03 g, 72%). LC-MS m/z 248 (M+H)+, 1.15 min (ret time).
Intermediate 32
2-fluoro-5-(l,3-thiazol-2-yl)benzonitrile
Figure imgf000051_0002
To a mixture of 2-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- benzonitrile (100 mg, 0.405 mmol), 2-bromothiazole (0.11 mL, 1.221 mmol), and K2CO3 (168 mg, 1.214 mmol) in 1,4-dioxane (1.5 mL) and water (0.5 mL) was added Pd(Ph3P)4 (5 mg, 0.004 mmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 100 0C for 30 min followed by an additional 2 h at 100 0C. Water (4 mL) and EtOAc (5 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 3 mL). The combined organic layers were washed with brine (3 x 1 mL) and concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-20% EtOAc/hexanes) afforded the title compound (2.4 mg, 2.9%). LC-MS m/z 205 (M+H)+, 0.87 min (ret time).
Intermediate 33
1 , 1-dimethylethyl ((3S)- 1- [2-cyano-4-(l ,3-thiazol-2-yl)phenyl] -3-pyrr olidinyl}- carbamate
Figure imgf000052_0001
To a solution of 1, 1-dimethylethyl (35)-3-pyrrolidinylcarbamate (2.2 mg, 0.012 mmol) and 2-fluoro-5-(l,3-thiazol-2-yl)benzonitrile (2.4 mg, 0.012 mmol) in DMF (0.5 mL) was added K2CO3 (1.7 mg, 0.012 mmol). The reaction vial was capped and heated to 100 0C for 15 h. Upon cooling, water (3 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-70% EtOAc/hexanes) afforded the title compound (2.8 mg, 64%). LC-MS m/z 371 (M+H)+, 1.06 min (ret time).
Intermediate 34
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(l,3-thiazol-2-yl)benzonitrile hydrochloride
Figure imgf000052_0002
A solution of 1, 1-dimethylethyl {(3S)-l-[2-cyano-4-(l,3-thiazol-2-yl)phenyl]-3- pyrrolidinyl} carbamate (2.8 mg, 7.56 μmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. Washing the solid with Et2O (2 x 0.5 mL), followed by dissolving in 1 :1 MeOHZCH2Cl2 and concentrating under a stream of nitrogen at 50 0C afforded the title compound (2 mg, 86%). LC-MS m/z 111 (M+H)+, 0.60 min (ret time). Intermediate 35
l,l-dimethylethyl {(15)-l-[({(3S)-l-[2-cyano-4-(l,3-thiazol-2-yl)phenyl]-3-pyrrolidin- yl} amino)carbonyl] propyl} carbamate
Figure imgf000053_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(l,3-thiazol-2-yl)benzonitrile hydrochloride (2 mg, 6.52 μmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (1.4 mg, 6.89 μmol), and HATU (2.5 mg, 6.57 μmol) in CH2Cl2 (0.5 mL) was added Et3N (0.003 mL, 0.022 mmol). The reaction mixture was stirred at RT for 20 min and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-70% EtOAc/hexanes) afforded the title compound (3 mg, >100%, contained some residual solvent). LC-MS m/z 456 (M+H)+, 1.03 min (ret time).
Intermediate 36
2-fluoro-5-(2-pyrimidinyl)benzonitrile
Figure imgf000053_0002
To a mixture of 2-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- benzonitrile (100 mg, 0.405 mmol), 2-bromopyrimidine (194 mg, 1.221 mmol), and K2CO3 (168 mg, 1.214 mmol) in 1,4-dioxane (1.5 mL) and water (0.5 mL) was added Pd(Ph3P)4 (47 mg, 0.041 mmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 100 0C for 30 min. Water (4 mL) and EtOAc (5 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 3 mL). The combined organic layers were washed with brine (3 x 1 mL) and concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-20% EtOAc/hexanes) afforded the title compound (30.6 mg, 38%). LC-MS m/z 200 (M+H)+, 0.88 min (ret time). Intermediate 37
1,1-dimethylethyl {(3S)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3-pyrrolidinyl}- carbamate
Figure imgf000054_0001
To a solution of 1 , 1 -dimethylethyl (35)-3-pyrrolidinylcarbamate (29 mg, 0.156 mmol) and 2-fluoro-5-(2-pyrimidinyl)benzonitrile (30.6 mg, 0.154 mmol) in DMF (0.5 niL) was added K2CO3 (22 mg, 0.159 mmol). The reaction vial was capped and heated to 100 0C for 15 h. Upon cooling, water (3 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-100% EtOAc/hexanes) afforded the title compound (7.4 mg, 13%). LC-MS m/z 366 (M+H)+, 1.03 min (ret time).
Intermediate 38
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(2-pyrimidinyl)benzonitrile hydrochloride
Figure imgf000054_0002
A solution of 1,1-dimethylethyl {(35)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3- pyrrolidinyl} carbamate (7.4 mg, 0.020 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h 45 min. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. Washing the solid with Et2O (2 x 0.5 mL), followed by dissolving in 1 : 1 MeOH/CH2Cl2 and concentrating under a stream of nitrogen at 50 0C afforded the title compound (5.3 mg, 87%). LC-MS m/z 266 (M+H)+, 0.61 min (ret time).
Intermediate 39
l,l-dimethylethyl {(15)-l-[({(3S)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3- pyrrolidinyl}amino)carbonyl]propyl}carbamate
Figure imgf000055_0001
To a solution of 2- [(35)-3 -amino- 1 -pyrrolidinyl] -5 -(2-pyrimidinyl)benzonitrile hydrochloride (5.3 mg, 0.018 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (3.6 mg, 0.018 mmol), and HATU (6.7 mg, 0.018 mmol) in CH2Cl2 (0.5 mL) was added Et3N (0.0073 mL, 0.053 mmol). The reaction mixture was stirred at RT for 20 min and then concentrated under a stream of nitrogen at 50 0C onto Isolute®.
Purification via flash column chromatography (20-70% EtOAc/hexanes) afforded the title compound (6.5 mg, 82%). LC-MS m/z 451 (M+H)+, 0.99 min (ret time).
Intermediate 40
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-l-
(cyclopropylmethyl)-2-oxoethyl]carbamate
Figure imgf000055_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride
(75 mg, 0.279 mmol), 3-cyclopropyl-Λ/-{[(l,l-dimethylethyl)oxy]-carbonyl}-L-alanine dicyclohexylamine (115 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (145 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 444 (M+H)+, 1.04 min (ret time).
Intermediate 41
1,1-dimethylethyl [(lS)-l-({[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}- carbonyl)-3-butyn-l-yl] carbamate
Figure imgf000056_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)-4-pentynoic acid (59.5 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h.
Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (124 mg, >100%, contained some residual solvent) as a yellow residue. LC-MS m/z 428 (M+H)+, 1.01 min (ret time).
Intermediate 42
1,1-dimethylethyl {(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- carbamate
Figure imgf000056_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (197 mg, 1.058 mmol) and 2-fluoro-5-(trifluoromethyl)benzonitrile (200 mg, 1.058 mmol) in DMF (4.0 mL) was added K2CO3 (146 mg, 1.058 mmol). The reaction vial was capped and heated to 100 0C for 45 min. Upon cooling, water (15 mL) and EtOAc (15 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 5 mL). The combined organic layers were washed with brine (3 x 5 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-40% EtOAc/hexanes) afforded the title compound (139 mg, 37%). LC-MS m/z 356 (M+H)+, 1.18 min (ret time).
Intermediate 43
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride
Figure imgf000057_0001
A solution of 1,1-dimethylethyl {(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl} carbamate (139 mg, 0.391 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (2 x 2 mL), followed by dissolving in MeOH and concentrating under a stream of nitrogen at 50 0C afforded the title compound (109 mg, 96%). LC-MS m/z 256 (M+H)+, 0.72 min (ret time).
Intermediate 44
1,1-dimethylethyl [(lS)-2-({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidin- yl}amino)-l-methyl-2-oxoethyl] carbamate
Figure imgf000057_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (54.5 mg, 0.187 mmol), jV-(te/t-butoxycarbonyl)-L-alanine (36 mg, 0.190 mmol), and HATU (71.0 mg, 0.187 mmol) in CH2Cl2 (0.6 mL) and MeOH (0.3 mL) was added Et3N (0.078 mL, 0.561 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-70% EtOAc/hexanes) afforded the title compound (75 mg, 94%). LC-MS m/z 427 (M+H)+, 1.08 min (ret time). Intermediate 45
l,l-dimethylethyl {(15)-l-[({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)carbonyl]propyl}carbamate
Figure imgf000058_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (54.5 mg, 0.187 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (38.0 mg, 0.187 mmol), and HATU (71.0 mg, 0.187 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.078 mL, 0.561 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®.
Purification via flash column chromatography (0-60% EtOAc/hexanes) afforded the title compound (82 mg, 100%). LC-MS m/z 441 (M+H)+, 1.11 min (ret time). Intermediate 46
1,1-dimethylethyl [(3S)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl] carbamate
Figure imgf000058_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (93 mg, 0.500 mmol) and 5-bromo-2-fluorobenzonitrile (100 mg, 0.500 mmol) in DMF (2.0 mL) was added K2CO3 (70 mg, 0.506 mmol). The reaction vial was capped and heated to 100 0C for 1 h 45 min. Upon cooling, water (5 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 2 mL). The combined organic layers were washed with brine (3 x 1 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (0-50% EtOAc/hexanes) afforded the title compound (140 mg, 76%). LC-MS m/z 366/368 (M+H)+, 1.17 min (ret time). Intermediate 47
2-[(3S)-3-amino-l-pyrrolidinyl]-5-bromobenzonitrile hydrochloride
Figure imgf000059_0001
A solution of 1,1-dimethylethyl [(35)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl] carbamate (140 mg, 0.382 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 rnL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (2 x 2 mL), followed by dissolving in MeOH and concentrating under a stream of nitrogen at 50 0C afforded the title compound (105 mg, 91%). LC-MS m/z 266/268 (M+H)+, 0.68 min (ret time).
Intermediate 48
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000059_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-bromobenzonitrile
hydrochloride (55 mg, 0.182 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (50 mg, 0.184 mmol), and HATU (70 mg, 0.184 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.076 mL, 0.545 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (89 mg, 94%). LC- MS m/z 519/521 (M+H)+, 1.21 min (ret time). Intermediate 49
1,1-dimethylethyl ((lS)-2-{[(3S)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000060_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-bromobenzonitrile
hydrochloride (50 mg, 0.165 mmol), Λ/-(tert-butoxycarbonyl)-L-alanine (32 mg, 0.169 mmol), and HATU (63 mg, 0.166 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.069 mL, 0.496 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (70 mg, 97%). LC-MS m/z 437/439
(M+H)+, 1.07 min (ret time).
Intermediate 50
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000060_0002
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-3-biphenylcarbonitrile hydrochloride (39 mg, 0.130 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (36 mg, 0.133 mmol), and HATU (49 mg, 0.129 mmol) in CH2Cl2 (0.8 mL) and MeOH (0.4 mL) was added Et3N (0.054 mL, 0.390 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (20.3 mg, 30%). LC-MS m/z 517 (M+H)+, 1.30 min (ret time).
Intermediate 51
2-chloro- lH-indole-3-carbaldehyde oxime
Figure imgf000061_0001
A solution of 2-chloro- lH-indole-3-carbaldehyde (500 mg, 2.78 mmol), hydroxylamine hydrochloride (195 mg, 2.81 mmol), and pyridine (0.225 mL, 2.81 mmol) in EtOH (5 mL) was heated to reflux for 1 h. The reaction mixture was concentrated under a stream of nitrogen at 50 0C and dried under high vacuum overnight to afford the title compound (541 mg, 100%). LC-MS m/z 195 (M+Η)+, 0.70 min (ret time).
Intermediate 52
2-chloro- lH-indole-3-carbonitrile
Figure imgf000061_0002
A solution of 2-chloro- lH-indole-3-carbaldehyde oxime (541 mg, 2.78 mmol) in Ac2O (5 mL, 52.9 mmol) was heated to reflux for 1 h. The reaction mixture was poured into water (30 mL) with stirring. After 3 min of stirring, the aqueous solution was decanted off, and the remaining precipitate was recrystallized from ~2: 1 MeOHZH2O. The solid was filtered and aspirated dry overnight. The precipitate and filtrate were combined, dissolved in EtOAc, and concentrated in vacuo onto Isolute®. Purification via flash column chromatography (0-20% EtOAc/hexanes) afforded the title compound (107 mg), along with l-acetyl-2-chloro-lH-indole-3-carbonitrile (139 mg). To a solution of 1- acetyl-2-chloro-lH-indole-3-carbonitrile (139 mg, 0.636 mmol) in MeOH (1.0 mL) was added a solution of 6 M aq. NaOH (0.5 mL, 3.00 mmol). The reaction mixture was stirred at RT for 2 h and then concentrated under a stream of nitrogen at 50 0C to remove the MeOH. A solution of 6 M aq. HCl (1 mL) and EtOAc (1 niL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo to afford additional title compound which was combined with the first batch (203 mg total, 41%). LC-MS m/z 111 (M+H)+, 0.96 min (ret time).
Intermediate 53
2-chloro- 1-methyl- lH-indole-3-carbonitrile
Figure imgf000062_0001
To a mixture of 2-chloro- lH-indole-3-carbonitrile (95 mg, 0.538 mmol) and K2CO3 (75 mg, 0.543 mmol) in acetone (3.0 mL) was added MeI (0.034 mL, 0.538 mmol). The reaction mixture was heated to 55 0C for 2 h and then concentrated under a stream of nitrogen at 50 0C. The residue was taken up in water (2 mL) and EtOAc (2 mL).
The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL).
The combined organic layers were washed with brine (1 mL), dried over Na2SO4, filtered, and concentrated under a stream of nitrogen at 50 0C to afford the title compound (96 mg,
94%). LC-MS m/z 191 (M+Η)+, 0.97 min (ret time).
Intermediate 54
l^-dimethylethyl lCS^-l-CS-cyano-l-methyl-lH-indol-l-ylJ-S-pyrrolidinyl]- carbamate
Figure imgf000062_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (94 mg, 0.504 mmol) and 2-chloro- 1 -methyl- lH-indole-3-carbonitrile (96 mg, 0.504 mmol) in DMF (2.0 mL) was added K2CO3 (70 mg, 0.506 mmol). The reaction vial was capped and heated to 150 0C for 4.5 h. Upon cooling, water (5 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (3 x 1 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (0-40% EtOAc/hexanes) afforded the title compound (68 mg, 40%). LC-MS m/z 341 (M+H)+, 1.08 min (ret time).
Intermediate 55
2- [(3S)-3-amino- 1-pyrrolidinyl] - 1-methyl- lH-indole-3-carbonitrile hydrochloride
Figure imgf000063_0001
A solution of 1,1-dimethylethyl [(3S)- l-(3-cyano- 1-methyl- lH-indol-2-yl)-3- pyrrolidinyl] carbamate (68 mg, 0.200 mmol) in ΗCI (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (5 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (2 x 2 mL), followed by dissolving in MeOH, concentrating under a stream of nitrogen at 50 0C, and drying under high vacuum afforded the title compound (48 mg, 87%). LC-MS m/z 241 (M+Η)+, 0.68 min (ret time).
Intermediate 56
1,1-dimethylethyl ((l^-l-IICSSJ-l-CS-cyano-l-methyl-lH-indoH-yO-S-pyrrolidin- yl]amino}-l-methyl-2-oxoethyl)carbamate
Figure imgf000063_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-l-methyl-lH-indole-3- carbonitrile hydrochloride (24 mg, 0.087 mmol), JV-(tert-butoxycarbonyl)-L-alanine (17 mg, 0.090 mmol), and ΗATU (33.0 mg, 0.087 mmol) in CH2Cl2 (0.6 mL) and MeOH (0.3 mL) was added Et3N (0.036 mL, 0.260 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (33 mg, 92%). LC-MS m/z 412 (M+H)+, 1.02 min (ret time). Intermediate 57
l,l-dimethylethyl [(15)-2-{[(3S)-l-(3-cyano-l-methyl-lH-indol-2-yl)-3-pyrrolidinyl]- amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000064_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-l-methyl-lH-indole-3- carbonitrile hydrochloride (24 mg, 0.087 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3- (2-thienyl)-L-alanine (24 mg, 0.088 mmol), and ΗATU (33.0 mg, 0.087 mmol) in CH2Cl2 (0.6 mL) and MeOH (0.3 mL) was added Et3N (0.036 mL, 0.260 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (2 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were washed with brine (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (37.5 mg, 88%). LC-MS m/z 494 (M+H)+, 1.14 min (ret time).
Intermediate 58
1,1-dimethylethyl [(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]carbamate
Figure imgf000064_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (201 mg, 1.078 mmol) and 2,5-difluorobenzonitrile (150 mg, 1.078 mmol) in DMF (2.0 mL) was added K2CO3 (149 mg, 1.078 mmol). The reaction vial was capped and heated to 100 0C for 5 h. Upon cooling, water (5 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 2 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-50% EtOAc/hexanes) afforded the title compound (143 mg, 43%). LC-MS m/z 306 (M+H)+, 1.09 min (ret time). Intermediate 59
2-[(3S)-3-amino-l-pyrrolidinyl]-5-fluorobenzonitrile hydrochloride
Figure imgf000065_0001
A solution of 1,1-dimethylethyl [(35)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]- carbamate (143 mg, 0.468 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (2 x 2 mL), followed by dissolving in MeOH, concentrating under a stream of nitrogen at 50 0C, and drying under high vacuum afforded the title compound (102 mg, 90%). LC-MS m/z 206 (M+H)+, 0.59 min (ret time).
Intermediate 60
1,1-dimethylethyl ((15)-2-{[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000065_0002
To a solution of 2-[(35)-3-amino- 1 -pyrrolidinyl]-5-fluorobenzonitrile
hydrochloride (34 mg, 0.141 mmol), jV-(te/t-butoxycarbonyl)-L-alanine (27 mg, 0.143 mmol), and HATU (54 mg, 0.142 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.058 mL, 0.422 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (34.2 mg, 65%). LC-MS m/z 311 (M+H)+, 0.96 min (ret time). Intermediate 61
1,1-dimethylethyl [(lS)-l-({[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000066_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-fluorobenzonitrile
hydrochloride (34 mg, 0.141 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (29 mg, 0.143 mmol), and HATU (54 mg, 0.142 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.058 mL, 0.422 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (53.7 mg, 98%). LC-MS m/z 391 (M+H)+, 1.03 min (ret time).
Intermediate 62
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000066_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-fluorobenzonitrile
hydrochloride (34 mg, 0.141 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (39 mg, 0.144 mmol), and HATU (54 mg, 0.142 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.058 mL, 0.422 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 1 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (60.4 mg, 94%). LC-MS m/z 459 (M+H)+, 1.12 min (ret time).
Intermediate 63
l,l-dimethylethyl [(15)-2-{[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000067_0001
To a mixture of 1,1-dimethylethyl [(lS)-2-{[(3S)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (52 mg, 0.100 mmol), (4- fluorophenyl)boronic acid (15 mg, 0.107 mmol), and K2CO3 (42 mg, 0.304 mmol) in 1,4- dioxane (0.75 mL) and water (0.25 mL) was added PdCl2(dppf) (7 mg, 9.57 μmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 140 0C for 10 min. Water (5 mL) and CH2Cl2 (2 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with CH2Cl2 (3 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C. The crude product was dissolved in DMSO (1 mL) and purified by reverse phase
HPLC (YMC C18 S-5 μm/12 nm 75 x 30 mm preparatory column), eluting at 35 mL/min with a linear gradient running from 20% CH3CN/H2O (0.1% TFA) to 70% CH3CN/H2O (0.1% TFA) over 15 min to afford the title compound (40.5 mg, 76%). LC-MS m/z 535 (M+H)+, 1.30 min (ret time). Intermediate 64
1,1-dimethylethyl [(15)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-2- oxo-l-(3-thienylmethyl)ethyl] carbamate
Figure imgf000067_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), Λ/-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(3-thienyl)-L-alanine (76 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (112 mg, 83%) as a light yellow solid. LC-MS m/z 486 (M+H)+, 1.06 min (ret time).
Intermediate 65
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-7V2-{[(l,l-dimethylethyl)oxy]- carbonyl}-L-norleucinamide
Figure imgf000068_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), JV-(te/t-butoxycarbonyl)-L-norleucine (64.6 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (112.6 mg, 91%) as a yellow residue. LC-MS m/z AAβ (M+H)+, 1.08 min (ret time).
Intermediate 66
l,l-dimethylethyl ((15)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-l- cyclopropyl-2-oxoethyl)carbamate
Figure imgf000068_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), (2S)-cyclopropyl({[(l,l-dimethylethyl)oxy]- carbonyl}amino)ethanoic acid (60.1 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30- 100% EtOAc/hexanes) to afford the title compound as a yellow residue. LC-MS m/z 430 (M+H)+, 0.99 min (ret time).
