# Patent

## Title
Substituted 2-quinolyl-oxazoles useful as PDE4 inhibitors

## Abstract

The invention claims compounds of the formula




wherein





    
    
        is a 5-membered heteroaryl;
        X is S or O;
        R1 is H, alkyl, cycloalkyl, cylcoalkylalkyl-, —CH2F, —CHF2, —CF3, —C(O)alkyl or —C(O)NR18R19;
        R3 and R4 H, alkyl, hydroxyalkyl or —C(O)Oalkyl;
        R5 and R6 are H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH2F, —CHF2, —CF3, —C(O)OH or —C(O)Oalkyl;
        R7 is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R17-phenyl)alkyl or —CH2—C(O)—O-alkyl; and R8 comprises alkyl, heteroaryl, phenyl or cycloalkyl, or heterocycloalkyl, all optionally substituted, or a cycloalkyl- or heterocycloalkyl-substituted amide; or
        R7 and R8 and the nitrogen to which they are attached together form an optionally substituted ring; and the remaining variables are as defined in the specification. Also claimed are pharmaceutical compositions, the use of the compounds as PDE4 inhibitors, and combinations with other actives.

## Background
CROSS REFERENCE TO RELATED APPLICATION
This application claims the benefit of U.S. Provisional Application 60/572,266, filed May 18, 2004.


FIELD OF THE INVENTION
The present invention relates to substituted 2-quinolyl-oxazoles, thiazoles, imidazoles and pyrazoles, pharmaceutical compositions comprising them, and their use as PDE4 inhibitors for treating a variety of diseases such as allergic and inflammatory diseases, CNS diseases and diabetes. Combinations with other agents useful in the treatment of several diseases are also claimed.
BACKGROUND
Phosphodiesterases are known to regulate cyclic AMP, and phosphodiesterase 4 (PDE4) has been shown to be the predominant regulator of cyclic AMP in respiratory smooth muscle and inflammatory cells. Inhibitors of PDE4 are useful in treating a variety of diseases, including allergic and inflammatory diseases, diabetes, central nervous system diseases, pain, and viruses that produce TNF.
Amino-substituted quinolyl PDE4 inhibitors are disclosed in U.S. Pat. No. 5,804,588; sulfonamide-substituted quinolyl PDE4 inhibitors are disclosed in U.S. Pat. No. 5,834,485; and (benzo-fused)heteroaryl-substituted PDE4 inhibitors are disclosed in U.S. Pat. No. 6,069,151.
SUMMARY OF THE INVENTION
The present invention relates to a compound having the structural formula I




or a pharmaceutically acceptable salt or solvate thereof, wherein





R is H or alkyl;
X is O or S;
R1 is H, alkyl, cycloalkyl, cycloalkyl(C1-C4)alkyl-, —CH2F, —CHF2, —CF3, —C(O)alkyl or —C(O)NR18R19;
R3 and R4 are independently selected from the group consisting of H, alkyl, hydroxyalkyl and —C(O)Oalkyl;
R5 and R6 are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH2F, —CHF2, —CF3, —C(O)OH, —C(O)Oalkyl and —C(O)NR43R44;
t is 1 or 2;
R7 is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R17-phenyl)alkyl or —CH2—C(O)—O-alkyl;
R8 is H, alkyl, alkenyl, alkoxy, alkoxyalkyl, hydroxyalkyl, dihydroxyalkyl, alkyl-NR18R19, cyanoalkyl, haloalkyl, R23-heteroaryl, R23-heteroarylalkyl, R36-heterocycloalkyl, (R36-heterocycloalkyl)alkyl, R17-phenyl, (R17-phenyl)alkyl, R17-naphthyl, (R17-naphthyl)alkyl, R17-benzyloxy, -alkyl-C(O)—NR18R19, -alkyl-C(O)—N(R30)—(R23-heteroaryl), -alkyl-C(O)—(R17-phenyl), -alkyl-C(O)—(R36-heterocycloalkyl); -alkyl-N(R30)—C(O)Oalkyl, -alkyl-N(R30)—C(O)—NR18R19, -alkyl-N(R30)—C(O)alkyl, -alkyl-N(R30)—C(O)-(fluoroalkyl), -alkyl-N(R30)—C(O)—(R39-cycloalkyl), -alkyl-N(R30)—C(O)—(R17-phenyl), -alkyl-N(R30)—C(O)—(R23-heteroaryl), -alkyl-N(R30)—C(O)-alkylene-(R23-heteroaryl), -alkyl-NH—SO2—NR18R19, -alkyl-N(R30)—(R17-phenyl), -alkyl-N(R30)—(R23-heteroaryl), -alkyl-O—(R17-phenyl), -alkyl-O—(R23-heteroaryl), -alkyl-N(R30)—SO2-alkyl, alkylthioalkyl-, alkyl-SO2-alkyl-, (R35-phenylalkyl)-S-alkyl-, (hydroxyalkyl)-S-alkyl-, (alkoxyalkyl)-S-alkyl-, -alkyl-CO2-alkyl, R45-hydroxyalkyl, dihydroxyalkyl substituted by R17-benzyloxy, dihydroxyalkyl substituted by R17-phenyl, alkoxyalkyl substituted by R17-phenyl, (R17-phenyl)alkyl substituted by —CO2alkyl, (R17-phenyl)alkyl substituted by —C(O)N(R30)2, alkyl substituted by (R23-heteroaryl) and —C(O)NR37R38, haloalkyl substituted by CO2alkyl, R12-cycloalkyl, (R12-cycloalkyl)alkyl,







or R7 and R8 and the nitrogen to which they are attached together form a ring system selected from the group consisting of







comprises an R35-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl or pyrrolidinyl ring;

p is 0 or 1;
q is 0 or 1;
the dotted line represents an optional double bond;
R9 is H, halo, alkyl, cycloalkyl, —CH2F, —CHF2 or CF3;
R10, R11, and R13 are independently selected from the group consisting of H and halo;
R12 is 1-3 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl, —(CH2)n—N(R30)—C(O)-cycloalkyl, —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)—C(O)Oalkyl, —(CH2)n—N(R30)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)—NR18R19, —(CH2)n—C(O)—NR18R19, R17-phenyl, R35-heteroarylalkyl, R35-heteroaryloxy, —C(O)-heterocycloalkyl, —O—C(O)-heterocycloalkyl, —O—C(O)—NR18R19, —NH—SO2-alkyl, —NH—C(═NH)NH2, and




or two R12 substituents on the same carbon form ═O, ═NOR30 or ═CH2;

R14 is 1 or 2 substituents independently selected from the group consisting of H, OH, halo, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, —CF3, CN, R17-phenyl, (R17-phenyl)alkyl, —NR18R19, alkyl-NR18R19, —(CH2)n—C(O)OH, —(CH2)n—C(O)Oalkyl, —(CH2)n—C(O)alkyl, —(CH2)n—C(O)(R35-phenyl), —(CH2)n—C(O)(R23-heteroaryl), —(CH2)n—C(O)NR18R19, —(CH2)n—C(O)N(R30)—(CH2)n—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)—C(O)-(fluoroalkyl), —(CH2)n—N(R30)—C(O)-(cycloalkyl), —(CH2)n—N(R30)—C(O)(R35-phenyl), —(CH2)n—N(R30)—C(O)(R23-heteroaryl), —(CH2)n—N(R30)C(O)NR18R19, —(CH2)n—N(R30)—C(O)Oalkyl, —(CH2)n—N(R30)cycloalkyl, —(CH2)n—N(R30)(R17-phenyl), —(CH2)n—N(R30)(R23-heteroaryl), —(CH2)n—N(R18)SO2alkyl, —(CH2)n—N(R20)SO2—(R17-phenyl), —(CH2)n—N(R30)SO2—CF3, —CH2S(O)0-2(R35-phenyl), —(CH2)n—OC(O)N(R30)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, (R23-heteroaryl)oxy, (R23-heteroaryl)amino, —CH(OH)—(R17-phenyl), —CH(OH)—(R23-heteroaryl), —C(═NOR30)—(R17-phenyl), —C(═NOR30)—(R23-heteroaryl), morpholinyl, thiomorpholinyl,




w is 0 or 1;
or two R14 substituents and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
each n is independently 0, 1, 2 or 3;
R15 is H, alkyl, cycloalkyl, (cycloalkyl)alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, —C(O)Oalkyl, —C(O)O(R30-cycloalkyl), -alkyl-C(O)O-alkyl, —C(O)O-alkylene-(R35-phenyl), R17-phenyl, (R17-phenyl)alkyl, —CH—(R17-phenyl)2, R23-heteroaryl, —(CH2), —C(O)NR18R19, —SO2-alkyl, —SO2-cycloalkyl, —SO2—CF3, —SO2—(R35-phenyl), —SO2—NR18R19, —C(O)alkyl, —C(O)-(fluoroalkyl), —C(O)—C(CH3)(CF3)2, —C(O)—(R17-phenyl), —C(O)—(R23-heteroaryl), —C(O)-hydroxyalkyl, —C(O)-alkoxyalkyl, —C(O)—(R39-cycloalkyl), —C(O)-alkylene-(R17-phenyl), —C(O)-alkylene-(R23-heteroaryl), —C(O)-alkylene-S—C(O)alkyl, —C(═S)—(R17-phenyl), hydroxyalkyl substituted by R17-phenyl, hydroxyalkyl substituted by R23-heteroaryl, alkoxyalkyl substituted by R17-phenyl, alkoxyalkyl substituted by R23-heteroaryl,




wherein z is 0, 1 or 2;

R16 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, R17-phenyl, (R17-phenyl)alkyl, (R23-heteroaryl)alkyl, hydroxyalkyl, alkoxyalkyl and —C(O)Oalkyl, or two R16 groups and the carbon to which they are both attached form —C(O)—;
R17 is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, cycloalkyl, —OH, hydroxyalkyl, alkoxy, alkoxyalkyl, —CN, —CF3, —OCF3, —OCHF2, —OCH2F, —C(O)OH, —C(O)Oalkyl, —C(O)O—(R35-phenyl), —C(O)alkyl, —C(O)—(R35-phenyl), —SOalkyl, —SO2alkyl, —SO2—CF3, alkylthio, —NR43R44, -alkyl-NR43R44, R35-phenyl, R35-phenoxy, R35-heteroaryl, R35-heteroaryloxy, R36-heterocycloalkyl, —C(O)—(R36-heterocycloalkyl), hydroxyalkyl-NH—, —C(O)N(R30)2, —N(R43)—(R35-cycloalkyl) and —C(═NOR30); or two R17 substituents on adjacent carbon atoms together form —O—CH2—O—, —O—(CH2)2—O—, —(CH2)2—O— or —O—CH2—O—CH2—;
R18 and R19 are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, R17-phenyl, (R17-phenyl)alkyl, naphthyl and cycloalkyl;
R20 is H, alkyl, or cycloalkyl;
R22 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF3, —NH2 and R35-phenyl;
R23 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF3, —NR18R19, —CN, —C(O)Oalkyl, —SO2-alkyl, —NHSO2-alkyl, R35-phenyl, R35-heteroaryl, morpholinyl, and —(CH2)n—C(O)—N(R30)2;
R24 is H, OH or alkoxy; or when the optional double bond is present, R24 and the adjacent carbon atom form the double bond;
R25 is H or R35-phenyl;
R27 is 1 to 3 substituents independently selected from the group consisting of H, halo, OH, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, haloalkyl, —CN, —C(O)OH, —C(O)Oalkyl, —C(O)N(R30)(R18), —C(O)—(R36-hetercycloalkyl), R17-phenyl, (R17-phenyl)-alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, (R23-heteroaryl)oxy, (R23-heteroaryl)amino NR18R19, NR18R19-alkyl, —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)—C(O)-(fluoroalkyl), —(CH2)n—N(R30)—C(O)alkoxyalkyl, —(CH2)n—N(R30)—C(O)(cycloalkyl), —(CH2)n—N(R30)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)O-alkyl, —(CH2)n—N(R30)—C(O)O—(CF3-alkyl), —(CH2)n—N(R30)—C(O)O—(R39-cycloalkyl), —(CH2)n—N(R30)—C(O)O-alkylene-cycloalkyl, —(CH2)n—N(R30)—C(O)—N(R30)(R20), —(CH2)n—N(R30)—SO2-alkyl, —(CH2)—N(R30)—SO2—CF3, —(CH2)n—N(R30)—SO2—N(R30)2 and




or two R27 groups and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
R28 is H, alkyl, R35-benzyl or -alkyl-C(O)O-alkyl;
R29 is alkyl, haloalkyl, —C(O)Oalkyl, —C(O)alkyl, —C(O)CF3, —C(O)—(R12-cycloalkyl), —C(O)—(R17-phenyl), —C(O)—(R23-heteroaryl), —C(O)—(R36-hetercycloalkyl), —SO2-alkyl, —SO2—(R35-phenyl), —C(O)NR18R19, R35-phenyl, (R35-phenyl)alkyl or R23-heteroaryl;
R30 is independently selected from the group consisting of H, alkyl, R35-benzyl and R35-phenyl;
R31 is H, alkyl, R35-benzyl or phenoxyalkyl;
R33 is H, OH or alkoxy;
R34 is H, alkyl, hydroxyalkyl, alkoxyalkyl or —C(O)Oalkyl;
R35 is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, OH, —CF3, alkoxy, —CO2alkyl and —N(R43)(R44);
R36 is 1 or 2 substituents independently selected from the group consisting of H, alkyl, R17-phenyl, —OH, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl and —NR18R19; or two R36 groups and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
R37 and R38 are independently selected from the group consisting of H and alkyl, or R37 and R38 together are —(CH2)3— or —(CH2)4—, and together with the nitrogen to which they are attached, form a ring;
R39 is H, OH, alkyl, alkoxy, or CF3;
R40 is —OR30 or —NHC(O)alkyl;
R41 is H or —SO2alkyl;
R42 is —(CH2)n—(R35-phenyl), —(CH2)n—(R23-heteroaryl), —C(O)Oalkyl or —C(O)alkyl;
R43 and R44 are independently selected from the group consisting of H and alkyl; and
R45 is 1 or 2 substituents independently selected from the group consisting of halo, alkoxyalkyl, —CO2alkyl, R17-phenyl, R23-heteroaryl and cycloalkyl.
This invention also provides a method of treating diseases mediated by PDE 4, including allergic and inflammatory diseases, CNS diseases, and diabetes comprising administering an effective amount of at least one compound of formula I to a patient in need of such treatment.
In particular, this invention also provides a method of treating a PDE4 mediated disease or condition selected from the group consisting of: pain (e.g., acute pain, acute inflammatory pain, chronic inflammatory pain, and neuropathic pain), acute inflammation, chronic inflammation, rheumatoid arthritis, psoriasis, atopic dermatitis, asthma, COPD, adult respiratory disease, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke, ischemia reperfusion injury, renal reperfusion injury, glomerulonephritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment (MCI), depression, anxiety, graft vs. host reaction (i.e., graft vs. host disease), allograft rejections (e.g., acute allograft rejection, and chronic allograft rejection), acute respiratory distress syndrome, delayed type hypersensitivity reaction, atherosclerosis, cerebral ischemia, osteoarthritis, multiple sclerosis, angiogenesis, osteoporosis, gingivitis, respiratory viruses, herpes viruses, hepatitis viruses, HIV, Kaposi's sarcoma associated virus (i.e., Kaposi's sarcoma), meningitis, cystic fibrosis, pre-term labor, cough, pruritis, multi-organ dysfunction, psoriatic arthritis, herpes, encephalitis, traumatic brain injury, CNS tumors, interstitial pneumonitis, hypersensitivity, crystal induced arthritis, acute pancreatitis, chronic pancreatitis, acute alcoholic hepatitis, necrotizing enterocolitis, chronic sinusitis, ocular inflammation, corneal neovascularization, polymyositis, acne, esophagitis, glossitis, airflow obstruction, airway hyperresponsiveness (i.e., airway hyperreactivity), bronchiectasis, bronchiolitis, bronchiolitis obliterans (i.e., bronchiolitis obliterans syndrome), chronic bronchitis, dyspnea, emphysema, hypercapnea, hyperinflation, hypoxemia, hyperoxia-induced inflammations, hypoxia, pulmonary fibrosis, pulmonary hypertension, peritonitis associated with continuous ambulatory peritoneal dialysis (CAPD), granulocytic ehrlichiosis, sarcoidosis, small airway disease, ventilation-perfusion mismatching, wheeze, colds, gout, alcoholic liver disease, lupus, periodontitis, cancer, transplant reperfusion injury, early transplantation rejection (e.g., acute allograft rejection), airway hyperreactivity, allergic contact dermatitis, allergic rhinitis, alopecia areata, autoimmune deafness (including, for example, Meniere's disease), autoimmune hemolytic syndromes, autoimmune hepatitis, autoimmune neuropathy, autoimmune ovarian failure, autoimmune orchitis, autoimmune thrombocytopenia, chronic inflammatory demyelinating polyneuropathy, cirrhosis, dermatomyositis, diabetes, drug-induced autoimmunity, endometriosis, fibrotic diseases, gastritis, Goodpasture's syndrome, Graves' disease, Gullain-Barre disease, Hashimoto's thyroiditis, hepatitis-associated autoimmunity, HIV-related autoimmune syndromes and hematologic disorders, hypophytis, interstitial cystitis, juvenile arthritis, Langerhans' cell histiocytitis, lichen planus, metal-induced autoimmunity, myocarditis (including viral myocarditis), myositis, neuropathies (including, for example, IgA neuropathy, membranous neuropathy and idiopathic neuropathy), nephritic syndrome, optic neuritis, pancreatitis, post-infectious autoimmunity, primary biliary cirrhosis, reactive arthritis, ankylosing spondylitis, Reiter's syndrome, reperfusion injury, scleritis, scleroderma, secondary hematologic manifestation of autoimmune diseases (such as anemias), silicone implant associated autoimmune disease, Sjogren's syndrome, systemic lupus erythematosus, transverse myelitis, tubulointerstitial nephritis, uveitis, and vitiglio in a patient in need of such treatment comprising administering to said patient an effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.
Compounds of formula I are preferably useful in treating pain (e.g., acute pain, acute inflammatory pain, chronic inflammatory pain, and neuropathic pain), acute inflammation, chronic inflammation, rheumatoid arthritis, psoriasis, atopic dermatitis, asthma, COPD, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke, ischemia reperfusion injury, glomerulonephritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment, depression, anxiety, graft vs. host reaction (i.e., graft vs. host disease), allograft rejections (e.g., acute allograft rejection, and chronic allograft rejection), delayed type hypersensitivity reaction, osteoarthritis, multiple sclerosis, angiogenesis, osteoporosis, HIV, cough, psoriatic arthritis, CNS tumors, necrotizing enterocolitis, airflow obstruction, airway hyperresponsiveness (i.e., airway hyperreactivity), bronchiolitis, chronic bronchitis, emphysema, pulmonary fibrosis, pulmonary hypertension, small airway disease, wheeze, lupus, cancer, transplant reperfusion injury, early transplantation rejection (e.g., acute allograft rejection), airway hyperreactivity, allergic contact dermatitis, allergic rhinitis, diabetes, juvenile arthritis, reactive arthritis, ankylosing spondylitis, reperfusion injury, and systemic lupus erythematosus.
More preferably, compounds of formula I are useful for treating COPD, asthma, IBD, dermatitis, MS, arthritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment, depression and anxiety.
Preferred veterinary uses for compounds of formula I include the treatment of dermatitis in dogs and the treatment of recurrent airway disease in horses.
This invention also provides a method of treating a PDE4 mediated disease or condition in a patient in need of such treatment comprising administering to said patient at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one other medicament (e.g., a drug, agent or therapeutic) selected from the group consisting of:

    
    
        a) disease modifying antirheumatic drugs;
        b) nonsteroidal anitinflammatory drugs;
        c) COX-2 selective inhibitors;
        d) COX-1 inhibitors;
        e) immunosuppressives;
        f) steroids;
        g) biological response modifiers;
        h) other anti-inflammatory agents or therapeutics useful for the treatment of chemokine mediated diseases; and
        i) other agents or therapeutics useful for the treatment of depression, anxiety, Alzheimer's Disease or Parkinson's Disease.
    