Intermediate 67
2-bromo-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile
Figure imgf000069_0001
To a slurry of CuBr2 (752 mg, 3.37 mmol) and tert-bvΛy\ nitrite (0.57 mL, 4.33 mmol) in CH3CN (10.0 mL) at 0 0C was added 2-amino-4,5,6,7-tetrahydro-l- benzothiophene-3-carbonitrile (500 mg, 2.80 mmol) portionwise over ~ 1 min. The ice bath was removed, and the reaction was stirred at RT for 15 min. A solution of 9:1 saturated aq. NH4Cl/saturated aq. NH4OH (20 mL) and EtOAc (30 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 20 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated in vacuo. Purification via flash column chromatography (0-20%
EtOAc/hexanes) afforded the title compound (325.7 mg, 48%) as an orange oil which solidified upon standing. LC-MS m/z 242/244 (M+H)+, 1.18 min (ret time). Intermediate 68
1,1-dimethylethyl [(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3- pyrrolidinyl] carbamate
Figure imgf000069_0002
A mixture of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (274 mg, 1.469 mmol), 2-bromo-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile (325.7 mg, 1.130 mmol), Pd2(dba)3 (10.35 mg, 0.011 mmol), racemic BINAP (21.11 mg, 0.034 mmol), and Cs2CO3 (374 mg, 1.148 mmol) in toluene (10 mL) was degassed by bubbling argon through for ~ 2 min and then heated to 150 0C in an Emrys Optimizer® microwave for 2 h. Additional Pd2(dba)3 (10.35 mg, 0.011 mmol) and Pd(OAc)2 (2.54 mg, 0.011 mmol) were added, the mixture was again degassed, and the mixture was heated at 150 0C in a Biotage Initiator® microwave for 2 h. The reaction mixture was diluted with EtOAc (30 mL) and washed with water (50 mL). The aqueous layer was extracted with EtOAc (30 mL), and the combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. Purification via flash column chromatography (0-50% EtOAc/hexanes) afforded the title compound (323.6 mg, 82%) as an off white solid. LC-MS m/z 348 (M+H)+, 1.24 min (ret time).
Intermediate 69
2-[(3S)-3-amino-l-pyrrolidinyl]-4,5,6,7-tetrahydro-l-benzothiophene-3-carbonitrile hydrochloride
Figure imgf000070_0001
A solution of 1,1-dimethylethyl [(35)-l- -(3-cya"n'o-t4,5>,6,7-tetrahydro-l-benzo-thien- 2-yl)-3-pyrrolidinyl] carbamate (690.5 mg, 1.987 mmol) in HCl (4 M solution in 1,4- dioxane, 10.0 mL, 40.0 mmol) was stirred at 30 0C for 45 min. The reaction mixture was diluted with Et2O (10 mL) and the resulting precipitate was collected by filtration.
Washing the solid with Et2O (10 mL) afforded the title compound (535.7 mg, 95%). LC- MS m/z 248 (M+H)+, 0.72 min (ret time).
Intermediate 70
1,1-dimethylethyl [(15)-2-{[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3- pyrrolidinyl] amino}-2-oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000070_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5,6,7-tetrahydro-l-benzo- thiophene-3-carbonitrile hydrochloride (179 mg, 0.631 mmol), N-{[(l,l-dimethylethyl)- oxy]carbonyl}-3-(2-thienyl)-L-alanine (171 mg, 0.631 mmol), and HATU (305 mg, 0.802 mmol) in CH2Cl2 (10.0 mL) was added Et3N (0.316 mL, 2.270 mmol). The reaction mixture was stirred at RT for 1 h. Water was added, the layers were separated, and the organic layer was concentrated. The residue was dissolved in EtOAc (30 mL) and washed with saturated aq. NaHCO3 (10 mL) and brine (10 mL), and the aqueous layers were extracted with EtOAc (2 x 20 mL). The combined organic layers were concentrated in vacuo and the residue was purified via flash column chromatography (20-70%
EtOAc/hexanes) to afford the title compound (374.7 mg, >100%, contained some residual solvent). LC-MS m/z 501 (M+H)+, 1.26 min (ret time).
Intermediate 71
l,l-dimethylethyl [(15)-l-({[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3- pyrrolidinyl] amino}carbonyl)propyl] carbamate
Figure imgf000071_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5,6,7-tetrahydro-l-benzo- thiophene-3-carbonitrile hydrochloride (179 mg, 0.631 mmol), (25)-2-({[(l,l- dimethylethyl)oxy]carbonyl}amino)butanoic acid (128 mg, 0.631 mmol), and HATU (305 mg, 0.802 mmol) in CH2Cl2 (10.0 mL) was added Et3N (0.316 mL, 2.270 mmol). The reaction mixture was stirred at RT for 1 h. Water was added, the layers were separated, and the organic layer was concentrated. The residue was dissolved in EtOAc (30 mL) and washed with saturated aq. NaHCO3 (10 mL) and brine (10 mL), and the aqueous layers were extracted with EtOAc (2 x 20 mL). The combined organic layers were concentrated in vacuo and the residue was purified via flash column chromatography (20-70%
EtOAc/hexanes) to afford the title compound (371.8 mg, >100%, contained some residual solvent). LC-MS m/z 433 (M+H)+, 1.15 min (ret time). Intermediate 72
1,1-dimethylethyl [(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl] carbamate
Figure imgf000072_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (667 mg, 3.58 mmol) and 3-chloro-2-pyrazinecarbonitrile (500 mg, 3.58 mmol) in THF (5.0 mL) was added Et3N (1.00 mL, 7.17 mmol). The reaction vial was capped and heated to 80 0C for 15 h. Upon cooling, the reaction mixture was diluted with EtOAc (~20 mL) and washed with brine (~30 mL). The aqueous layer was extracted with EtOAc (~ 20 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo to afford the title compound (1.07 g, >100%, contained some residual solvent) as a tan solid. LC-MS m/z 290 (M+H)+, 0.91 min (ret time).
Intermediate 73
3-[(3S)-3-amino-l-pyrrolidinyl]-2-pyrazinecarbonitrile hydrochloride
Figure imgf000072_0002
A solution of 1 , 1 -dimethylethyl [(3S)- 1 -(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]- carbamate (1.037 g, 3.58 mmol) in HCl (4 M solution in 1,4-dioxane, 10.0 mL, 40.0 mmol) was stirred at RT for 1 h. Additional HCl (4 M solution in 1,4-dioxane, 4.0 mL, 16.0 mmol) was added and the mixture was stirred at RT for another 0.5 h. The reaction mixture was filtered and the isolated solid was washed with Et2O to afford the title compound (1.05 g, >100%, contained some residual solvent) as a bright yellow solid. LC- MS m/z 190 (M+H)+, 0.45 min (ret time).
Intermediate 74
1,1-dimethylethyl ((15)-2-{[(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000072_0003
To a solution of 3-[(35)-3-amino-l-pyrrolidinyl]-2-pyrazinecarbonitrile
hydrochloride (100 mg, 0.443 mmol), JV-(tert-butoxycarbonyl)-L-alanine (84 mg, 0.443 mmol), and HATU (214 mg, 0.564 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.222 mL, 1.595 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (81.6 mg, 51 %). LC-MS m/z 361 (M+H)+, 0.79 min (ret time).
Intermediate 75
l,l-dimethylethyl [(15)-l-({[(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000073_0001
To a solution of 3-[(35)-3-amino- 1 -pyrrolidinyl]-2-pyrazinecarbonitrile
hydrochloride (100 mg, 0.443 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (90 mg, 0.443 mmol), and HATU (214 mg, 0.564 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.222 mL, 1.595 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (94.3 mg, 57%). LC-MS m/z 375 (M+H)+, 0.86 min (ret time). Intermediate 76
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000074_0001
To a solution of 3-[(35)-3-amino- 1 -pyrrolidinyl]-2-pyrazinecarbonitrile hydrochloride (100 mg, 0.443 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (120 mg, 0.443 mmol), and HATU (214 mg, 0.564 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.222 mL, 1.595 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (141.5 mg, 72%). LC-MS m/z 443 (M+H)+, 1.00 min (ret time).
Intermediate 77
1,1-dimethylethyl [(3S)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl] carbamate
Figure imgf000074_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (0.763 g, 4.10 mmol) and 2-fluoro-3-pyridinecarbonitrile (0.500 g, 4.10 mmol) in DMF (5.0 mL) was added K2CO3 (1.132 g, 8.19 mmol). The reaction mixture was heated to 80 0C for 15 h. Upon cooling, the reaction mixture was diluted with water (~30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over Na2SO4, filtered, and concentrated in vacuo to afford the title compound (1.71 g, >100%, contained some residual solvent) as a cream colored solid. LC-MS m/z 289 (M+H)+, 0.91 min (ret time). Intermediate 78
2-[(3S)-3-amino-l-pyrrolidinyl]-3-pyridinecarbonitrile hydrochloride
Figure imgf000075_0001
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl]- carbamate (1.18 g, 4.09 mmol) in HCl (4 M solution in 1,4-dioxane, 10.0 rnL, 40.0 mmol) was stirred at RT for 1 h. The reaction mixture was filtered and washed with Et2O to afford the title compound (1.03 g, >100%, contained some residual solvent) as an off white solid. LC-MS m/z 189 (M+H)+, 0.52 min (ret time).
Intermediate 79
1,1-dimethylethyl [(15)-l-({[(3S)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000075_0002
To a solution of 2-[(3S)-3-amino-l-pyrrolidinyl]-3-pyridinecarbonitrile hydrochloride (100 mg, 0.445 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (90 mg, 0.445 mmol), and HATU (215 mg, 0.566 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.223 mL, 1.602 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (145.7 mg, 88%). LC-MS m/z 374 (M+H)+, 0.86 min (ret time). Intermediate 80
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl]amino}-2-oxo- l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000076_0001
To a solution of 2-[(3S)-3-amino-l-pyrrolidinyl]-3-pyridinecarbonitrile
hydrochloride (100 mg, 0.445 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (121 mg, 0.445 mmol), and HATU (215 mg, 0.566 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.223 mL, 1.602 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (175.9 mg, 90%). LC-MS m/z 442 (M+H)+, 0.99 min (ret time).
Intermediate 81
1,1-dimethylethyl ((15)-2-{[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- amino}-l-methyl-2-oxoethyl)carbamate
Figure imgf000076_0002
To a mixture of 1,1-dimethylethyl ((15)-2-{[(35)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (41 mg, 0.094 mmol), (4-fluoro- phenyl)boronic acid (14 mg, 0.100 mmol), and K2CO3 (40 mg, 0.289 mmol) in 1,4- dioxane (0.75 mL) and water (0.25 mL) was added PdCl2(dppf) (7 mg, 9.57 μmol). The vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 140 0C for 10 min. Water (5 mL) and CH2Cl2 (2 mL) were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted with CH2Cl2 (3 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C. The crude product was dissolved in DMSO (1 rnL) and purified by reverse phase HPLC (YMC C18 S-5 μm/12 nm 75 x 30 mm preparatory column), eluting at 35 mL/min with a linear gradient running from 20% CH3CN/H2O (0.1% TFA) to 70% CH3CN/H2O (0.1% TFA) over 15 min to afford the title compound (17.6 mg, 41.5%). LC-MS m/z 453 (M+H)+, 1.16 min (ret time).
Intermediate 82
1,1-dimethylethyl [(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl] carbamate
Figure imgf000077_0001
A mixture of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (260 mg, 1.396 mmol), 3-bromo-2-thiophenecarbonitrile (200 mg, 1.064 mmol), Pd2(dba)3 (128 mg, 0.140 mmol), racemic BINAP (26 mg, 0.042 mmol), and Cs2CO3 (460 mg, 1.411 mmol) in toluene (7 mL) was degassed by bubbling argon through for 4 min and then heated to 100 0C for 22 h 45 min. The reaction mixture was diluted with EtOAc (7 mL) and washed with water (5 mL). The aqueous layer was extracted with EtOAc (2 x 5 mL), and the combined organic layers were washed with brine (4 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (0-30% EtOAc/hexanes) afforded the title compound (246 mg, 60%). LC-MS m/z 294 (M+H)+, 1.04 min (ret time).
Intermediate 83
3-[(3S)-3-amino-l-pyrrolidinyl]-2-thiophenecarbonitrile hydrochloride
Figure imgf000077_0002
A solution of 1,1-dimethylethyl [(35)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]- carbamate (246 mg, 0.838 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1 h 30 min. The reaction mixture was diluted with Et2O (5 mL) and the resulting yellow precipitate was collected by filtration. Washing the solid with Et2O (3 mL), followed by dissolving in MeOH (5 mL) and concentrating under a stream of nitrogen at 50 0C afforded the title compound (175 mg, 91%). LC-MS m/z 194 (M+H)+, 0.66 min (ret time). Intermediate 84
l,l-dimethylethyl ((lS)-2-{[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000078_0001
To a solution of 3-[(35)-3-amino- 1 -pyrrolidinyl]-2-thiophenecarbonitrile hydrochloride (38 mg, 0.165 mmol), N-(tert-butoxycarbonyl)-L-alanine (31.3 mg, 0.165 mmol), and HATU (69.2 mg, 0.182 mmol) in CH2Cl2 (1.0 mL) was added Et3N (0.07 mL, 0.502 mmol). The reaction mixture was stirred at RT for 3 h 20 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-70% EtOAc/hexanes) afforded the title compound (15.6 mg, 26%). LC-MS m/z 365 (M+H)+, 0.92 min (ret time).
Intermediate 85
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]amino}-2-oxo-l- (2-thienylmethyl)ethyl] carbamate
Figure imgf000078_0002
To a solution of 3-[(35)-3-amino-l-pyrrolidinyl]-2-thiophenecarbonitrile hydrochloride (38 mg, 0.165 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (44.9 mg, 0.165 mmol), and HATU (69.2 mg, 0.182 mmol) in CH2Cl2 (1.0 mL) was added Et3N (0.07 mL, 0.502 mmol). The reaction mixture was stirred at RT for 3 h 20 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-70% EtOAc/hexanes) afforded the title compound (43.5 mg, 59%). LC-MS m/z 447 (M+H)+, 1.02 min (ret time). Intermediate 86
1,1-dimethylethyl [(lS)-l-({[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000079_0001
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-3-biphenylcarbonitrile
hydrochloride (36 mg, 0.120 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (24.4 mg, 0.120 mmol), and HATU (49 mg, 0.129 mmol) in CH2Cl2 (0.8 mL) and MeOH (0.4 mL) was added Et3N (0.05 mL, 0.360 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (39 mg, 72%). LC-MS m/z 449 (M+H)+, 1.21 min (ret time). Intermediate 87
1,1-dimethylethyl {(3S)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- carbamate
Figure imgf000079_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (197 mg, 1.058 mmol) and 2-fluoro-4-(trifluoromethyl)benzonitrile (200 mg, 1.058 mmol) in DMF (4.0 mL) was added K2CO3 (146 mg, 1.058 mmol). The reaction vial was capped and heated to 100 0C for 1 h 45 min. Upon cooling, water (10 mL) and EtOAc (5 mL) were added.
The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 5 mL).
The combined organic layers were washed with brine (3 x 2 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-40% EtOAc/hexanes) afforded the title compound (247 mg, 66%). LC-MS m/z 356 (M+H)+, 1.23 min (ret time). Intermediate 88
2-[(3S)-3-amino-l-pyrrolidinyl]-4-(trifluoromethyl)benzonitrile hydrochloride
Figure imgf000080_0001
A solution of 1,1-dimethylethyl {(35)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3- pyrrolidinyl} carbamate (247 mg, 0.695 mmol) in HCl (4 M solution in 1,4-dioxane, 3.0 rnL, 12.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (10 mL) and the resulting precipitate was collected by filtration. The solid was washed with Et2O (2 x 5 mL) and collected. Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (187 mg, 92%). LC-MS m/z 256 (M+H)+, 0.72 min (ret time).
Intermediate 89
1,1-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-l-methyl-2-oxoethyl]carbamate
Figure imgf000080_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4-(trifluoromethyl)benzonitrile hydrochloride (60 mg, 0.206 mmol), JV-(te/t-butoxycarbonyl)-L-alanine (39 mg, 0.206 mmol), and HATU (78 mg, 0.206 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.086 mL, 0.617 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (68 mg, 78%). LC-MS m/z All (M+H)+, 1.09 min (ret time). Intermediate 90
l^-dimethylethyl Kl^-l-ICKS^-l-Il-cyano-S-CtrifluoromethyOphenyll-S- pyrrolidinyl}amino)carbonyl]propyl}carbamate
Figure imgf000081_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4-(trifluoromethyl)benzonitrile hydrochloride (60 mg, 0.206 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (42 mg, 0.207 mmol), and HATU (78 mg, 0.206 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.086 mL, 0.617 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (76 mg, 84%). LC-MS m/z 441 (M+H)+, 1.14 min (ret time). Intermediate 91
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000081_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4-(trifluoromethyl)benzonitrile hydrochloride (62 mg, 0.213 mmol), N-{[(\ ,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (58 mg, 0.214 mmol), and HATU (81 mg, 0.213 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.088 mL, 0.638 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 rnL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (76 mg, 70%). LC-MS m/z 509 (M+H)+, 1.26 min (ret time).
Intermediate 92
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000082_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (134 mg, 0.459 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (125 mg, 0.459 mmol), and HATU (175 mg, 0.459 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.191 mL, 1.378 mmol). The reaction mixture was stirred at RT for 20 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (223 mg, 95%). LC-MS m/z 509 (M+H)+, 1.28 min (ret time).
Intermediate 93
1,1-dimethylethyl [(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]carbamate
Figure imgf000082_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (231 mg, 1.239 mmol) and 2-fluorobenzonitrile (150 mg, 1.239 mmol) in DMF (2.5 mL) was added K2CO3 (171 mg, 1.239 mmol). The reaction vial was capped and heated to 100 0C overnight. Upon cooling, water (5 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (249 mg, 70%). LC-MS m/z 288 (M+H)+, 1.08 min (ret time).
Intermediate 94
2-[(3S)-3-amino-l-pyrrolidinyl]benzonitrile hydrochloride
Figure imgf000083_0001
A solution of 1,1-dimethylethyl [(3S)- l-(2-cyanophenyl)-3-pyrrolidinyl] -carbamate (249 mg, 0.867 mmol) in HCl (4 M solution in 1,4-dioxane, 4.0 mL, 16.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (7 mL) and the resulting precipitate was collected by filtration. The solid was washed with Et2O (2 x 3 mL) and collected. Residual solid was dissolved in MeOH, concentrated under a stream of nitrogen at 50 0C, and combined with the main batch to afford the title compound (177 mg, 91%). LC-MS m/z 188 (M+H)+, 0.61 min (ret time).
Intermediate 95
1,1-dimethylethyl ((lS)-2-{[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]amino}-l-methyl- 2-oxoethyl)carbamate
Figure imgf000083_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]benzonitrile hydrochloride (54 mg, 0.241 mmol), JV-(ter£-butoxycarbonyl)-L-alanine (46 mg, 0.243 mmol), and HATU (92 mg, 0.241 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.100 mL, 0.724 mmol). The reaction mixture was stirred at RT for 20 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (93 mg, >100%, contained some residual solvent). LC-MS m/z 359 (M+H)+, 0.97 min (ret time). Intermediate 96
1,1-dimethylethyl [(lS)-l-({[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]amino}carbonyl)- propyl] carbamate
Figure imgf000084_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]benzonitrile hydrochloride (54 mg, 0.241 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)butanoic acid (50 mg, 0.246 mmol), and HATU (92 mg, 0.241 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.100 mL, 0.724 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (89.5 mg, 100%). LC-MS m/z 373 (M+H)+, 1.02 min (ret time).
Intermediate 97
1,1-dimethylethyl [(15)-2-{[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]amino}-2-oxo-l-(2- thienylmethyl)ethyl] carbamate
Figure imgf000084_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]benzonitrile hydrochloride (54 mg, 0.241 mmol), Λ/-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)-L-alanine (66 mg, 0.243 mmol), and HATU (92 mg, 0.241 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.100 mL, 0.724 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (103 mg, 97%). LC-MS m/z 441 (M+H)+, 1.11 min (ret time).
Intermediate 98
1,1-dimethylethyl [(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]carbamate
Figure imgf000085_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (178 mg, 0.955 mmol) and 2,4,5-trifluorobenzonitrile (150 mg, 0.955 mmol) in DMF (2.5 mL) was added K2CO3 (132 mg, 0.955 mmol). The reaction vial was capped and heated to 100 0C overnight. Upon cooling, water (5 mL) and EtOAc (3 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (4 x 1 mL). The combined organic layers were washed with brine (3 x 1 mL) and then concentrated onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (227 mg, 74%). LC-MS m/z 324 (M+H)+, 1.12 min (ret time).
Intermediate 99
2-[(3S)-3-amino-l-pyrrolidinyl]-4,5-difluorobenzonitrile hydrochloride
Figure imgf000085_0002
A solution of 1,1-dimethylethyl [(35)-l-(2-cyano-4,5-difluorophenyl)-3- pyrrolidinyl] carbamate (227 mg, 0.702 mmol) in HCl (4 M solution in 1,4-dioxane, 3.5 mL, 14.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (7 mL) and the resulting precipitate was collected by filtration. The solid was washed with Et2O (2 x 3 mL) and collected. Residual solid was dissolved in MeOH, combined with the main batch, and concentrated under a stream of nitrogen at 50 0C to afford the title compound (169 mg, 93%). LC-MS m/z 224 (M+H)+, 0.63 min (ret time). Intermediate 100
l,l-dimethylethyl ((15)-2-{[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]- amino}-l-methyl-2-oxoethyl)carbamate
Figure imgf000086_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5-difluorobenzonitrile hydrochloride (50 mg, 0.193 mmol), Λ/-(tert-butoxycarbonyl)-L-alanine (37 mg, 0.196 mmol), and HATU (74 mg, 0.195 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.080 mL, 0.578 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with
EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (80 mg, >100%, contained some residual solvent). LC-MS m/z 395 (M+H)+, 1.02 min (ret time). Intermediate 101
l,l-dimethylethyl [(15)-l-({[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]- amino}carbonyl)propyl] carbamate
Figure imgf000086_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5-difluorobenzonitrile hydrochloride (50 mg, 0.193 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)- butanoic acid (40 mg, 0.197 mmol), and HATU (74 mg, 0.195 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.080 mL, 0.578 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (85 mg, >100%, contained some residual solvent). LC-MS m/z 409 (M+H)+, 1.05 min (ret time).
Intermediate 102
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]- amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000087_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-4,5-difluorobenzonitrile hydrochloride (50 mg, 0.193 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)- L-alanine (53 mg, 0.195 mmol), and HATU (74 mg, 0.195 mmol) in CH2Cl2 (1.5 mL) and MeOH (0.75 mL) was added Et3N (0.080 mL, 0.578 mmol). The reaction mixture was stirred at RT for 15 min and then concentrated under a stream of nitrogen at 50 0C. Water (3 mL) and EtOAc (2 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 x 1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (83 mg, 90%). LC-MS m/z All (M+H)+, 1.15 min (ret time).
Intermediate 103
1,1-dimethylethyl ((lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]amino}-l- cyclopentyl-2-oxoethyl)carbamate
Figure imgf000087_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-nitrobenzonitrile hydrochloride (75 mg, 0.279 mmol), (25)-cyclopentyl({[(l,l-dimethylethyl)oxy]- carbonyl}amino)ethanoic acid dicyclohexylamine (119 mg, 0.279 mmol), and HATU (135 mg, 0.355 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.14 mL, 1.004 mmol). The reaction mixture was stirred at RT for 2 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (133.7 mg, >100%, contained some residual solvent). LC-MS m/z 458 (M+H)+, 1.09 min (ret time).
Intermediate 104
l,l-dimethylethyl [(15)-l-({[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000088_0001
HCl
To a solution of 3-[(35)-3-amino-l-pyrrolidinyl]-2-thiophenecarbonitrile hydrochloride (101 mg, 0.440 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (89 mg, 0.440 mmol), and HATU (184 mg, 0.484 mmol) in CH2Cl2 (2.0 mL) was added Et3N (0.184 mL, 1.319 mmol). The reaction mixture was stirred at RT for 1 h 40 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification twice via flash column chromatography (20-80% then 20-100% EtOAc/hexanes) afforded the title compound (156 mg, 94%). LC-MS m/z 379 (M+H)+, 0.85 min (ret time).
Intermediate 105
2-nitro-5-(phenyloxy)benzonitrile
Figure imgf000088_0002
To a slurry of NaH (16 mg, 0.667 mmol) in THF (3.0 mL) were added 5-fluoro-2- nitrobenzonitrile (100 mg, 0.602 mmol) and phenol (57 mg, 0.606 mmol). The reaction vial was capped and the reaction mixture was stirred at RT for 50 min. Water (3 mL) and EtOAc (5 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 3 rnL). The combined organic layers were washed with brine (3 rnL) and then concentrated under a stream of nitrogen at 50 0C to afford the title compound (173 mg, 63%). LC-MS m/z 241 (M+H)+, 1.08 min (ret time).
Intermediate 106
2-amino-5-(phenyloxy)benzonitrile
Figure imgf000089_0001
A mixture of 2-nitro-5-(phenyloxy)benzonitrile (173 mg, 0.720 mmol) and
Pd(OH)2 (20 wt. % on carbon, 25 mg, 0.036 mmol) in MeOH (3 mL) was purged with hydrogen for 15 sec and the reaction was stirred at RT under an atmosphere of hydrogen. After 3 h 30 min, the reaction mixture was filtered through a plug of Celite® and washed through with MeOH (~10 mL). The filtrate was concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-20%
EtOAc/hexanes) afforded the title compound (80 mg, 53%). LC-MS m/z 211 (M+H)+, 1.02 min (ret time). Intermediate 107
2-bromo-5-(phenyloxy)benzonitrile
Figure imgf000089_0002
To a slurry of CuBr2 (102 mg, 0.457 mmol) and tert-hvXy\ nitrite (0.068 mL, 0.574 mmol) in CH3CN (2.0 mL) at 0 0C was added a solution of 2-amino-5- (phenyloxy)benzonitrile (80 mg, 0.381 mmol) in CH3CN (1.0 mL). The ice bath was removed, and the reaction was stirred at RT for 10 min. 6 M aq. HCl (2 mL), brine (1 mL), and EtOAc (10 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were washed with 6 M aq. HCl (2 mL) and brine (2 mL), and concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-20% EtOAc/hexanes) afforded the title compound (62 mg, 59%). LC-MS m/z 2141216 (M+H)+, 1.21 min (ret time).
Intermediate 108
1,1-dimethylethyl {(3S)-l-[2-cyano-4-(phenyloxy)phenyl]-3-pyrrolidinyl}carbamate
Figure imgf000090_0001
A mixture of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (67 mg, 0.360 mmol), 2-bromo-5-(phenyloxy)benzonitrile (62 mg, 0.226 mmol), Pd2(dba)3 (33 mg, 0.036 mmol), racemic BINAP (7 mg, 0.011 mmol), and Cs2CO3 (118 mg, 0.364 mmol) in toluene (1 mL) was degassed by bubbling argon through for 4 min and then heated to 100 0C for 19 h 30 min. The reaction mixture was diluted with EtOAc (2 mL) and washed with water (1 mL). The aqueous layer was extracted with EtOAc (2 x 2 mL), and the combined organic layers were washed with brine (1 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (0-30%
EtOAc/hexanes) afforded the title compound (62 mg, 43%). LC-MS m/z 380 (M+H)+, 2.18 min (ret time).
Intermediate 109
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(phenyloxy)benzonitrile hydrochloride
Figure imgf000090_0002
A solution of 1,1-dimethylethyl {(35)-l-[2-cyano-4-(phenyloxy)phenyl]-3- pyrrolidinyl} carbamate (62 mg, 0.163 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (3 mL) and the resulting white precipitate was collected by filtration through a plug of cotton. Washing the solid with Et2O (1 mL), followed by dissolving in MeOH (3 mL) and concentrating under a stream of nitrogen at 50 0C afforded the title compound (49.7 mg, 96%). LC-MS m/z 280 (M+H)+, 0.83 min (ret time). Intermediate 110
1,1-dimethylethyl [(lS)-2-({(3S)-l-[2-cyano-4-(phenyloxy)phenyl]-3-pyrrolidinyl}- amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000091_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(phenyloxy)benzonitrile hydrochloride (49.7 mg, 0.157 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (49 mg, 0.181 mmol), and HATU (75 mg, 0.197 mmol) in CH2Cl2 (1.0 mL) was added Et3N (0.074 mL, 0.531 mmol). The reaction mixture was stirred at RT for 2 h 15 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-80% EtOAc/hexanes) afforded the title compound (61 mg, 63%). LC-MS m/z 533 (M+H)+, 1.32 min (ret time).
Intermediate 111
1,1-dimethylethyl [(3S)-l-(3-cyano-4-pyridinyl)-3-pyrrolidinyl] carbamate
Figure imgf000091_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (470 mg, 2.53 mmol) and 4-chloro-3-pyridinecarbonitrile (350 mg, 2.53 mmol) in DMF (5.0 mL) was added K2CO3 (698 mg, 5.05 mmol). The reaction vial was capped and heated to 100 0C for 15 h. Upon cooling, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (2 x 20 mL). The combined organic layers were washed with brine (3 x 30 mL), dried over Na2SO4, filtered, and concentrated in vacuo to afford the title compound (890 mg, >100%, contained some residual solvent) as a yellowish, orange oil. LC-MS m/z 289 (M+H)+, 0.65 min (ret time). Intermediate 112
4-[(3S)-3-amino-l-pyrrolidinyl]-3-pyridinecarbonitrile hydrochloride
Figure imgf000092_0001
HCI
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-4-pyridinyl)-3-pyrrolidinyl]- carbamate (728 mg, 2.52 mmol) in HCl (4 M solution in 1,4-dioxane, 10.0 mL, 40.0 mmol) was stirred at RT for 1 h. The reaction mixture was filtered and washed with Et2O to afford the title compound (547 mg, 96%) as an off white solid. LC-MS m/z 189 (M+H)+, 0.14 min (ret time).
Intermediate 113
1,1-dimethylethyl [(15)-2-{[(3S)-l-(3-cyano-4-pyridinyl)-3-pyrrolidinyl]amino}-2-oxo- l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000092_0002
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-3-pyridinecarbonitrile
hydrochloride (100 mg, 0.445 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (121 mg, 0.445 mmol), and HATU (215 mg, 0.566 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.223 mL, 1.602 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (127 mg, 65%). LC-MS m/z 442 (M+H)+, 0.88 min (ret time). Intermediate 114
1,1-dimethylethyl [(3S)-l-(2-cyano-3-pyridinyl)-3-pyrrolidinyl] carbamate
Figure imgf000093_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (0.763 g, 4.10 mmol) and 3-fluoro-2-pyridinecarbonitrile (0.500 g, 4.10 mmol) in DMF (5.0 mL) was added K2CO3 (1.132 g, 8.19 mmol). The reaction mixture was heated to 100 0C for 15 h. Upon cooling, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (3 x 30 mL) and concentrated in vacuo. Purification via flash column chromatography (30-100% EtOAc/hexanes) afforded the title compound (1.36 g, >100%, contained some residual solvent) as a colorless oil. LC-MS m/z 289 (M+H)+, 1.36 min (ret time).
Intermediate 115
3-[(3S)-3-amino-l-pyrrolidinyl]-2-pyridinecarbonitrile hydrochloride
Figure imgf000093_0002
HCI
A solution of 1 , 1 -dimethylethyl [(3S)- 1 -(2-cyano-3 -pyridinyl)-3 -pyrrolidinyl] - carbamate (1.36 g, 4.72 mmol) in HCl (4 M solution in 1,4-dioxane, 10.0 mL, 40.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O and the resulting precipitate was collected by filtration to afford the title compound (941 mg, 89%) as a bright yellow solid. LC-MS m/z 189 (M+H)+, 0.51 min (ret time). Intermediate 116
1,1-dimethylethyl ((l^-l-ilCSSJ-l-Cl-cyano-S-pyridinyO-S-pyrrolidinyllamino}-!- methyl-2-oxoethyl)carbamate
Figure imgf000093_0003
HCl
To a solution of 3-[(35)-3-amino-l-pyrrolidinyl]-2-pyridinecarbonitrile
hydrochloride (100 mg, 0.445 mmol), JV-(tert-butoxycarbonyl)-L-alanine (84 mg, 0.445 mmol), and HATU (215 mg, 0.566 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.223 mL, 1.602 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound. LC-MS m/z 360 (M+H)+, 1.13 min (ret time).
Intermediate 117
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-3-pyridinyl)-3-pyrrolidinyl]amino}-2-oxo- l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000094_0001
HCI
To a solution of 3-[(35)-3-amino-l-pyrrolidinyl]-2-pyridinecarbonitrile
hydrochloride (100 mg, 0.445 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (121 mg, 0.445 mmol), and HATU (215 mg, 0.566 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.223 mL, 1.602 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (169 mg, 86%). LC-MS m/z 442 (M+H)+, 1.48 min (ret time).
Intermediate 118
1,1-dimethylethyl [(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl] carbamate
Figure imgf000094_0002
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (0.494 g, 2.65 mmol) and 2-chloro-3-quinolinecarbonitrile (0.500 g, 2.65 mmol) in DMF (5.0 mL) was added K2CO3 (0.733 g, 5.30 mmol). The reaction mixture was heated to 100 0C for 15 h. Upon cooling, the reaction mixture was diluted with water. The resulting yellow precipitate was collected by filtration and washed with water. Drying of the solid for 2 h in vacuo followed by an additional 15 h of drying in vacuo at 45 0C afforded the title compound (1.35 g, 97%). LC-MS m/z 339 (M+H)+, 0.97 min (ret time).
Intermediate 119
2-[(3S)-3-amino-l-pyrrolidinyl]-3-quinolinecarbonitrile hydrochloride
Figure imgf000095_0001
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]- carbamate (897 mg, 2.65 mmol) in HCl (4 M solution in 1,4-dioxane, 10.0 mL, 40.0 mmol) was stirred at RT for 1 h. The reaction mixture was concentrated under a stream of nitrogen at 50 0C. The resulting residue was rinsed three times with Et2O and dried in vacuo to afford the title compound (772 mg, >100%, contained some residual solvent) as a tan solid. LC-MS m/z 239 (M+H)+, 0.71 min (ret time).
Intermediate 120
1,1-dimethylethyl ((lS)-2-{[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]amino}-l- methyl-2-oxoethyl)carbamate
Figure imgf000095_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-3-quinolinecarbonitrile (100 mg, 0.420 mmol), JV-(tert-butoxycarbonyl)-L-alanine (79 mg, 0.420 mmol), and HATU (203 mg, 0.534 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.211 mL, 1.511 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (95 mg, 55%) as a bright yellow solid. LC-MS m/z 410 (M+H)+, 1.27 min (ret time).
Intermediate 121
l,l-dimethylethyl [(15)-l-({[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]amino}- carbonyl)propyl] carbamate
Figure imgf000096_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-3-quinolinecarbonitrile (100 mg, 0.420 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}amino)butanoic acid (85 mg, 0.420 mmol), and HATU (203 mg, 0.534 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.211 mL, 1.511 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 mL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (95 mg, 54%) as a bright yellow solid. LC-MS m/z 424 (M+H)+, 1.36 min (ret time).
Intermediate 122
1,1-dimethylethyl [(lS)-2-{[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]amino}-2- oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000096_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-3-quinolinecarbonitrile (100 mg, 0.420 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2-thienyl)-L-alanine (114 mg, 0.420 mmol), and HATU (203 mg, 0.534 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.211 mL, 1.511 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the aqueous layer was extracted with CH2Cl2. The combined organic layers were concentrated in vacuo and the residue was dissolved in EtOAc (15 rnL) and washed with saturated aq. NaHCO3 (10 mL), brine (10 mL), and water (20 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30-100% EtOAc/hexanes) to afford the title compound (113 mg, 55%) as a bright yellow solid. LC-MS m/z 492 (M+H)+, 1.60 min (ret time).
Intermediate 123
1,1-dimethylethyl [(3S)-l-(3-cyano-2-thienyl)-3-pyrrolidinyl] carbamate
Figure imgf000097_0001
A mixture of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (650 mg, 3.49 mmol), 2-bromo-3-thiophenecarbonitrile (500 mg, 2.66 mmol), Pd2(dba)3 (320 mg, 0.349 mmol), racemic BINAP (65 mg, 0.104 mmol), and Cs2CO3 (1.15 g, 3.53 mmol) in toluene (15 mL) was degassed by bubbling argon through for 4 min and then heated to 100 0C overnight. The reaction mixture was diluted with EtOAc (15 mL) and washed with water (10 mL). The aqueous layer was extracted with EtOAc (2 x 10 mL), and the combined organic layers were washed with brine (8 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification twice via flash column chromatography (0-30% EtOAc/hexanes) afforded the title compound (324 mg, 32%) as a bright yellow solid. LC- MS m/z 294 (M+H)+, 1.07 min (ret time).
Intermediate 124
2-[(3S)-3-amino-l-pyrrolidinyl]-3-thiophenecarbonitrile hydrochloride
Figure imgf000097_0002
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-2-thienyl)-3-pyrrolidinyl]- carbamate (324 mg, 1.104 mmol) in HCl (4 M solution in 1,4-dioxane, 1.1 mL, 4.40 mmol) was stirred at RT for 1 h 40 min. The reaction mixture was diluted with Et2O (5 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (3 mL), followed by dissolving in MeOH (5 mL) and concentrating under a stream of nitrogen at 50 0C afforded the title compound (318 mg, >100%, contained some residual solvent) as a bright yellow solid. LC-MS m/z 194 (M+H)+, 0.64 min (ret time).
Intermediate 125
l,l-dimethylethyl [(15)-2-{[(3S)-l-(3-cyano-2-thienyl)-3-pyrrolidinyl]amino}-2-oxo-l-
(2-thienylmethyl)ethyl]carbamate
Figure imgf000098_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-3-thiophenecarbonitrile hydrochloride (110 mg, 0.479 mmol), 7V-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (122 mg, 0.450 mmol), and HATU (190 mg, 0.500 mmol) in CH2Cl2 (2.0 mL) was added Et3N (0.18 mL, 1.291 mmol). The reaction mixture was stirred at RT for 18 h 20 min. Water (2 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification twice via flash column chromatography (20-80% then 20-100% EtOAc/hexanes) afforded the title compound (5.4 mg, 2.5%) as a bright yellow solid. LC-MS m/z 447 (M+H)+, 0.93 min (ret time).
Intermediate 126
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl] carbamate
Figure imgf000098_0002
HCi
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (100 mg, 0.343 mmol), Λ/-(ter^butoxycarbonyl)-JV-methyl-L-alanine (70 mg, 0.344 mmol), and HATU (130 mg, 0.343 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.15 mL, 1.082 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (4 mL) and
EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 niL). The combined organic layers were washed with saturated aq. NH4Cl (2 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (140 mg, 93%). LC-MS m/z 441 (M+H)+, 1.29 min (ret time).
Intermediate 127
1,1-dimethylethyl {(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}carbamate
Figure imgf000099_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrr (1.24 g, 6.66 mmol) and 2-fluoro-5-(methyloxy)benzonitrile (1.00 g, 6.62 mmol) in DMF (15.0 mL) was added K2CO3 (0.92 g, 6.66 mmol). The reaction mixture was heated to 100 0C for 120 h. Upon cooling, water (40 mL) and EtOAc (20 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3 x 15 mL), dried over Na2SO4, filtered, and concentrated onto Isolute®. Purification via flash column chromatography (0-80%
EtOAc/hexanes) afforded the title compound (0.460 g, 22%). LC-MS m/z 318 (M+H)+, 1.19 min (ret time).
Intermediate 128
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(methyloxy)benzonitrile hydrochloride
Figure imgf000099_0002
A solution of 1 , 1 -dimethylethyl {(35)- 1 -[2-cyano-4-(methyloxy)phenyl]-3- pyrrolidinyl} carbamate (444 mg, 1.339 mmol) in HCl (4 M solution in 1,4-dioxane, 7.0 mL, 28.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (20 mL) and the resulting precipitate was collected by filtration. Washing the solid with Et2O (2 x 5 mL), followed by dissolving in MeOH, concentrating under a stream of nitrogen at 50 0C, and drying under high vacuum afforded the title compound (318 mg, 90%). LC- MS m/z 218 (M+H)+, 0.67 min (ret time). Intermediate 129
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}- amino)-l-methyl-2-oxoethyl]carbamate
Figure imgf000100_0001
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(methyloxy)benzonitrile hydrochloride (100 mg, 0.394 mmol), JV-(tert-butoxycarbonyl)-L-alanine (75 mg, 0.396 mmol), and HATU (150 mg, 0.394 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.17 mL, 1.226 mmol). The reaction mixture was stirred at RT for 40 min and then concentrated under a stream of nitrogen at 50 0C. Water (4 mL) and EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with
EtOAc (3 x 2 mL). The combined organic layers were washed with saturated aq. NH4Cl (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®.
Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (111 mg, 73%). LC-MS m/z 389 (M+H)+, 1.13 min (ret time). Intermediate 130
l,l-dimethylethyl {(15)-l-[({(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}- amino)carbonyl] propyl} carbamate
Figure imgf000100_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(methyloxy)benzonitrile hydrochloride (100 mg, 0.394 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (80 mg, 0.394 mmol), and HATU (150 mg, 0.394 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.17 mL, 1.226 mmol). The reaction mixture was stirred at RT for 40 min and then concentrated under a stream of nitrogen at 50 0C. Water (4 mL) and EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with saturated aq. NH4Cl (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (108 mg, 68%). LC-MS m/z 403 (M+H)+, 1.17 min (ret time).
Intermediate 131
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}- amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate
Figure imgf000101_0001
HCl
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(methyloxy)benzonitrile hydrochloride (118 mg, 0.465 mmol), N-{[(l,l-dimethylethyl)oxy]carbonyl}-3-(2- thienyl)-L-alanine (126 mg, 0.465 mmol), and HATU (177 mg, 0.465 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.20 mL, 1.443 mmol). The reaction mixture was stirred at RT for 40 min and then concentrated under a stream of nitrogen at 50 0C. Water (4 mL) and EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with saturated aq. NH4Cl (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (0-80%
EtOAc/hexanes) afforded the title compound (194 mg, 89%). LC-MS m/z 471 (M+H)+, 1.23 min (ret time).