    


This invention also provides a method of treating a pulmonary disease (e.g., COPD, asthma or cystic fibrosis) in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of: glucocorticoids, 5-lipoxygenase inhibitors, β-2 adrenoceptor agonists, muscarinic M1 antagonists, muscarinic M3 antagonists, muscarinic M2 agonists, NK3 antagonists, LTB4 antagonists, cysteinyl leukotriene antagonists, bronchodilators, PDE4 inhibitors, PDE inhibitors, elastase inhibitors, MMP inhibitors, phospholipase A2 inhibitors, phospholipase D inhibitors, histamine H1 antagonists, histamine H3 antagonists, dopamine agonists, adenosine A2 agonists, NK1 and NK2 antagonists, GABA-b agonists, nociceptin agonists, expectorants, mucolytic agents, decongestants, antioxidants, anti-IL-8 anti-bodies, anti-IL-5 antibodies, anti-IgE antibodies, anti-TNF antibodies, IL-10, adhesion molecule inhibitors, and growth hormones.
This invention also provides a method of treating multiple sclerosis in a patient in need of such treatment comprising administering to said patient, a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of glatiramer acetate, glucocorticoids, methotrexate, azothioprine, mitoxantrone, chemokine inhibitors, and CB2-selective agents.
This invention also provides a method of treating multiple sclerosis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of: methotrexate, cyclosporin, leflunimide, sulfasalazine, β-methasone, β-interferon, glatiramer acetate, and prednisone.
This invention also provides a method of treating rheumatoid arthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of COX-2 inhibitors, COX inhibitors, immunosuppressives (e.g., methotrexate, cyclosporin, leflunimide and sulfasalazine), steroids (e.g., betamethasone, cortisone and dexamethasone), anti-TNF-α compounds, MMP inhibitors, glucocorticoids, chemokine inhibitors, CB2-selective inhibitors, and other classes of compounds indicated for the treatment of rheumatoid arthritis.
This invention also provides a method of treating stroke and ischemia reperfusion injury in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of thrombolitics (e.g., tenecteplase, TPA, alteplase), antiplatelet agents (e.g., gpIIb/IIIa), antagonists (e.g., abciximab and eftiifbatide), anticoagulants (e.g., heparin), and other compounds indicated for the treatment of rheumatoid arthritis.
This invention also provides a method of treating stroke and ischemia reperfusion injury in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of tenecteplase, TPA, alteplase, abciximab, eftiifbatide, and heparin.
This invention also provides a method of treating psoriasis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of immunosuppressives (e.g., methotrexate, cyclosporin, leflunimide and sulfasalazine), steroids (e.g., β-methasone) and anti-TNF-α compounds (e.g., etonercept and infliximab).
This invention also provides a method of treating COPD in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.
This invention also provides a method of treating arthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.
This invention also provides a method of treating osteoarthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.
This invention also provides a method of treating acute pain, acute inflammatory pain, chronic inflammatory pain, or neuropathic pain in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.
This invention also provides a method of treating pain in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and administering a therapeutically effective amount of at least one medicament selected from the group consisting of: NSAIDs, COXIB inhibitors, anti-depressants, and anti-convulsants.
This invention also provides a pharmaceutical composition comprising at least one (e.g., 1-3, usually 1) compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier. Preferred are oral dosage forms and dosage forms suitable for inhalation.
This invention also provides a pharmaceutical composition comprising at least one (e.g., 1-3, usually 1) compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and at least one (e.g., 1-3, usually 1) other agent, medicament, antibody and/or inhibitor disclosed above, and a pharmaceutically acceptable carrier.


DETAILED DESCRIPTION
Preferred compounds of formula I are those wherein the quinolyl portion has the structure




More preferred are compounds wherein R10, R11 and R13 are each H. Also preferred are compounds wherein R1 is H, alkyl, cycloalkyl or CF3; more preferably, R1 is alkyl, especially methyl. Also preferred are compounds wherein R9 is H, alkyl or —CF3, more preferably —CF3.

In compounds of formula I, X is preferably O.
In compounds of formula I,




is preferably oxazolyl, more preferably





In compounds of formula I, R3 is preferably H, alkyl, hydroxalkyl or —C(O)Oalkyl, and R4 is H or alkyl. More preferably, R3 and R4 are each independently H or alkyl.
In compounds of formula I, R5 is preferably H, and R6 is preferably H, alkyl or hydroxyalkyl. When R6 is alkyl, it is preferably methyl, ethyl or isopropyl, more preferably methyl; when it is hydroxyalkyl, it is preferably hydroxymethyl or hydroxyethyl (i.e., —(CH2)2OH or —CH(OH)CH3). In a more preferred embodiment, R5 is H and R6 is H, methyl or hydroxymethyl. Preferably, t is 1. When t is 2, preferably both R5 substituents and one R6 substituent are H and one R6 substituent is H or methyl.
Preferably, R5 and R6 have the following stereochemistry (i.e., R6 is “S”):




When R7 and R8 do not form a ring, the following definitions are preferred.
R7 is preferably H, alkyl, cycloalkyl, hydroxyalkyl or alkoxyalkyl. More preferably, R7 is H, alkyl, hydroxyalkyl, especially wherein alkyl is methyl or ethyl, and hydroxyalkyl is hydroxyethyl. Especially preferred are compounds wherein R7 is H or alkyl, especially H, methyl or ethyl, with H being most preferred.
R8 is preferably R12-cycloalkyl, (R12-cycloalkyl)alkyl, R45-hydroxyalkyl, R17-phenyl, (R17-phenyl)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, -alkyl-N(R30)—C(O)—NR18R19, -alkyl-N(R30)—C(O)alkyl, -alkyl-N(R30)—C(O)—(R17-phenyl), -alkyl-N(R30)—C(O)—(R23-heteroaryl), -alkyl-N(R30)—(R23-heteroaryl),




More preferably, R8 is R12-cycloalkyl, R45-hydroxyalkyl, (R17-phenyl)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, -alkyl-N(R30)—(R23-heteroaryl), -alkyl-N(R30)—C(O)alkyl,





(especially where p is 0) or





(especially where p is 1). Especially preferred are compounds wherein R8 is R12-cycloalkyl, R45-hydroxyalkyl, (R17-phenyl)alkyl, (R23-heteroaryl)alkyl, -alkyl-N(R30)—C(O)-alkyl, -alkyl-N(R30)—(R23-heteroaryl) or





When R8 comprises R12-cycloalkyl, R12 is preferably OH, —(CH2)n—N(R30)—C(O)-cycloalkyl or —(CH2)n—N(R30)—(R23-heteroaryl), especially OH. When R8 is




n is preferably 0 and R29 is preferably heteroaryl, —C(O)alkyl or —C(O)cycloalkyl. When R8 is R45-hydroxyalkyl, R45 is preferably R17-phenyl.

R30 is preferably H.
Preferred heteroaryl groups include pyrimidyl, benzothienyl, benzofuranyl, indolyl, pyridyl and pyrazinyl.
Especially preferred are compounds of formula I wherein R7 is H and R8 is (R17-phenyl)alkyl, (R23-heteroaryl)alkyl, R45-hydroxyalkyl, -alkyl-N(R30)—(R23-heteroaryl) or




R17 is preferably 1-3 substituents selected from the group consisting of halogen, OH, alkoxy and alkyl; R23 is preferably 1 or 2 substituents independently selected from the group consisting of H, alkyl, alkoxy and halogen; R45 is preferably R17-phenyl, wherein R17 is 1-3 substituents selected from the group consisting of halogen, OH, alkoxy and alkyl; heteroaryl is pyrimidyl, benzothienyl, benzofuranyl, indolyl, pyridyl or pyrazinyl, and R30 is H.

Also preferred are compounds of formula I wherein R7 and R8 and the nitrogen to which they are attached form




In the preferred compounds where R7 and R8 form




the optional double bond preferably is not present (i.e., a single bond is present). R14 is preferably selected from H, OH, alkoxy, —(CH2)n—N(R30)(R23-heteroaryl), R23-heteroaryl or (R23-heteroaryl)-alkyl. In a further preferred embodiment, one R14 is OH and the other R14 is R23-heteroaryl; in another embodiment, one R14 is H and the other is (R23-heteroaryl)-alkyl or —(CH2)n—N(R30)(R23-heteroaryl) (especially wherein n is 1).

In the preferred compounds where R7 and R8 form




q is preferably 1. R27 is preferably 1-3 substituents independently selected from the group consisting of H, OH, alkyl, alkoxy, alkoxyalkyl, R17-phenyl, —C(O)OH, —C(O)Oalkyl, R23-heteroaryl, (R23-heteroaryl)amino and —(CH2)n—N(R30)—C(O)(cycloalkyl), wherein n is 0.

In the preferred compounds where R7 and R8 form




R15 is preferably alkyl, R17-phenyl, R23-heteroaryl, —C(O)alkyl, —C(O)(fluoroalkyl), —C(O)—(R23-heteroaryl), —C(O)-alkoxyalkyl, —C(O)—(R38-cycloalkyl), —SO2-alkyl, —SO2—NR18R19 or





R16 is preferably H, alkyl, or two R16 groups and the carbon to which they are attached form —C(O)—.

In the preferred compounds where R7 and R8 form




preferably p is 0, R34 is hydrogen, and R35 is 1 or 2 substituents independently selected from H, OH, halo and alkyl.

In the preferred compounds where R7 and R8 form




preferably p is 0, ring B is a pyrazolyl or thiazolyl ring, and R35 is 1 or 2 substituents independently selected from H and alkyl.

As used above, and throughout the specification, the following terms, unless otherwise indicated, shall be understood to have the following meanings:
“Patient” includes both humans and animals.
“Mammal” means humans and other mammalian animals.
“Alkyl” means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl and n-pentyl.
“Alkenyl” means an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched and comprising about 2 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkenyl chain. Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl and n-pentenyl.
“Alkylene” means a difunctional group obtained by removal of a hydrogen atom from an alkyl group that is defined above. Non-limiting examples of alkylene include methylene (i.e., —CH2—), ethylene (i.e., —CH2—CH2—) and branched chains such as —CH(CH3)—CH2—.
“Heteroaryl” means a single ring, bicyclic or benzofused heteroaromatic group of 5 to 10 atoms comprised of 2 to 9 carbon atoms and 1 to 4 heteroatoms independently selected from the group consisting of N, O and S, provided that the rings do not include adjacent oxygen and/or sulfur atoms. N-oxides of the ring nitrogens are also included. Examples of single-ring heteroaryl groups are pyridyl, oxazolyl, isoxazolyl, oxadiazolyl, furanyl, pyrrolyl, thienyl, imidazolyl, pyrazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrazinyl, pyrimidyl, pyridazinyl and triazolyl. Examples of bicyclic heteroaryl groups are naphthyridyl (e.g., 1,5 or 1,7), imidazopyridyl, pyridopyrimidinyl and 7-azaindolyl. Examples of benzofused heteroaryl groups are indolyl, quinolyl, isoquinolyl, phthalazinyl, benzothienyl (i.e., thianaphthenyl), benzimidazolyl, benzofuranyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl and benzofurazanyl. All positional isomers are contemplated, e.g., 2-pyridyl, 3-pyridyl and 4-pyridyl. The term R23-heteroaryl refers to such groups wherein substitutable ring carbon atoms have a substituent as defined above. When the heteroaryl group is a benzofused ring, the substituents can be attached to either or both the phenyl ring portion and the heteroaromatic ring portion, and the heteroaryl group can be attached to the rest of the molecule either through the phenyl ring portion or the heteroaromatic ring portion.
“Cycloalkyl” means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 3 to about 6 carbon atoms. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1-decalin, norbornyl, adamantyl and the like. Monocyclic rings are preferred.
“Halo” means fluoro, chloro, bromo, or iodo groups. Preferred are fluoro, chloro or bromo, and more preferred are fluoro and chloro.
“Haloalkyl” means an alkyl as defined above wherein one or more hydrogen atoms on the alkyl is replaced by a halo group defined above; in particular, fluoroalkyl refers to an alkyl chain substituted by one or more fluoro atoms.
“Aminoalkyl” means an alkyl as defined above wherein a hydrogen atom on the alkyl is replaced by an amino (i.e., —NH2) group.
“Heterocycloalkyl” means a non-aromatic saturated monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more, preferably 1, 2, 3 or 4, of the atoms in the ring system is independently selected from an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and/or sulfur atoms present in the ring system. Preferred heterocycloalkyls contain 5 to 6 ring atoms. The prefix aza, oxa or thia before the heterocycloalkyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom. The nitrogen or sulfur atom of the heterocyclyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S-dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. The heterocycloalkyl group can be attached to the parent moiety through a ring carbon or a ring nitrogen.
“(Heterocycloalkyl)alkyl” means a heterocycloalkyl-alkyl group in which the heterocycloalkyl and alkyl groups are as defined above. The bond to the parent is through the alkyl.
“(Heteroaryl)alkyl” means a heteroaryl-alkyl- group in which the heteroaryl and alkyl are as previously described. Non-limiting examples of suitable heteroarylalkyl groups include pyridylmethyl, 2-(furan-3-yl)ethyl and quinolin-3-ylmethyl. The bond to the parent moiety is through the alkyl.
“(Phenyl)alkyl and “(naphthyl)alkyl similarly mean phenyl-alkyl and naphthyl-alkyl groups wherein the bond to the parent moiety is through the alkyl.
“Hydroxyalkyl” means a HO-alkyl- group in which alkyl is as previously defined. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl. Similarly, “dihydroxyalkyl” refers to a straight or branched alkyl chain substituted by two hydroxy groups.
“Alkoxy” means an alkyl-O— group in which the alkyl group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond to the parent moiety is through the ether oxygen.
“Alkylthio” means an alkyl-S— group in which the alkyl group is as previously described. Non-limiting examples of suitable alkylthio groups include methylthio, ethylthio and isopropylthio. The bond to the parent moiety is through the sulfur.
“Heteroarylamino” means an heteroaryl-NH— group in which the heteroaryl group is as previously described. Non-limiting examples of suitable heteroarylamino groups include pyrimidinyl-amino and pyrazinyl-amino. The bond to the parent moiety is through the amino nitrogen.
“Heteroaryloxy” means an heteroaryl-O— group in which the heteroaryl group is as previously described. Non-limiting examples of suitable heteroaryloxy groups include pyrimidinyl-O— and pyrazinyl-O—. The bond to the parent moiety is through the ether oxygen.
The term “hydroxyalkyl substituted by CO2alkyl” means an alkyl chain substituted by a hydroxy group and a CO2alkyl group. Similarly, terms such as “hydroxyalkyl substituted by R17-phenyl” means an alkyl chain substituted by a hydroxy group and a R17-phenyl group; “hydroxyalkyl substituted by R17-phenyl and alkoxy” means an alkyl group substituted by a hydroxy group, a R17-phenyl, and an alkoxy group. In each of these substituents and other similar substituents listed in the definitions, the alkyl chains can be branched.
Examples of moieties formed when two adjacent R17 groups form a ring with the carbons on the phenyl ring to which they are attached are:




When R7 and R8 together form




the dotted line indicates an optional double bond as defined above. When the double bond is absent, i.e., when a single bond is present, the one or two R14 substituents can be attached to the same or different ring carbons. When the double bond is present, only one R14 substituent can be attached to a carbon that is part of the double bond.

When R7 and R8 together form




the dotted line indicates an optional double bond as defined above. When the double bond is absent, i.e., when a single bond is present, R24 can be H, OH or alkoxy and R25 can be H or R35-phenyl, but when the double bond is present, R24 forms the double bond with the adjacent carbon and R25 is H or R35-phenyl. That is, the moiety has the structural formula





When R7 and R8 together form




it means that an optionally substituted fused bicyclic ring is formed, wherein the





portion comprises an R35-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl ring.

Examples are:





The term “optionally substituted” means optional substitution with the specified groups, radicals or moieties, in available position or positions.
With reference to the number of moieties (e.g., substituents, groups or rings) in a compound, unless otherwise defined, the phrases “one or more” and “at least one” mean that there can be as many moieties as chemically permitted, and the determination of the maximum number of such moieties is well within the knowledge of those skilled in the art.
As used herein, the term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
The wavy line  as a bond generally indicates a mixture of, or either of, the possible isomers, e.g., containing (R)- and (S)-stereochemistry. For example,




Lines drawn into the ring systems, such as, for example:




indicate that the indicated line (bond) may be attached to any of the substitutable ring carbon atoms.

As well known in the art, a bond drawn from a particular atom wherein no moiety is depicted at the terminal end of the bond indicates a methyl group bound through that bond to the atom, unless stated otherwise. For example:




It should also be noted that any carbon or heteroatom with unsatisfied valences in the text, schemes, examples, structural formulae, and any Tables herein is assumed to have the hydrogen atom or atoms to satisfy the valences.
Prodrugs and solvates of the compounds of the invention are also contemplated herein. The term “prodrug”, as employed herein, denotes a compound that is a drug precursor which, upon administration to a subject, undergoes chemical conversion by metabolic or chemical processes to yield a compound of formula I or a salt and/or solvate thereof. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) Volume 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, both of which are incorporated herein by reference thereto.
“Solvate” means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. “Hydrate” is a solvate wherein the solvent molecule is H2O.
“Effective amount” or “therapeutically effective amount” is meant to describe an amount of compound or a composition of the present invention effective in inhibiting PDE4 and thus producing the desired therapeutic effect in a suitable patient.
The compounds of formula I form salts which are also within the scope of this invention. Reference to a compound of formula I herein is understood to include reference to salts thereof, unless otherwise indicated. The term “salt(s)”, as employed herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a compound of formula I contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions (“inner salts”) may be formed and are included within the term “salt(s)” as used herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful. Salts of the compounds of the formula I may be formed, for example, by reacting a compound of formula I with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization. Acids (and bases) which are generally considered suitable for the formation of pharmaceutically useful salts from basic (or acidic) pharmaceutical compounds are discussed, for example, by S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York; in The Orange Book (Food & Drug Administration, Washington, D.C. on their website); and P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (2002) Int'l. Union of Pure and Applied Chemistry, pp. 330-331. These disclosures are incorporated herein by reference thereto.
Exemplary acid addition salts include acetates, adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides, hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates, methanesulfonates, methyl sulfates, 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pamoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates, sulfonates (such as those mentioned herein), tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) undecanoates, and the like.
Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, aluminum salts, zinc salts, salts with organic bases (for example, organic amines) such as benzathines, diethylamine, dicyclohexylamines, hydrabamines (formed with N,N-bis(dehydroabietyl)ethylenediamine), N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, piperazine, phenylcyclohexylamine, choline, tromethamine, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quarternized with agents such as lower alkyl halides (e.g. methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g. dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g. benzyl and phenethyl bromides), and others.
All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.
Compounds of formula I, and salts, solvates and prodrugs thereof, may exist in their tautomeric form (for example, as an amide or imino ether). All such tautomeric forms are contemplated herein as part of the present invention.
All stereoisomers (for example, geometric isomers, optical isomers and the like) of the present compounds (including those of the salts, solvates and prodrugs of the compounds as well as the salts and solvates of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention. Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms “salt”, “solvate” “prodrug” and the like, is intended to equally apply to the salt, solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers, racemates or prodrugs of the inventive compounds.
Polymorphic forms of the compounds of Formula I, and of the salts, solvates and prodrugs of the compounds of Formula I, are intended to be included in the present invention.
This invention also includes the compounds of this invention in isolated and pure form.
Compounds of formula I can be prepared by known methods from starting materials either known in the art or prepared by methods known in the art. Non-limiting examples of suitable methods are illustrated in the following schemes.
In the schemes, the quinolyl portion is shown as the preferred structure, but those skilled in the art will recognize that other substitutions on the quinolyl portion can be made by these procedures. Also, one skilled in the art will recognize that the schemes show the significant steps of the procedures, and that the synthesis of compounds of formula I may require the need for the protection of certain functional groups during the preparation of the compounds; the synthesis of compounds also may require the reduction of a reducible functional group or the oxidation of an oxidizable functional group.




Step a

Formation of the oxazole ring can be accomplished by a number of methods including, but not limited to the following.










Using the appropriate starting materials, both the amine and ester functional groups can be incorporated when the oxazole ring is synthesized.
Step b
Introduction of the ester moiety COOR2 can be accomplished stepwise by reaction with phosphorous oxychloride and subsequent oxidation of the intermediate aldehyde to the carboxylic acid and further esterification. Alternatively, reaction at this position can proceed with a Lewis acid such as zinc triflate and an acid chloride.
Step c
Introduction of the R moiety can be accomplished by deprotonation with a strong base such as n-butyl lithium, sec-butyl lithium, lithium diisopropylamine or lithium hexamethyldisilazide, followed by addition of an aldehyde or alkyl halide. This reaction can use a variety of solvents including diethyl ether, THF, dioxane, hexane, toluene, HMPA, DMPU and TMEDA.
Step d
Activation of the R moiety in (3) can be accomplished by several different methods. If R is an alkyl moiety, halogenation, for example with bromine or N-bromo-succinimide and an initiator such as benzoyl peroxide, AIBN or light in carbon tetrachloride as the solvent provides (5) as a halide. If R incorporates an ester or alcohol functional group, through appropriate oxidation or reduction reactions, the aldehyde or ketone functional group can be obtained for further reaction in Step e through a reductive amination reaction. If R incorporates an ester, ketone, or aldehyde functional group, appropriate reduction reaction with a hydride such as NaBH4, LiBH4, LiAlH4, or diisobutylaluminum hydride will provide the alcohol moiety. This alcohol can be activated by conversion, for example, to the corresponding mesylate, tosylate, chloride, bromide or iodide.
Step e
Introduction of the amine moiety in (6) can be accomplished by an alkylation reaction on (5) if X is a leaving group such as chloride, bromide, mesylate or tosylate. This reaction can use a variety of bases including TEA, DIPEA, N-methyl morpholine, pyridine, dimethylaminopyridine, imidazole, K2CO3, Cs2CO3, potassium t-butoxide, and NaOH, and can be done in a variety of solvents including DMF, dimethylacetamide, THF, dioxane, CH3CN, toluene, CH2Cl2 and dichloroethane. Alternatively, if the —C(X)(R5)(R6) moiety incorporates a ketone or aldehyde functional group, the amine moiety can be introduced through a reductive amination reaction. Suitable reducing reagents for this reaction include NaBH3CN, sodium triacetoxyborohydride in a mixture of solvents including THF, dioxane, CH3CN, toluene, CH2Cl2, dichloroethane, methanol, ethanol, trifluoroethanol. The reductive amination reaction may require the addition of a drying agent such as sieves or MgSO4, or azeotropic removal of water or the addition of a Lewis acid such as titanium isopropoxide. In addition, the ketone or aldehyde moiety can be converted into an oxime with hydroxylamine and a variety of bases such as pyridine, TEA, sodium acetate, and Na2CO3. The oxime can be reduced to an amine.
Step f
Hydrolysis of ester (6) to acid (7) can be accomplished with a suitable base such as NaOH, LiOH, sodium methoxide, sodium ethoxide, K2CO3, Cs2CO3, BCl3, potassium t-butoxide, TEA, DBU and DIPEA in a mixture of solvents including water, methanol, ethanol, isopropanol, CH2Cl2, THF, diethyl ether and dioxane.
Step g
Amide bond formation to obtain (8) can be accomplished by formation of the acid chloride, a mixed anhydride, or activated ester and addition of the appropriate amine. A variety of suitable amide bond coupling reagents such as HATU, CDI, EDC, DCC, PyBOP, polymer supported CDI, polymer supported EDC and the like, with or without HOBt, can be used. These coupling reagents can be used with a suitable base such as TEA, DIPEA, N-methyl morpholine, pyridine, dimethylaminopyridine, DBU, imidazole and the like in a mixture of solvents including DMF, dimethylacetamide, THF, dioxane, CH3CN, N-methylpyrrolidine, CH2Cl2, and dichloroethane.
Abbreviations used in the above general schemes and in the following examples, as well as throughout the specification, are as follows: Me (methyl); Bu (butyl); Et (ethyl); Ac (acetyl); Boc or BOC (t-butoxycarbonyl); DMF (dimethyl-formamide); THF (tetrahydrofuran); DIPEA (diisopropylethylamine); RT (room temperature); HOBt (hydroxybenzotriazole); TFA (trifluoroacetic acid); TEA (triethyl amine); KHMDS (potassium bis(trimethylsilyl)amide); TLC (thin layer chromatography); EDC (1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride); HMPA (hexamethylphosphoramide); DMPU (1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone); TMEDA (N,N,N′,N′-tetramethyletheylenediamine); HATU (O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyl uranium hexafluoro-phosphate); NBS (N-bromosuccinimide); DCC (1,3-dicyclohexylcarbodiimide); DEC (1,2-diethylaminoethyl chloride hydrochloride); TMSCN (trimethylsilylcyanide); CDI (carbonyldiimidazole); PyBOP (benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate).
EXAMPLE 1




Step 1: SOCl2 (26.7 ml, 367 mmol) was added to a mixture of compound 1 (40 g, 147 mmol) in dry toluene (300 ml) and DMF (0.4 ml). The mixture was heated at 70° C. for 2 h, then the excess of SOCl2 and solvents were evaporated to dryness to obtain compound 2 as an off white-solid (41 g).
Step 2: A solution of compound 2 (41 g, 141 mmol) in CH2Cl2 (200 ml) was added slowly to a solution of L-threonine methyl ester HCl salt (29 g, 170 mmol) in CH2Cl2 (200 ml) and DIPEA (38 g, 296 mmol) at 0° C. The solution was stirred at 0° C., then warmed to RT over 3 h. After 3 h at RT, the mixture was washed with aqueous NH4Cl solution, then the solid was precipitated in the organic layer and filtered off to give compound 3 (54 g) as a white solid. MS: C17H17F3N2O5 [M+1]+387.1.
Step 3: SOCl2 (76.8 ml, 645 mmol) was added through a syringe to a suspension of compound 3 (50 g, 129 mmol) in dry CH2Cl2 (500 ml) cooled to −45 0° C. The mixture was stirred at −45° C. for 1 h, then warmed up to RT slowly. After the reaction was complete, solvent and excess SOCl2 were evaporated. The residue was dissolved in CH2Cl2 (800 ml) and washed with saturated NaHCO3 solution (3×600 ml), dried (Na2SO4), filtered and concentrated to give compound 4 as a beige solid (43 g, 120 mmol, 93%). MS: C17H15F3N2O4 [M+1]+369.1.
Step 4: DBU (13.9 ml, 93 mmol) was added via a syringe to a solution of compound 4 (31 g, 84 mmol) in dry CH2Cl2 (300 ml) at 0° C., followed by the addition of BrCCl3 (9.1 ml, 93 mmol). The mixture was stirred at 0° C. for 2 h, then at RT overnight. The reaction was quenched with 0.15 N HCl (400 ml) and extracted with CH2Cl2 (2×100 ml). The organic layer was dried with Na2SO4, filtered and concentrated to give crude title compound 5 (35 g). The crude material was triturated with MeOH (200 ml) and 23.5 g of compound 5 was collected as a pale yellow solid. MS: C17H13F3N2O4 [M+1]+367.1.
EXAMPLE 2




Step 1: NBS (23.5 g, 132 mmol) and benzoyl peroxide (1.4 g, 5.75 mmol) were added to a mixture of compound 5 (42 g, 115 mmol) in dry CCl4 (550 ml). The mixture was refluxed for 3 h, then concentrated by evaporating off most of the solvent. Saturated NH4Cl solution was added and the product was extracted from the aqueous layer with CH2Cl2 (2×300 ml). The organic fractions were combined, dried (Na2SO4), filtered and evaporated. The crude material was triturated with MeOH to give compound 6 as a white solid (49.5 g, 110 mmol, 96%). MS: C17H12F3BrN2O4 [M+1]+Br79,81 445.1, 447.1.
Step 2: Potassium phthalimide (20.6 g, 111 mmol) was added to a solution of compound 6 (49.5 g, 111 mmol) in dry DMF (650 ml) at RT. After stirring at RT for 2 h, the reaction mixture was poured into an ice water bath (1.5 L). The resultant yellow precipitate was collected, washed with water, and dried at 45° C. under vacuum to give compound 7 as a yellow solid (56 g, 110 mmol). MS: C25H16F3N3O6 [M+1]+512.0.
Step 3: BCl3 (1 M in CH2Cl2, 78 ml, 78 mmol) solution was added to a solution of compound 7 (10 g, 19.57 mmol) in dry CH2Cl2 (400 ml) at −15° C. After the addition of BCl3, the mixture turned yellow and precipitate started to form. The reaction was warmed to 0° C. After the reaction was complete (checked by TLC), the mixture was poured into ice-water (600 ml). The yellow precipitate was filtered, washed with water, and dried (Na2SO4) to give the title compound 8 as a yellow solid (8.5 g, 17.1 mmol, 87%). MS: C24H14F3N3O6 [M+1]+498.1.
EXAMPLE 3




Step 1: To a suspension of compound 8 (0.35 g, 0.7 mmol) in dry DMF (16 ml), compound 9 (3-aminomethyl benzothiophene) (0.11 g, 0.7 mmol), DI PEA (0.18 g, 1.4 mmol) and HATU (0.53 g, 1.4 mmol) were added at RT. After 30 ml, the reaction mixture was poured into cold water (30 ml). The precipitate was filtered, washed with water and dried under vacuum to give crude compound 10 (0.45 g, 0.7 mmol) as a yellow solid.
Step 2: The crude material of compound 10 (0.45 g, 0.7 mmol) was treated with absolute EtOH (15 ml) and 98% hydrazine (0.22 g, 7 mmol) at RT overnight. The reaction mixture was evaporated and purified on a Biotage (40 M) system, eluting with 3% NH4OH:CH3OH (1:9)/97% CH2Cl2. Compound 11 was obtained as a pure yellow solid (0.22 g, 0.43 mmol, 61% yield) which was converted to its HCl salt by treatment with 1.2 equivalent of 4 N HCl/dioxane in CH2Cl2. Compound 12 was obtained by evaporating off solvents and excess acid. MS (M+1): m/e. 513.
EXAMPLE 4
A series of aromatic or heteroaromatic amide analogs (compound 13) was made by methods analogous to those described for compound 12 in Example 3 or via an alternative coupling method by treatment of compound 8 (0.2 mmol) either with aromatic or heteroaromatic amine reagent (0.2 mmol), DEC (0.24 mmol), HOBT (0.24 mmol), and TEA (0.24 mmol) in DMF (2.5 ml) at RT overnight. Water (3 ml) was added to the reaction and precipitate was collected, rinsed with water, and vacuum dried at 40° C. The phthalamido protecting group of the coupled product was removed with 98% hydrazine in EtOH (as in step 2 of Example 3) and purified by silica gel chromatography [5% NH4OH—CH3OH (1:9) in 95% CH2Cl2] or by preparative Gilson Prep column (XTerra RP C18, 5 μm) chromatography, gradient eluted with 0.5% TFA in (9:1) (H2O—CH3CN) to 0.5% TFA in CH3CN:H2O (8:2). Compound 13 was obtained as free form or as a TFA salt, depending on the method of purification. The free form of compound 13 was treated with 1.2 equivalent of HCl to give compound 13 as a HCl salt. The data for compound 13 analogs are listed as follows:












13








 









Cpd.

MS


No.
Structure
(M + 1)





13-1 




487  


 


13-2 




471  


 


13-3 




471  


 


13-4 




475  


 


13-5 




489  


 


13-6 




487  


 


13-7 




491  


 


13-8 




525  


 


13-9 




515  


 


13-10 




515  


 


13-11 




515  


 


13-12 




501  


 


13-13 




525  


 


13-14 




475  


 


13-15 




509  


 


13-16 




525  


 


13-17 




491  


 


13-18 




475  


 


13-19 




491  


 


13-20 




525  


 


13-21 




509  


 


13-22 




493  


 


13-23 




493  


 


13-24 




493  


 


13-25 




535  


 


13-26 




487  


 


13-27 




517  


 


13-28 




509  


 


13-29 




541  


 


13-30 




523  


 


13-31 




487  


 


13-32 




505  


 


13-33 




483  


 


13-34 




539  


 


13-35 




513  


 


13-36 




469  


 


13-37 




457  


 


13-38 




487  


 


13-39 




537  


 


13-40 




517  


 


13-41 




497  


 


13-42 




525  


 


13-43 




499  


 


13-44 




547  


 


13-45 




485  


 


13-46 




483  


 


13-47 




485  


 


13-48 




501  


 


13-49 




501  


 


13-50 




485  


 


13-51 




523  


 


13-52 




526  


 


13-53 




541  


 


13-54 




525  


 


13-55 




507  


 


13-56 




535  


 


13-57 




515  


 


13-58 




499  


 


13-59 




533  


 


13-60 




503  


 


13-61 




483  


 


13-62 




473  


 


13-63 




521  


 


13-64 




521  


 


13-65 




521  


 


13-66 




507  


 


13-67 




499  


 


13-68 




499  


 


13-69 




485  


 


13-70 




485  


 


13-71 




485  


 


13-72 




485  


 


13-73 




538  


 


13-74 




500  


 


13-75 




487  


 


13-76 




565  


 


13-77 




505  


 


13-78 




505  


 


13-79 




505  


 


13-80 




523  


 


13-81 




527  


 


13-82 




513  


 


13-83 




527  


 


13-84 




527  


 


13-85 




555  


 


13-86 




555  


 


13-87 




509  


 


13-88 




515  


 


13-89 




508  


 


13-90 




482  


 


13-91 




499  


 


13-92 




473  


 


13-93 




515  


 


13-94 




503  


 


13-95 




458  


 


13-96 




488  


 


13-97 




487  


 


13-98 




499  


 


13-99 




519  


 


13-100




527  


 


13-101




547  


 


13-102




531  


 


13-103




571  


 


13-104




515  


 


13-105




545  


 


13-106






 


13-107




496  


 


13-108




511  


 


13-109




523  


 


13-110




527  


 


13-111




550  


 


13-112




536  


 


13-113




561  


 


13-114




567  


 


13-115




581  


 


13-116




595  


 


13-117




525  


 


13-118




532  


 


13-119




539  


 


13-120




569  


 


13-121




533  


 


13-122




546  


 


13-123




555  


 


13-124




540  


 


13-125




541  


 


13-126




536  


 


13-127




633  


 


13-128




617  


 


13-129




664  


 


13-130




539  


 


13-131




533  


 


13-132




568  


 


13-133




554  


 


13-134




437  


 


13-135




453  


 


13-136




497  


 


13-137




591  


 


13-138




635  


 


13-139




629  


 


13-140




526  


 


13-141




511  


 


13-142




582  


 


13-143




569  


 


13-144




574  


 


13-145




629  


 


13-146




447  


 


13-147




538  


 


13-148




589  


 


13-149




573  


 


13-150




579  


 


13-151




579  


 


13-152




576  


 


13-153




512  


 


13-154




580  


 


13-155




553  


 


13-156




535  


 


13-157




619  


 


13-158




605  


 


13-159




513  


 


13-160




501  


 


13-161




501  


 


13-162




636  


 


13-163




604  


 


13-164




473.5


 


13-165




501  


 


13-166




501  


 


13-167




518  


 


13-168




487  


 


13-169




487  


 


13-170




601  


 


13-171




645  


 


13-172




580  


 


13-173




552  


 


13-174




541  


 


13-175




568  


 


13-176




584  


 


13-177




636  


 


13-178




652  


 


13-179




478  


 


13-180




478  


 


13-181




542  


 


13-182




542  


 


13-183




568  


 


13-184




585  


 


13-185




515  


 


13-186




637  


 


13-187




542  


 


13-188




530  


 


13-189




540  


 


13-190




510  


 


13-191




609  


 


13-192




629  


 


13-193




541  


 


13-194




482  


 


13-195




514  


 


13-196




512  


 


13-197




517  


 


13-198




518  


 


13-199




529  


 


13-200




546  


 


13-201




568  


 


13-202




584  


 


13-203




543  


 


13-204




543  


 


13-205




561  


 


13-206




582  


 


13-207




517  


 


13-208




541  


 


13-209




536  


 


13-210




624  


 


13-211




541  


 


13-212




554  


 


13-213




550  


 


13-214




550  


 


13-215




526  


 


13-216




529  


 


13-217




576  


 


13-218




573  


 


13-219




521  


 


13-220




566  


 


13-221




471  


 


13-222




485  


 


13-223




522  


 


13-224




536  


 


13-225




543  


 


13-226




522  


 


13-227




505  


 


13-228




510  


 


13-229




545  


 


13-230




467  


 


13-231




519  


 


13-232




536  


 


13-233




453  


 


13-234




514  


 


13-235




534  


 


13-236




590  


 


13-237




513  


 


13-238




520  


 


13-239




467  


 


13-240




581  


 


13-241




581  


 


13-242




544  


 


13-243




529  


 


13-244




477  


 


13-245




529  


 


13-246




520  


 


13-247




534  


 


13-248




569  


 


13-249




542  


 


13-250




507  


 


13-251




625  


 


13-252




577  


 


13-253




521  


 


13-254




522  


 


13-255




604  


 


13-256




593  


 


13-257




556  


 


13-258




575  


 


13-259




602  


 


13-260




465  


 


13-261




590  


 


13-262




590  


 


13-263




594  


 


13-264




593  


 


13-265




542  


 


13-266




560  


 


13-267




587  


 


13-268




527  


 


13-269




526  


 


13-270




541  


 


13-271




541  


 


13-272




599  


 


13-273




602  


 


13-274




520  


 


13-275




527  


 


13-276




580  


 


13-277




529  


 


13-278




528  


 


13-279




529  


 


13-280




542  


 


13-281




500  


 


13-282




505  


 


13-283




501  


 


13-284




536  


 


13-285




557  


 


13-286




558  


 


13-287




559  


 


13-288




524  


 


13-289




544  


 


13-290




541  


 


13-291




616  


 


13-292




464  


 


13-293




518  


 


13-294




472  


 


13-295




502  


 


13-296




502  


 


13-297




507  


 


13-298




586  


 


13-299




516  


 


13-300




502  


 


13-301




516  


 


13-302




497  


 


13-303




517  


 


13-304




527  


 


13-305




527  


 


13-306




498  


 


13-307




522  


 


13-308




535  


 


13-309




556  


 


13-310













EXAMPLE 5







Step 1: Glycine ethyl ester hydrochloride (14 g, 100 mmol) was mixed with TEA (29 ml, 200 mmol) in dry CH2Cl2 and cooled in an ice-water bath. Compound 2 (80 mmol) (see Example 1) in dry CH2Cl2 (150 ml) was transferred by cannulation into the above cooled solution. The resulting mixture was allowed to warm to RT slowly. After 2 h, reaction was complete and water (300 ml) was added to dissolve the TEA salt. The organic layer was separated, washed with 5% HCl solution, then water, dried (Na2SO4), filtered and concentrated to give a crude solid, Compound 14, which was used in the next step without further purification.
Step 2: Compound 14 (7.2 g, 20 mmol) was mixed with Lawesson's reagent (5.5 g, 13.6 mmol) in anhydrous THF (100 ml) and heated to 78° C. for 40 min. After cooling to RT, THF was removed and product was purified by silica chromatography, eluting with 100% CH2Cl2 to 5% EtOAc in CH2Cl2, to give compound 15 as a yellow product.
Step 3: Compound 15 (3.9 g, 10 mmol) was dissolved in dry CH2Cl2 (40 ml) and cooled to −78° C. Trimethyloxonium tetrafluoroborate (1.6 g, 11 mmol) was added in one portion. The resulting mixture was then stirred in an ice-water bath for 2 h. NaHCO3 solution was added to quench the reaction. The organic layer was separated, washed with H2O, dried (Na2SO4), and evaporated to give compound 16 as a crude solid which was used in the next reaction without purification.
Step 4a:




A neat liquid of cyanuric fluoride (3.4 ml, 40 mmol) was added dropwise to a cooled solution of N-[(1,1-dimethylethoxy)carbonyl]-L-alanine (compound 17) (3.90 g, 20 mmol) in pyridine (1.78 ml, 22 mmol) and dry CH2Cl2 (50 ml) at −40° C. The reaction was kept at −30° C. to −10° C. for 2 h. After 2 h, crushed ice and CH2Cl2 (100 ml) were added. After stirring for 5 min, the mixture was filtered twice, first with a coarse, then with a medium glass filter funnel. The clear solution was separated and the organic phase was washed with H2O, dried (Na2SO4), filtered and concentrated at RT to give compound 18 as a white solid (3.59 g, 18.7 mmol) with a 94% yield.
Step 5: Compound 18 (2.87 g, 15 mmol) was added to a solution of compound 16 (5.0 g, 12.5 mmol) in dry THF (60 ml). The reaction mixture was cooled to −78° C. and KHMDS (0.5 M in toluene) (52.5 ml, 26.25 mmol) was added dropwise over 40 min. During the addition of the first equivalent of base, the reaction mixture turned a deep blue color, which disappeared immediately. A deep brown color was formed when the second equivalent of base was added. The reaction solution was kept at −78° C. for 1 h then gradually warmed to RT. After completion of the reaction (checked by TLC), ice-cold 0.5 M HCl solution (70 ml) was added. The organic layer was separated and the aqueous layer was extracted with EtOAc (70 ml). The combined organic layer were washed with NaHCO3 solution and brine, dried (Na2SO4), filtered and concentrated to give a crude product which was purified by silica gel chromatograph to yield compound 19 as a solid (3.5 g, 6.88 mmol, yield 55%). Alternative method for the preparation of compound 19:
Step 4b:




A mixture of BOC-L-alanine (17b) (2.9 g, 15.4 mmol), p-nitrophenol (3.3 g, 15.4 mmol) and DCC (3.3 g, 16.2 mmol) in EtOAc (60 ml) was stirred at RT for 2 h. A white precipitate was formed; the solid was filtered off and the filtrate was evaporated. The crude material was purified on a Biotage silica column, eluting with 20% hexane in CH2Cl2 to give compound 18b (2.8 g, 9 mmol, 58.4% yield) as a yellow solid. LCMS C14H18N2O6 [M+1]+311.1.
Step 5b: By a method analogous to that described in Example 5, Step 5, using compound 18b in place of compound 18, compound 19 was prepared.
Step 6: At 0° C., LiOH solution (150 mg, 6 mmol, in 15 ml of H2O) was added to a solution of compound 19 (1.02 g, 2 mmol) in THF (37 ml). After 1 h at 0° C., the reaction was gradually warmed to RT and stirred at RT overnight. After the reaction was complete, EtOAc (50 ml) and H2O (5 ml) were added, followed by the addition of 1 N HCl to acidify the mixture. The organic phase was separated, dried (Na2SO4), filtered and concentrated to give the title compound 20 as a white solid (0.94 g, 1.95 mmol, 98% yield). MS: C22H22F3N3O6 [M+1]+482.1.
EXAMPLE 6




Step 1: A mixture of 4-chlorobenzaldehyde (21) (0.79 g, 5.45 mmol), 2-hydroxyethyl amine (22) (0.34 ml, 5.45 mmol) and Na2SO4 (1.44 g, 10.9 mmol) in dichloroethane (40 ml) was stirred at RT for 40 min. To this mixture, NaBH(OAc)3 (3.12 g, 14.72 mmol) and AcOH (0.82 ml, 13.67 mmol) were added. After stirring at RT overnight, the reaction was quenched with saturated NaHCO3 solution. The mixture was diluted with brine (200 ml) and extracted with CH2Cl2 (100 ml, 3×), combined and washed with brine (100 ml, 2×), dried (MgSO4), filtered and evaporated to give crude compound 23 as an oil. The oil was purified with flash grade silica gel (100 g), eluting with 5% (1:9) (NH4OH/CH3OH)/95% CH2Cl2 to yield compound 23 (0.3 g, 1.62 mmol, 30% yield).
Step 2: A mixture of compound 20 (0.241 g, 0.5 mmol), compound 23 (92.8 mg, 0.5 mmol), HATU (285 mg, 0.75 mmol) and DIPEA (0.131 ml, 0.75 mmol) in dry DMF (3.0 ml) was stirred at RT for 4 h. After the reaction was complete, water (3 ml) was added to quench the reaction and the mixture was stirred for 10 min. Solid was collected, rinsed with water, and redissolved in CH2Cl2 (10 ml), dried (Na2SO4), filtered and evaporated. Product was purified by flash grade silica gel (100 g), eluting with 4.5% (1:9) (NH4OH/CH3OH)/95% CH2Cl2 to give pure compound 24 (0.18 g, 0.28 mmol, 56% yield) as a solid.
4 N HCl-dioxane solution (0.8 ml, 3.2 mmol) and CH3OH (1 ml) were added to a solution of compound 24 (0.18 g, 0.33 mmol) in CH2Cl2 (2 ml). The mixture was stirred at RT overnight. Solvents were evaporated and product was triturated with CH2Cl2, filtered and dried under high vacuum to give title compound 25 as a HCl salt. LCMS: C26H24F3N4O4Cl. HCl [M+1]+549.1
EXAMPLE 7




By employing methods analogous to those described in Example 6, Step 2, the following compounds were prepared using an appropriate aromatic or heteroaromatic amine coupled with compound 20 either by HATU or DEC (Example 4). The data for compounds of formula 26 are as follows:












Cpd.