Intermediate 132
4,4'-difluoro-3-biphenylcarbonitrile
Figure imgf000101_0002
To a mixture of 5-bromo-2-fluorobenzonitrile (1.00 g, 5.00 mmol), (4- fluorophenyl)boronic acid (0.70 g, 5.00 mmol), and K2CO3 (2.073 g, 15.00 mmol) in 1,4- dioxane (12.0 mL) and water (4.0 mL) was added PdCl2(dppf) (10 mg, 0.014 mmol). After degassing with nitrogen for 2 min, the vial was capped and the reaction mixture was heated in a Biotage Initiator® microwave reactor to 140 0C for 30 min. An identical reaction on the same scale was run in a separate vial and the crude reaction mixtures were combined. The combined reaction mixtures were diluted with EtOAc (50 mL) and washed with water (50 mL) and brine (2 x 50 mL). The organic layer was filtered through a pad of silica gel and washed through with EtOAc. Isolute® Si-TMT (~ 0.5 g) was added and the mixture stirred at RT for 30 min, followed by filtration, and concentration in vacuo. Purification via flash column chromatography (0-20% EtOAc/hexanes) afforded the title compound (1.82 g, 85%) as a white solid. LC-MS m/z 216 (M+H)+, 1.20 min (ret time).
Intermediate 133
1,1-dimethylethyl [(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl] carbamate
Figure imgf000102_0001
To a solution of 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (1.575 g, 8.46 mmol) and 4,4'-difluoro-3-biphenylcarbonitrile (1.82 g, 8.46 mmol) in DMF (20.0 mL) was added K2CO3 (1.169 g, 8.46 mmol). The reaction mixture was heated to 100 0C for 15 h. Upon cooling, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with water (3 x 50 mL) and brine (2 x 50 mL), dried over Na2SO4, filtered, and concentrated in vacuo.
Purification via flash column chromatography (0-60% EtOAc/hexanes) afforded the title compound (3.23 g, 52%) as a white solid. LC-MS m/z 382 (M+H)+, 1.40 min (ret time).
Intermediate 134
4-[(3S)-3-amino-l-pyrrolidinyl]-4'-fluoro-3-biphenylcarbonitrile hydrochloride
Figure imgf000102_0002
HCi
A solution of 1,1-dimethylethyl [(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3- pyrrolidinyl] carbamate (1.66 g, 4.35 mmol) in HCl (4 M solution in 1,4-dioxane, 15.0 mL, 60.0 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (30 mL) and the resulting precipitate was collected by filtration. The solid was washed with Et2O and dried in vacuo to afford the title compound (1.55 g, >100%, contained some residual solvent) as an off white solid. LC-MS m/z 282 (M+H)+, 0.93 min (ret time). Intermediate 135
l,l-dimethylethyl [(15)-l-({[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- amino}carbonyl)propyl] carbamate
Figure imgf000103_0001
HCI
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-4'-fluoro-3-biphenylcarbonitrile hydrochloride (517 mg, 1.464 mmol), (25)-2-({[(l,l-dimethylethyl)oxy]carbonyl}- amino)butanoic acid (298 mg, 1.464 mmol), and HATU (708 mg, 1.862 mmol) in CH2Cl2 (10.0 mL) was added Et3N (0.735 mL, 5.27 mmol). The reaction mixture was stirred at RT for 1 h and then concentrated. The residue was dissolved in EtOAc (50 mL) and washed with water (50 mL), saturated aq. NH4Cl (50 mL), saturated aq. NaHCO3 (50 mL), and brine (2 x 100 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (20-100% EtOAc/hexanes) to afford the title compound (564 mg, 82%). LC-MS m/z 467 (M+H)+, 1.32 min (ret time).
Intermediate 136
l,l-dimethylethyl (25)-2-({[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- amino}carbonyl)-l-azetidinecarboxylate
Figure imgf000103_0002
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-4'-fluoro-3-biphenylcarbonitrile hydrochloride (150 mg, 0.472 mmol), (25)-l-{[(l,l-dimethylethyl)oxy]carbonyl}-2- azetidinecarboxylic acid (95 mg, 0.472 mmol), and Et3N (0.197 mL, 1.416 mmol) in CH2Cl2 (2.0 mL) at 0 0C was added 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphorinane 2,4,6-trioxide (50 wt. % solution in EtOAc, 0.365 mL, 0.614 mmol). The reaction mixture was stirred at 0 0C for 3 h followed by RT for 4 h 20 min. CH2Cl2 (2 mL) was added and the solution was washed with saturated aq. NaHCO3 (2 x 1 mL) and 10% aq. citric acid (2 x 1 mL). The organic solution was dried over Na2SO4, filtered, and concentrated under a stream of nitrogen at 50 0C onto Isolute®. Purification via flash column chromatography (20-70% EtOAc/hexanes) afforded the title compound (114 mg, 87%). LC-MS m/z 465 (M+H)+, 1.20 min (ret time).
Intermediate 137
l,l-dimethylethyl (2S)-2-({[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3- pyrrolidinyl]amino}carbonyl)-l-pyrrolidinecarboxylate
Figure imgf000104_0001
To a solution of 4-[(35)-3-amino-l-pyrrolidinyl]-4'-fluoro-3-biphenylcarbonitrile hydrochloride (100 mg, 0.315 mmol), Λ/-(tert-butoxycarbonyl)-L-proline (68 mg, 0.316 mmol), and HATU (120 mg, 0.315 mmol) in CH2Cl2 (1.0 mL) and MeOH (0.5 mL) was added Et3N (0.131 mL, 0.944 mmol). The reaction mixture was stirred at RT for 30 min and then concentrated under a stream of nitrogen at 50 0C. Water (5 mL) and EtOAc (3 mL) were added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with saturated aq. NH4Cl (1 mL) and then concentrated under a stream of nitrogen at 50 0C onto Isolute®.
Purification via flash column chromatography (0-80% EtOAc/hexanes) afforded the title compound (72 mg, 48%). LC-MS m/z 479 (M+H)+, 1.23 min (ret time).
Intermediate 138
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-l-methyl-2-oxoethyl]carbamate-</i
Figure imgf000104_0002
HCi
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (75 mg, 0.257 mmol), N-(tert-butoxycarbonyl)-L-alanine-Ji (48.9 mg, 0.257 mmol), and HATU (124 mg, 0.327 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.129 mL, 0.926 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the organic layer was concentrated in vacuo. The residue was dissolved in EtOAc (10 mL) and washed with saturated aq. NH4Cl (10 mL), saturated aq. NaHCO3 (10 mL), and brine (10 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30- 100% EtOAc/hexanes) to afford the title compound (87 mg, 79%) as a white solid. LC- MS m/z 428 (M+H)+, 1.21 min (ret time).
Intermediate 139
l,l-dimethylethyl [(15)-2-({(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3- pyrrolidinyl}amino)-l-methyl-2-oxoethyl]carbamate-</4
Figure imgf000105_0001
HCI
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-(trifluoromethyl)benzonitrile hydrochloride (75 mg, 0.257 mmol), N-(tert-butoxycarbonyl)-L-alanine-<i4 (49.7 mg, 0.257 mmol), and HATU (124 mg, 0.327 mmol) in CH2Cl2 (5.0 mL) was added Et3N (0.129 mL, 0.926 mmol). The reaction mixture was stirred at RT for 1 h. Water (5 mL) was added, the layers were separated, and the organic layer was concentrated in vacuo. The residue was dissolved in EtOAc (10 mL) and washed with saturated aq. NH4Cl (10 mL), saturated aq. NaHCO3 (10 mL), and brine (10 mL). The organic solution was concentrated in vacuo and the residue was purified via flash column chromatography (30- 100% EtOAc/hexanes) to afford the title compound (89 mg, 80%) as a white solid. LC- MS m/z 431 (M+H)+, 1.21 min (ret time).
Intermediate 140
3-Bromo-2-nitro-l-benzothiophene
Figure imgf000105_0002
NO2 »BF4 (1015 mg, 7.64 mmol) was added portionwise over 10 min to a solution of 3-bromo-l-benzothiophene (1 mL, 7.64 mmol) in CH2Cl2 (10 mL) at 0 0C (bath temp). Stirring was continued at 0 0C. After 2 h, water (5 mL) was added, and the mixture was filtered. The layers of the filtrate were separated. The organic layer was concentrated, and the crude product was purified by flash column chromatography to afford the title compound (446 mg, 23%). LC-MS m/z 258 (M)+, 1.14 min (ret time). Intermediate 141
l-Nitro-l-benzothiophene-S-carbonitrile
Figure imgf000106_0001
CuCN (185 mg, 2.069 mmol) was added to a solution of 3-bromo-2-nitro-l- benzothiophene (445 mg, 1.724 mmol) in DMF (10 mL). The reaction mixture was heated at 140 0C (bath temp). After 1 h, the reaction mixture was taken up in a solution of 9:1 NH4CI/NH4OH (20 mL) and EtOAc (10 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL) and were concentrated. The crude product was purified by flash column chromatography to afford the title compound (134 mg, 38%). LC-MS m/z 205 (M+H)+, 0.95 min (ret time).
Intermediate 142
l-Amino-l-benzothiophene-S-carbonitrile
Figure imgf000106_0002
A mixture of 2-nitro-l-benzothiophene-3-carbonitrile (134 mg, 0.656 mmol) and
10% Pd/C (70 mg, 0.066 mmol) was taken up in MeOH (3mL) under nitrogen. The flask was purged with hydrogen, and the reaction was stirred under an atmosphere of hydrogen for 24 h. The reaction mixture was filtered through a pad of Celite®, and the pad was washed with MeOH (10 mL) and EtOAc (10 mL). The filtrate was concentrated under reduced pressure and dried under high vacuum to afford the title compound (109 mg, 95%). LC-MS m/z 175 (M+H)+, 0.77 min (ret time).
Intermediate 143
2-bromo-l-benzothiophene-3-carbonitrile
Figure imgf000106_0003
90 wt% tert-butyl nitrite (0.12 mL, 0.911 mmol) was added to a solution of CuBr2 (168 mg, 0.751 mmol) in CH3CN (3 mL) at 0 0C (bath temp). 2-Amino-l- benzothiophene-3-carbonitrile (109 mg, 0.626 mmol) was added portionwise over 30 sec. The ice bath was removed, and the reaction was stirred at RT for 5 min. A solution of 9: 1 saturated aq. NH4CI/NH4OH (3 mL) and EtOAc (2 rnL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were washed with brine (1 x 2 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (71 mg, 48%). LC-MS m/z 238 (M)+, 1.02 min (ret time).
Intermediate 144
1,1-dimethylethyl [(3S)-l-(3-cyano-l-benzothien-2-yl)-3-pyrrolidinyl] carbamate
Figure imgf000107_0001
Pd2(dba)3 (27 mg, 0.029 mmol), BINAP (55 mg, 0.088 mmol), Cs2CO3 (96 mg, 0.294 mmol) and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (71 mg, 0.381 mmol) were added to a 2 dram vial containing a solution of 2-bromo-l-benzothiophene-3- carbonitrile (70 mg, 0.294 mmol) in degassed toluene (1.5 mL) (degassed by bubbling argon through it for 10 min) under argon. The vial was capped, and the reaction mixture was heated at 100 0C (bath temp) for 18 h. Water (3 mL) and EtOAc (2 mL) were added to the reaction, and the organic layer was filtered through a Stratosphere PL-Thio MP SPE cartridge (0.5 g). The aqueous layer was extracted with EtOAc (3 x 2 mL), and each of these extractions was filtered through the Thiol cartridge. The eluent was concentrated, and the crude product was purified by flash column chromatography to afford the title compound (104 mg, 100%). LC-MS m/z 344 (M+H)+, 1.21 min (ret time).
Intermediate 145
2-[(3S)-3-amino-l-pyrrolidinyl]-l-benzothiophene-3-carbonitrile hydrochloride
Figure imgf000107_0002
1 , 1 -Dimethylethyl [(3S)- 1 -(3 -cyano- 1 -benzothien-2-yl)-3 -pyrrolidinyl] carbamate (104 mg, 0.303 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (1.5 mL, 49.4 mmol). The reaction was stirred at RT. After 30 min, Et2O (5 mL) was added. The mixture was scraped and agitated with a spatula to assist in solid formation. The cloudy mixture was filtered and washed with Et2O (2 x 2 mL). The solid was set aside, and the residual product in the funnel and reaction vial was transferred to a vial with MeOH. The solid residue was added to the MeOH solution, and the mixture was concentrated under a stream of nitrogen at 50 0C to afford the title compound (70 mg, 82%). LC-MS m/z 244 (M+H)+, 0.67 min (ret time).
Intermediate 146
2-bromo-4-(l-pyrrolidinyl)benzonitrile
Figure imgf000108_0001
K2CO3 (0.691 g, 5.00 mmol) was added to a solution of 2-bromo-4- fluorobenzonitrile (1 g, 5.00 mmol) and pyrrolidine (0.42 mL, 5.08 mmol) in DMSO (10.00 mL) in a vial. The vial was sealed, and the reaction was stirred at 100 0C (bath temp) for 17.5 h. The reaction mixture was diluted with water (50 mL) and EtOAc (25 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 25 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over Na2SO4, filtered, concentrated, and dried under high vacuum to afford the title compound (1.21 g, 96%). LC-MS m/z 251 (M)+, 1.11 min (ret time). Intermediate 147
2-bromo-5-(l-pyrrolidinyl)benzonitrile
Figure imgf000108_0002
A mixture of 2-bromo-5-fluorobenzonitrile (500 mg, 2.500 mmol), pyrrolidine (0.21 mL, 2.54 mmol), and K2CO3 (346 mg, 2.500 mmol) was taken up in DMSO (5 mL) in a 1 dram vial. The vial was sealed, and the reaction was heated at 100 0C (bath temp) for 16.5 showed. Water (15 mL) and EtOAc (10 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (411 mg, 66%). LC-MS m/z 251 (M)+, 1.16 min (ret time).
Intermediate 148
1,1-dimethylethyl {(35)-l-[2-cyano-5-(l-pyrrolidinyl)phenyl]-3- pyrrolidinyl}carbamate
Figure imgf000109_0001
Pd2(dba)3 (182 mg, 0.199 mmol), BINAP (372 mg, 0.597 mmol), Cs2CO3 (649 mg, 1.991 mmol), and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (482 mg, 2.59 mmol) were added to a vial containing a solution of 2-bromo-4-(l-pyrrolidinyl)benzonitrile (500 mg, 1.991 mmol) in degassed toluene (10 mL) (degassed by bubbling argon through it for 10 min) under argon. The vial was capped, and the reaction mixture was heated at 100 0C (bath temp) for 16.5 h. Water (15 mL) and EtOAc (10 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (659 mg, 93%). LC-MS m/z 357 (M+H)+, 1.21 min (ret time).
Intermediate 149
1,1-dimethylethyl {(3S)-l-[2-cyano-4-(l-pyrrolidinyl)phenyl]-3- pyrrolidinyl}carbamate
Figure imgf000109_0002
Pd2(dba)3 (141 mg, 0.154 mmol), BINAP (288 mg, 0.462o mmol), Cs2CO3 (502 mg, 1.541 mmol), and 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate (373 mg, 2.003 mmol) were added to a 20 mL vial containing a solution of 2-bromo-5-(l- pyrrolidinyl)benzonitrile (387 mg, 1.541 mmol) in degassed toluene (8 mL) (degassed by bubbling argon through it for 10 min) under argon. The vial was capped, and the reaction mixture was heated at 100 0C (bath temp) for 14.5 h. Water (10 mL) was added, and the layers were separated. The aqueous layer was extracted with EtOAc (3 x 5 mL), and the combined organic layers were filtered through a Stratosphere PL-Thio MP SPE cartridge (0.5 g). The eluent was concentrated, and the crude product was purified by flash column chromatography to afford the title compound (447 mg, 81%). LC-MS m/z 357 (M+H)+, 0.98 min (ret time).
Intermediate 150
2-[(3S)-3-amino-l-pyrrolidinyl]-4-(l-pyrrolidinyl)benzonitrile hydrochloride
Figure imgf000110_0001
1,1 -dimethylethyl {(35)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3- pyrrolidinyl} carbamate (659 mg, 1.849 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (10 mL, 40.0 mmol). The reaction mixture was stirred at RT for 45 min and then Et2O (50 mL) was added. The cloudy mixture was filtered and washed with Et2O (2 x 10 mL). The solid was set aside, and the residual product in the funnel and reaction flash was transferred to a 2 dram vial with MeOH. The solid residue was added to the MeOH solution, and the mixture was concentrated under a stream of nitrogen at 50 0C to afford the title compound (466.5 mg, 86%). LC-MS m/z 257 (M+H)+, 0.76 min (ret time).
Intermediate 151
2-[(3S)-3-amino-l-pyrrolidinyl]-5-(l-pyrrolidinyl)benzonitrile hydrochloride
Figure imgf000110_0002
1 , 1 -Dimethylethyl {(35)- 1 -[2-cyano-4-( 1 -pyrrolidinyl)phenyl]-3- pyrrolidinyl} carbamate (442 mg, 1.240 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (6 mL, 24.00 mmol) in a 2 dram vial. The reaction mixture was stirred at RT. After 0.5 h, Et2O (30 mL) was added. The cloudy mixture was filtered through a medium porosity glass fritted filter and washed with Et2O (2 x 10 mL). The solid was set aside, and the residual product in the funnel and reaction flash was transferred to a 2 dram vial with MeOH. The solid residue was added to the MeOH solution, and the mixture was concentrated under a stream of nitrogen at 50 0C to afford the title compound (304 mg, 80%). LC-MS m/z 257 (M+H)+, 0.62 min (ret time).
Intermediate 152
2-br omo-5,6-dihydr o-4H-cyclopenta [b] thiophene-3-carbonitrile
Figure imgf000111_0001
2-Amino-5,6-dihydro-4H-cyclopenta[b]thiophene-3-carbonitrile (500 mg, 3.04 mmol) was added to slurry of CuBr2 (816 mg, 3.65 mmol) and 90 wt% tert-butyl nitrite (0.4 mL, 3.04 mmol) in CH3CN (15 mL) at 0 0C (bath temp) in a 6 dram vial. The ice bath was removed, and the reaction mixture was stirred at RT for 5 min. A solution of 9:1 saturated aq. NH4CI/NH4OH (15 mL) and EtOAc (10 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 5 mL). The combined organic layers were washed with brine (1 x 10 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (86 mg, 12%). LC-MS m/z 228 (M)+, 1.13 min (ret time).
Intermediate 153
1 , 1-dimethylethyl [(3S)- l-(3-cyano-5,6-dihydro-4H-cyclopenta [b] thien-2-yl)-3- pyrrolidinyl] carbamate
Figure imgf000111_0002
A mixture of 2-bromo-5,6-dihydro-4H-cyclopenta[δ]thiophene-3-carbonitrile (86 mg, 0.377 mmol), Pd2(dba)3 (35 mg, 0.038 mmol), BINAP (71 mg, 0.114 mmol), Cs2CO3 (123 mg, 0.377 mmol), and 1, 1-dimethylethyl (35)-3-pyrrolidinylcarbamate (91 mg, 0.490 mmol) was taken up in degassed toluene (2 mL) (degassed by bubbling argon through it for 10 min) in a 2 dram vial under argon. The vial was capped, and the reaction mixture was heated at 100 0C (bath temp) for 15 h. Water (3 mL) was added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were concentrated, and the crude product was purified by flash column chromatography to afford the title compound (103 mg, 82%). LC-MS m/z 334 (M+Η)+, 1.23 min (ret time). Intermediate 154
2- [(3S)-3-amino- 1-pyrr olidinyl] -5,6-dihydro-4H-cyclopenta [b] thiophene-3- carbonitrile hydrochloride
Figure imgf000112_0001
HCI
1 , 1 -Dimethylethyl [(3S)- 1 -(3-cyano-5,6-dihydro-4H-cyclopenta[δ]thien-2-yl)-3- pyrrolidinyl] carbamate (103 mg, 0.309 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (1.5 mL, 6.00 mmol) in a 2 dram vial. The reaction mixture was stirred at RT. After 40 min, Et2O (5 mL) was added. The cloudy mixture was filtered through a medium porosity glass fritted filter and washed with Et2O (2 x 2 mL). The solid was set aside, and the residual product in the funnel and reaction flash was transferred to a haystack vial with MeOH. The solid residue was added to the MeOH solution, and the mixture was concentrated under a stream of nitrogen at 50 0C to afford the title compound (69 mg, 83%). LC-MS m/z 234 (M+Η)+, 0.72 min (ret time).
Intermediate 155
2-bromo-5,6,7,8-tetrahydro-4H-cyclohepta[6]thiophene-3-carbonitrile
Figure imgf000112_0002
90 wt% tert-bvXy\ nitrite (0.51 mL, 3.87 mmol) was added to a solution of CuBr2 (697 mg, 3.12 mmol) in CH3CN ( 15 mL) at 0 0C (bath temp) in a 100 mL round-bottomed flask. 2-Amino-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thiophene-3-carbonitrile (500 mg, 2.60 mmol) was added portionwise over 30 sec. The ice bath was removed, and the reaction was stirred at RT for 5 min. A solution of 9:1 saturated aq. NΗ4C1/NΗ4OΗ (15 mL) and EtOAc (10 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (1 x 5 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (239 mg, 36%). LC-MS m/z 256 (M)+, 1.26 min (ret time).