MS


No.
Structure
(M + 1)







26-1




505


 


26-2




539


 


26-3




515


 


26-4




501


 


26-5




501


 


26-6




529


 


26-7




529


 


26-8




505


 


26-9




539


 


26-10




507


 


26-11




507


 


26-12




507


 


26-13




523


 


26-14




519


 


26-15




540


 


26-16




523


 


26-17




583


 


26-18




519


 


26-19




519


 


26-20




519


 


26-21




537


 


26-22




541


 


26-23




555


 


26-24




489


 


26-25




489


 


26-26




569


 


26-27




569


 


26-28




537


 


26-29




537


 


26-30




527


 


26-31




541


 


26-32




555


 


26-33




527


 


26-34




513


 


26-35




529


 


26-36




541


 


26-37




523


 


26-38




522


 


26-39




511


 


26-40




511


 


26-41




511


 


26-42




513


 


26-43




487


 


26-44




513


 


26-45




513


 


26-46




500


 


26-47




517


 


26-48




529


 


26-49




517


 


26-50




512


 


26-51




512


 


26-52




536


 


26-53




513


 


26-54




513


 


26-55




513


 


26-56




488


 


26-57




529


 


26-58




513


 


26-59




529


 


26-60




513


 


26-61




473


 


26-62




527


 


26-63




527


 


26-64




545


 


26-65




585


 


26-66




541


 


26-67




529


 


26-68




559


 


26-69




478


 


26-70




537


 


26-71




478


 


26-72




487


 


26-73




491


 


26-74




486


 


26-75




529


 


26-76




521


 


26-77




521


 


26-78




510


 


26-79




518


 


26-80




492


 


26-81




529


 


26-82




487


 


26-83




501


 


26-84




541


 


26-85




518


 


26-86




458


 


26-87




488


 


26-88




472


 


26-89




472


 


26-90




472


 


26-91




522


 


26-92




511


 


26-93




515


 


26-94




514


 


26-95




525


 


26-96




537


 


26-97




620


 


26-98




465


 


26-99




461


 


26-100




511


 


26-101




524


 


26-102




572


 


26-103




572


 


26-104




583


 


26-105




557


 


26-106




583


 


26-107




557


 


26-108




546


 


26-109




546


 


26-110




494


 


26-111




494


 


26-112




575


 


26-113




559


 


26-114




581


 


26-115




541


 


26-116




541


 


26-117




595


 


26-118






 


26-119




575


 


26-120




451


 


26-121




509


 


26-122




492


 


26-123




492


 


26-124




451


 


26-125




550


 


26-126




594


 


26-127




528


 


26-128




523


 


26-129




425


 


26-130




381


 


26-131




529


 


26-132




529


 


26-133




555


 


26-134




637


 


26-135




673


 


26-136




588


 


26-137




556


 


26-138




566


 


26-139




588


 


26-140




556


 


26-141




494


 


26-142




425


 


26-143




541


 


26-144




381


 


26-145




528


 


26-146




563


 


26-147




479


 


26-148




479


 


26-149




536


 


26-150




549


 


26-151




572


 


26-152




583


 


26-153




601


 


26-154




558


 


26-155




554


 


26-156




639


 


26-157




589


 


26-158




604


 


26-159




618


 


26-160




541


 


26-161




541


 


26-162




541


 


26-163




542


 


26-164




604


 


26-165




604


 


26-166




541


 


26-167




555


 


26-168




584


 


26-169




521


 


26-170




535


 


26-171




594


 


26-172




543


 


26-173




608


 


26-174




536


 


26-175




586


 


26-176




543


 


26-177




542


 


26-178




507


 


26-179




507


 


26-180




507


 


26-181




509


 


26-182




543


 


26-183




556


 


26-184




562


 


26-185




514


 


26-186




589


 


26-187




519


 


26-188




556


 


26-189




493


 


26-190




562


 


26-191




515


 


26-192




558


 


26-193




546


 


26-194




526


 


26-195




618


 


26-196




560


 


26-197




572


 


26-198




508


 


26-199




533


 


26-200




523


 


26-201




571


 


26-202




572


 


26-203




556


 


26-204




583


 


26-205




538


 


26-206




522


 


26-207




573


 


26-208




550


 


26-209




555


 


26-210




550


 


26-211




556


 


26-212




536


 


26-213




525


 


26-214




525


 


26-215




509


 


26-216




608


 


26-217




583


 


26-218




583


 


26-219




560


 


26-220




540


 


26-221




473


 


26-222




571


 


26-223




507


 


26-224




605


 


26-226




620


 


26-227




577


 


26-228




557


 


26-229




568


 


26-230




569


 


26-231




556


 


26-232




542


 


26-234




558


 


26-235




558


 


26-236




548


 


26-237




572


 


26-238




543


 


26-239




538


 


26-240




614


 


26-241




615


 


26-242




520


 


26-243




630


 


26-244




571


 


26-245




543


 


26-246




534


 


26-247




552


 


26-248




527


 


26-249




537


 


26-250




582


 


26-251




556


 


26-252




547


 


26-253




533


 


26-254




586


 


26-255




516


 


26-256




502


 


26-257




545


 


26-258




564


 


26-259




557


 


26-260




540


 


26-261




544


 


26-262




595


 


26-263




542


 


26-264




555


 


26-265




546


 


26-266




557


 


26-267




546


 


26-268




542


 


26-269




556


 


26-270




537


 


26-271




551


 


26-272




571


 


26-273




527


 


26-274




439


 


26-275




552


 


26-276




548


 


26-277




552


 


26-278




586


 


26-279




583


 


26-280




561


 


26-281




532


 


26-282




439


 


26-283




532


 


26-264




533


 


26-285




608


 


26-286




620


 


26-287




560


 


26-288




546


 


26-289




512


 


26-290




494


 


26-291




508


 


26-292




474


 


26-293




518


 


26-294




439


 


26-295




585


 


26-296




546


 


26-297




546


 


26-298




569


 


26-299




594


 


26-300




564


 


26-301




561


 


26-302




583


 


26-303




572


 


26-304




530


 


26-305




529


 


26-306




506


 


26-307




530


 


26-308




556


 


26-309




542


 


26-310




453


 


26-311




506


 


26-312




556


 


26-313




508


 


26-314




554


 


26-315




532


 


26-316




506


 


26-317




507


 


26-318




586


 


26-319




552


 


26-320




560


 


26-321




506


 


26-322




492


 


26-323




516


 


26-324




642


 


26-325




532


 


26-326




466


 


26-327




537


 


26-328




600


 


26-329




531


 


26-330




480


 


26-331




466


 


26-332




453


 


26-333




466


 


26-334




522


 


26-335




516


 


26-336




551


 


26-337




514


 


26-338




632


 


26-339




519


 


26-340




516


 


26-341




508


 


26-342




508


 


26-343




516


 


26-344




496(M +! +H)


 


26-345




439


 


26-346




574


 


26-347




590


 


26-348




536


 


26-349




542


 


26-350




627


 


26-351




582


 


26-352




534


 


26-353




542


 


26-354




542


 


26-355




542


 


26-356




532


 


26-357




532


 


26-358




600


 


26-359




600


 


26-360




550


 


26-361




556


 


26-362




542


 


26-363




528


 


26-364




528


 


26-365




528


 


26-366




570


 


26-367




542


 


26-368




532


 


26-369




532


 


26-370




533


 


26-371




533


 


26-372




614


 


26-373




560


 


26-374




492


 


26-375




548


 


26-376




548


 


26-377




534


 


26-378




532


 


26-379




654


 


26-380




521


 


26-381




590


 


26-382




522


 


26-383




562


 


26-384




530


 


26-385




586


 


26-386




514


 


26-387




521


 


26-388




605


 


26-389




592


 


26-390




606


 


26-391




494


 


26-392




494


 


26-393




542


 


26-394




544


 


26-395




530


 


26-396




575


 


26-397




629


 


26-398




520


 


26-399




615


 


26-400




542


 


26-401




592


 


26-402




516


 


26-403




600


 


26-404




564


 


26-405




542


 


26-406




571


 


26-407




536


 


26-408




536


 


26-409




506


 


26-410




492


 


26-411




492


 


26-412




478


 


26-413




554


 


26-414




600


 


26-415




542


 


26-416




465


 


26-417




465


 


26-418




536


 


26-419




536


 


26-420




509


 


26-421




520


 


26-422




509


 


26-423




520


 


26-424




506


 


26-425




451


 


26-426




504


 


26-427




530


 


26-428




506


 


26-429




504


 


26-430




520


 


26-431




547


 


26-432




528


 


26-433




518


 


26-434




532


 


26-435




592


 


26-436




536


 


26-437




551


 


26-438




550


 


26-439




518


 


26-440




528


 


26-441




614


 


26-442




521


 


26-443




520


 


2-444




506


 


26-445




619


 


26-446




580


 


26-447




542


 


26-448




557


 


26-449




590


 


26-450




506


 


26-451




518


 


26-452




522


 


26-453




481


 


26-454




481


 


26-455




467


 


26-456




467


 


26-457




473


 


26-458




481


 


26-459




530


 


26-460




522


 


26-461




588


 


26-462




534


 


26-463




520


 


26-464




541


 


26-465




535


 


26-466




541


 


26-467




549


 


26-468




501


 


26-469




548


 


26-470




507


 


26-471




509


 


26-472




511


 


26-473




531


 


26-474




557


 


26-475




534


 


26-476




520


 


26-477




590


 


26-478




525


 


26-479




537


 


26-480




589


 


26-481




540


 


26-482




473


 


26-483




502


 


26-484




587


 


26-485




520


 


26-486




543


 


26-487




551


 


26-488




527


 


26-489




423


 


26-490




437


 


26-491




421


 


26-492




435


 


26-493




449


 


26-494




435


 


26-495




479


 


26-496




435


 


26-497




463


 


26-498




465


 


26-499




466


 


26-500




487


 


26-501




511


 


26-502




 541,543


 


26-503




531


 


26-504




492


 


26-505




483


 


26-506




512


 


26-507




557


 


26-508




455


 


26-509




549


 


26-510




518


 


26-511




520


 


26-512




475


 


26-513




461


 


26-514




541


 


26-515




524


 


26-516




501


 


26-517




549


 


26-518




461


 


26-519




503


 


26-520




517


 


26-521




549


 


26-522




546


 


26-523




527


 


26-524




521


 


26-525




528


 


26-526




528


 


26-527




463


 


26-528




451


 


26-529




451


 


26-530




437


 


26-531




465


 


26-532




451


 


26-533




497


 


26-534




552


 


26-535




532


 


26-536




546


 


26-537




556


 


26-538




526


 


26-539




536


 


26-540




534


 


26-541




518


 


26-542




520


 


26-543




479


 


26-544




479


 


26-545




451


 


26-546




451


 


26-547




437


 


26-548




521


 


26-549




528


 


26-550




529


 


26-551




497


 


26-552




511


 


26-553




502


 


26-554




479


 


26-555




445


 


26-556




449


 


26-557




423


 


26-558




437


 


26-559




479


 


26-560




409


 


26-561




479


 


26-562




562


 


26-563




501


 


26-564




479


 


26-565




493


 


26-566




395


 


26-567




409


 


26-568




537


 


26-569




555


 


26-570




529


 


26-571




487


 


26-572




559


 


26-573




549


 


26-574




549


 


26-575




478


 


26-576




572


 


26-577




499


 


26-578




499


 


26-579




477


 


26-580




423


 


26-581




546


 


26-582




534


 


26-583




492


 


26-584




492


 


26-585




548


 


26-586




569


 


26-587




583


 


26-588




501


 


26-589




437


 


26-590




451


 


26-591




437


 


26-592




437


 


26-593




481


 


26-594




451


 


26-595




481


 


26-596




520


 


26-597




485


 


26-598




546


 


26-599




463


 


26-600




492


 


26-601




506


 


26-602




478


 


26-603




492


 


26-604




506


 


26-605




492


 


26-606




506


 


26-607




518


 


26-608




569


 


26-609




547


 


26-610




545


 


26-611




545


 


26-612




529


 


26-613




457


 


26-614




501


 


26-615




443


 


26-616




492


 


26-617




545


 


26-618




532


 


26-619




532


 


26-620




504


 


26-621




492


 


26-622




531


 


26-623




585


 


26-624




534


 


26-625




534


 


26-626




479


 


26-627




479


 


26-628




535


 


26-629




508


 


26-630




457


 


26-631




439


 


26-632




453


 


26-633




492


 


26-634




520


 


26-635




465


 


26-636




489


 


26-637




613


 


26-638




522


 


26-639




463


 


26-640




449


 


26-641




463


 


26-642




538


 


26-643




538


 


26-644




447


 


26-645




463


 


26-646




627


 


26-647




588


 


26-648




554


 


26-649




554


 


26-650




451


 


26-651




465


 


26-652




527


 


26-653




560


 


26-654




560


 


26-655




476


 


26-656




526


 


26-657




518


 


26-658




528


 


26-659




517


 


26-660




506


 


26-661




506


 


26-662




508


 


26-663




524


 


26-664




541


 


26-665




548


 


26-666




520


 


26-667




554


 


26-668




529


 


26-669




464


 


26-670




465


 


26-671




478


 


26-672




492


 


26-673




395


 


26-674




508


 


26-675




546


 


26-676




532


 


26-677




571


 


26-678




514


 


26-679




588


 


26-680




519


 


26-681




570


 


26-682




514


 


26-683




518


 


26-684




528


 


26-685




540


 


26-686




511


 


26-687




520


 


26-688




548


 


26-689




546


 


26-690




528


 


26-691




528


 


26-692




521


 


26-693




520


 


26-694




506


 


26-695




453


 


26-696




565


 


26-697




504


 


26-698




520


 


26-699




534


 


26-700




503


 


26-701




544


 


26-702




421


 


26-703




547


 


26-704




492


 


26-705




467


 


26-706




451


 


26-707




570


 


26-708




542


 


26-709




484


 


26-710




569


 


26-711




574


 


26-712




491


 


26-713




572


 


26-714




558


 


26-715




548


 


26-716




564


 


26-717




565


 


26-718




551


 


26-719




497


 


26-720




542


 


26-721




492


 


26-722




506


 


26-723




532


 


26-724




546


 


26-725




518


 


26-726




534


 


26-727




565


 


26-728




564


 


26-729




564


 


26-730




506


 


26-731




532M + Na


 


26-732




535


 


26-733




557


 


26-734




520


 


26-735




568


 


26-736




554


 


26-737




467


 


26-738




540


 


26-739




506


 


26-740




518


 


26-741




558


 


26-742




498


 


26-743




498


 


26-744




493


 


26-745




465


 


26-746




527


 


26-747




507


 


26-748




556


 


26-749




556


 


26-750




572


 


26-751




556


 


26-752




695


 


26-753




556


 


26-754




594


 


26-755




533


 


26-756




547


 


26-757




549


 


26-758




527


 


26-759




554


 


26-760




636


 


26-761




619


 


26-762




699


 


26-763




549


 


26-764




527


 


26-765




506


 


26-766




518


 


26-767




526


 


26-768




526


 


26-769




592


 


26-770




654


 


26-771




496


 


26-772




598


 


26-773




510


 


26-774




529


 


26-775




487


 


26-776




485


 


26-777




584


 


26-778




584


 


26-779




508


 


26-780




563


 


26-781




596


 


26-782




473


 


26-783




557


 


26-784




570


 


26-785




564


 


26-786




534


 


26-787




548


 


26-788




540


 


26-789




530


 


26-790




529


 


26-791




522


 


26-792




536


 


26-793




550


 


26-794




562


 


26-795




479


 


26-796




532


 


26-797




548


 


26-798




562


 


26-799




530


 


26-800




478


 


26-801




570


 


26-802




554


 


26-803




522


 


26-804




536


 


26-805




548


 


26-806




576


 


26-807




554


 


26-808




546


 


26-809




550


 


26-810




520


 


26-811




598


 


26-812




598


 


26-813




572


 


26-814




492


 


26-815




492


 


26-816




497


 


26-817




506


 


26-818




489


 


26-819




539


 


26-820




570


 


26-821




570


 


26-822




575


 


26-823




531


 


26-824




580


 


26-825




558


 


26-826




506


 


26-827




515


 


26-828




557


 


26-829




465


 


26-830




566


 


26-831




507


 


26-832




514


 


26-833




531


 


26-834




496


 


26-835




510


 


26-836




580


 


26-837




518


 


26-838




532


 


26-839




527


 


26-840




514


 


26-841




510


 


26-842




497


 


26-843




555


 


26-844




517


 


26-845




532


 


26-846




606


 


26-847




584


 


26-848




584


 


26-849




518


 


26-850




532


 


26-851




462


 


26-852




476


 


26-853




463


 


26-854




536


 


26-855




534


 


26-856




512


 


26-857




519


 


26-858




524


 


26-859




538


 


26-860




522


 


26-861




584


 


26-862




584


 


26-863




569


 


26-864




569


 


26-865




546


 


26-866




551


 


26-867




573


 


26-868




558


 


26-869




559


 


26-870




586


 


26-871




552


 


26-872




479


 


26-873




546


 


26-874




478


 


26-875




506


 


26-876




527


 


26-877




558


 


26-878




514


 


26-879




544


 


26-880




544


 


26-881




572


 


26-882




534


 


26-883




516


 


26-884




584


 


26-885




518


 


26-886




534


 


26-887




465


 


26-888




529


 


26-889




496


 


26-890




572


 


26-891




522


 


26-892




478


 


26-893




493


 


26-894




580


 


26-895




493


 


26-896




527


 


26-897




558


 


26-898




558


 


26-899




490


 


26-890




487


 


26-891




518


 


26-892




544


 


26-893




594


 


26-894




612


 


26-895




560


 


26-896




528


 


26-897




540


 


26-898




487


 


26-899




515


 


26-900




501


 


26-901




511


 


26-902




529


 


26-903




545


 


26-904




551


 


26-905




617


 


26-906




599


 


26-907




560


 


26-908




574


 


26-909




501


 


26-910




529


 


26-911




564


 


26-912




578


 


26-913




499









EXAMPLE 8


    Step 1:






Using N-[(1,1-dimethylethoxy)carbonyl]-O-(1,1-dimethylethyl)-L-serine (compound 27) as starting material, [1(S)-[(1,1-dimethylethoxy)methyl]-2-fluoro-2-oxoethyl] carbamic acid, 1,1-dimethylethyl ester (compound 28) was prepared by a method analogous to that in Example 5, step 4.

    Step 2:






0.5 M KHMDS in toluene (92.5 ml, 46.25 mmol) was added slowly via a syringe to a mixture of compound 16 (8.5 g, 22 mmol) and compound 28 (6.8 g, 25.8 mmol) in dry THF (90 ml) at −78° C. The mixture was slowly warmed to RT, then stirred at RT for 1 h. After the reaction was complete, it was quenched with 1 N HCl (80 ml)(cooled with ice-water bath), diluted with saturated NH4Cl solution (100 ml), extracted with EtOAc (200 ml×2), dried (Na2SO4), filtered and evaporated. Crude material was purified on Biotage with CH2Cl2 (4 L) and 5% EtOAc/CH2Cl2 (4 L) to give compound 29 as a light yellow solid (6.5 g, 11.8 mmol, 52%). MS C28H34F3N3O7 [M+1]+582.1.

EXAMPLE 9




Compound 29 (13.5 g, 23.24 mmol) was treated with THF:H2O (2:1) (200 ml) and LiOH.H2O (0.95 g, 39.6 mmol) (dissolved in 10 ml of H2O). After stirring at RT for 2 h, the suspension was not dissolved. Additional THF:H2O (2:1) (100 ml) and LiOH.H2O (0.95 g, 39.6 mmol) was added. It was stirred at RT overnight. After completion, the reaction was neutralized with 1 N HCl. The mixture was extracted with CH2Cl2 (100 ml×3), combined, washed with brine (100 ml), dried (Na2SO4), filtered and evaporated to give the title compound 30 as a yellow solid (11.8 g, 21.3 mmol, 92%). LCMS: C26H3F3N3O7 [M+1]+554.1.
EXAMPLE 10


    Step 1:






By a method analogous to Example 2, using 5-fluoro-3-methyl-benzo[B]-thiophene (31) as starting material, compound 33 was obtained. It was treated with 10 equivalents of 98% hydrazine in absolute EtOH and CH2Cl2 (1:1) to give compound 34, which was purified by treatment with a slight excess of 4 N HCl/dioxane solution to give compound 35 as a HCl salt. FABMS: C9H8FNS. HCl [M+1]+182.0

    Step 2:






By methods analogous to those described in Example 3, using compound 35 as a starting material, compound 36 was obtained.

    Step 3:






The protecting groups on compound 36 were removed by treatment with HCl-dioxane/CH2Cl2 or CF3COOH. The title compound 37 was obtained directly as a HCl salt or as a TFA salt depending on the acid treatment. The TFA salt was neutralized with NH4OH and converted to HCl salt with 1.0 equivalent of HCl. HRMS C26H20F4N4O4S. HCl calculated [M+1]+561.1220, Found 561.1230.

EXAMPLE 11




By employing analogous methods to those described in Example 10, the following compounds were obtained as HCl salts using compound 30 coupled with the appropriate primary or secondary amine, followed by removal of the protecting group as described for Example 10, step 3.












Cpd.

MS


No.
Structure
(M + 1)







38-1




521


 


38-2




557


 


38-3




523


 


38-4




539


 


38-5




523


 


38-6




523


 


38-7




555


 


38-8




539


 


38-9




513


 


38-10




529


 


38-11




535


 


38-12




505


 


38-13




505


 


38-14




537


 


38-15




529


 


38-16




543


 


38-17




601


 


38-18




561


 


38-19




529


 


38-20




541


 


38-21




545


 


38-22




553


 


38-23




532


 


38-24




558


 


38-25




527


 


38-26




604


 


38-27




557


 


38-28




636


 


38-29




557


 


38-30




397


 


38-31




590


 


38-32




557


 


38-33




573


 


38-34




553


 


38-35




548


 


38-36




562









EXAMPLE 12




By employing methods analogous to those described for Example 5, using compound 18c in place of compound 18, compound 42 was obtained, which was treated with LiOH.H2O to give the title compound 43.
EXAMPLE 13




By employing methods analogous to those described in Example 6, using compound 43 in place of compound 20 and 4-chlorobenzylamine in place of compound 23 in the coupling reaction, compound 44 was obtained. After removal of the t-BOC group of compound 44 with HCl, the title compound 45 was obtained as a HCl salt. MS: C25H22ClF3N4O3. HCl [M+1]+519.1.
Using a procedure similar to that described for compound 45, the following compounds were prepared:












Cpd.

MS


No.
Structure
(M + 1)







45-1




562


 


45-2




479


 


45-3




557


 


45-4




541


 


45-5




576


 


45-6




569


 


45-7




506


 


45-8




479


 


45-9




449


 


45-10




435


 


45-11




506









EXAMPLE 14




By employing methods analogous to those described in Example 6, using compound 43 in place of compound 20 and compound 9 in place of compound 23 compound 46 was obtained. After removal of the t-BOC group of compound 46 with HCl, the title compound 47 was obtained as a HCl salt. MS C27H23F3N4O3S .HCl [M+1]+541.1.
EXAMPLE 15




By employing methods analogous to those described in Example 5, using compound 18d in place of compound 18, compound 48 was obtained, which was treated with LiOH.H2O to obtain the title compound 49.
EXAMPLE 16




By employing methods analogous to those described in Example 6, using compound 50 in place of compound 20 and 2,4-diflurobenzylamine in place compound 23, compound 51 was obtained. After removal of the t-BOC group of compound 51 with HCl, the title compound 52 was obtained as a HCl salt. MS: C26H23F5N4O3. HCl [M+1]+535.
Using similar procedures and the appropriate staring materials, the following compounds were also prepared:












Cpd.

MS


No.
Structure
(M + 1)







47-1




542


 


47-2




541


 


47-3




519


 


52-1




556









EXAMPLE 17




By employing methods analogous to those described in Example 5, using compound 18e in place of compound 18, compound 53 was obtained, which was treated with LiOH.H2O to yield the title compound 54.
EXAMPLE 18




By employing methods analogous to those described in Example 10, using compound 55 in place of compound 30 and 1-amino-2-hydroxyethyl benzene in place of compound 35, compound 56 was obtained. After purification and removal of the t-BOC group of compound 56 with HCl, compound 57 was obtained as a HCl salt. MS: C26H25F3N4O5. HCl [M+1]+567.1.
EXAMPLE 19




Using the appropriate aromatic or heteroaromatic amine reagent coupled with compound 55 according to the procedure described for Example 10, steps 2 and 3, the desired compound 58 was obtained as a hydrochloride salt.












Cpd.

MS


No.
Structure
(M + 1)







58-1




537


 


58-2




564


 


58-3




543









EXAMPLE 20




    Step 1: DBU (1.7 g, 11 mmol) was added to a mixture of 4-chromanol (59) (1.5 g, 10 mmol) and diphenyl-phosphoryl azide (DPPA) (3.0 g, 11 mmol) in CH2Cl2 (10 ml) at RT. The mixture immediately turned brown (a water bath was used to cool the reaction temperature). The solution was stirred at RT overnight. After completion of the reaction, the reaction mixture was diluted with ether/EtOAc (1:1) (100 ml) and washed with saturated NaHCO3, 5% HCl and brine. The organic layer was dried (Na2CO3), filtered and concentrated to give a residue which was purified by column chromatography, eluting with 30% CH2Cl2/hexane to give compound 60 (1.45 g, 0.83 mmol) with a 83% of yield.
    Step 2: 4 N HCl/dioxane (2 ml) and 10% Pd/C (0.5 g) were added to a solution of compound 60 (1.3 g, 7.4 mmol) in MeOH (50 ml). The mixture was stirred under a H2 balloon at RT for 46 h. After the reaction was complete, the solid was filtered off. The filtrate was concentrated and to obtain the desired amine (61) as a light yellow HCl salt. LCMS C9H11NO. HCl [M+1]+149.0.


EXAMPLE 21




By employing methods analogous to those described for Example 20, replacing compound 59 with 62 and 60 with 63, the title compound 64 was obtained as an amine HCl salt. LCMS C9H11O2N. HCl [M+1]+166.0
EXAMPLE 22




NaBH4 (0.7 g, 18.5 mmol) at RT was added to a solution of compound 65 (0.6 g, 3.57 mmol) in MeOH (20 ml) cooled with a water bath. After 10 min, solvent was removed. The residue was treated with 5% NaHCO3 and the product was extracted with CH2Cl2, then with EtOAc. The combined organic solution was washed with brine, dried (Na2SO4), filtered and concentrated to give compound 66 as a white solid. Using a method similar to Example 27, the title compound 67 was obtained as a HCl salt. LCMS C7H11N3O2.HCl [M+1]+170.0
EXAMPLE 23




Solid 1,3,5-triazine (2.4 g, 30 mmol) (68) was mixed with 1-(pyrrolidino)-1-cyclohexene (69) in a pressure tube (15 ml) and heated at 93° C. (bath temperature) with stirring for 22 h. After completion of the reaction, the reaction mixture was concentrated and dissolved in CH2Cl2, washed with saturated NaHCO3 solution, dried (Na2SO4), then purified by column chromatography to give compound 70 as a solid.
The title compound 73 was prepared according to the procedure described for Example 2, and step 2 of Example 3.
EXAMPLE 24




A mixture containing 3-acetylthianaphthene (74) (0.5 g, 2.8 mmol), allylamine (0.42 g, 5.6 mmol), NaBH(OAc)3 (1.2 g, 5.6 mmol), and HOAc (0.15 ml) in dichloroethane (15 ml) was stirred at RT overnight. After completion, the reaction mixture was quenched with NaHCO3 and extracted with CH2Cl2. The organic solvent was dried (Na2SO4), filtered and evaporated. The crude material was purified by column chromatography to give compound 75 (0.43 g) as an oil.
N,N-dimethylbarbituric acid (0.65 g, 4.14 mmol) and tetrakis-(triphenyl-phosphine)palladium (16 mg, 0.0138 mmol) were added to a solution of compound 75 (0.3 g, 1.38 mmol) in CH2Cl2 (30 ml). The mixture was stirred at 40° C. for 2 h, then at RT overnight. After completion of the reaction, the mixture was diluted with CH2Cl2 and washed with saturated Na2CO3 solution. The organic layer was separated and the aqueous layer was re-extracted with CH2Cl2. The organic fractions were combined and concentrated. The crude material was purified by silica gel chromatography to give the title compound 76 as an oil.
EXAMPLE 25




    Step 1: MgO (0.4 g, 10 mmol) and 10% Pd/C (0.5 g) were added to a solution of compound 77 (1.0 g, 5.4 mmol) in EtOH:MeOH (1:1) (100 ml). The mixture was stirred at RT overnight. After completion of the reaction, MgO and Pd/C were filtered off and the filtrate was concentrated to dryness. The residue was dissolved in EtOAc and washed with water, dried (Na2SO4), filtered and evaporated to give a solid compound 78.
    Step 2: Using the procedure described in Example 2, Step 1, the bromo derivative 79 was obtained. The crude material was used in the next reaction without purification.
    Step 3: Sodium azide was mixed with DMSO and stirred at RT until all solid was dissolved. Compound 78 was added at RT, and after stirring at RT for 1 h, ice-water was added. The product was extracted with EtOAc:ether (1:1). The combined organic layers were washed with water, dried (Na2SO4), filtered and evaporated to obtain the azido derivative 80, as an oil.
    Step 4: By employing methods analogous to Example 20, Step 2, the azido derivative 80 was converted to the title compound 81 as a hydrochloride salt.


EXAMPLE 26




    Step 1: Znl2 (0.64 g, 2 mmol) was added in one portion at RT under N2 to a mixture of 2,4-dichlorobenzaldehyde (82) (3.5 g, 20 mmol) and TMSCN (2.6 g, 26 mmol). After 15 min, 7 N NH3 solution in MeOH (20 ml) was added and the mixture was stirred at 40° C. for 2 h. Solvents were evaporated and the residue was re-dissolved in Et2O, washed with water, dried (MgSO4) and filtered. HCl gas was bubbled through the filtrate to give an off-white solid of compound 83 (3.5 g, 74%). MS: C8H6N2Cl2, [M+1]+202.
    Step 2: A stream of HCl gas was bubbled through a solution of compound 83 (3.5 g, 14.8 mmol) in MeOH (85 ml) for 4 h. Water (2 ml) was added and the reaction mixture was concentrated to provide an off-white solid of compound 84 (3.4 g, 90%) as a HCl salt. MS: C8H8N2OCl2, [M+1]+219


EXAMPLE 27




    Step 1: Compound 85 (2.4 g, 14.4 mmol) was suspended in phosphorus oxychloride (30 ml) and heated to reflux for 15 h. The reaction mixture was cooled to RT, and saturated (aq) NaHCO3 (250 ml) was added carefully with vigorous stirring at 0° C., followed by the addition of Et2O (150 ml). The aqueous layer was separated and extracted with Et2O. The organic layers were combined, washed with brine, filtered and concentrated to give compound 86 as a yellow oil (2.14 g, 81%). MS: C7H5ClN2S [M+1] 185; [M+2], 186
    Step 2: Compound 87 was obtained as a white solid (93% yield) according to the procedure described for the Example 25, Step 1. MS: C7H6N2S [M+1]+151.
    Step 3: Compound 88 [MS: C7H5BrN2S [M+1]+Br79 229, Br81 231] was synthesized from compound 88 according to the method described in Example 2, Step 1. The bromo-derivative 88 was converted to its azido-derivative-89 [MS: C7H5N5S [M+1]+192] according to the procedure described for Example 25, Step 3. The title compound 90 was obtained as a HCl salt by hydrogenation of compound 89 according to the procedure described for Example 20. MS: C7H7N3S [M+1]+166.


EXAMPLE 28




    Step 1: Compound 91 (1.47 g, 7.99 mmol) was treated with a solution of 0.5 M NaOMe in MeOH (32 ml) under N2. The suspension was stirred at RT overnight. The solvent was removed and the resultant residue partitioned between EtOAc (75 ml) and water (75 ml). The aqueous layer was separated and extracted with EtOAc. The organic extracts were combined, dried (MgSO4), filtered and concentrated in vacuo to give 1.41 g (98%) of white solid, compound 92. MS: C8H8N2OS [M+1]+181.
    Step 2: The title compound 93 was obtained from compound 92 using methods similar to those described for Example 27, Step 3. MS: C8H9N3SO [M+1]+196.


EXAMPLE 29




Compound 94 was prepared according to the literature (Tetrahedron Letters 41, 8661-8664, (2000)). Compound 94 (3.0 g, 22.5 mmol) was mixed with NaI (3.37 g, 22.5 mmol) and NaN3 (1.9 g, 29 mmol) in CH2Cl2/acetone (1:1, 250 ml) and refluxed for 36 h. After completion of the reaction, the reaction mixture was filtered and the filtrate concentrated to dryness. Crude compound 95 was purified on Biotage system, eluting with 2% MeOH in CH2Cl2, to obtain pure compound 95 as a white solid. Compound 95 (2.46 g, 17.6 mmol) was dissolved in MeOH (100 ml) and formic acid (0.81 ml, 17.6 mmol), and 10% Pd/C (490 mg) was added. The mixture was stirred at RT under a H2 balloon overnight. The solids were filtered off and the filtrate was evaporated to give the title compound, 96, as a formic acid salt. MS: C3H6N4O [M+1]+115.
EXAMPLE 30




    Step 1: To a solution of compound 97 (280 mg, 0.462 mmol) dissolved in THF (10 ml) was added Lawesson reagent (467 mg, 1.15 mmol). The reaction mixture was heated at reflux for 24 h then cooled to RT. The solvent was evaporated. Purification by silica gel chromatography (eluant: 1%-3% EtOAc—CH2Cl2) gave 166 mg (0.267 mmol, 58%) of the product 98 as a yellow foam. MS (M+1): m/e 623.
    Step 2: To a solution of compound 98 (273 mg, 0.438 mmol) dissolved in CH2Cl2 (4 ml) was added TFA (1 ml). The reaction mixture was stirred at RT for 4 h. The solvent was evaporated, and the crude product was dissolved in 10 ml of 1:1 CH2Cl2: MeOH and diethylaminomethylpolystyrene resin (0.50 g, from Fluka) was added. The resulting mixture was stirred for 15 min, filtered, and the resin was washed with MeOH. The filtrate was evaporated. Purification by silica gel chromatography (eluant: 2%-3% MeOH—CH2Cl2) gave 169 mg (0.323 mmol, 74%) of the product 99 as a yellow foam. MS (M+1): m/e 523.


EXAMPLE 31




    Step 3: To a solution of trans-4-benzoyl-cyclohexylamine (2.06 g, 9.4 mmol) dissolved in dry CH2Cl2 (50 ml) was added 3A sieves (3 g), Et3N (2.38 g, 3.3 ml, 23.5 mmol), [4-(N-BOC-aminomethyl)phenyl]boronic acid 100A (3.00 g, 11.9 mmol), and copper acetate (2.16 g, 11.9 mmol). The reaction mixture was stirred at RT for 24 h. 2 N aqueous NH4OH (50 ml) was added, and the reaction mixture was filtered to remove the sieves which were washed with additional CH2Cl2 and 2 N aqueous NH4OH. The layers of the filtrate were separated, and the aqueous layer was extracted width CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 5%-10% EtOAc—CH2Cl2) gave 0.73 g (1.72 mmol, 18%) of the product 101A as a yellow foam. MS (M+1): m/e 425.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







101B




277


 


101C




371


 


101D




349


 


101E




321


 


101F




335


 


101G




335


 


101H




425


 


101I




277


 


101J




371


 


101K




411


 


101L




335


 


101M




335









    Step 2: To a solution of compound 101A (0.72 g, 1.70 mmol) dissolved in THF (6 ml) MeOH (6 ml), and water (3 ml) was added LiOH (0.36 g, 8.48 mmol). The reaction mixture was stirred at RT for 4 h. The solvent was evaporated, saturated NH4Cl (25 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 3%-5% MeOH—CH2Cl2) gave 0.48 g (1.50 mmol, 89%) of the product 102A as a white foam. MS (M+1): m/e 321.


The following intermediates were synthesized by using a similar procedure:














MS


Number
Compound
(M + 1)







102B




267


 


102C




307


 


102D




279


 


102E




293


 


102F




293


 


102G




321


 


102H




267


 


102I




307


 


102J




293


 


102K




293









    Step 3: To a solution of compound 102A (475 mg, 1.48 mmol) dissolved in 1:1 CH2Cl2:MeOH (10 ml) was added 4 N HCl in dioxane (3.0 ml, 11.9 mmol). The reaction mixture was stirred at RT for 3 h. The solvent was evaporated to give 429 mg (1.46 mmol, 99%) of the product 103A as a white solid. MS (M+1): m/e 221.


To a solution of compound 102D (0.73 g, 2.62 mmol) suspended in CH2Cl2 (18 ml) was added TFA (3 ml). The reaction mixture was stirred at RT for 3 h. The solvent was evaporated, and the TFA salt of the product was dissolved in MeOH (20 ml). Diethylaminomethylpolystrene resin (4 g, Fluka) was added and stirred at RT for 20 min. The resin was removed by filtration and washed with MeOH. The filtrate was concentrated to give 0.47 g (2.62 mmol, 100%) of the product 103B as a yellow solid. MS (M+1-OH): m/e 162.
The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







103C




150(M + 1-OH)


 


103D




207


 


103E




176M + 1-OH


 


103F




176M + 1-OH


 


103G




221


 


103H




167


 


103I




207


 


103J




193


 


103K




193









EXAMPLE 32




    Step 1: To a solution of methyl 4-(BOC-aminomethyl)-benzoate 104A (4.15 g, 15.6 mmol) dissolved in THF (20 ml), MeOH (20 ml), and water (10 ml) was added LiOH (0.72 g, 17.2 mmol). The reaction mixture was stirred at RT for 24 h. The solvent was evaporated to give 4.02 g (15.6 mmol, 100%) of the product 105A as a white solid. MS (M+2-tBu for acid COOH): m/e 196.
    Step 2: To a solution of 4-hydroxypiperidine (0.41 g, 4.08 mmol) dissolved in dry DMF (20 ml) was added 3A sieves (1.0 g) and the mixture was stirred at RT for 15 min. HOBT (0.55 g, 4.08 mmol), EDCI (0.78 g, 4.08 mmol), compound 105A (0.70 g, 2.72 mmol), and Et3N (0.55 g, 0.76 ml, 5.44 mmol) were then added. The reaction mixture was stirred at RT for 20 h. The solvent was evaporated, 0.2 N NaOH (40 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 5%-10% MeOH—CH2Cl2) gave 0.78 g (2.33 mmol, 86%) of the product 106A as a white foam. MS (M+1): m/e 335.


The following intermediates were synthesized by using a similar procedure:














MS


Number
Compound
(M + 1)







106B




307


 


106C




321


 


106D




321


 


106E




335


 


106F




307


 


106G




321


 


106H




321


 


106I




266


 


106J




308


 


106K




306


 


106L




322









    Step 3: Using the procedure of step 3 from Example 31, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







107A




235


 


107B




207


 


107C




221


 


107D




221


 


107E




235


 


107F




207


 


107G




221


 


107H




221


 


107I




166


 


107J




208


 


107K




206


 


107L




222









    Step 4: To a solution of compound 104A (2.47 g, 9.31 mmol) dissolved in Et2O (50 ml) was added LiBH4 (0.81 g, 37.2 mmol) then MeOH (1.19 g, 1.5 ml, 37.2 mmol). The reaction mixture was heated at reflux for 5 h and then cooled to RT. The solvent was evaporated. Water (50 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 5%-8% MeOH —CH2Cl2) gave 2.15 g (9.06 mmol, 97%) of the product 108A as a white solid. MS (M+1): m/e 238.


The following intermediate was synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







108B




182M + 2-tBu









    Step 5: To a solution of oxalyl chloride (1.43 g, 0.98 ml, 11.3 mmol) dissolved in dry CH2Cl2 (20 ml) and cooled to −78° C. under a N2 atmosphere was added DMSO (1.76 g, 1.6 ml, 22.5 mmol) dissolved in CH2Cl2 (5 ml) dropwise via addition funnel. The reaction mixture was stirred at −78° C. for 15 min then compound 108A (2.14 g, 9.02 mmol) dissolved in CH2Cl2 (25 ml) was added dropwise via addition funnel. The reaction mixture was stirred at −78° C. for 60 min, then Et3N (2.74 g, 3.8 ml, 27.0 mmol) was added. The reaction mixture was stirred at −78° C. for 20 min, then warmed to RT. Water (75 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 2%-3% MeOH —CH2Cl2) gave 2.12 g (9.02 mmol, 100%) of the product 109A as a white solid. MS (M+1): m/e 236.


The following intermediate was synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







109B




180M + 2-tBu









    Step 6: To a solution of compound 109A (0.50 g, 2.12 mmol) dissolved in 10% water by volume in EtOH (20 ml) was added sodium acetate (1.05 g, 12.7 mmol) and hydroxylamine hydrochloride (0.59 g, 8.50 mmol). The reaction mixture was heated at reflux for 4 h and then cooled to RT. The solvent was evaporated. Water (30 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated to give 0.53 g (2.12 mmol, 100%) of the product 110A as a white solid. MS (M+1): m/e 251.