I l l Intermediate 156
l,l-dimethylethyl [(35)-l-(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[6]thien-2-yl)-3- pyrrolidinyl] carbamate
Figure imgf000113_0001
A mixture of 2-bromo-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thiophene-3- carbonitrile (239 mg, 0.933 mmol), Pd2(dba)3 (85 mg, 0.093 mmol), BINAP (174 mg, 0.280 mmol), Cs2CO3 (304 mg, 0.933 mmol), and 1,1-dimethylethyl (35)-3- pyrrolidinylcarbamate (226 mg, 1.213 mmol) was taken up in degassed toluene (5 mL) (degassed by bubbling argon through it for 10 min) in a 2 dram vial under argon. The vial was capped, and the reaction was heated at 100 0C (bath temp) for 15 h. Water (3 mL) was added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 2 mL). The combined organic layers were filtered through a Stratosphere PL-Thio MP SPE cartridge (0.5 g). The cartridge was rinsed with EtOAc (5 mL). The eluent was concentrated, and the crude product was purified by flash column chromatography to afford the title compound (231 mg, 69%). LC-MS m/z 362 (M+Η)+, 1.38 min (ret time).
Intermediate 157
2- [(3 S)-3-amino- 1-pyrr olidinyl] -5,6,7,8-tetrahydro-4H-cyclohepta [b] thiophene-3- carbonitrile hydrochloride
Figure imgf000113_0002
1 , 1 -Dimethylethyl [(3S)- 1 -(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thien-2- yl)-3-pyrrolidinyl] carbamate (231 mg, 0.639 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (3 mL, 12.00 mmol) in a 2 dram vial. The reaction was stirred at RT.
After 1 h, 15 min, Et2O (5 mL) was added. The cloudy mixture was filtered through a medium porosity glass fritted filter and washed with Et2O (2 x 2 mL). The solid was set aside, and the residual product in the funnel and reaction flash was transferred to a haystack vial with MeOH. The solid residue was added to the MeOH solution, and the mixture was concentrated under a stream of nitrogen at 50 0C to afford the title compound (178 mg, 94%). LC-MS m/z 262 (M+Η)+, 0.84 min (ret time). Intermediate 158
2-br omo-4,7-dihydro-5H-spiro [ l-benzothiophene-6,2 '- [ 1 ,3] dioxolane] -3-carbonitrile
Figure imgf000114_0001
2-Amino-4,7-dihydro-5H-spiro[l-benzothiophene-6,2'-[l,3]dioxolane]-3- carbonitrile (500 mg, 2.116 mmol) was added portionwise over 1.5 min to slurry of CuBr2 (567 mg, 2.54 mmol) and tert-butyl nitrite (0.418 mL, 3.17 mmol) in CH3CN (10 mL) at 0 0C (bath temp) in a 250 mL round-bottomed flask. The ice bath was removed, and the reaction mixture was stirred at RT for 5 min. A solution of 9:1 saturated aq.
NH4CI/NH4OH (45 mL) and EtOAc (25 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 25 mL). The combined organic layers were washed with brine (1 x 20 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (178 mg, 28%). LC-MS m/z 300 (M)+, 0.98 min (ret time).
Intermediate 159
1,1-dimethylethyl [(3S)-l-(3-cyano-4,7-dihydro-5H-spiro[l-benzothiophene-6,2'-
[ 1 ,3] dioxolan] -2-yl)-3-pyrrolidinyl] carbamate
Figure imgf000114_0002
A mixture of 2-bromo-4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'-
[1,3] dioxolane] -3-carbonitrile (109.1 mg, 0.363 mmol), 1,1-dimethylethyl (3<S)-3- pyrrolidinylcarbamate (67.7 mg, 0.363 mmol), Pd2(dba)3 (33.3 mg, 0.036 mmol), BINAP (67.9 mg, 0.109 mmol), and Cs2CO3 (178 mg, 0.545 mmol) in toluene (2 mL)(degassed by bubbling argon through it for 20 min) was stirred for 16 h at 100 0C. Water (5 mL) was then added. The mixture was extracted with EtOAc (3 x 5 mL). The combined organic layers were concentrated, and the crude product was purified by flash chromatography. The desired fractions were combined, concentrated, and further purified by reverse phase HPLC to afford the title compound (98 mg, 66%). LC-MS m/z 406 (M+H)+, 1.10 min (ret time).
Intermediate 160
2-[(3S)-3-amino-l-pyrrolidinyl]-4,7-dihydro-5H-spiro[l-benzothiophene-6,2'-
[l,3]dioxolane]-3-carbonitrilehydrochloride
Figure imgf000115_0001
1 , 1 -dimethylethyl [(3S)- 1 -(3-cyano-4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'- [l,3]dioxolan]-2-yl)-3-pyrrolidinyl]carbamate (97.7 mg, 0.241 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (1325 μL, 5.30 mmol) in a 2 dram vial. After 2.5 h, the mixture was concentrated under a stream of nitrogen at 50 0C. The crude product was dissolved in DMSO (2 mL) and purified by ΗPLC. The desired fractions were concentrated under a stream of nitrogen at 50 0C to afford the title compound (46 mg, 56%). LC-MS m/z 306 (M+Η)+, 0.58 min (ret time).
Intermediate 161
2-chloro-l-{[(phenylmethyl)oxy]methyl}-lH-indole-3-carbonitrile
Figure imgf000115_0002
To a suspension of NaH (28.6 mg, 1.132 mmol) in DMF (2 mL) at RT was added 2-chloro-lH-indole-3-carbonitrile (100 mg, 0.566 mmol). The reaction mixture was stirred at RT for 5 min. Chloromethyl phenylmethyl ether (0.086 mL, 0.623 mmol) was added to the reaction mixture. After 1 h, water (2 mL) was added. The layers were separated, and the aqueous layer was extracted with Et2O (3 x 2 mL). The combined organic layers were washed with brine (2 mL) and concentrated. The crude product was purified by flash column chromatography to afford the title compound (144 mg, 86%). LC-MS m/z 297 (M+Η)+, 1.25 min (ret time). Intermediate 162
l,l-dimethylethyl [(35)-l-(3-cyano-l-{[(phenylmethyl)oxy]methyl}-lH-indol-2-yl)-3- pyrrolidinyl] carbamate
Figure imgf000116_0001
K2CO3 (74.0 mg, 0.536 mmol) was added to a solution of 2-chloro-l- {[(phenylmethyl)oxy]methyl}-lH-indole-3-carbonitrile (144.5 mg, 0.487 mmol) and 1,1- dimethylethyl (35)-3-pyrrolidinylcarbamate (91 mg, 0.487 mmol) in DMSO (4 mL) in a 2 dram vial. The vial was capped, and the reaction mixture was heated at 100 0C (bath temp). After 48 h, the reaction mixture was allowed to cool to RT. Water (3 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (3 x 2 mL) and concentrated. The crude product was purified by flash column
chromatography to afford the title compound (133 mg, 61%). LC-MS m/z 447 (M+Η)+, 1.35 min (ret time) .
Intermediate 163
1,1-dimethylethyl [(SSH-^-cyano-lH-indol-l-ylJ-S-pyrrolidinyljcarbamate
Figure imgf000116_0002
A mixture of 1,1-dimethylethyl [(35)-l-(3-cyano-l-{[(phenylmethyl)oxy]methyl}- lH-indol-2-yl)-3-pyrrolidinyl]carbamate (133 mg, 0.298 mmol), 10% Pd/C (95 mg, 0.089 mmol), and ammonium formate (100 mg, 1.579 mmol) in EtOH (2 mL) was heated at reflux under Argon until the reaction was complete as determined by LC-MS. The mixture was filtered and the filtrate was purified by flash column chromatography to afford the title compound (34.8 mg, 36%). LC-MS m/z 327 (M+H)+, 1.02 min (ret time).
Intermediate 164
2-[(3S)-3-amino-l-pyrrolidinyl]-lH-indole-3-carbonitrile
Figure imgf000117_0001
1 , 1 -Dimethylethyl [(3S)- 1 -(3-cyano- lH-indol-2-yl)-3-pyrrolidinyl]carbamate (43.6 mg, 0.134 mmol) was taken up in a solution of 4 M HCl in 1,4-dioxane (0.8 mL, 3.20 mmol) in a 2 dram vial. After 45 min, the mixture was concentrated under a stream of nitrogen at 50 0C and dried under high vacuum to afford the title compound (38 mg, 96%). LC-MS m/z 227 (M+Η)+, 0.63 min (ret time).
Intermediate 165
l,l-dimethylethyl (2S)-2-({[(3S)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl]amino}carbonyl)-l-pyrrolidinecarboxylate
Figure imgf000117_0002
To a solution of 2-[(35)-3-amino-l-pyrrolidinyl]-5-bromobenzonitrile
hydrochloride (0.926 g, 3.06 mmol), l-{[(l,l-dimethylethyl)oxy]carbonyl}-L-proline (0.660 g, 3.07 mmol), and Et3N (1.30 mL, 9.33 mmol) in CH2Cl2 (16 mL) was added a solution of 50% wt% ®T3P in EtOAc (1.2 mL, 4.03 mmol) at 0 0C. The reaction was allowed to warm to RT and stirred for 7 days. The reaction mixture was diluted with CH2Cl2 (40 mL), and then washed with NaHCO3 (2 x 2OmL) followed by 10% citric acid (20 mL). The layers were separated, and the organic layer was concentrated to afford the title compound (1.032 g, 73 %). LC-MS m/z 463 (M+H)+, 1.10 min (ret time). COMPOUNDS OF FORMULA (I)
Example 1
7V1-[(3S)-l-(3-chloro-4-cyano-5-isothiazolyl)-3-pyrrolidinyl]-L-alaninamide
hydrochloride
Figure imgf000118_0001
HCI
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-chloro-4-cyano-5- isothiazolyl)-3-pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (31 mg, 0.078 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (2 mL) and the resultant white precipitate was filtered through a plug of cotton. Residual solid was dissolved in MeOH (3 mL) and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in MeOH (2 mL) and CH2Cl2 (1 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (23.3 mg, 87%). 1H NMR (400 MHz, MeOD) δ ppm 4.56 - 4.62 (m, 1 H), 3.92 (q, J=6.9 Hz, 2 H), 3.65 - 3.83 (m, 2 H), 3.50 - 3.60 (m, 1 H), 2.41 (dd, J=13.3, 5.8 Hz, 1 H), 2.17 - 2.23 (m, 1 H), 1.50 (d, J=7.0 Hz, 3 H). LC-MS m/z 300 (M+H)+, 0.65 min (ret time).
Example 2
TV1- [(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl] -L-alaninamide hydrochloride
Figure imgf000118_0002
HCl
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (190 mg, 0.471 mmol) in HCl (4 M solution in 1,4-dioxane, 3.1 mL, 12.40 mmol) was stirred at RT for 3 h 30 min. The reaction mixture was diluted with Et2O (5 mL) and the resultant yellow precipitate was filtered and washed with Et2O (5 mL). The precipitate was dissolved partially in MeOH (10 mL) and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (75 mg, 47%). LC-MS m/z 304 (M+H)+, 0.71 min (ret time). Example 3
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]glycinamide hydrochloride
Figure imgf000119_0001
HCI
A solution of 1,1-dimethylethyl (2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl] amino }-2-oxoethyl)carbamate (42.4 mg, 0.109 mmol) in HCl (4 M solution in 1,4-dioxane, 0.72 mL, 2.88 mmol) was stirred at RT for 17 h. The reaction mixture was diluted with Et2O (2 mL) and the resultant yellow precipitate was filtered and washed with Et2O (2 mL). The precipitate was dissolved partially in MeOH (5 mL) and concentrated under a stream of nitrogen at 50 0C. 1,4-Dioxane (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X, followed by the addition of water (2 mL) and lyophilization to afford the title compound (29 mg, 82%). LC-MS m/z 290 (M+H)+, 0.64 min (ret time).
Example 4
7V1-[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-L- alaninamide hydrochloride
Figure imgf000119_0002
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l- benzothien-2-yl)-3-pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (7.5 mg, 0.018 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (5 mL) and the cloudy mixture was filtered through a plug of cotton. The precipitate was dissolved in MeOH and
concentrated under a stream of nitrogen at 50 0C to afford the title compound (2.1 mg, 31%). 1H NMR (400 MHz, MeOD) δ ppm 4.49 - 4.55 (m, 1 H), 3.88 (q, J=7.0 Hz, 1 H), 3.82 (dd, J=10.3, 6.0 Hz, 1 H), 3.68 - 3.74 (m, 1 H), 3.59 - 3.66 (m, 1 H), 3.48 (dd, J=10.3, 3.8 Hz, 1 H), 2.51 (t, J=4.9 Hz, 2 H), 2.46 (t, J=5.0 Hz, 2 H), 2.31 - 2.38 (m, 1 H), 2.06 - 2.13 (m, 1 H), 1.78 - 1.84 (m, 4 H), 1.48 (d, J=7.3 Hz, 3 H). LC-MS m/z 319 (M+H)+, 1.01 min (ret time). Example 5
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-valinamide hydrochloride
Figure imgf000120_0001
To a solution of N1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-Λ/2-{[(l,l- dimethylethyl)oxy]carbonyl}-L-valinamide (131 mg, 0.304 mmol) in 1,4-dioxane (2.0 niL) was added HCl (4 M solution in 1,4-dioxane, 0.175 mL, 0.700 mmol). The reaction mixture was stirred at RT for 15 min. Additional HCl (4 M solution in 1,4-dioxane, 0.10 mL, 0.40 mmol) was added, and the reaction mixture was heated to 45 0C for 1 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL) and the residue was scraped until a yellow solid formed, which was collected by filtration and washed with Et2O (2 x 3 mL). The solid was dried in vacuo for 15 h. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (55.5 mg, 50%) as a fluffy yellow solid. LC-MS m/z 332 (M+H)+, 0.64 min (ret time).
Example 6
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]butanamide
hydrochloride
Figure imgf000120_0002
To a solution of 1,1-dimethylethyl [( 1 S)-I -({ [(3S)-I -(2-cyano-4-nitrophenyl)-3 - pyrro lidinyl] amino }carbonyl)propyl] carbamate (110 mg, 0.263 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.40 mL, 1.60 mmol). The reaction mixture was heated to 45-50 0C for 5 h. Additional HCl (4 M solution in 1,4-dioxane, 0.10 mL, 0.40 mmol) was added, and the reaction mixture was stirred at RT for 17 h. The reaction mixture was diluted with Et2O (6 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (62.1 mg, 67%). 1H NMR (DMSO-d6) δ ppm 8.85 (d, J= 6.5 Hz, IH), 8.43 (d, J= 2.8 Hz, IH), 8.13 - 8.20 (m, 4H), 6.92 (d, J= 9.8 Hz, IH), 4.46 (m, IH), 3.98 (m, IH), 3.81 - 3.88 (m, 2H), 3.60-3.63 (m, 2H), 2.23 (m, IH), 2.07 (m, IH), 1.69 - 1.76 (m, 2H), 0.85 (t, J= 7.4 Hz, 3H). LC-MS m/z 318 (M+H)+, 0.65 min (ret time).
Example 7
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-norvalinamide hydrochloride
Figure imgf000121_0001
HCI
To a solution of N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-Λ/2-{[(l,l- dimethylethyl)oxy]carbonyl}-L-norvalinamide (127 mg, 0.294 mmol) in 1,4-dioxane (2.0 rnL) was added HCl (4 M solution in 1,4-dioxane, 0.30 rnL, 1.20 mmol). The reaction mixture was heated to 45-50 0C for 5 h. Upon cooling, the reaction mixture was diluted with Et2O (6 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (72.8 mg, 67%). LC-MS m/z 332 (M+H)+, 0.68 min (ret time).
Example 8
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-leucinamide hydrochloride
Figure imgf000121_0002
HCI
To a solution of N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-N2-{[(l,l- dimethylethyl)oxy]carbonyl}-L-leucinamide (121.7 mg, 0.273 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol). The reaction mixture was stirred at RT for 18 h and then heated to 50 0C for 2 h. Upon cooling, the reaction mixture was diluted with Et2O (6 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (78.6 mg, 75%). LC-MS m/z 346 (M+H)+, 0.71 min (ret time). Example 9
7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-phenylalaninamide
hydrochloride
Figure imgf000122_0001
HCI
To a solution of N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-Λ/2-{[(l,l- dimethylethyl)oxy]carbonyl}-L-phenylalaninamide (139.6 mg, 0.291 mmol) in 1,4- dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 niL, 2.00 mmol). The reaction mixture was stirred at RT for 18 h and then heated to 50 0C for 2 h. Upon cooling, the reaction mixture was diluted with Et2O (6 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (83.8 mg, 76%). LC-MS m/z 380 (M+H)+, 0.74 min (ret time).
Example 10
7V1-[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000122_0002
HCI
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-4-biphenylyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (7.8 mg, 0.018 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT- 4X to afford the title compound (4.4 mg, 66%). LC-MS m/z 335 (M+H)+, 0.93 min (ret time). Example 11
7V1-[(3S)-l-(4-cyano-3-biphenylyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000123_0001
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(4-cyano-3-biphenylyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (41.6 mg, 0.096 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), the cloudy mixture was filtered through a plug of cotton, and the solid was washed with Et2O (1 mL). The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (29.7 mg, 79%). LC-MS m/z 335 (M+H)+, 0.85 min (ret time).
Example 12
ethyl 5-[(3S)-3-(L-alanylamino)-l-pyrrolidinyl]-4-cyano-2-thiophenecarboxylate hydrochloride
Figure imgf000123_0002
A solution of ethyl 4-cyano-5-{(35)-3-[(N-{[(l,l-dimethylethyl)oxy]carbonyl}-L- alanyl)amino]-l-pyrrolidinyl}-2-thiophenecarboxylate (18.1 mg, 0.041 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), the cloudy mixture was filtered through a plug of cotton, and the solid was washed with Et2O (2 x 1 mL). The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (15 mg, 97%). LC-MS m/z 337 (M+H)+, 0.70 min (ret time). Example 13
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-serinamide hydrochloride
Figure imgf000124_0001
HCl
To a solution of 1,1-dimethylethyl [(lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-l-(hydroxymethyl)-2-oxoethyl]carbamate (93.4 mg, 0.223 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 niL, 2.00 mmol). The reaction mixture was heated to 40 0C for 15 h. Additional HCl (4 M solution in 1,4- dioxane, 0.50 mL, 2.00 mmol) was added, and the reaction mixture was heated to 50 0C for 4 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (55.5 mg, 70%) as a yellow-orange solid. LC-MS m/z 320 (M+H)+, 0.63 min (ret time).
Example 14
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-3-(l,3-thiazol-4-yl)-L-alaninamide hydrochloride
Figure imgf000124_0002
HCl
To a solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(l,3-thiazol-4-ylmethyl)ethyl]carbamate (105 mg, 0.216 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol). The reaction mixture was heated to 40 0C for 15 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (90.8 mg, 96%) as a yellow-orange solid. LC-MS m/z 387 (M+H)+, 0.67 min (ret time). Example 15
(2S)-2-amino-3-cyano-7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]- propanamide hydrochloride
Figure imgf000125_0001
HCI
To a solution of 1,1-dimethylethyl ((15)-l-(cyanomethyl)-2-{[(35)-l-(2-cyano-4- nitrophenyl)-3 -pyrrolidinyl] amino }-2-oxoethyl)carbamate (134.7 mg, 0.314 mmol) in 1,4- dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 niL, 2.00 mmol). The reaction mixture was heated to 40 0C for 15 h. Additional HCl (4 M solution in 1,4- dioxane, 0.50 mL, 2.00 mmol) was added, and the reaction mixture was heated to 50 0C for 1 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (43.5 mg, 38%). LC-MS m/z 329 (M+H)+, 0.63 min (ret time).
Example 16
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000125_0002
HCI
To a solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (157 mg, 0.323 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol). The reaction mixture was heated to 40 0C for 15 h. Additional HCl (4 M solution in 1,4- dioxane, 0.50 mL, 2.00 mmol) was added, and the reaction mixture was heated to 50 0C for 1 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL) and the resultant yellow solid was collected by filtration. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (77.4 mg, 58%). LC-MS m/z 386 (M+H)+, 0.73 min (ret time). Example 17
(l^-l-amino-TV-iCS^-l-Il-cyano^-Cl^-thiazol-l-ylJphenyll-S-pyrrolidinyl}- butanamide hydrochloride
Figure imgf000126_0001
HCI
A solution of 1,1-dimethylethyl {(15)-l-[({(35)-l-[2-cyano-4-(l,3-thiazol-2- yl)phenyl]-3-pyrrolidinyl}amino)carbonyl]propyl} carbamate (3 mg, 6.59 μmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT- 4X to afford the title compound (2 mg, 77%). LC-MS m/z 356 (M+H)+, 0.70 min (ret time).