The following intermediate was synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







110B




195M + 2-tBu









    Step 7: Using the procedure of step 3 from Example 31, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







111A




151


 


111B




151









EXAMPLE 33




    Step 1: 3-Methylphthalic anhydride 112A (5.00 g, 30.8 mmol) and urea (1.85 g, 30.8 mmol) were combined and heated at 320-350° C. with stirring for 5 min, then cooled to RT. The brown solid was triturated with water and filtered. The solid was washed with water and dried to give 4.80 g (29.8 mmol, 97%) of the product 113A as a pink solid. MS (M+1): m/e 162.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







113B




204


 


113C




164


 


113D




165 forM+


 


113E




184









    Step 2: To compound 113A (4.80 g, 29.8 mmol) was added 1 M borane in THF (104 ml, 0.104 mol) under a N2 atmosphere. The reaction mixture was heated at reflux for 16 h and then cooled to 0° C. EtOH (80 ml) and K2CO3 (9.20 g, 66 mmol) were added carefully. The resulting mixture was heated at reflux for 16 h and then cooled to RT. (tBOC)2O (10.00 g, 45.8 mmol) was added, and the reaction mixture was stirred at RT for 3 h. The solvent was evaporated. Water (200 ml) was added, and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 5% EtOAc—CH2Cl2) gave 4.50 g (19.3 mmol, 64%) of the product 114A as a beige foam. MS (M+2-tBu): m/e 178.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







114B




220M + 2-tBu


 


114C




180M + 2-tBu


 


114D




182M + 2-tBu


 


114E




182M + 2-tBu


 


114F




198M + 2-tBu


 


114G




200M + 2-tBu


 


114H




232M + 2-tBu


 


114I




235


 


114J




235









    Step 3: Using the procedure of step 3 from Example 31, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







115A




134


 


115B




176


 


115C




136


 


115D




138


 


115E




138


 


115F




154


 


115G




156


 


115H




188


 


115I




135


 


115J




135









EXAMPLE 34




    Step 1: Using the procedure of step 6 from Example 32, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







117A




178


 


117B




192









    Step 2: To a solution of compound 117A (1.08 g, 6.09 mmol) dissolved in EtOH (20 ml) was added 10% palladium on carbon catalyst (0.25 g) and 1.73 M HCl in EtOH (10.6 ml, 18.3 mmol). The reaction mixture was shaken on a Parr shaker under 50 psi of hydrogen pressure for 16 h. The catalyst was removed by filtration through celite and washed with EtOH. The filtrate was concentrated to give 1.14 g (5.71 mmol, 93%) of the product 118A as a beige solid. MS (M−NH2): m/e 147.


The following intermediate was synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







118B




161M-NH2









EXAMPLE 35




    Step 1: To a solution of compound 119 (3.00 g, 14.9 mmol) dissolved in dry DMF (60 ml) under a N2 atmosphere was added NaH (60 wt % in oil, 1.19 g, 29.8 mmol). The reaction mixture was stirred at RT for 30 min, then 2-chloropyrimidine (3.41 g, 29.8 mmol) was added. The reaction mixture was heated at 80° C. for 16 h and then cooled to RT. The solvent was evaporated. Water (75 ml) was added and the aqueous solution was extracted with CH2Cl2. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1%-4% MeOH—CH2Cl2) gave 2.49 g (8.91 mmol, 60%) of the product 120A as a yellowish-orange solid. MS (M+1): m/e 280.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







120B




280


 


120C




310









    Step 2: Using the procedure of step 3 from Example 31, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







121A




180


 


121B




180


 


121C




210









EXAMPLE 36




    Step 1: Trans-4-hydroxy-L-proline 122A (10.48 g, 80 mmol) was refluxed in a 5-6 M solution of HCl in 2-propanol (200 ml) for 2 h. The solvent was evaporated to give 16.33 g of the product 123A as a white solid (97% yield). MS (M+1) 174.
    Step 2: 123A (16.33 g, 78.1 mmol) was suspended in dichloromethane (460 ml). Et3N (30 ml) and di-tert-butyl dicarbonate (20.33 g) were added, and the mixture was stirred for 20 h at RT. The reaction mixture was washed twice with equivolume 1 N HCl, once with saturated NaHCO3, and once with saturated NaCl. The organic solution was dried (anhydrous Na2SO4), filtered, and concentrated to give the product 124A as an amber oil (19.7 g, 92% yield). MS (M+1): m/e 274.
    Step 3: Using the procedure of step 5 from Example 32, compound 125A was synthesized. MS (M+1): m/e 272.
    Step 4: A 5-gram vial of CeCl3 from Aldrich was cracked open and quickly added to a flame-dried, 125-ml, round-bottomed flask under an Ar atmosphere. Anhydrous THF was added, and the mixture was sonicated for 1 h and stirred an additional 1 h. The ketone 125A was dissolved in dry THF (5 ml) and added to the CeCl3/THF mixture and stirred at RT for 1 h. In a separate round bottom flask, 2-pyrimidyl tri-n-butylstannane was dissolved in dry THF (18 ml) under an Ar atmosphere and cooled to −78° C. A 2.5M solution of n-butyllithium in hexanes (4 ml) was added dropwise to the pyrimidyl stannane, and the mixture turned thick and brown. After stirring for 1 h at −78° C., this cold mixture was transferred via cannula to the ketone 125A/CeCl3 mixture also cooled to −78° C. The resulting reaction mixture was stirred at −78° C. for 3 h and then stirred at −50° C. for 30 min. The mixture was again cooled to −78° C. and quenched dropwise with 1 M citric acid (200 ml). The aqueous solution was extracted with hexane and then Et2O. The combined organic extract was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography gave 0.95 g of the product 126A (27% yield). MS (M+1): m/e 352.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS (M + 1)







126B




351


 


126C




280









    Step 5: Using the procedure of step 3 from Example 31, the following intermediates were synthesized:














Number
Compound
MS (M + 1)







127A




252


 


127B




251


 


127C




180









EXAMPLE 37




2,4,6-Trifluorobenzylamine 128 was prepared according to the literature procedure of A. Marfat et al, WO 9845268.
EXAMPLE 38




    Step 1: Compound 129 (0.24 g, 0.5 mmol) was mixed with Et3N (0.1 ml, 0.7 mmol) in dry THF (4 ml) and cooled to −78° C. Trimethylacetyl chloride (0.08 ml, 0.6 mmol) was added, and the resulting mixture was stirred at 0° C. for 20 min, then cooled to −78° C. again. In a separate flask, oxazolidinone (0.07 g, 0.8 mmol) was dissolved in dry THF (2 ml), cooled to −78° C., and 1.6 ml of a 1.6 M n-BuLi solution in hexane was added. After stirring at −78° C. for 15 min, the mixture was cannulated into the above mixed anhydride solution. The resulting solution was then slowly warmed up to RT. The reaction mixture was quenched with saturated NH4Cl solution (2 ml). EtOAc (50 ml) was added, and the organic solution was washed with 1 N HCl solution, saturated NaHCO3 solution, and brine. The organic solution was dried (Na2SO4), filtered, and concentrated. Purification by flash chromatography gave the product which was treated with 2 N HCl in ether (50 ml) at RT overnight. The precipitate was collected by filtration and dried in a vacuum oven at 50° C. overnight to give 0.15 gram of the product 130 as the HCl salt. MS (M+1): m/e 451.


EXAMPLE 39




    Step 1: Compound 131 (1.15 g, 6 mmol) was mixed with Et3N (0.9 ml, 6.4 mmol) in dry THF (20 ml), and cooled to −30° C. Trimethylacetyl chloride (0.75 ml, 6 mmol) was added, and the resulting mixture was stirred at −10° C. for 20 min. Pyrrolidine (0.85 ml, 10 mmol) was added, and the resulting mixture was stirred at 0° C. for 30 min. The reaction mixture was diluted with EtOAc (150 ml) and washed with 1 N HCl solution, saturated NaHCO3 solution, and brine. The organic solution was dried (MgSO4), filtered, and concentrated to give the product 132.
    Step 2: Using the procedure of step 3 from Example 31, intermediate 133 was synthesized.
    Step 3: To a solution of compound 129 (0.48 g, 1 mmol) dissolved in DMF (4 ml) and CH2Cl2 (10 ml) at RT was added DIPEA (1 ml) and HATU (0.6 g). After 5 min, compound 133 (HCl salt, 0.23 g, 1.3 mmol) was added. The reaction mixture was stirred at RT for 30 min, then the mixture was diluted with EtOAc (75 ml) and washed with 1 N HCl (50 ml), saturated NaHCO3 (50 ml), and brine. The organic solution was dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography gave the product 134.
    Step 4: Using the procedure of step 3 from Example 31, compound 135 was synthesized. MS (M+1): m/e 506.


EXAMPLE 40




    Step 1: Using the procedure for step 3 from Example 39, compound 136 was synthesized.
    Step 2: To a solution of compound 136 (0.2 g, 0.35 mmol) dissolved in dry CH2Cl2 (6 ml) was added DAST (0.1 ml, 0.7 mmol). The reaction mixture was stirred at RT for 2 days, then quenched with saturated NaHCO3 (2 ml). The mixture was diluted with CH2Cl2 (75 ml) and washed with water then 1 N HCl solution. The organic solution was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography gave the product 137.
    Step 3: Using the procedure of step 3 from Example 31, compound 138 was synthesized. MS (M+1): m/e 467


EXAMPLE 41




    Step 1: Using the procedure of step 2 from Example 32, intermediate 140 was synthesized. MS (M+1): m/e 281.
    Step 2: Using the procedure of step 3 from Example 31, intermediate 141 was synthesized. MS (M+1): m/e 181.
    Step 3: To a solution of compound 141 (1.9 g, 8.2 mmol, a TFA salt) and Et3N (2.5 g, 24.6 mmol) in CH2Cl2 (32 ml) at 0° C. was added a solution of 2-nitrophenylsulphonyl chloride (1.99 g, 9 mmol) in CH2Cl2 (8 ml) over a period of 5 min. The reaction mixture was stirred at 0° C. for 2 h, and then saturated NaHCO3 solution was added. The product was extracted with CH2Cl2, washed with brine (1×70 ml), dried over Na2SO4, filtered, and concentrated to give an oily residue. Purification by silica gel chromatography (Biotage System, eluant: 40:1 CH2Cl2:MeOH) gave 3.11 g (8.4 mmol, 100%) of the product 142 as an off white solid. MS (M+1): m/e 366.
    Step 4: The combined reaction mixture of compound 142 (730 mg, 2 mmol), K2CO3 (2.76 g, 20 mmol) and 1,2-dibromoethane (3.74 g, 20 mmol) in DMF (6 ml) was heated at 60° C. for 17 h, and then quenched with water. The product was extracted with EtOAc (3×30 ml), and the combined extract was washed with brine (3×60 ml), dried over Na2SO4, filtered, and concentrated to give an oily residue. Purification by preparative silica gel chromatography (eluant: EtOAc) gave 640 mg (1.64 mmol, 82%) of the product 143 as an oil. MS (M+1): m/e 392.
    Step 5: To a solution of compound 143 (640 mg, 1.64 mmol) in CH3CN (13 ml) was added Cs2CO3 (1.6 g, 4.92 mmol) and PhSH (216 mg, 1.97 mmol). The reaction mixture was stirred at RT for 1 h, filtered, and the solid was washed with CH2Cl2. The filtrate was concentrated to give a yellow oil. Purification by silica gel chromatography (Biotage System, eluant: 20:1 CH2Cl2:MeOH (with 4% NH3) gave 210 mg (1 mmol, 61%) of the product 144A as a colorless oil. MS (M+1): m/e 207.


The following intermediates were synthesized by using a similar procedure:












Number
Compound
MS







144B




242


 


144C




209









EXAMPLE 42




    Step 1: To a solution of amine 145 (400 mg, 2 mmol) and Et3N (202 mg, 2 mmol) in EtOH (10 ml) was added 1-bromo-2-fluoroethane (1.27 g, 10 mmol). The reaction mixture, charged in a pressurized tube, was heated at 70° C. for 3 days. Mass spectroscopy was used to monitor the reaction. The reaction mixture was concentrated, and then water was added. The product was extracted with CH2Cl2 (3×40 ml), washed with brine (3×50 ml), dried over Na2SO4, filtered, and concentrated to give of the product 146 (1.75 mmol, 87%) as an oil, which was used without further purification. MS (M+1): m/e 247.
    Step 2: Using the procedure of step 3 from Example 31, the following compounds were synthesized:


EXAMPLE 43




    Step 1: To a solution of amide 148 (1.4 g, 7 mmol) in DMF (28 ml) was added NaH (554 mg, 23.1 mmol, 60% in oil) in portions over a period of 8 min. The reaction mixture was stirred at RT for 50 min, then Etl (3.28 g, 21 mmol) was added over a period of 2 min. The reaction mixture was stirred at RT for 15 h and then quenched with ice-water. The product was extracted with EtOAc/CH2Cl2, washed with brine (3×30 ml), dried over Na2SO4, filtered, and concentrated to give an oily residue. Purification by silica gel chromatography (Biotage System, eluant: 100:1 CH2Cl2:MeOH) gave 1.19 g of the product 149 (5.2 mmol, 74%) as an oil. MS (M+1): m/e 229.
    Step 2: Using the procedure of step 3 from Example 31, the following compounds were synthesized:














Number
Compound
MS







150A




129


 


150B




115


 


150C




143


 


150D




157


 


150E




191


 


150F




143


 


150G




155


 


150H




159


 


150I




247









EXAMPLE 44




    Step 1: Using the procedure of step 1 from Example 2, intermediate 152 was synthesized.
    Step 2: Using the procedure of step 2 from Example 2, intermediate 153 was synthesized.
    Step 3: Using the procedure of step 2 from Example 3, intermediate 154 was synthesized. MS (M+1): m/e 174.


EXAMPLE 45




    Step 1: 3,5-Dichloro-4-pyridinecarboxaldehyde (155, 0.44 g, 2.5 mmol) was mixed with allylamine (0.56 ml, 7.5 mmol), NaB(OAc)3H (1.1 g, 5 mmol) and HOAc (0.15 ml) in 1,2-dichloroethane (10 ml). The reaction mixture was stirred at RT for 20 h and then poured into saturated NaHCO3 solution (10 ml). The resulting-mixture was stirred at RT for 30 min, and the product was extracted with ether (3×40 ml). The combined organic extract was dried (Na2SO4), filtered, concentrated, and then purified by silica gel chromatography to give 0.46 g of the product 156 as an oil. MS (M+1): m/e 217.
    Step 2: Compound 156 (0.32 g, 1.47 mmol) was mixed with tetrakis (triphenylphosphine) palladium (0) (20 mg) and N,N-dimethylbarbituric acid (0.73 g, 4.4 mmol) in CH2Cl2 (35 ml). The reaction mixture was heated at reflux for 15 h. CH2Cl2 (35 ml) was added, and the organic solution was washed with saturated NaHCO3 solution, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography gave 0.25 g of the product 157 as an oil. MS (M+1): m/e 177.


EXAMPLE 46




    Step 1: To a solution of N-Boc-L-hydroxyproline ethyl ester 158 (7.0 g, 27 mmol) dissolved in CH2Cl2 (20 ml) was added 15% Dess-Martin reagent in CH2Cl2 solution (112 g). The reaction mixture was stirred at RT for 15 h. CH2Cl2 (100 ml) was added, and the organic solution was washed with 6% NaHCO3 solution, dried, filtered, and concentrated. Purification by silica gel chromatography gave 6.5 g of the product 159 as an oil. MS (M+1): m/e 258.
    Step 2: To a solution of compound 159 (1.1 g, 4.28 mmol) dissolved in dry THF (25 ml) and cooled to −78° C. was added CH3MgBr solution (3.7 ml, 1.7 M in toluene/THF) dropwise. The reaction mixture was stirred at −78° C. for 1 h, then slowly warmed up to −25° C. The reaction was quenched by the addition of 5% HCl solution and then warmed up to RT. The resulting mixture was extracted with EtOAc (2×40 ml). The combined organic extract was dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography gave 0.30 g of the product 160 as an oil. MS (M+1): m/e 274.
    Step 3: Using the procedure of step 3 from Example 31, compound 161 was synthesized. MS (M+1): m/e 174.


EXAMPLE 47




Anhydrous CeCl3 (1.85 g, 7.5 mmol) was suspended in THF (25 ml) under N2 and stirred overnight at RT. EtMgBr (2.5 ml of 3.0 M in THF, 7.5 mmol) was added dropwise, and the reaction mixture was stirred at RT for 1 h. A solution of ketone 162 (463 mg, 2.5 mmol) dissolved in THF (5 ml) was added dropwise to the suspension, and the resulting mixture was stirred at RT for 2 h. The reaction mixture was treated with EtOAc (5 ml) for 30 min at 20° C. and 2 M HCl, respectively, followed by extraction with EtOAc (2×100 ml). The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the crude intermediate. This intermediate was dissolved in minimal EtOAc and HCl (10 ml of 2 M in Et2O) was added. The reaction mixture was stirred at RT overnight to give the product 163 as a precipitate. The precipitate was filtered, washed with EtOAc, and dried in vacuo to give the product 163 as brown solid (276 mg, 73%). (M+1): m/e 116.
EXAMPLE 48




    Step 1: Compound 164 was synthesized according to the procedure of Cowden, Organic Letters (2003), 5(23), 4497-4499.
    Step 2: Using the procedure of step 3 from Example 31, compound 166 was synthesized. MS (M+1): m/e 178.


EXAMPLE 49




    Step 1: To a suspension of compound 164 (9.95 g, 107 mmol) in H2O (100 ml) and EtOH (10 ml) was added KOH (28 g, 500 mmol). The resulting mixture was heated at reflux for 3.5 h and then cooled to 0° C. Concentrated HCl (50 ml) was added with caution. The resulting mixture was concentrated, and the remaining aqueous layer was extracted with Et2O. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the product 165 (12.6 g, 88%) as a yellow liquid.
    Step 2: To a solution of compound 165 (10.2 g, 89.2 mmol) dissolved in t-BuOH (150 ml) was added Et3N (14 ml, 100.6 mmol) and diphenylphosphoryl azide (21 ml, 97.4 mmol). The reaction mixture was heated at reflux overnight and then cooled to RT. The resulting mixture was concentrated, diluted with EtOAc, washed with 1 N HCl (150 ml), satd. NaHCO3 (50 ml) and brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:6 EtOAc:hexane) gave the product 166 (4.05 g, 25%) as a white solid.
    Step 3: To a solution of compound 166 (3.0 g, 16.4 mmol) dissolved in MeOH (200 ml), and cooled to −78° C. was bubbled ozone until the light blue color persisted. Triphenyl phosphine (9.3 g, 35.5 mmol) was added, and the reaction mixture was stirred at RT overnight. The resulting mixture was concentrated. Purification by silica gel chromatography (eluant: 1:3 EtOAc:hexane) gave the product 167 (2.95 g, 97%) as a white solid. MS (M+1): m/e 186.
    Step 4: To a solution of compound 167 (3.4 g, 18.4 mmol) dissolved in THF (70 ml), and cooled to −78° C. was added L-selectride (1.0 M in THF, 22.4 ml, 22.4 mmol) dropwise. The reaction mixture was stirred at −78° C. for 2 h. Water was added, and the resulting mixture was warmed up to RT. The solution was concentrated, and water was added. The aqueous solution was extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:1 EtOAc:hexane) gave the product 168 (2.74 g, 80%) as a white foam. MS (M+1): m/e 188.
    Step 5: To a solution of compound 168 (1.0 g, 5.35 mmol) and p-nitrobenzoic acid (0.98 g, 5.88 mmol) dissolved in THF (25 ml) was added triphenylphosphine (2.1 g, 8.0 mmol) and DEAD (1.27 ml, 8.0 mmol) sequentially. The reaction mixture was stirred at RT overnight. The resulting solution was concentrated. Purification by silica gel chromatography (eluant: 1:5 EtOAc:hexane) gave the product 169 (1.53 g, 85%) as a white solid. MS (M+1): m/e 237.
    Step 6: To a solution of compound 169 (1.53 g, 4.55 mmol) dissolved in MeOH (30 ml) at 0° C. was added K2CO3 (0.24 g, 1.8 mmol). The resulting suspension was stirred at 0° C. for 2 h and then concentrated. Purification by silica gel chromatography (eluant: 1:5 EtOAc:hexane) gave the product 170 (0.71 g, 83%) as a white solid. MS (M+Na+): m/e 210.
    Step 7: To a solution of compound 170 (0.85 g, 4.5 mmol) dissolved in CH2Cl2 (40 ml) at 0° C. was added Et3N (0.94 ml g, 6.7 mmol) and mesyl chloride (0.45 ml, 5.8 mmol). The resulting solution was stirred at 0° C. for 2 h. Water was added, and the resulting mixture warmed up to RT. The aqueous layer was extracted with CH2Cl2. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the product 171 (1.0 g, 83%) as a white solid. MS (M+Na+): m/e 288.
    Step 8: To a solution of compound 171 (1.0 g, 3.8 mmol) dissolved in DMF (4 ml) was added NaN3 (378 mg, 5.8 mmol). The reaction mixture was heated at 85° C. overnight. The resulting solution was cooled to RT, concentrated, and water was added. The aqueous layer was extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the product 172 (0.78 g, 98%) as a white solid. MS (M+H+): m/e 213.
    Step 9: To a solution of compound 172 (0.78 g, 3.8 mmol) dissolved in THF (30 ml) and H2O (3 ml) was added triphenylphosphine (3.86 g, 14.7 mmol). The resulting solution was heated at reflux for 2 h, cooled to RT, and then concentrated. Purification by silica gel chromatography (eluant: 1:10 4% NH3MeOH:CH2Cl2 then 1:2 4% NH3-MeOH:CH2Cl2) gave the product 173 (0.68 g, 100%) as a white foam. MS (M+1): m/e 187.


EXAMPLE 50




    Step 1: To compound 166 (1.0 g, 5.46 mmol was added 9-BBN (0.5 N in THF, 16.4 ml, 8.2 mmol) dropwise. The reaction mixture was stirred at RT overnight. The resulting mixture was cooled to 0° C., and 2-bromopyrimidine (1.3 g, 9.2 mmol), Pd(dppf)2Cl2 (446 mg, 0.55 mmol), K2CO3 (1.13 g, 8.19 mmol), DMF (6 ml), and water (0.44 ml) were added. The reaction mixture was stirred at RT overnight. 0.5 N NaOH (50 ml) was added and the mixture extracted with CH2Cl2. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:3 EtOAc:hexane) gave a 4:1 mixture of 174A and 174B (0.8 g, 56%) as a white solid.
    Step 2: Using the procedure of step 3 from Example 31, compounds 175A and 175B were synthesized.