Example 18
(2S)-2-amino-7V-{(3S)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3-pyrrolidinyl}- butanamide hydrochloride
Figure imgf000126_0002
HCl
A solution of 1,1-dimethylethyl {(15)-l-[({(35)-l-[2-cyano-4-(2- pyrimidinyl)phenyl]-3-pyrrolidinyl}amino)carbonyl]propyl}carbamate (5.3 mg, 0.012 mmol) in HCl (4 M solution in 1,4-dioxane, 0.003 mL, 0.012 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (4.5 mg, 99%). LC-MS m/z 351 (M+H)+, 0.68 min (ret time). Example 19
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-3-cyclopropyl-L-alaninamide hydrochloride
Figure imgf000127_0001
HCl
To a solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-l-(cyclopropylmethyl)-2-oxoethyl]carbamate (145 mg, 0.327 mmol) in 1,4-dioxane (2.0 niL) was added HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol). The reaction mixture was heated to 40 0C for 15 h. Upon cooling, the reaction mixture was diluted with Et2O (5 mL). The resultant solid was collected by filtration and purified by reverse phase HPLC (OBD 5 μm 19 x 100 mm preparatory column), eluting at 20 mL/min (2-100% CH3CN/H2O). Following concentration of the desired fractions at 45 0C under a stream of nitrogen, water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (42.6 mg, 34%). LC-MS m/z 344 (M+H)+, 0.69 min (ret time). Example 20
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-4-pentynamide hydrochloride
Figure imgf000127_0002
HCl
To a solution of 1,1-dimethylethyl [(I S)-I -({ [(3S)-I -(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}carbonyl)-3-butyn-l-yl]carbamate (124 mg, 0.290 mmol) in 1,4- dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol). The reaction mixture was heated to 50 0C for 1 h. Upon cooling, the reaction mixture was diluted with Et2O (6 mL), and the resultant solid was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (66.9 mg, 63%). 1U NMR (DMSO-d6) δ ppm 9.00 (d, J= 6.5 Hz, IH), 8.42 (br. s., 4H), 8.18 (dd, J= 9.5, 2.8 Hz, IH), 6.90 (d, J = 9.8 Hz, IH), 4.47 (br. s., IH), 3.97 (dd, J= 11.3, 5.8 Hz, IH), 3.83 - 3.91 (m, 3H), 3.66 (m, IH), 2.96 (t, J= 2.5 Hz, IH), 2.83-2.77 (m, IH), 2.71-2.65 (m, IH), 2.23 (m, IH), 2.05 (m, IH). LC-MS m/z 328 (M+H)+, 0.65 min (ret time).
Example 21
7V1-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-L-alaninamide hydrochloride
Figure imgf000128_0001
HCI
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4- (trifluoromethyl)phenyl]-3-pyrrolidinyl} amino)- l-methyl-2-oxoethyl]carbamate (75 mg, 0.176 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (5 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (44 mg, 69%). LC-MS m/z 327 (M+H)+, 0.77 min (ret time).
Example 22
(2S)-2-amino-7V-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- butanamide hydrochloride
Figure imgf000128_0002
HCI
A solution of 1,1-dimethylethyl {(15)-l-[({(35)-l-[2-cyano-4-
(trifluoromethyl)phenyl]-3-pyrrolidinyl}amino)carbonyl]propyl} carbamate (82 mg, 0.186 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (5 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (57 mg, 81%). LC-MS m/z 341 (M+H)+, 0.81 min (ret time).
Example 23
7V1-[(3S)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000129_0001
HCI
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (30 mg, 0.058 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was washed with Et2O (2 x 1 mL), dissolved in MeOH, and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (21.7 mg, 82%). 1H NMR (400 MHz, MeOD) δ ppm 7.60 (d, J=2.5 Hz, 1 H), 7.51 (dd, J=9.3, 2.5 Hz, 1 H), 7.18 (d, J=4.3 Hz, 1 H), 6.92 (d, J=3.0 Hz, 1 H), 6.87 (dd, J=5.0, 3.5 Hz, 1 H), 6.68 (d, J=9.3 Hz, 1 H), 4.42 - 4.47 (m, 1 H), 4.00 (t, J=7.0 Hz, 1 H), 3.80 (dd, J=10.4, 5.6 Hz, 1 H), 3.65 - 3.72 (m, 1 H), 3.60 (td, J=8.9, 4.5 Hz, 1 H), 3.40 (dd, J=10.5, 3.0 Hz, 1 H), 3.35 (d, J=7.3 Hz, 2 H), 2.22 - 2.29 (m, 1 H), 2.02 - 2.07 (m, 1 H). LC-MS m/z 419/421 (M+H)+, 0.87 min (ret time).
Example 24
7V1-[(3S)-l-(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000129_0002
HCl
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(4-bromo-2-cyanophenyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (23 mg, 0.053 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was washed with Et2O (2 x 1 mL), dissolved in MeOH, and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (11.9 mg, 61%). LC-MS m/z 337/339 (M+H)+, 0.75 min (ret time).
Example 25
7V1-[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000130_0001
HCl
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-4-biphenylyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (20 mg, 0.039 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (12.8 mg, 73%). 1H NMR (400 MHz, MeOD) δ ppm 7.72 - 7.76 (m, 2 H), 7.58 (d, J=I.3 Hz, 1 H), 7.56 (s, 1 H), 7.39 - 7.45 (m, 2 H), 7.28 - 7.33 (m, 1 H), 7.18 (dd, J=5.1, 1.1 Hz, 1 H), 6.93 - 6.94 (m, 1 H), 6.86 - 6.89 (m, 1 H), 6.84 - 6.86 (m, 1 H), 4.45 - 4.50 (m, 1 H), 4.01 (t, J=7.2 Hz, 1 H), 3.86 (dd, J=10.5, 5.8 Hz, 1 H), 3.72 - 3.79 (m, 1 H), 3.63 - 3.70 (m, 1 H), 3.46 - 3.49 (m, 1 H), 3.35 - 3.37 (m, 2 H), 2.30 (ddd, J=12.9, 8.0, 2.1 Hz, 1 H), 2.03 - 2.10 (m, 1 H). LC-MS m/z All (M+H)+, 0.98 min (ret time).
Example 26
7V1-[(3S)-l-(3-cyano-l-methyl-lH-indol-2-yl)-3-pyrrolidinyl]-L-alaninamide
hydrochloride
Figure imgf000130_0002
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-l-methyl-lH-indol-2- yl)-3-pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (33 mg, 0.080 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® ΗT-4X to afford the title compound (18.6 mg, 63%). LC-MS m/z 312 (M+H)+, 0.72 min (ret time).
Example 27
^-[(S^-l-CS-cyano-l-methyl-lH-indol-l-yO-S-pyrrolidinylJ-S-Cl-thienyO-L- alaninamide hydrochloride
Figure imgf000131_0001
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-l-methyl-lH-indol-2- yl)-3-pyrrolidinyl]amino}-2-oxo-l -(2 -thienylmethyl)ethyl] carbamate (37.5 mg, 0.076 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® ΗT-4X to afford the title compound (25.1 mg, 75%). 1H NMR (400 MHz, MeOD) δ ppm 7.34 - 7.38 (m, 1 H), 7.28 - 7.33 (m, 1 H), 7.14 - 7.20 (m, 2 H), 7.06 (dd, J=5.1, 1.1 Hz, 1 H), 6.93 (d, J=3.3 Hz, 1 H), 6.78 - 6.82 (m, 1 H), 4.46 - 4.52 (m, 1 H), 4.01 - 4.06 (m, 1 H), 3.94 - 3.98 (m, 1 H), 3.90 - 3.92 (m, 1 H), 3.65 - 3.72 (m, 4 H), 3.35 - 3.42 (m, 3 H), 2.34 (ddd, J=13.4, 7.2, 6.8 Hz, 1 H), 2.05 (ddd, J=16.8, 7.8, 4.3 Hz, 1 H). LC-MS m/z 394 (M+H)+, 0.81 min (ret time). Example 28
7V1-[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000132_0001
HCl
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-4-fluorophenyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (34 mg, 0.090 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (21.2 mg, 75%). LC-MS m/z 277 (M+H)+, 0.66 min (ret time).
Example 29
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]butanamide
hydrochloride
Figure imgf000132_0002
HCI
A solution of 1,1-dimethylethyl [( 1 S)-I -({ [(3S)-I -(2-cyano-4-fluorophenyl)-3 - pyrro lidinyl] amino }carbonyl)propyl] carbamate (53.7 mg, 0.138 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (4 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (39 mg, 87%). LC-MS m/z 291 (M+H)+, 0.68 min (ret time). Example 30
7V1-[(3S)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000133_0001
HCI
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4-fluorophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (60 mg, 0.131 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (44.4 mg, 82%). LC-MS m/z 359 (M+H)+, 0.77 min (ret time).
Example 31
TV1- [(3S)- l-(3-cyano-4 '-fluoro-4-biphenylyl)-3-pyrrolidinyl] -3-(2-thienyl)-L- alaninamide hydrochloride
Figure imgf000133_0002
HCl
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)- 3-pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (40.5 mg, 0.076 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (30.8 mg, 85%). 1H NMR (400 MHz, MeOD) δ ppm 7.68 - 7.74 (m, 2 H), 7.59 (ddd, J=12.1, 5.3, 3.0 Hz, 2 H), 7.12 - 7.19 (m, 3 H), 6.93 (d, J=2.3 Hz, 1 H), 6.87 (dd, J=5.3, 3.5 Hz, 1 H), 6.84 (d, J=8.8 Hz, 1 H), 4.45 - 4.50 (m, 1 H), 4.01 (t, J=7.1 Hz, 1 H), 3.86 (dd, J=10.4, 5.7 Hz, 1 H), 3.72 - 3.79 (m, 1 H), 3.66 (td, J=8.9, 4.3 Hz, 1 H), 3.45 - 3.49 (m, 1 H), 3.36 (d, J=7.1 Hz, 2 H), 2.25 - 2.32 (m, 1 H), 2.03 - 2.10 (m, 1 H). LC-MS m/z 435 (M+H)+, 0.98 min (ret time). Example 32
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-3-(3-thienyl)-L-alaninamide hydrochloride
Figure imgf000134_0001
HCI
To a solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(3-thienylmethyl)ethyl]carbamate (112 mg, 0.231 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 1.0 rnL, 4.00 mmol). The reaction mixture was stirred at RT for 1 h and then diluted with Et2O (6 mL). The precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (76.4 mg, 79%). LC-MS m/z 386 (M+H)+, 0.72 min (ret time).
Example 33
7V1-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-norleucinamide hydrochloride
Figure imgf000134_0002
To a solution of N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-Λ/2-{[(l,l- dimethylethyl)oxy]carbonyl}-L-norleucinamide (112.9 mg, 0.253 mmol) in 1,4-dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol). The reaction mixture was stirred at RT for 1 h and then diluted with Et2O (6 mL). The precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (66.8 mg, 60%). LC-MS m/z 346 (M+H)+, 0.74 min (ret time). Example 34
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopropylethan- amide hydrochloride
Figure imgf000135_0001
A solution of 1,1-dimethylethyl ((lS)-2-{[(3S)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyljaminol-l-cyclopropyl^-oxoethy^carbamate (120 mg, 0.279 mmol) in HCl (4 M solution in 1,4-dioxane, 1.50 niL, 6.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (68.3 mg, 67%). LC-MS m/z 330 (M+H)+, 0.67 min (ret time).
Example 35
7V1-[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-3-(2- thienyl)-L-alaninamide hydrochloride
Figure imgf000135_0002
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l- benzothien-2-yl)-3-pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (316 mg, 0.631 mmol) in HCl (4 M solution in 1,4-dioxane, 3.0 mL, 12.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (10 mL), the slurry was mixed with a spatula, and the cloudy mixture was filtered and washed with Et2O (2 x 5 mL).
Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X, dried under high vacuum, dissolved in MeOH (5 mL), concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum. Water (5 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (192 mg, 70%). 1U NMR (400 MHz, MeOD) δ ppm 7.24 (dd, J=5.0, 1.3 Hz, 1 H), 6.90 - 6.95 (m, 2 H), 4.43 - 4.48 (m, 1 H), 3.99 (t, J=I 2 Hz, 1 H), 3.73 (dd, J=10.5, 5.8 Hz, 1 H), 3.56 - 3.61 (m, 2 H), 3.37 - 3.41 (m, 1 H), 3.34 - 3.36 (m, 2 H), 2.53 (t, J=4.8 Hz, 2 H), 2.48 (t, J=4.9 Hz, 2 H), 2.28 - 2.34 (m, 1 H), 2.04 (d, J=10.8 Hz, 1 H), 1.79 - 1.86 (m, 4 H). LC-MS m/z 401 (M+H)+, 0.93 min (ret time). Example 36
(2S)-2-amino-7V-[(3S)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidin- yl]butanamide hydrochloride
Figure imgf000136_0001
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l- benzothien-2-yl)-3-pyrrolidinyl]amino}carbonyl)propyl]carbamate (273 mg, 0.631 mmol) in HCl (4 M solution in 1,4-dioxane, 3.0 rnL, 12.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (10 mL), the slurry was mixed with a spatula, and the cloudy mixture was filtered and washed with Et2O (2 x 5 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (179 mg, 77%). LC-MS m/z 333 (M+H)+, 0.85 min (ret time).
Example 37
7V1-[(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000136_0002
Hci
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-2-pyrazinyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (81.6 mg, 0.226 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (48.1 mg, 72%). LC-MS m/z 261 (M+H)+, 0.54 min (ret time). Example 38
(2S)-2-amino-7V-[(3S)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]butanamide
hydrochloride
Figure imgf000137_0001
HCI
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(3-cyano-2-pyrazinyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (94.3 mg, 0.252 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (37.9 mg, 54%). LC-MS m/z 275 (M+H)+, 0.56 min (ret time).
Example 39
^-[(S^-l-CS-cyano-l-pyrazinylJ-S-pyrrolidinyll-S-Cl-thienylJ-L-alaninamide hydrochloride
Figure imgf000137_0002
HCI
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-2-pyrazinyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (141.5 mg, 0.320 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (39.3 mg, 46%). LC-MS m/z 343 (M+H)+, 0.62 min (ret time). Example 40
(2S)-2-amino-7V-[(3S)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl]butanamide
hydrochloride
Figure imgf000138_0001
HCI
A solution of 1,1-dimethylethyl [(lS)-l-({[(3S)-l-(3-cyano-2-pyridinyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (145.7 mg, 0.390 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (112.6 mg, >100%, contained some residual solvent). LC-MS m/z 21 A (M+H)+, 0.58 min (ret time).
Example 41
TV^^SH-β-cyano-l-pyridinylJ-S-pyrroMinyll-S-Cl-thienylJ-L-alaninamide hydrochloride
Figure imgf000138_0002
HCI
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-2-pyridinyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (175.9 mg, 0.398 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (109.1 mg, 81%). LC- MS m/z 342 (M+H)+, 0.64 min (ret time). Example 42
TV1- [(3S)- l-(3-cyano-4 '-fluoro-4-biphenylyl)-3-pyrrolidinyl] - L-alaninam ide
hydrochloride
Figure imgf000139_0001
HCl
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-
3-pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (17.6 mg, 0.039 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL), and the cloudy mixture was filtered and washed with Et2O (2 x 2 mL). Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (13.2 mg, 87%). LC-MS m/z 353 (M+H)+, 0.90 min (ret time).
Example 43
7V1-[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000139_0002
HCI
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-3-thienyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (15.6 mg, 0.043 mmol) in HCl (4 M solution in 1,4-dioxane, 0.035 mL, 0.140 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (2 mL), and the resultant white precipitate was filtered through a plug of cotton and washed with Et2O (1 mL). The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (12 mg, 93%). LC-MS m/z 265 (M+H)+, 0.62 min (ret time). Example 44
7V1-[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide
hydrochloride
Figure imgf000140_0001
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-3-thienyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (43.5 mg, 0.097 mmol) in HCl (4 M solution in 1,4-dioxane, 0.08 mL, 0.320 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (5 mL), and the resultant white precipitate was filtered through a plug of cotton and washed with Et2O (3 mL). The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT -4X, dissolved in MeOH (3 mL), and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (30 mg, 80%). 1H NMR (400 MHz, MeOD) δ ppm 7.58 (d, J=5.5 Hz, 1 H), 6.92 (s, 1 H), 6.88 - 6.91 (m, 2 H), 6.62 (d, J=5.5 Hz, 1 H), 4.41 - 4.46 (m, 1 H), 3.97 (t, J=7.2 Hz, 1 H), 3.78 (dd, J=10.5, 5.5 Hz, 1 H), 3.63 - 3.66 (m, 1 H), 3.42 - 3.48 (m, 2 H), 3.33 - 3.35 (m, 1 H), 2.23 - 2.32 (m, 1 H), 1.97 - 2.05 (m, 1 H). LC-MS m/z 347 (M+H)+, 0.72 min (ret time).
Example 45
(2S)-2-amino-7V-[(3S)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]butanamide
hydrochloride
Figure imgf000140_0002
HCl
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(3-cyano-4-biphenylyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (39 mg, 0.087 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (4 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 rnL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (18.2 mg, 54%). LC-MS m/z 349 (M+H)+, 0.93 min (ret time).
Example 46
^-{(SSVl-β-cyano-S-^rifluoromethylJphenyll-S-pyrroMinylJ-L-alaninamide hydrochloride
Figure imgf000141_0001
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-5- (trifluoromethyl)phenyl]-3-pyrrolidinyl} amino)- 1 -methyl-2-oxoethyl]carbamate (68 mg, 0.159 mmol) in HCl (4 M solution in 1 ,4-dioxane, 1.0 niL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (44.8 mg, 77%). LC-MS m/z 327 (M+H)+, 0.84 min (ret time).
Example 47
(l^-l-amino-TV-KS^-l-Il-cyano-S-CtrifluoromethylJphenyll-S-pyrrolidinyl}- butanamide hydrochloride
Figure imgf000141_0002
A solution of 1,1-dimethylethyl {(15)-l-[({(35)-l-[2-cyano-5-
(trifluoromethyl)phenyl]-3-pyrrolidinyl}amino)carbonyl]propyl} carbamate (76 mg, 0.173 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac HT-4X to afford the title compound (21.9 mg, 34%). LC-MS m/z 341 (M+H)+, 0.86 min (ret time).
Example 48
^-{(S^-l-Il-cyano-S-CtrifluoromethylJphenyll-S-pyrrolidinylJ-S-Cl-thienylJ-L- alaninamide hydrochloride
Figure imgf000142_0001
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-5-(trifluoromethyl)- phenyl]-3-pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (76 mg, 0.149 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (44.9 mg, 68%). LC-MS m/z 409 (M+H)+, 0.92 min (ret time).
Example 49
7V1-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-3-(2-thienyl)-L- alaninamide hydrochloride
Figure imgf000142_0002
HCl
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(trifluoromethyl)- phenyl]-3-pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (223 mg, 0.438 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (133.7 mg, 69%). LC- MS m/z 409 (M+H)+, 0.94 min (ret time).
Example 50
7V1-[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000143_0001
HCl
A solution of 1,1-dimethylethyl ((15)-2-{[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]- amino}-l-methyl-2-oxoethyl)carbamate (93 mg, 0.259 mmol) in HCl (4 M solution in 1,4- dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT -4X to afford the title compound (68.2 mg, 89%). LC-MS m/z 259 (M+H)+, 0.67 min (ret time).
Example 51
(2S)-2-amino-7V-[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]butanamide hydrochloride
Figure imgf000143_0002
HCi
A solution of 1,1-dimethylethyl [( IS)- 1-(([(3S)-I -(2-cyanophenyl)-3 - pyrro lidinyl] amino }carbonyl)propyl] carbamate (89.5 mg, 0.240 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (62.6 mg, 84%). LC-MS m/z 273 (M+H)+, 0.72 min (ret time). Example 52
7V1-[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide
hydrochloride
Figure imgf000144_0001
A solution of 1,1-dimethylethyl [(15)-2-{[(3S)-l-(2-cyanophenyl)-3-pyrrolidinyl]- amino}-2-oxo-l -(2 -thienylmethyl)ethyl] carbamate (103 mg, 0.234 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the solvent was decanted off. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (3 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (41.1 mg, 47%). 1H NMR (400 MHz, MeOD) δ ppm 7.47 (dd, J=7.8, 1.3 Hz, 1 H), 7.43 (ddd, J=8.6, 7.2, 1.8 Hz, 1 H), 7.15 (dd, J=5.2, 1.1 Hz, 1 H), 6.92 (dd, J=3.5, 1.0 Hz, 1 H), 6.85 (dd, J=5.2, 3.4 Hz, 1 H), 6.74 - 6.78 (m, 2 H), 4.42 - 4.47 (m, 1 H), 4.01 (t, J=7.2 Hz, 1 H), 3.80 (dd, J=IOA, 5.7 Hz, 1 H), 3.66 - 3.72 (m, 1 H), 3.57 - 3.63 (m, 1 H), 3.42 (dd, J=10.3, 2.8 Hz, 1 H), 3.34 - 3.36 (m, 2 H), 2.22 - 2.30 (m, 1 H), 2.00 - 2.08 (m, 1 H). LC-MS m/z 341 (M+H)+, 0.80 min (ret time).