EXAMPLE 51




    Step 1: To a solution of compound 168 (374 mg, 2.0 mmol) dissolved in DMF (5 ml) was added NaH (60% dispersion in mineral oil, 0.2 g, 5 mmol). The reaction mixture was stirred at RT for 30 min. 2-Bromopyrimidine (350 g, 2.2 mmol) was added, and the resulting solution was stirred at RT for 4 h. The reaction mixture was concentrated, and EtOAc and satd. NaHCO3 (aq) were added. The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:1 EtOAc:hexane) gave the product 176 (0.25 g, 47%) as a white solid. MS (M+1): m/e 266.
    Step 2: Using the procedure of step 3 from Example 31, compound 177 was synthesized.


EXAMPLE 52




    Step 1: To a solution of compound 168 (374 mg, 2.0 mmol) dissolved in CH3CN (8 ml) was added N,N′-disuccinimidyl carbonate (769 mg, 3.0 mmol) and Et3N (0.84 ml, 6.0 mmol). The reaction mixture was heated at 85° C. for 1 h. The resulting solution was concentrated, and EtOAc and satd. NaHCO3 (aq) were added. The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the product 178 (0.25 g, 47%) as a white solid. MS (M+Na+): m/e 288.
    Step 2: To a solution of compound 178 (164 mg, 0.5 mmol) dissolved in CH3CN (8 ml) was added methylamine hydrochloride salt (68 mg, 1.0 mmol), Et3N (0.45 ml, 3.3 mmol), and DMAP (2 mg). The reaction mixture was stirred at RT overnight. EtOAc and satd. NaHCO3 (aq) were added. The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:50 MeOH:CH2Cl2) gave the product 179 (60 mg, 49%) as a white solid. MS(M+H+−100): m/e 145.
    Step 3: Using the procedure of step 3 from Example 31, compound 177 was synthesized.


EXAMPLE 53




    Step 1: To a suspension of compound 165 (6.22 g, 55.5 mmol) in DMF (60 ml) was added Etl (26.0 g, 166 mmol) and Cs2CO3 (36 g, 111 mmol). The reaction mixture was stirred at RT overnight, then diluted with Et2O (200 ml) and washed with water (60 ml×3). The aqueous layer was extracted with Et2O. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated to give the product 181 (7.2 g, 93%) as a light yellow oil.
    Step 2: Using the procedure for step 3 from Example 49, intermediate 182 was synthesized.
    Step 3: Using the procedure for step 1 from Example 45, intermediate 183 was synthesized. Purification of 183 by silica gel chromatography (eluant: 1:20 EtOAc:hexane) gave the product 183A, cis-isomer (2.07 g, 29%) as a colorless liquid; a mixture of cis and trans isomer (183A and 183B) (2.54 g, 35%) as a colorless liquid.


MS (M+1): m/e 233.

    Step 4: To a solution of compound 183A (2.0 g, 6.5 mmol) dissolved in EtOH (25 ml) was added 4 N HCl in dioxane (0.25 ml) and Pd(OH)2 catalyst (1.1 g). The reaction mixture was placed on a Parr shaker under 50 psi of hydrogen pressure overnight. The resulting mixture was filtered through celite. The filtrate was concentrated to give the amine HCl salt (2.3 g). The amine HCl salt (1.04 g) was suspended in CH2Cl2 (20 ml), and Et3N (3.2 ml, 23.2 mmol) and Boc2O (0.76 g, 3.48 mmol) were added. The resulting mixture was stirred at RT overnight, diluted with EtOAc and washed with 1 N HCl. The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:6 EtOAc:hexane) gave the product 184A (0.34 g, 48%, two steps) as a white solid. MS (M+Na+): m/e 266.
    Step 5: To a solution of compound 184A (0.15 g, 0.64 mmol) dissolved in CH2Cl2 (6 ml), and cooled to −78° C. was added a solution of DIBAL (1.0 M in CH2Cl2, 1.6 ml, 1.6 mmol) dropwise. The reaction mixture was stirred at −78° C. to −40° C. for 2 h. The resulting solution was warmed to RT, 10% potassium sodium tartrate solution (4 ml) was added, and stirred for 30 min. The mixture was filtered, and the filter cake was washed with CH2Cl2. The filtrate was washed with brine, dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 1:6 EtOAc:hexane) gave the product 185A (60 mg, 47%) as a colorless film. MS (M+Na+): m/e 222.
    Step 6: Using the procedure for step 6 from Example 32, intermediate 186A was synthesized. MS (M+1): m/e 215.
    Step 7: To a solution of compound 186A dissolved in THF (2 ml) was added 1 M. LAH (0.3 ml, 0.3 mmol) dropwise under a N2 atmosphere. The reaction mixture was stirred at RT overnight. The resulting solution was cooled to 0° C., and H2O (50 μl), 15% NaOH (aq) (30 μl), and H2O (0.5 ml) were added. The resulting slurry was stirred at RT for 30 min and filtered through a pad of celite. The filtrate was diluted with CH2Cl2 and washed with brine, dried (MgSO4), filtered, and concentrated to give the product 187A (34 mg, 97%) as a white solid. MS (M+1): m/e 201.


EXAMPLE 54




    Step 1: Compound 188 (8.80 g, 32 mmol) and ethyl 2-chloroacetoacetate (27.2 g, 23 ml, 160 mmol) were mixed together and heated at 180° C. for 7 h. Excess ethyl 2-chloroacetoacetate was removed by vacuum distillation. The residue was suspended in MeOH (200 ml) and stirred at 60° C. for 40 min, then at RT overnight. The solid was collected by vacuum filtration, washed with MeOH, and dried under vacuum to give 8.5 g (74%) of the product 189 as a beige solid. MS (M+1): m/e 381.
    Step 2: Using the procedure for step 1 from Example 2, intermediate 190 was synthesized. MS (M+1): m/e 459.
    Step 3: Compound 190 (0.20 g, 0.44 mmol) was suspended in 7 M NH3 in MeOH (10 ml) and heated at 55° C. for 16 h. The reaction mixture was cooled to RT and concentrated. Purification by reverse phase chromatography gave 35 mg (22%) of the title compound 191. MS (M+1): m/e 367.


EXAMPLE 55




    Step 1: Using the procedure for step 1 from Example 50, intermediate 192 was synthesized. MS (M+1): m/e 381.
    Step 2: Using the procedure for step 1 from Example 2, intermediate 193 was synthesized.
    Step 3: To a solution of compound 193 (2.0 g, 4 mmol) dissolved in DMSO (20 ml) was added NaN3 (0.29 g, 4.4 mmol). The reaction mixture was stirred at RT for 24 h. Water was added and a precipitate formed. The solid was collected by vacuum filtration, washed with water, and dried under vacuum to give 1.7 g (92%) of the product 194. MS (M+1): m/e 422.
    Step 4: To a solution of compound 194 (1.7 g, 4 mmol) dissolved in toluene (30 ml) was added trimethylphosphine (1 M in toluene, 4.4 ml, 4.4 mmol). The reaction mixture was stirred at RT for 1 h and then cooled to −20° C. 2-(tert-butoxycarbonyl-oxyimino)-2-phnylacetonitrile (BOC-ON) (1.18 g, 4.8 mmol) was added. The reaction mixture was warmed to RT and stirred for 16 h. CH2Cl2 was added and the organic solution was washed with water. The organic solution was dried (MgSO4), filtered, and concentrated. Purification by silica gel chromatography gave 1.26 g (64%) of the product 195. MS (M+1): m/e 496.
    Step 5: Using the procedure for step 1 from Example 32, intermediate 196 was synthesized. MS (M+1): m/e 482.
    Step 6 and Step 7: Using the procedure for step 2 from Example 32 and then step 3 from Example 31, the following compounds were synthesized.














Number
Compound
MS (M + 1)







197A




541


 


197B




575









EXAMPLE 56




    Step 1: Using the procedure for step 1 from Example 30, the compound 198 was synthesized.
    Step 2: To compound 198 (1.6 g, 5.6 mmol) dissolved in EtOH (50 ml) was added ethyl 2-chloroacetoacetate (2.7 g, 2.3 ml, 16.8 mmol). The reaction mixture was heated at 65° C. for 16 h and then cooled to RT. The solid was collected by vacuum filtration and washed with MeOH. Purification by silica gel chromatography gave the product 199.
    Step 3: Using the procedure for step 1 from Example 2, intermediate 200 was synthesized. MS (M+1): m/e 477.
    Step 4: Using the procedure for step 3 from Example 54, title compound 201 was synthesized. MS (M+1): m/e 383.


EXAMPLE 57




    Step 1: Using the procedure for step 1 from Example 54, compound 202 was synthesized.
    Step 2: Using the procedure for step 1 from Example 2, compound 203 was synthesized.
    Step 3: Using the procedure for step 3 from Example 55, compound 204 was synthesized.
    Step 4: Using the procedure for step 4 from Example 55, compound 205 was synthesized.
    Step 5: Using the procedure for step 1 from Example 32, intermediate 206 was synthesized.
    Step 6 and Step 7: Using the procedure for step 2 from Example 32 and then step 3 from Example 31, the following compounds were synthesized.














Number
Compound
MS (M + 1)







207A




557


 


207B




591


 


207C




611


 


207D




397









EXAMPLE 58




    Step 1: To a suspension of threonine-OMe-HCl (10.2 g, 0.06 mol) in CH2Cl2 (200 ml) was added Hunig's base (14.1 g, 19 ml, 0.11 mol) and the mixture cooled to 0° C. Compound 2 (15.0 g, 0.05 mol) dissolved in CH2Cl2 (150 ml) was added dropwise via addition funnel. The reaction mixture was stirred at 0° C. for 15 min, then at RT for 60 min. The solvent was evaporated and dilute HCl solution was added. The solid was collected by vacuum filtration and washed with MeOH. A second crop was collected by vacuum filtration of the filtrate. The combined solid was dried under vacuum to give 19.3 g (100%) of the product 208.
    Step 2: To a solution of compound 208 (7.7 g, 20 mmol) dissolved in DMSO (50 ml) and toluene (50 ml) and cooled to 0° C. was added EDCI (9.6 g, 50 mmol) and dichloroacetic acid (3.3 g, 2.1 ml, 25 mmol). The reaction mixture was stirred at 0° C. for 5 min, then at RT for 45 min. Na2S2O3 (7 g) dissolved in water (600 ml) and hexane (300 ml) was added. The reaction mixture was stirred at RT for 15 min. The solid was collected by vacuum filtration and washed with water, 1:1 water:MeOH, and then 1:1 ether:hexane. The filtrate was filtered to give additional solid. The combined solid was dried under vacuum to give 7.2 g (94%) of the product 209.
    Step 3: Using the procedure for step 1 from Example 30, the compound 210 was synthesized.
    Step 4: Using the procedure for step 1 from Example 2, the compound 211 was synthesized.
    Step 5: Using the procedure for step 2 from Example 2, the compound 212 was synthesized.
    Step 6: Using the procedure for step 3 from Example 2, the compound 213 was synthesized.
    Step 7 and Step 8: Using the procedures for step 1 and step 2 from Example 3, the following compounds were synthesized.














Number
Compound
MS (M + 1)







214A




543


 


214B




577


 


214C




383









EXAMPLE 59




    Step 1: To a solution of compound 215 (1.89 g, 10 mmol) in anhydrous CH2Cl2 (25 ml) at −20° C. was added pyridine (790 mg, 10 mmol), followed by the addition of cyanuric fluoride (3.6 ml, 40 mmol) over a period of 5 min. After 2 h at −20° C., the reaction mixture was quenched with ice-water and extracted with CH2Cl2. The combined extract was washed with brine, dried over Na2SO4, filtered, and concentrated to give 1.15 g (6 mmol, 60%) of the product 216 as a colorless liquid.
    Step 2: To a solution of compound 217 (1.54 g, 3.98 mmol) and 216 (920 mg, 4.81 mmol) in anhydrous THF (16 ml) at −78° C. was added KN(TMS)2 (20 ml, 20 mmol) over a period of 5 min. After 1 h at −78° C., the cold bath was removed and the reaction mixture was stirred for another 30 min, quenched with water, and extracted with EtOAc. The combined extract was washed with brine, dried over Na2SO4, filtered, and concentrated to give an oily residue. Purification by silica gel chromatography (Biotage System, eluant: 3:1 hexane:EtOAc) gave 0.89 g (2.1 mmol, 54%) of the product 218 as a white powder. MS (M+1): m/e 510.
    Step 3: Using the procedure for step 1 from Example 32, intermediate 219 was synthesized. MS (M+1): m/e 482.
    Step 4 and Step 5: Using the procedure for step 2 from Example 32 and then step 3 from Example 31, the following compounds were synthesized:














Number
Compound
MS







220A




507


 


220B




558


 


220C




605


 


220D




572


 


220E




537









EXAMPLE 60




    Step 1: To a solution of compound 221 (6.01 g, 42.9 mmol) dissolved in DMF (50 ml) was added N-iodosuccinimide (10.27 g, 45.6 mmol). The solution was heated at 40° C. overnight. The reaction was followed by taking 1H NMR of small amounts of the reaction mixture. Additional N-iodosuccinimide (1.34 g, 5.96 mmol) was added, and the resulting solution was stirred at RT for 2 days. The solution was diluted with EtOAc (150 ml) and washed with 0.5 N Na2S2O3 (50 ml×2). The combined aqueous wash was extracted with EtOAc (100 ml×2). The combined organic extract was dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:20 EtOAc:hexane) to give the product 222 (8.35 g, 73%) as a light yellow liquid. MS (M+1): m/e 367.
    Step 2 and Step 3: To a solution of compound 222 (8.33 g, 31.3 mmol) dissolved in CCl4 (100 ml) was added NBS (11.1 g, 62.3 mmol) and benzoylperoxide (1.3 g, 5.36 mmol). The reaction mixture was heated at reflux for 16 h and then cooled to RT. CH2Cl2 was added (400 ml) and the organic solution was washed with 0.5 N Na2S2O3 (150 ml×2). The aqueous washes were combined and extracted with CH2Cl2 (100 ml×3). The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was dissolved in acetone (300 ml) and water (150 ml), and Ag2CO3 (10.3 g, 37.4 mmol) was added. The reaction mixture was heated at reflux overnight and then cooled to RT. The mixture was filtered through a pad of celite. The filtrate was concentrated, and the remaining aqueous solution was extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:3 EtOAc:hexane) to give the product 224 (5.25 g, 60%) as a light yellow liquid. MS (M+1): m/e 283.
    Step 4: To a solution of compound 224 (4.57 g, 16.2 mmol) dissolved in CH2Cl2 (100 ml) was added Dess-Martin reagent (14 g, 33 mmol). The reaction mixture was stirred at RT overnight. The resulting solution was washed with 1 N NaOH (150 ml). The aqueous layer was separated and extracted with CH2Cl2. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:5 EtOAc:hexane) to give the product 225 (5.25 g, 60%) as a white solid. MS (M+1): m/e 281.
    Step 5: To a solution of compound 226 (14 g, 51.7 mmol) dissolved in quinoline (100 ml) was added copper (17 g, 268 mmol). The reaction mixture was heated at 180° C. for 6 h and then cooled to RT. The resulting mixture was filtered through a pad of celite and the filter cake was washed with EtOAc. The filtrate was washed with 4 N HCl (800 ml). The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:10 EtOAc:hexanes) to give the product 227 (9.25 g, 79%) as a white solid. MS (M+1): m/e 228.
    Step 6: To a solution of compound 227 (9.1 g, 40.0 mmol) dissolved in MeOH (200 ml) was added bromine (2.1 ml, 41.0 mmol). The reaction mixture was heated at 40° C. for 2 h and then cooled to RT and concentrated. The residue was purified by silica gel chromatography (eluant: 1:6 EtOAc:hexane) to give the product 228 (12.1 g, 99%) as a white solid. MS (M+1): m/e 306.
    Step 7: Pd2(dba)3 (1.69 g, 1.85 mmol) and 1.0 M PCy3 in THF (3.87 ml, 3.87 mmol) were added to a 500 ml three-neck reaction flask (evacuated and backfilled with N2). Dioxane (200 ml) was added and the mixture was evacuated and refilled with N2 again. The resulting mixture was stirred at RT for 30 min. Bromide 228 (5.91 g, 19.4 mmol), bis(pinocolo)diboron (6.88 g, 27.1 mmol), and KOAc (6.89 g, 70.0 mmol) were added sequentially. The reaction mixture was heated at 85° C. overnight and then cooled to RT. The resulting mixture was filtered through a pad of celite and the filter cake was washed with EtOAc. The filtrate was washed with H2O (100 ml). The aqueous layer was separated and extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:15 EtOAc:hexane) to give the product 229 (5.35 g, 78%) as a white solid. MS (M+1): m/e 354.
    Step 8: Boronic ester 229 (5.35 g, 15.15 mmol), 2-iodofuran 225 (4.27 g, 1.5.25 mmol), palladium acetate (172 mg, 0.77 mmol), S-Phos (682 mg, 1.65 mmol), and K3PO4 (12.5 g, 54.3 mmol) were combined in a 100 ml round bottom flask. The mixture was suspended in THF (100 ml), degassed, and refilled with N2. Water (0.55 ml, 30 mmol) was added. The resulting mixture was stirred at RT under a N2 atmosphere overnight. The reaction mixture was filtered through celite and the filter cake was washed with EtOAc. The filtrate was concentrated, and the residue was purified by silica gel chromatography (eluant 1:3 EtOAc:hexane) to give the product 230 (3.00 g, 46%) as a yellow solid. MS (M+1): m/e 433.
    Step 9: To a solution of compound 230 (1.1 g, 2.90 mmol) dissolved in CH3CN (60 ml) and CH2Cl2 (15 ml) was added BocNH2 (1.02 g, 8.71 mmol), Et3SiH (1.4 ml, 8.76 mmol), and TFA (0.43 ml, 5.79 mmol) sequentially. The reaction mixture was stirred at RT overnight. The resulting solution was diluted with CH2Cl2 and washed with 1 N NaOH (40 ml). The aqueous layer was separated and extracted with CH2Cl2. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (eluant: 1:3 EtOAc:hexane) to give the product 231 (0.95 g, 68%) as a yellow solid. MS (M+1): m/e 481.
    Step 10: Using the procedure for step 1 from Example 32, compound 232 was synthesized. MS (M+1): m/e 467.
    Step 11 and Step 12: Using the procedure for step 2 from Example 32 and for step 3 from Example 31, the following compounds were synthesized:














Number
Compound
MS







233A




492


 


233B




525


 


233C




496


 


233D




512


 


233E




506


 


233F




508


 


233G




520


 


233H




482









EXAMPLE 61







    Step 1: Starting aldehyde 230 (1.21 g, 2.79 mmol), t-(R)-butanesulfinylamide (400 mg, 3.30 mmol), and titanium ethoxide (5.6 ml, 27 mmol) were mixed in dry THF (40 ml), degassed, and refluxed under a N2 atmosphere overnight. The reaction mixture was cooled to RT and poured into brine (40 ml) with vigorous stirring. The resulting mixture was filtered through celite. The filtrate was extracted with EtOAc. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (Biotage, 40S+, eluant: 1:3 EtOAc:hexane) to give the product 234 (1.05 g, 76%) as a yellow solid. MS (M+1): m/e 497.
    Step 2: To a solution of compound 234 (0.60 g, 1.2 mmol) dissolved in dry THF (40 ml) under a N2 atmosphere and cooled to −40° C. was added a solution of MeMgBr (3 M in Et2O, 0.5 ml, 1.5 mmol) dropwise. The reaction mixture was stirred at −40° C. for 5 h and warmed up overnight. The mixture was diluted with EtOAc, poured into saturated NH4Cl (aq) and filtered through celite. The aqueous layer was separated and extracted with CH2Cl2. The combined organic extract was washed with brine, dried (MgSO4), filtered, and concentrated. The residue was purified by silica gel chromatography (Biotage, 40S+, eluant: 1:1 EtOAc:hexane) to give the separated isomer 235A (0.41 g, 66%) as a yellow solid MS (M+1): m/e 513, and isomer 235B (0.10 g, 16%) as a yellow solid MS: (M+1): m/e 513.