Example 53
7V1-[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]-L-alaninamide
hydrochloride
Figure imgf000144_0002
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-4,5-difiuorophenyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (80 mg, 0.203 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the cloudy mixture was filtered. Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (47.9 mg, 71%). LC-MS m/z 295 (M+H)+, 0.71 min (ret time).
Example 54
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]butanamide hydrochloride
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(2-cyano-4,5-difiuorophenyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (85 mg, 0.208 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the cloudy mixture was filtered. Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 mL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (47.7 mg, 67%). LC-MS m/z 309 (M+H)+, 0.75 min (ret time).
Example 55
7V1-[(3S)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000145_0002
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-4,5-difiuorophenyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (83 mg, 0.174 mmol) in HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol) was stirred at RT for 2 h. The reaction mixture was diluted with Et2O (5 mL), the slurry was mixed with a spatula, and the cloudy mixture was filtered. Residual solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. All solids were combined and dissolved in water (3 niL). The mixture was lyophilized with a Genevac® HT-4X to afford the title compound (47.1 mg, 66%). LC-MS m/z 377 (M+H)+, 0.82 min (ret time).
Example 56
(2S)-2-amino-7V-[(3S)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopentylethan- amide hydrochloride
Figure imgf000146_0001
HCl
To a solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-4-nitrophenyl)-3- pyrrolidinyl]amino}-l-cyclopentyl-2-oxoethyl)carbamate (133.7 mg, 0.292 mmol) in 1,4- dioxane (2.0 mL) was added HCl (4 M solution in 1,4-dioxane, 1.0 mL, 4.00 mmol). The reaction mixture was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL). The resultant solid was collected by filtration, washed with Et2O (6 mL), and purified by reverse phase HPLC (OBD 5 μm 19 x 100 mm preparatory column), eluting at 15 mL/min (5-40% CH3CN/H2O). Following concentration of the desired fractions at 45 0C under a stream of nitrogen, water (1 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (38.8 mg, 34%) as a fluffy yellow solid. LC-MS m/z 358 (M+H)+, 0.97 min (ret time).
Example 57
(2S)-2-amino-7V-[(3S)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]butanamide hydrochloride
Figure imgf000146_0002
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(2-cyano-3-thienyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (156 mg, 0.412 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 35 min. The reaction mixture was diluted with Et2O (2 mL) and the cloudy mixture was filtered through a plug of cotton. The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT- 4X to afford the title compound (108 mg, 83%). LC-MS m/z 279 (M+H)+, 0.90 min (ret time).
Example 58
7V1-{(3S)-l-[2-cyano-4-(phenyloxy)phenyl]-3-pyrrolidinyl}-3-(2-thienyl)-L- alaninamide hydrochloride
Figure imgf000147_0001
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(phenyloxy)phenyl]- 3-pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (61 mg, 0.115 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (2 mL), and the resultant white precipitate was filtered through a plug of cotton and washed with Et2O (1 mL). The solid was dissolved in MeOH (2 mL) and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (44 mg, 82%). 1H NMR (400 MHz, MeOD) δ ppm 7.31 - 7.36 (m, 2 H), 7.23 (dd, J=5.1, 1.1 Hz, 1 H), 7.13 - 7.20 (m, 2 H), 7.08 (t, J=7.4 Hz, 1 H), 6.89 - 6.95 (m, 4 H), 6.81 (d, J=9.3 Hz, 1 H), 4.43 - 4.50 (m, 1 H), 4.01 (t, J=7.0 Hz, 1 H), 3.79 (dd, J=10.4, 5.9 Hz, 1 H), 3.67 - 3.74 (m, 1 H), 3.58 (td, J=8.8, 4.8 Hz, 1 H), 3.35 - 3.42 (m, 3 H), 2.24 - 2.33 (m, 1 H), 2.00 - 2.07 (m, 1 H). LC-MS m/z 433 (M+H)+, 1.23 min (ret time).
Example 59
7V1-[(3S)-l-(3-cyano-4-pyridinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide
hydrochloride
Figure imgf000147_0002
HCI
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-4-pyridinyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (127 mg, 0.288 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1.5 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (82.9 mg, 75%). LC-MS m/z 342 (M+H)+, 0.56 min (ret time).
Example 60
7V1-[(3S)-l-(2-cyano-3-pyridinyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000148_0001
HCI
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(2-cyano-3-pyridinyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (138 mg, 0.383 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (47.7 mg, 42%). LC-MS m/z 260 (M+H)+, 0.61 min (ret time).
Example 61
7V1-[(3S)-l-(2-cyano-3-pyridinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000148_0002
HCl
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(2-cyano-3-pyridinyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (169 mg, 0.383 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (119 mg, 82%). LC-MS m/z 342 (M+H)+, 0.63 min (ret time). Example 62
7V1-[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]-L-alaninamide hydrochloride
Figure imgf000149_0001
A solution of 1,1-dimethylethyl ((15)-2-{[(35)-l-(3-cyano-2-quinolinyl)-3- pyrrolidinyl]amino}-l-methyl-2-oxoethyl)carbamate (95 mg, 0.232 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (23.3 mg, 29%). LC-MS m/z 310 (M+H)+, 0.70 min (ret time).
Example 63
(2S)-2-amino-7V- [(3 S)- l-(3-cyano-2-quinolinyl)-3-pyrr olidinyl] butanamide
hydrochloride
Figure imgf000149_0002
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(3-cyano-2-quinolinyl)-3- pyrro lidinyl] amino }carbonyl)propyl] carbamate (95 mg, 0.224 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (47.8 mg, 57%). LC-MS m/z 324 (M+H)+, 0.72 min (ret time). Example 64
7V1-[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide hydrochloride
Figure imgf000150_0001
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-2-quinolinyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (113 mg, 0.230 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL), and the precipitate was collected by filtration and washed with Et2O (6 mL). Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (10 mg, 10%). LC-MS m/z 392 (M+H)+, 0.79 min (ret time).
Example 65
^-[(S^-l-CS-cyano-l-thienylJ-S-pyrrolidinyll-S-Cl-thienylJ-L-alaninamide hydrochloride
Figure imgf000150_0002
A solution of 1,1-dimethylethyl [(15)-2-{[(35)-l-(3-cyano-2-thienyl)-3- pyrrolidinyl]amino}-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (5.4 mg, 0.012 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 50 min. The reaction mixture was diluted with Et2O (2 mL), and the resultant blue precipitate was filtered through a plug of cotton and washed with Et2O (1 mL). The solid was dissolved in MeOH (2 mL) and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (4.5 mg, 97%). LC-MS m/z 347 (M+H)+, 0.81 min (ret time). Example 66
7V1-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-7V2-methyl-L- alaninamide hydrochloride
Figure imgf000151_0001
HCI
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(trifluoromethyl)- phenyl]-3-pyrrolidinyl} amino)- l-methyl-2-oxoethyl]methylcarbamate (140 mg, 0.318 mmol) in HCl (4 M solution in 1,4-dioxane, 1.50 mL, 6.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (8 mL), filtered, and washed with Et2O (2 x 2 mL). The solid was dissolved in MeOH (4 mL), concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (53.4 mg, 45%). LC-MS m/z 341 (M+H)+, 0.91 min (ret time).
Example 67
7V1-{(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}-L-alaninamide
hydrochloride
Figure imgf000151_0002
HCl
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(methyloxy)phenyl]- 3-pyrrolidinyl} amino)- l-methyl-2-oxoethyl]carbamate (111 mg, 0.286 mmol) in HCl (4 M solution in 1,4-dioxane, 1.50 mL, 6.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (6 mL), filtered, and washed with Et2O (2 x 2 mL). The solid was dissolved in MeOH (4 mL), concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (58.6 mg, 63%). LC-MS m/z 289 (M+H)+, 0.91 min (ret time). Example 68
(2S)-2-amino-7V-{(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}butanamide hydrochloride
Figure imgf000152_0001
A solution of 1,1-dimethylethyl {(15)-l-[({(35)-l-[2-cyano-4-(methyloxy)- phenyl]-3-pyrrolidinyl}amino)carbonyl]propyl} carbamate (108 mg, 0.268 mmol) in HCl (4 M solution in 1,4-dioxane, 1.50 mL, 6.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (6 mL), filtered, and washed with Et2O (2 x 2 mL). The solid was dissolved in MeOH (4 mL), concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (63.5 mg, 70%). LC-MS m/z 303 (M+H)+, 0.89 min (ret time).
Example 69
7V1-{(3S)-l-[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl}-3-(2-thienyl)-L- alaninamide hydrochloride
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(methyloxy)phenyl]- 3-pyrrolidinyl}amino)-2-oxo-l-(2-thienylmethyl)ethyl]carbamate (194 mg, 0.412 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 45 min. The reaction mixture was diluted with Et2O (6 mL), filtered, and washed with Et2O (2 x 2 mL). The solid was dissolved in MeOH (4 mL), concentrated under a stream of nitrogen at 50 0C, and dried under high vacuum. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (109.7 mg, 65%). 1H NMR (400 MHz, MeOD) δ ppm 7.20 (dd, J=5.1, 1.1 Hz, I H), 7.11 (dd, J=9.3, 3.0 Hz, 1 H), 7.03 (d, J=3.0 Hz, 1 H), 6.94 (d, J=2.8 Hz, 1 H), 6.88 (dd, J=5.0, 3.5 Hz, 1 H), 6.78 (d, J=9.3 Hz, 1 H), 4.41 - 4.47 (m, 1 H), 4.03 (t, J=7.0 Hz, 1 H), 3.76 (s, 3 H), 3.71 (dd, J=10.2, 5.9 Hz, 2 H), 3.66 (d, J=9.5 Hz, 1 H), 3.49 (dt, J=9.3, 4.1 Hz, 1 H), 3.35 - 3.37 (m, 2 H), 2.23 - 2.30 (m, 1 H), 1.99 - 2.05 (m, 1 H). LC-MS m/z 371 (M+H)+, 0.93 min (ret time).
Example 70
(2S)-2-amino-7V- [(3S)- l-(3-cyano-4 '-fluoro-4-biphenylyl)-3-pyr rolidinyl] butenamide hydrochloride
Figure imgf000153_0001
HCI
A solution of 1,1-dimethylethyl [(15)-l-({[(35)-l-(3-cyano-4'-fluoro-4- biphenylyl)-3-pyrrolidinyl]amino}carbonyl)propyl]carbamate (564 mg, 1.209 mmol) in HCl (4 M solution in 1,4-dioxane, 5.0 mL, 20.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (6 mL) and the supernatant was carefully removed by pipet. Water (6 mL) was added and the mixture was lyophilized with a Genevac® HT- 4X to afford the title compound (28 mg, 4.8%). 1H NMR (DMSO-d6) δ ppm 8.78 (d, J = 6.3 Hz, IH), 8.12 (br. s., 3H), 7.81 (d, J= 2.3 Hz, IH), 7.75 (dd, J= 9.0, 2.3 Hz, IH), 7.65 - 7.71 (m, 2H), 7.24 (t, J= 8.9 Hz, 2H), 6.88 (d, J= 9.0 Hz, IH), 4.43 (m, IH), 3.85 (dd, J = 10.3, 5.8 Hz, IH), 3.75 (m, IH), 3.61 - 3.68 (m, 2H), 3.48 (m, IH), 2.20 (m, IH), 2.01 (m, IH), 1.72 (m, 2H), 0.84 (t, J= 7.4 Hz, 3H). LC-MS m/z 367 (M+H)+, 1.00 min (ret time).
Example 71
(2S)-7V-[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-2-azetidine- carboxamide trifluoroacetate
Figure imgf000153_0002
To a solution of 1,1-dimethylethyl (2S)-2-({[(3S)-l-(3-cyano-4'-fluoro-4- biphenylyl)-3-pyrrolidinyl]amino}carbonyl)-l-azetidinecarboxylate (114 mg, 0.245 mmol) in CH2Cl2 (1.0 mL) was added TFA (1.0 mL, 12.98 mmol). The reaction mixture was stirred at RT for 3 h and then concentrated under a stream of nitrogen at 50 0C. The residue was dissolved in DMSO (2 rnL), filtered through a 0.45 μm Acrodisc® filter, and purified by reverse phase HPLC (YMC Cl 8 S-5 μm/12 nm 50 x 20 mm preparatory column), eluting at 20 mL/min with a linear gradient running from 20% CH3CN/H2O (0.1 % TFA) to 60% CH3CN/H2O (0.1 % TFA) over 20 min. The desired fractions were concentrated under a stream of nitrogen at 50 0C, dissolved in 1,4-dioxane (2 mL), and lyophilized on a Genevac® HT-4X to afford the title compound (47 mg, 40%). LC-MS m/z 365 (M+H)+, 0.93 min (ret time).
Example 72
7V-[(3S)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-L-prolinami(ie
hydrochloride
Figure imgf000154_0001
A solution of 1,1-dimethylethyl (25)-2-({[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)- 3-pyrrolidinyl]amino}carbonyl)-l-pyrrolidinecarboxylate (72 mg, 0.150 mmol) in HCl (4 M solution in 1,4-dioxane, 0.50 mL, 2.00 mmol) was stirred at RT for 1 h 25 min. The reaction mixture was diluted with Et2O (2 mL), and the resultant precipitate was filtered through a plug of cotton and washed with Et2O (1 mL). The solid was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (48 mg, 77%). LC-MS m/z 379 (M+H)+, 0.95 min (ret time).
Example 73
7V1-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-L-alaninamide-</i hydrochloride
Figure imgf000154_0002
HCI
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(trifluoromethyl)- phenyl]-3-pyrrolidinyl}amino)-l-methyl-2-oxoethyl]carbamate-(ii (87 mg, 0.204 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL) and then partially concentrated under a stream of nitrogen. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford the title compound (64.8 mg, 87%) as an off white solid. LC- MS m/z 328 (M+H)+, 0.88 min (ret time).
Example 74
7V1-{(3S)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-L-alaninamide-</4 hydrochloride
Figure imgf000155_0001
HCl
A solution of 1,1-dimethylethyl [(15)-2-({(35)-l-[2-cyano-4-(trifluoromethyl)- phenyl]-3-pyrrolidinyl}amino)-l-methyl-2-oxoethyl]carbamate-(i4 (89 mg, 0.207 mmol) in HCl (4 M solution in 1,4-dioxane, 2.0 mL, 8.00 mmol) was stirred at RT for 1 h. The reaction mixture was diluted with Et2O (5 mL) and then partially concentrated under a stream of nitrogen. Water (2 mL) was added and the mixture was lyophilized with a Genevac® HT-4X to afford an oily residue. Water (2 mL) was added and the mixture was lyophilized a second time to afford the title compound (59.5 mg, 78%) as an off white solid. LC-MS m/z 331 (M+H)+, 0.91 min (ret time).
Examples 75-120
General Experimental for Table 1
The compounds in Table 1 were prepared according to the general procedures described below, from previously described intermediates (where indicated) and commercially available Boc-protected α-amino acids: General Procedure A:
Figure imgf000156_0001
An appropriate aryl bromide (1 equiv.) was coupled with an appropriate aryl boronic acid or boronic ester (1 equiv.) in the presence of PdCl2(dppf) (0.01 - 0.1 equiv.) and K2CO3 (3 equiv.) in 3:1 1 ,4-dioxane/water in a microwave reactor at 140 0C (high absorption) for 1 h. Water and EtOAc were added to the reaction mixture. The layers were separated, and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed thrice with brine, dried over Na2SO4, and concentrated. The crude product was purified by flash column chromatography to afford the desired compound II.
General Procedure B:
Figure imgf000156_0002
A mixture of a biaryl fluoride, 1,1-dimethylethyl (35)-3-pyrrolidinylcarbamate, and K2CO3 in DMSO was heated at 100 0C (bath temp) for 2 - 7O h until the reaction was determined to be complete by LC-MS. Water and EtOAc were added. The layers were separated, and the aqueous layer was extracted thrice with EtOAc. The combined organic layers were washed thrice with brine and concentrated. The crude product was purified by flash column chromatography to afford the desired compound III.
General Procedure C:
Figure imgf000156_0003
A solution of 4 M HCl/l,4-dioxane (excess) was added to compound III, and the mixture was stirred at RT until the reaction was determined to be complete by LC-MS (typically 15 min - 5 h). Et2O was added, causing a precipitate to form. This precipitate was filtered and washed with Et2O. The precipitate was dissolved in MeOH and concentrated under a stream of nitrogen at 50 0C to afford the desired compound IV.
General Procedure D:
Figure imgf000157_0001
A solution of 50 wt% ®T3P in EtOAc (1.5 equiv.) was added to a solution of the amine HCl salt IV (1 equiv.), an appropriate Boc-protected α-amino acid (1 equiv.), and Et3N (3 equiv.) in CH2Cl2 at 0 0C (bath temp). The ice bath was removed, and the reaction was stirred at RT until complete as determined by LC-MS. The reaction was washed twice each with saturated aq. NaHCO3 and 10% citric acid. The organic layer was dried over Na2SO4 and concentrated. The crude product was purified by flash column chromatography to afford the desired compound V.
General Procedure E:
Figure imgf000157_0002
PdCl2(dppf) (0.01 - 0.1 equiv.) was added to a mixture of 1 , 1 -dimethylethyl (2S)-2-
({[(3 S)-I -(4-bromo-2-cyanophenyl)-3 -pyrrolidinyl] amino } carbonyl)- 1 - pyrrolidinecarboxylate (1 equiv.), an appropriate boronic acid (1.05 equiv.), and K2CO3 (3 equiv.) in 1,4-dioxane and water in a microwave vial. The vial was capped and the reaction mixture was heated in a microwave reactor at 140 0C (high absorbance) for 15 min. The reactions were transferred into 20 mL vials, and the 1,4-dioxane was blown off using nitrogen at 55 0C. Water and CH2Cl2 were added to the reaction mixture. The mixture was transferred to a hydrophobic frit and the layers were separated. The aqueous layer was extracted with CH2Cl2. The solvent was evaporated using nitrogen at 50 0C, the residue was dissolved in 1.0 mL DMSO and purified by reverse phase HPLC to afford the desired compound VI.
General Procedure F:
Figure imgf000158_0001
V VII
A solution of 4 M HCl/l,4-dioxane (excess) was added to compound V, and the mixture was stirred at RT until the reaction was determined to be complete by LC-MS (typically 15 min - 5 h). Et2O was added, causing a precipitate to form. This precipitate was filtered and washed with Et2O. 1,4-Dioxane was added, and the mixture was lyophilized in a Genevac HT -4X to afford the desired compound VII.
General Procedure G:
Figure imgf000158_0002
V VIII
Alternatively, TFA was added to a solution of compound V in CH2Cl2. The reaction was stirred at 0 0C for 15 min - 5 h. The reaction mixture was concentrated under reduced pressure. The crude product was purified by HPLC. The desired fractions were concentrated under a stream of nitrogen at 50 0C to afford the desired compound VIII.
Table 1
Figure imgf000159_0001
Figure imgf000160_0001
Figure imgf000161_0001
Figure imgf000162_0001
Figure imgf000163_0001
Biological Background:
Biological Assay(s)
The compounds according to Formula (I) are cathepsin C inhibitors, which indirectly inhibit the activity of serine proteases that are activated by cathepsin C, such as NE. The compounds according to Formula (I), therefore, are useful in the treatment of COPD and other conditions involving cathepsin C and/or such serine proteases. 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 cathepsin C inhibitor or for determining the ability of a candidate compound to prevent the cathepsin C mediated activation of certain serine proteases, as well as suitable tissue and in vivo models. All examples were found to be cathepsin C inhibitors.
A. Transpeptidation of Leucine-Leucine-O-Methyl (LLOM) cell-based Luminescence Viability Assay
Principle:
Cathepsin C has been shown to catalyze the transpeptidation of dipeptidyl methyl-
O -esters within the lysosomes of cells from the monocytic lineage such as HL60, U937 or THPl causing a membranolytic effect that results in cell death (DL. Thiele, P. Lipsky
PNAS 1990 Vol. 87, pp. 83-87). This mechanism was used to assess Cathepsin C in cells activity in the presence of the compounds of the invention.
Frozen HL-60 cells were resuspended at 1.25 x 105 cells/mL in fresh prewarmed
Iscove's modified Dulbeccos' medium (IMDM, contains 25 mM glutamine) with 20 % FBS. This suspension was dispensed (8 μL) into white low volume 384 well plates.
Plates were previously stamped with 100 nL of compound at a top concentration of 2.5 mM and serially diluted 1 :3. Control and blank wells contained 100 nL of DMSO. Each well then received 2 μL of a fresh 1.25 mM solution of leucine-leucine-OMethyl (LLOM,
Bachem) in IMDM plus 25 mM HEPES (final concentration LLOM 250 μM). The plates were covered and incubated for 4 h at 37 0C in a 5% CO2 incubator, then removed and equilibrated to room temperature for 10 min. Cell viability was determined with a CellTiter-Glo luminescent assay (Promega) according to the manufacturer's instructions. Cell viability was compared to controls containing no LLOM (100 %).
B. Human Neutrophil Cathepsin C Assay
Neutrophils (PMN) were isolated from human peripheral blood using standard methods. In brief, 25 mL blood was layered over 15 mL Ficol-Paque Plus (Amersham Biosciences) and centrifuged at 400 g at room temperature for 30 min. The red blood cell pellets were resuspended in 35 mL phosphate-buffered saline without Ca2+ or Mg2+ (PBS). Dextran T-500 (Pharmacia, 6 % solution in PBS) was added to each tube (12 mL), tubes were mixed by inversion, and allowed to stand at room temperature for 40 min. The layer above the red cells was collected, centrifuged at 800 g, and gently resuspended ~3 mL. Red blood cells were lysed by addition of 18 mL sterile water for 30 sec, followed immediately by addition of 2 mL 1OX PBS. Cells were recollected and resuspended to 2 x 105 cells/mL in PBS with 0.1 % gelatin.
Compounds were prepared in serial three-fold dilutions starting with a top concentration of 10 mM solution in DMSO. PMN were then were plated in wells in 96-well flat-bottom tissue culture plates in (20,000 cells in 100 μL). Compound was added (1 μL each concentration) to the wells in triplicate, plates were mixed for 5 min on a plate shaker and then incubated for 30 min at 37 0C, 5% CO2. Substrate (H-Gly-Arg)2 Rl 10 was added (5 μL of a 0.5 mM solution in PBS) and plates incubated for a further 3 h. The cleavage of substrate was measured at using an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Compounds were compared to controls containing DMSO only and ICso's were determined using non-linear regression curve fit analysis (GraphPad Prism).
C. Recombinant Cathepsin C in vitro assay:
The activity of recombinant human cathepsin C was measured by the cleavage of a fluorogenic substrate, H-Ser-Tyr-AMC. Briefly, 24 pM cathepsin C was incubated with test compound (e.g. inhibitor) in a buffer consisting of 50 mM sodium acetate, 30 mM sodium chloride, 1 mM CHAPS, 1 mM dithiothreitol, 1 mM EDTA, pH 5.5 at room temperature for one hour. After one hour of incubating test compound with cathepsin C, the activity assay was initiated by the addition of an equal volume of 0.010 mM H-Ser- Tyr-AMC in the same buffer. After one hour, the activity assay was stopped by the addition of 1/5 volume of 100 μM E-64. The reaction product was measured on a fluorescence reader set at an excitation wavelength of 360 nm and emission wavelength of 460 nm and equipped with a 400 nm dichroic mirror.
The compounds of the invention (Examples 1-120) exhibit 50% cathepsin C inhibition (as determined using the above method) at concentrations of from
approximately 8,000 nM to approximately 0.01 nM. For instance, the compound of Example 62 exhibited 50% cathepsin C inhibition at a concentration of approximately 1,000 nM. Preferred compounds of the invention exhibit 50% inhibition at concentrations of from approximately 100 nM to approximately 0.01 nM. For instance, the compound of Example 26 exhibited 50% cathepsin C inhibition at a concentration of approximately 100 nM. More preferred compounds of the invention exhibit 50% inhibition at concentrations of from approximately 10 nM to approximately 0.01 nM.
The following Example numbers represent preferred compounds of this invention: 4, 6, 10, 11, 12, 16, 17, 18, 20, 21, 22, 24, 26, 29, 32, 39, 41, 43, 48, 49, 51, 57, 65, 67, 68, 73, 74, 77, 84, 90, 112, and 113, or a salt thereof. The following Example numbers represent the more preferred compounds of this invention: 23, 25, 27, 30, 31, 35, 36, 42, 44, 45, 52, 58, 69, 70 and 76, or a salt thereof. The compounds of the invention are believed to be useful in therapy as defined above and to not have unacceptable or untoward effects when used in compliance with a permitted therapeutic regime.
The foregoing examples and assay have been set forth to illustrate the invention, not limit it. What is reserved to the inventors is to be determined by reference to the claims.

Claims

What is claimed is:
1. A compound according to Formula (I) or a pharmaceutically acceptable salt thereof:
Figure imgf000167_0001
wherein:
A represents a 5- or 6-membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, (C3-Ce)cycloalkyl, heterocycloalkyl, hydroxyl, (Ci-C4)alkoxy, aryl, aryloxy, heteroaryl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl,
-SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said aryl, aryloxy, or heteroaryl is optionally substituted by -OCH2O- or is optionally substituted one to three times, independently, by halogen, -CF3, (d-C4)alkyl, formyl, -CO(Ci-C4)alkyl, -CO2(C i-C4)alkyl, hydroxyl, (Ci-C4)alkoxy, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 12-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 5- or 6-membered heterocycloalkyl group;
R1 is hydrogen, (Ci-C4)alkyl, (C2-C5)alkenyl, (C2-C5)alkynyl, (C3-C5)cycloalkyl, (C3-C4)cycloalkyl(Ci-C2)alkyl, cyano(Ci-C2)alkyl, hydroxy(Ci-C2)alkyl,
methoxy(Ci-C2)alkyl, aryl(Ci-C2)alkyl, or heteroaryl(Ci-C2)alkyl, wherein the heteroaryl moiety of said heteroaryl(Ci-C2)alkyl is a 5-membered aromatic ring containing one heteroatom which is oxygen or sulfur and optionally containing one or two nitrogen atoms; and
R2 is hydrogen or methyl; or R1 and R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted one or two times, independently, by halogen, -CF3, cyano, (Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, hydroxyl, (CrC4)alkoxy, or (d-C4)alkylthio-; wherein said ring is optionally fused to a (C3-Cs)cycloalkyl ring.
2. The compound or salt according to Claim 1, wherein A represents a 6- membered aromatic ring optionally containing one or two nitrogen atoms, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl,
imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3- dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano, -CO2(Ci-C4)alkyl, -CONH(C i-C4)alkyl,
-CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(Ci-C4)alkyl, -SO2NH(Ci-C4)alkyl,
-SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 11-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy.
3. The compound or salt according to Claim 1, wherein A represents a phenyl ring optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1,3- dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl,
-CON(Ci-C4)alkyl(Ci-C4)alkyl, -SO2(Ci-C4)alkyl, -SO2NH(Ci-C4)alkyl,
-SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino, (Ci-C4)alkylamino,
((Ci-C4)alkyl)((Ci-C4)alkyl)amino, or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy.
4. The compound or salt according to Claim 1, wherein A represents a 5- membered aromatic ring optionally containing one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein A is optionally substituted one to three times, independently, by halogen, (Ci-C4)alkyl, -CF3, hydroxyl, (Ci-C4)alkoxy, azetidinyl, pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, 1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3- dioxanyl, 1 ,4-dioxanyl, 1,3-oxathiolanyl, 1,3-oxathianyl, 1, 3 -dithianyl, phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pteridinyl, cyano, -CO2(C i-C4)alkyl, -CONH(C i-C4)alkyl, -CON(Ci-C4)alkyl(Ci-C4)alkyl,
-SO2(Ci-C4)alkyl, -SO2NH(C i-C4)alkyl, -SO2N(Ci-C4)alkyl(Ci-C4)alkyl, amino,
(Ci-C4)alkylamino, ((Ci-C4)alkyl)((Ci-C4)alkyl)amino, Or NO2, wherein said phenyl, phenyloxy, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-naphthyridinyl, 1 ,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8- naphthyridinyl, or pteridinyl is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy;
and wherein A is optionally fused to an aromatic or non-aromatic carbocyclic or heterocyclic ring moiety to form an 8- to 10-membered bicyclic group, wherein said aromatic carbocyclic or heterocyclic ring moiety is optionally substituted one to three times, independently, by halogen, -CF3, (Ci-C4)alkyl, hydroxyl, or (Ci-C4)alkoxy, and wherein said non-aromatic carbocyclic ring moiety is optionally substituted by a spiro-fused 1,3-dioxolanyl or 1,3-dioxanyl group.
5. The compound or salt according to any one of Claims 1-4, wherein R1 is (Ci-C4)alkyl or thienyl(Ci-C2)alkyl.
6. The compound or salt according to any one of Claims 1-5, wherein R1 is methyl, ethyl, n-propyl, isopropyl, or 2-thienylmethyl.
7. The compound or salt according to any one of Claims 1-6, wherein R1 is methyl.
8. The compound or salt according to any one of Claims 1-6, wherein R1 is 2-thienylmethyl.
9. The compound or salt according to any one of Claims 1-4, wherein R1 and
R2 taken together with atoms through which they are connected form a 4- to 6-membered saturated ring optionally substituted by F, Cl, -CF3, cyano, methyl, methoxy, or methylthio-.
10. A compound which is :
N1-[(35)-l-(3-chloro-4-cyano-5-isothiazolyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-alaninamide;
N1 -[(3S)-I -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] glycinamide;
N1-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-L- alaninamide;
N1 - [(35)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-valinamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]butanamide;
N1 - [(35)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-norvalinamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-leucinamide;
N- [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-phenylalaninamide;
N1-[(35)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(4-cyano-3-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
ethyl 5-[(3iS)-3-(L-alanylamino)- 1 -pyrrolidinyl]-4-cyano-2-thiophenecarboxylate;
N1 - [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -L-serinamide;
N1 -[(3S)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(1,3 -thiazol-4-yl)-L- alaninamide;
(25)-2-amino-3 -cyano-iV- [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl]- propanamide;
N1 - [(3S)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(l,3-thiazol-2-yl)phenyl]-3-pyrrolidinyl}- butanamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(2-pyrimidinyl)phenyl]-3-pyrrolidinyl}- butanamide;
N1- [(3S)- l-(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -cyclopropyl-L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-4-pentynamide; N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- butanamide; N1 - [(35)- 1 -(4-bromo-2-cyanophenyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide;
N1 - [(35)- 1 -(4-bromo-2-cyanophenyl)-3 -pyrrolidinyl] -L-alaninamide;
N1 - [(35)- 1 -(3 -cyano-4-biphenylyl)-3 -pyrrolidinyl]-3 -(2-thienyl)-L-alaninamide;
N1 - [(35)- 1 -(3 -cyano- 1 -methyl- lH-indol-2-yl)-3 -pyrrolidinyl] -L-alaninamide; N1-[(35)-l-(3-cyano-l-methyl-lH-indol-2-yl)-3-pyrrolidinyl]-3-(2-thienyl)-L- alaninamide;
N1 - [(35)- 1 -(2-cyano-4-fluorophenyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-fluorophenyl)-3-pyrrolidinyl]butanamide;
N1- [(35)- l-(2-cyano-4-fluorophenyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide; N1 -[(35)- 1 -(3 -cyano-4'-fluoro-4-biphenylyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L- alaninamide;
N1 - [(35)- 1 -(2-cyano-4-nitrophenyl)-3 -pyrrolidinyl] -3 -(3 -thienyl)-L-alaninamide;
N1-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-L-norleucinamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopropyl- ethanamide;
N1-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-3-(2- thienyl)-L-alaninamide;
(25)-2-amino-JV-[(35)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3- pyrrolidinyl]butanamide;
N1- [(35)- 1 -(3 -cyano-2-pyrazinyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]butanamide;
N1-[(35)-l-(3-cyano-2-pyrazinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
(25)-2-amino-N-[(35)-l-(3-cyano-2-pyridinyl)-3-pyrrolidinyl]butanamide;
N1 - [(35)- 1 -(3 -cyano-2-pyridinyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide; N1-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-L-alaninamide;
N1-[(35)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]-L-alaninamide;
N1- [(35)- l-(2-cyano-3-thienyl)-3 -pyrrolidinyl] -3 -(2 -thienyl)-L-alaninamide;
(25)-2-amino-N-[(35)-l-(3-cyano-4-biphenylyl)-3-pyrrolidinyl]butanamide;
N1- {(35)- 1 -[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide; (25)-2-amino-N-{(35)-l-[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl}- butanamide; N1- [(3S)- 1 -[2-cyano-5-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1- [(3S)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1-[(35)-l-(2-cyanophenyl)-3-pyrrolidinyl]-L-alaninamide;
(25)-2-amino-Λ/-[(35)-l-(2-cyanophenyl)-3-pyrrolidinyl]butanamide;
N1-[(35)-l-(2-cyanophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide;
N1 - [(35)- 1 -(2-cyano-4,5 -difluorophenyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]butanamide; N1-[(35)-l-(2-cyano-4,5-difluorophenyl)-3-pyrrolidinyl]-3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-[(35)-l-(2-cyano-4-nitrophenyl)-3-pyrrolidinyl]-2-cyclopentyl- ethanamide;
(25)-2-amino-Λ/-[(35)-l-(2-cyano-3-thienyl)-3-pyrrolidinyl]butanamide;
N1- {(35)- 1 -[2-cyano-4-(phenyloxy)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
N1 - [(35)- 1 -(3 -cyano-4-pyridinyl)-3 -pyrrolidinyl] -3 -(2-thienyl)-L-alaninamide; N1 - [(3S)- 1 -(2-cyano-3 -pyridinyl)-3 -pyrrolidinyl] -L-alaninamide;
N1 - [(3 S)- 1 -(2-cyano-3 -pyridinyl)-3 -pyrrolidinyl]-3 -(2-thienyl)-L-alaninamide; N1- [(3S)- 1 -(3 -cyano-2-quinolinyl)-3 -pyrrolidinyl] -L-alaninamide;
(25)-2-amino-N-[(3S)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]butanamide;
N1-[(35)-l-(3-cyano-2-quinolinyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide; N1-[(35)-l-(3-cyano-2-thienyl)-3-pyrrolidinyl]-3-(2-thienyl)-L-alaninamide; N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -N2-methyl-L- alaninamide;
N1- {(35)- 1 -[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(methyloxy)phenyl]-3- pyrrolidinyl}butanamide;
N1- {(35)- 1 -[2-cyano-4-(methyloxy)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]- butenamide; (25)-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-2-azetidine- carboxamide;
N- [(3S)- 1 -(3 -cyano-4'-fluoro-4-biphenylyl)-3 -pyrrolidinyl] -L-prolinamide;
N1-{(35)-l-[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl}-L-alaninamide-(ii; N1- {(35)- 1 -[2-cyano-4-(trifluoromethyl)phenyl]-3-pyrrolidinyl} -L-alaninamide-^;
N1- {(35)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
N1- {(35)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -3-(2-thienyl)-L- alaninamide;
(25)-2-amino-N-{(35)-l-[2-cyano-4-(4-pyridinyl)phenyl]-3- pyrrolidinyl}butanamide;
N1- {(35)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
N1- {(35)- 1 -[2-cyano-4-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -L-alaninamide;
(25)-JV- {(35)- 1 -[2-cyano-5-(l -pyrrolidinyl)phenyl]-3-pyrrolidinyl} -2- azetidinecarboxamide;
(2S)-N-[(3S)- 1 -(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-2-azetidinecarboxamide;
(2S)-N-[(3S)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -2- azetidinecarboxamide;
JV-[(35)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -L- prolinamide;
(2S)-N-[(3S)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-2- azetidinecarboxamide;
N- [(35)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3 -pyrrolidinyl] -L-prolinamide;
(2S)-N- {(35)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -2- azetidinecarboxamide;
N- {(35)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
(2S)-N-[(3S)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -2- piperidinecarboxamide;
(2S)-N-[(3S)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-2- piperidinecarboxamide;
(4S)-N-[(3S)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -4- fluoro-L-prolinamide; (4i?)-N-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-4- fluoro-L-prolinamide;
N-[QS)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3-pyrrolidinyl]-4,4- difluoro-L-prolinamide;
(3i?)-N-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-3-pyrrolidinyl]-3- hydroxy-L-prolinamide;
Λ/-[(35)-l-(3-cyano-5,6-dihydro-4H-cyclopenta[δ]thien-2-yl)-3-pyrrolidinyl]-L- prolinamide;
N-[(35)-l-(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thien-2-yl)-3- pyrrolidinyl] -L-prolinamide;
(25)-N-[(35)-l-(3-cyano-5,6,7,8-tetrahydro-4H-cyclohepta[δ]thien-2-yl)-3- pyrrolidinyl]-2-azetidinecarboxamide;
N- [QS)- 1 -(3 -cyano-4'-methyl-3 '-nitro-4-biphenylyl)-3 -pyrrolidinyl] -L- prolinamide;
ethyl 3'-cyano-4'-[(35)-3-(L-prolylamino)- 1 -pyrrolidinyl] -3 -biphenylcarboxylate;
N- [QS)- 1 -[4-(1 ,3-benzodioxol-5-yl)-2-cyanophenyl]-3-pyrrolidinyl} -L- prolinamide;
N- [QS)- 1 -[2-cyano-4-(5-formyl-2-thienyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
Λ/-[(35)-l-(3'-acetyl-3-cyano-4-biphenylyl)-3-pyrrolidinyl]-L-prolinamide;
N- [QS)- 1 -[2-cyano-4-(3-quinolinyl)phenyl]-3-pyrrolidinyl} -L-prolinamide;
N- [QS)- 1 -[3-cyano-4'-(methyloxy)-4-biphenylyl]-3-pyrrolidinyl} -L-prolinamide;
N- [QS)- 1 - [3 -cyano-3 '-fluoro-4'-(methyloxy)-4-biphenylyl] -3 -pyrrolidinyl} -L- prolinamide;
N- [Q S)- 1 -(3 '-amino-3-cyano-4-biphenylyl)-3 -pyrrolidinyl] -L-prolinamide;
JV-[(3IS)- 1 -(4-bromo-2-cyanophenyl)-3-pyrrolidinyl]-L-prolinamide;
N-[QS)- 1 -(3-cyano- lH-indol-2-yl)-3-pyrrolidinyl]-L-prolinamide;
N-[QS)- 1 -(3-cyano-4,7-dihydro-5H-spiro[ 1 -benzothiophene-6,2'-[ 1 ,3]dioxolan]-2- yl)-3-pyrrolidinyl]-L-prolinamide;
(4S)-N-[QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-4-methyl-L- prolinamide;
(4S)-N-[QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide; (4S)-N-[(3S)- 1 -(3 -cyano-4-biphenylyl)-3 -pyrrolidinyl] -4-fluoro-L-prolinamide;
(45)-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide;
(4S)-N- [(3S)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -4-fluoro-L- prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano-4,5,6,7-tetrahydro- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -4- methyl-L-prolinamide;
N-[(35)- 1 -(3-cyano- 1 -benzothien-2-yl)-3-pyrrolidinyl]-L-prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -benzothien-2-yl)-3-pyrrolidinyl]-4-fluoro-L- prolinamide;
(4S)-N-[(3S)- 1 -(3-cyano- 1 -benzothien-2-yl)-3 -pyrrolidinyl] -4-methyl-L- prolinamide;
(45)-N-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3-pyrrolidinyl]-4-methyl-L- prolinamide;
(4S)-N- [(3S)- 1 -[2-cyano-4-(3-thienyl)phenyl]-3-pyrrolidinyl} -4-methyl-L- prolinamide; or
(2S)-N- [(3S)- 1 -[2-cyano-4-(4-pyridinyl)phenyl]-3-pyrrolidinyl} -2- piperidinecarboxamide;
or a pharmaceutically acceptable salt thereof.
11. A compound which is N1-[(35)-l-(3-cyano-4'-fluoro-4-biphenylyl)-3- pyrrolidinyl]-L-alaninamide or a pharmaceutically acceptable salt thereof.
12. A compound which is N1-{(35)-l-[2-cyano-4-(4-pyridinyl)phenyl]-3- pyrrolidinyl} -3-(2-thienyl)-L-alaninamide or a pharmaceutically acceptable salt thereof.
13. A compound which is (25)-2-amino-JV-[(3iS)- 1 -(3-cyano-4-biphenylyl)-3- pyrrolidinyl]butanamide or a pharmaceutically acceptable salt thereof.
14. A compound which is (45)-N-[(35)-l-(3-cyano-4,5,6,7-tetrahydro-l- benzothien-2-yl)-3-pyrrolidinyl]-4-fluoro-L-prolinamide or a pharmaceutically acceptable salt thereof.
15. A compound which is (2S)-2-amino-N-[(3S)- 1 -(3-cyano-4'-fluoro-4- biphenylyl)-3-pyrrolidinyl]-butenamide or a pharmaceutically acceptable salt thereof.
16. A compound which is (2S)-N-[QS)- 1 -(3-cyano- 1 -methyl- lH-indol-2-yl)-3- pyrrolidinyl]-2-azetidinecarboxamide or a pharmaceutically acceptable salt thereof.
17. A pharmaceutical composition which comprises the compound or salt according to any one of Claims 1-16, and a pharmaceutically acceptable excipient.
18. A process for preparing the composition as defined in claim 17, the process comprising mixing the compound or salt according to any one of Claims 1-16 with a pharmaceutically acceptable excipient.
19. A method for treating chronic obstructive pulmonary disease comprising administering to a patient in need thereof an effective amount of the compound or salt according to any one of Claims 1-16.
20. A method for treating chronic obstructive pulmonary disease comprising administering to a patient in need thereof the pharmaceutical composition according to Claim 17.
PCT/US2010/046566 2009-08-26 2010-08-25 Cathepsin c inhibitors WO2011025799A1 (en)

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