Step 3: Using the procedure of step 1 from Example 32, the isomers 236A (from 235A) and 236B (from 235B) were synthesized. MS (M+1): m/e 485.
Step 4 and Step 5: Using the procedure of step 2 from Example 32 and then step 3 from Example 31, the compounds 237A (from 236A) and 237B (from 236B) were synthesized. MS (M+1): m/e 472.
EXAMPLE 62







Step 1: Solid t-BuOK (2.20 g, 20 mmol) was dissolved in dry THF (50 ml) and cooled to −78° C. Compound 239 (5.34 g, 20 mmol) dissolved in dry THF (20 ml) was added while the reaction mixture was maintained at −78° C. After stirring at −78° C. for 30 min, the solution was cannulated into a vigorously stirred solution of compound 238 (20 mmol) dissolved in dry THF (50 ml) and also cooled to −78° C. The reaction mixture was stirred at −78° C. for 30 min, then 3 N aqueous HCl solution (50 ml) was added, and the reaction mixture was stirred at RT for 1 h. The resulting mixture was concentrated, and the aqueous solution was washed with Et2O (2×75 ml). The aqueous solution was concentrated by co-evaporation with toluene at temperature <40° C. The residue was dried under vacuum overnight, then suspended in MeOH (500 ml) and stirred at RT. The insoluble salt was removed by filtration. The filtrate was concentrated, and dried in a vacuum oven at 50° C. overnight to give the product 240 (6.6 g, 76%, HCl salt) as a solid. MS (M+1): m/e 397.
Step 2: To a solution of compound 241 (10 mmol) dissolved in dry THF (60 ml) and cooled to −78° C. was added compound 240 (4.3 g, 10 mmol) dissolved in dry DMF (30 ml) and then Et3N (2.7 ml, 20 mmol). The reaction mixture was stirred at RT for 3 days. The resulting mixture was concentrated and the residue was dissolved in EtOAc/Et2O. The organic solution was washed with 1 N HCl, 10% NaHCO3, and brine, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography gave 242 (4.2 g, 65%) as pale solid. MS (M+1): m/e 650.
Step 3: Compound 242 (2.0 g, 3 mmol) was dissolved in dry p-xylene (60 ml) and 7 N NH3/MeOH (2 ml) and TFA (2.2 ml) was added. The reaction mixture was heated at 150° C. for 2 h and then 0.5 N NH3/dioxane (15 ml) and AcOH (2 ml) were added. The resulting mixture was heated at 160° C. with azeotropic removal of water overnight, cooled to RT, and concentrated. Purification by silica gel chromatography (20% EtOAc in CH2Cl2) gave the product 243 (0.51 g, 27%) as a light-yellow solid. MS (M+1): m/e 631.
Step 4: Compound 243 (0.46 g, 0.73 mmol) was dissolved in AcOH (20 ml) and concentrated HCl (10 ml) and was heated to reflux for 24 h. The resulting mixture was concentrated and water (50 ml) was added. The precipitate was collected by filtration, washed with water, and dried in a vacuum oven at 50° C. overnight, to give the product 244 (0.41 g, 93%). MS (M+1): m/e 603.
Step 5: To a solution of compound 244 (0.12 g, 0.2 mmol) dissolved in dry DMF (0.5 ml) and CH2Cl2 (3 ml) was added 2,4-difluorobenzylamine (0.05 ml, 0.4 mmol), DIPEA (0.07 ml, 0.4 mmol), and HATU (0.114 g, 0.3 mmol). The resulting mixture was stirred at RT overnight and then concentrated. The residue was dissolved in DMF (2 ml) and purified by Gilson reverse phase prep HPLC to give the product 245 (0.081 g, 56%). MS (M+1): m/e 728.
Step 6: Compound 245 (0.080 g, 0.11 mmol) was dissolved in Et2NH (2 ml) and CH3CN (2 ml) and stirred at RT for 30 min. The resulting mixture was concentrated, and the residue was purified by Gilson reverse phase prep HPLC. The product was treated with HCl in ether, then dried in a vacuum oven at 50° C. overnight to give the product 246 (0.052 g, 94%) as a di-HCl salt. MS (M+1): m/e 506.
EXAMPLE 63




Step 1: To a solution of compound 247 (38 g, 0.19 mol) dissolved in CH2Cl2 (450 ml) and cooled to 0° C. was added Et3N (35 ml, 0.25 mol) and t-Boc anhydride (54 g, 0.25 mol). The reaction mixture was stirred at RT overnight. The resulting mixture was diluted with CH2Cl2, washed with 1 N HCl solution, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography gave the product 248 (47 g, 96%). MS (M+1): m/e 260.
Step 2: To a solution of compound 248 (1.55 g, 6 mmol) dissolved in dry THF (60 ml) and cooled to 0° C. was added triphenylphosphine (2.0 g, 7.8 mmol), diethyl azodicarboxylate (1.3 ml, 7.8 mmol) dropwise, and then diphenylphosphoryl azide (1.7 ml, 7.8 mmol). The reaction mixture was stirred at RT overnight, then diluted with ether. The organic solution was washed with saturated NaHCO3 and brine, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography (eluant: 15-20% EtOAc in hexane) gave compound 249 (1.7 g, 100%). MS (M+1): m/e 285.
Step 3: To a solution of compound 249 (0.5 g, 1.76 mmol) dissolved in THF (40 ml) was added 10% Pd/C catalyst (0.25 g). The reaction mixture was stirred under H2 (1 atm) at RT overnight. The resulting mixture was filtered, and the filtrate was concentrated to give the product 250 (0.45 g, 100%). MS (M+1): m/e 259.
Step 4 and 5: To a solution of compound 250 (0.13 g, 0.5 mmol) dissolved in CH2Cl2 (1 ml) was added DIPEA (0.2 ml) and cyclopropanecarbonyl chloride (0.053 ml, 0.5 mmol). The reaction mixture was stirred at RT for 2 h. The resulting mixture was diluted with EtOAc. The organic solution was washed with 1 N HCl, saturated NaHCO3, and brine, dried, filtered, and concentrated. Purification by silica gel chromatography gave the product 251. Compound 251 was treated with 4 N HCl in dioxane at RT for 4 h. The resulting mixture was concentrated, and the residue was dried under vacuum for 2 days to give the product 252 as the HCl salt (0.1 g, 76%). MS (M+1): m/e 227.
Step 6: To a solution of compound 248 (3.1 g, 12 mmol) dissolved in dry THF (100 ml) and cooled to 0° C. was added triphenylphosphine (4.0 g, 15 mmol), DEAD (2.5 ml, 15 mmol) dropwise, and LiBr (5 g, 57 mmol). Within 2 min, all the LiBr dissolved. The resulting clear yellow solution was stirred at RT overnight. The reaction mixture was diluted with EtOAc, washed with water, dried (Na2SO4), filtered, and concentrated. Purification by silica gel chromatography gave the product 253 (2.15 g, 56%). MS (M+1): m/e 323.
Step 7: To a solution of compound 253 (2.1 g, 6.5 mmol) dissolved in DMSO (15 ml) was added NaN3 (0.46 g, 7 mmol). The resulting mixture was stirred at RT for 2 days. Water was added to the mixture and product was extracted with ether (3×40 ml). The combined organic layer was washed with brine, dried (Na2SO4), filtered, and concentrated to give the product 254.
Step 8: Using the procedure of step 3, compound 255 was synthesized. MS (M+1): m/e 259.
Step 9: To a solution of compound 255 (0.26 g, 1 mmol) dissolved in DMF (2 ml) was added Et3N (0.28 ml, 2 mmol) and 2-bromopyrimidine (0.16 g, 1 mmol). The reaction mixture was heated at 100° C. overnight then cooled to RT. The resulting mixture was diluted with DMSO (3 ml) and purification by reverse phase Gilson prep HPLC gave the product 256 (0.18 g, 54%). MS (M+1): m/e 337.
Step 10: Using the procedure of step 5, compound 257 was synthesized. MS (M+1): m/e 237.
EXAMPLE 64







Step 1: Compound 258 (4.14 g, 13.6 mmol), CuI (288 mg, 0.37 mmol), NaI (4.32 g, 28.8 mmol), and sym-dimethylethylenediamine (0.38 ml, 0.72 mmol) were suspended in toluene (12 ml). The reaction mixture was heated in a sealed tube at 125° C. for 48 h. The resulting mixture was cooled to RT and filtered through celite. The filtrate was concentrated, and the residue was purified by silica gel chromatography (eluant: 1:10 EtOAc:hexane) to give the product 259 (4.06 g, 85%) as a beige liquid. MS (M+1): m/e 354.
Step 2: Compound 259 (3.55 g, 10.0 mmol), pyrazole 260 (2.31 g, 15 mmol), trans-1,2-di(methylamine)cyclohexane (450 mg, 3.17 mmol), CuI (190 mg, 1.0 mmol), and K2CO3 (4.14 g, 30 mmol) were suspended in toluene (40 ml). The reaction mixture was heated in a sealed tube at 125° C. for 10 days. The resulting mixture was cooled to RT and filtered through celite. The filtrate was concentrated and the residue was purified by silica gel chromatography (eluant: 1:1 EtOAc:hexane) to give the starting compound 259 (2.1 g, 46%) and the product 261 (1.29 g, 45%) as a white solid. MS (M+1): m/e 380.
Steps 3, 4, and 5: Using procedures similar to that of step 1 from Example 2, step 3 from Example 55, and step 4 from Example 55, intermediate 262 was synthesized.
MS (M+1): m/e 495.
Step 6: Using a procedure similar to that of step 1 from Example 32, compound 263 was synthesized. MS (M+1): m/e 467.
Steps 7 and 8: Using procedures similar to that of step 2 from Example 32 and step 3 from Example 31, the following compounds were synthesized.












Number
Compound
MS







264A




492


 


264B




506









EXAMPLE 65




Using procedures from Examples 5 and 6, compound 266 was synthesized. MS (M+1): m/e 439.
The pharmacological activity of the compounds of the invention was measured by the following assays.
PDE4 Screening Assay
1. Human PDE4 Enzyme
The neutrophils were isolated from human blood using a standard procedure, then homogenized with a glass-glass homogenizer in a buffer containing 20 mM Tris/HCl (pH 8.0), protease inhibitor cocktail tablet (Cat. No. 1836145/Boehringer Mannheim), 2 mM EDTA, 1% Triton X-100 and 0.5% deoxycholate. After stirring for 2 h at 4° C., the samples were centrifuged at 100,000 g for 1 h. The supernatants were collected, filtered and applied to Mono Q column chromatography. The fractions containing the activity of hydrolyzing cAMP were determined and pooled as the enzymatic source of the PDE4 screening assay.
2. PDE4 Assay and Compound Screening
The PDE4 assays were performed using Phosphodiesterase [3H]cAMP SPA enzyme assay kits and its procedures (Cat. No. TRKQ 7090, Amersham). The assay procedures are described briefly as follows. The diluted PDE4 enzyme, 10× assay buffer and water were mixed at a ratio of 1:1:6 (10 μl/10 μl/60 μl). 80 μl aliquots of this mixture were added into the test wells of a 96-well Microlite plate (Cat. No. 7416, ThermoLabsystems). Enzyme dilution buffer, instead of the diluted enzyme, and water were added into the wells of negative control (background). 10 μl test compounds in 10% DMSO, standard inhibitor in 10% DMSO or 10% DMSO (for positive and negative controls) were added into the corresponding wells, respectively. After a 10 min incubation at RT, the reactions were initiated by addition of 10 μl pre-diluted [3H]cAMP into each well, then incubated at 30° C. for 30 min. The reactions were stopped by addition of 50 μl SPA beads into the test wells, then counted in a β-counter over 30 min  ̃24 hr.
10× Assay Buffer: 500 mM Tris/HCl pH 7.5, 83 mM MgCl2, 17 mM EGTA
[3H]cAMP: [3H] cAMP (40-60 Ci/mmol) is diluted at a 1:200 ratio with water. The final concentration is 0.005 μCi/μl
Yttrium SPA Beads: 500 mg of beads was reconstituted with 28 ml of water, stored at 4° C.
PDE10 and 11 Screening Assay
PDE10 (human recombinant PDE10A2, expressed in Sf9 insect cells by the baculovirus expression technique) was assayed using [3H]cGMP PDE SPA Assay kit (Amersham) at a final concentration of cGMP of 0.7 μM. PDE11 (human recombinant PDE11A3, expressed in Sf9 insect cells by the baculovirus expression technique) was assayed using [3H]cAMP PDE SPA Assay kit (Amersham) at a final concentration of cAMP of 0.0125 μM. Compounds were evaluated at 0.1-10,000 nM in 2% DMSO and 0.1% BSA from a stock solution of 4 mM in 100% DMSO. All assays were performed in duplicate, and each set of experiments was performed at least twice. Analysis of dose-response data and calculation of IC50 values were performed using GraphPad Prism.
PBMC (Peripheral Blood Mononuclear Cell) Preparation and TNF Inhibition Assay
This protocol was modified from Prabhaker et al. (Int. J. Immunopharmac, Vol 16, No 10 pp 805-816, 1994. Smithkline Beecham Pharmaceuticals).

    1. Human blood was collected from internal donors. The plasma was separated from red blood cells by mixing with 6% dextran (4 ml for a 15-ml blood) and a 40 min-incubation at 37° C.
    2. 10 ml plasma was then layered on 9 ml Ficoll-paque (Cat. No. 17-1440-03, Amersham) in a centrifuge tube.
    3. After a centrifugation at 1500 rpm for 45 min, PBMC was removed from the interface.
    4. PBMC was washed twice with PBS and counted.
    5. PBMC was suspended in RPMI medium containing 2.5% heat-inactivated FCS (Hyclone laboratories Inc. Logan, Utah, USA), Penicillin and streptomycin, and the cell volume was adjusted to 1×106 cell/ml.
    6. 0.5 ml cells were transferred into each well of a 24 well plate.
    7. After one hour incubation at 37° C., the cells were pre-treated for 1 h with 5 μl 10% DMSO (control) and 5 μl test compounds at various concentrations (100 fold stock solutions in 10% DMSO).
    8. LPS was added to stimulate TNF production at a final concentration of 100 ng/ml (E. coli 055:13S, SIGMA).
    9. The cells were stimulated for 14-16 h at 37° C.
    10. The supernatants were removed and transferred to new tubes. TNF alpha levels were assayed by ELISA using Human TNF-α ELISA kit (Cat. No. KHC3012, Biosource) and its procedures with an optimal dilution. (1:10→1:100 dilution).

In vivo TNFα Assay



C57BI/6 mice were injected with 25 ug of LPS (LPS O55-B5, Sigma: L2880) by the intraperitoneal route. One hour prior to injection of the LPS, mice were treated orally with the PDE4 compounds at the selected doses. Ninety min after the LPS challenge, the mice were euthanized, and blood was collected through a heparinized syringe tip into Capijet T-MGA tubes. The blood was centrifuged for 10 min in a microcentrifuge at maximum speed (−13,000 rpm), and the serum was collected and analyzed for TNFα protein using an R&D ELISA kit.
Lipopolysaccharide (LPS) in vivo Assay
Male Sprague/Dawley rats (200-250 g) were purchased from Charles River Laboratories. Prior to use, the animals were permitted unrestricted access to food and water. Test compounds were delivered by gavage 5 hours prior to LPS-challenge. Compounds were suspended in a 0.4% methylcellulose vehicle with the same vehicle being given to control animals.
LPS-treatment: Animals were anethesized by inhalation of isoflurane, supplemented with oxygen (flow rate 1.0 ml/min). Once anesthetized, animals were placed supine and the trachea visualized using a small laryngoscope. Animals then received either 0.1 ml of saline or 0.1 ml of a 100 μg/ml lipopolysaccharide solution (LPS; E. coli) in saline by use of a Penn-Century Microspray needle (Penn-Century, Philadelphia, Pa.). Animals were allowed to recover on a heat pad, returned to housing and permitted access to food and water ad libitium. Sixteen hours after LPS-challenge, animals were anesthetized with an intra-peritoneal injection of the combination of ketamine/xylazine (10:1, 200 mg/kg ketamine, 20 mg/kg xylazine). After reaching anesthesia, animals were surgically prepared for bronchial lavage by inserting a tracheal cannula. Animals were lavaged with 2×2 ml of phosphate buffered saline, pH 7.2 (PBS). Routine recovery of BAL fluids did not significantly differ between animals with >80% of instilled volume recovered. Afterwards, animals were euthanized by surgically opening the thoracic cavity and cutting the diaphragm to assure lung collapse. Bronchial lavage (BAL) fluid was analyzed for cellular contents as described below.
BAL samples: Bronchial lavage (BAL) fluid was spun at 350×g for 10 min at 4° C. One ml of supernatant was removed and stored at −20° C. until analyzed for cytokine levels. Remaining fluid was aspirated and the cell pellet lysed for residual erythrocytes and resuspended in PBS, pH 7.2 containing 10 ug/ml of DNase I. Afterwards, the cell suspension was centrifuged at 350×g for 10 mins at 4° C., the supernatant aspirated and the cell pellet resuspended in 1 ml of PBS with 10 ug/ml DNase 1 and 5% heat inactivated fetal bovine serum. Cytospin slide preparations were made and stained with Hema3TM staining system (Fisher Scientific, Springfield N.J.). Differential cell counts were performed using standard histological parameters and at least 200 cells were enumerated. Total cell counts were performed using a Neubauer chamber.
Assay Procedure for Testing of Dermatitis in Dogs:
Five dogs are selected for each treatment group. Administration of experimental medications begins and continues through the end of the animal phase of the experiment. After three days, all dogs are sedated using medetomadine intravenously. An approximately 5 cm by 13 cm area is shaved on the lateral thorax of each dog. 1 cc of lidocaine is injected subcutaneously, and then two 8 mm punch biopsies are taken to act as Time 0 controls. Biopsy sites are closed with simple interrupted sutures of 3-0 Nylon suture.
Ten intradermal injections are given (five rows of two injections)—two injections are of phosphate buffered saline (PBS), and the remaining eight injections are of rabbit IgG antibody to dog IgE. Each injection is 0.05 ml. The total dose of anti-IgE per injection is 7 μg, as previously determined to be optimal. After injection, sites are observed and sampled. After injection and between all future samples, all dogs wear a protective garment (Quick Cover incision cover, Four Flags over Aspen) to prevent disturbance of the injection and/or biopsy sites.
The test compounds are compounds of formula I; the negative control is phosphate buffered saline (PBS); the positive control is commercially available prednisone tablets. Tablets are given orally by placement in the back of the mouth. Liquids are given by syringe to place the test article toward the back of the mouth. The dog's mouth may be held closed to ensure that all of the test article is swallowed. Plasma samples are analyzed for the concentration of test compound from the dogs treated with the active compounds. Samples from the negative control and prednisone treated dogs need not be analyzed.
Anti-IgE Site Observations: Sites of anti-IgE injection are examined and evaluated for erythema and wheal formation. At the 20 min observation time, the two PBS sites and the two 6 hr. biopsy sites are measured. At the other post-injection times, the two PBS sites and the corresponding biopsy sites are measured. If the size of the reaction is not consistent across sites in the same dog, then all sites that have not been previously biopsied will be measured. Wheals will be measured by calipers in two orthogonal dimensions as well as measured for thickness.
Collection of Skin Samples: Two 8 mm punch biopsies are taken of the sites injected with anti-IgE. One biopsy is placed in RNA isolation buffer and the other biopsy is bisected. One half goes into a standard 10% formalin solution for routine histopathological analysis and the other is deposited in Optimal Cutting Temperature Medium and quick frozen in liquid nitrogen, then maintained at −70° C. for immunohistochemical staining with Luna's stain for eosinophils, and Alcian Blue with Nuclear Fast Red counterstain for mast cells. Using manual or computerized morphometric analysis, the extent of infiltration by the following specific leukocytes is quantitated: CD 1a+c, IgE, CD3, 4+8, TCR alpha/beta and gamma/delta, TNF alpha, and TSLP. Cytokine analysis is to determine the presence of the following: TNF alpha, IL4, IL13, IL2, IFN gamma, and Thymic stromal lymphopoietin.
Allergic Brown Norway (BN) Rat Model:
Inbred male BN rats weighing 150 to 200 g were obtained from Charles River Laboratory (Wilmington, Mass.). Prior to use, the animals were allowed food and water ad libitum. The test compounds were administered 5 h prior to antigen challenge either by oral or inhalational route, as detailed in the “delivery of test compounds” section.
Sensitization and Antigen Bronchoprovocation
The animals were divided into two main groups viz. an alum group and an antigen group. In the antigen group, animals were sensitized by an intra-peritoneal (i.p.) injection of 1 ml alum-precipitated antigen containing 20 μg of ovalbumin (OVA, grade III; Sigma chemical Co., St Louis, Mo.) and 8 mg of Al(OH)3 suspended in 0.9% saline vehicle. A booster injection of this alum-OVA mixture was given again 7 days later. Animals belonging to the alum group received injections containing alum only. Seven days after the second injection, animals were exposed to aerosolized antigen bronchoprovocation which was performed by placing the rats in an enclosed plexiglass chamber (21 liters) and exposing the rats to aerosolized OVA (1%) for 30 min. The aerosolized OVA was produced by an ultrasonic nebulizer (DeVilbiss, Somerset, Pa., USA; Model Ultra-Neb 99) at a flow rate of approximately 8 liters/min. Twenty four hours after aerosolized OVA challenge, the animals were euthanized with an overdose of pentobarbital sodium. The trachea was exteriorized and intubated, and the lungs were lavaged with two aliquots of 3 ml of physiological saline. The bronchoalveolar lavage fluid (BALF) thus collected was subjected to cell enumeration. Ten microliter of the BALF was utilized to manually enumerate the total white cells using a hemocytometer. One hundred microliter of BALF was used to prepare cytocentrifuge which was stained with Hema3TM staining system (Fisher Scientific, Springfield, N.J.) to identify and enumerate differential white blood cells such as eosinophils, neutrophils, mononuclear cells and epithelial cells. A total of 200 cells were enumerated from each cytocentrifuge. The ability of the compound to inhibit recruitment of inflammatory cells into the airways is reported.
Delivery of Test Compounds:
Oral administration: the compounds were dissolved in 0.4% methylcellulose and delivered to animals orally @3 ml/kg. An equivalent volume of 0.4% methylcellulose was given to both negative (alum group) and positive (antigen) control groups.
Intra-tracheal administration: the appropriate dose of the compound was mixed with lactose powder to achieve a final amount of 3 mg, which was delivered intra-tracheally to anesthetized animals. Animals were held in an upright position for 3-4 min and were allowed to recover from anesthesia before returning to their cages.
Using the procedures described above in the PDE 4, PDE10 and PDE11 screening assays, compounds of formula I were found to have IC50 values for PDE4 in a range of 0.01 to 500 nM, with preferred compounds having a range of 0.01 to 100 nM, more preferably 0.01 to 10 nM, and most preferably 0.01 to 3 nM. Compounds of formula I are preferably selective PDE4 inhibitors compared to PDE10 and PDE11: preferably the IC50 values for PDE10 and PDE 11 are 100 to 300 times the values for PDE4.
Representative compounds of formula I have the following IC50 values for PDE4:














Compound
IC50



No.
(nM)



















13-106
0.14



26-42
0.07



26-92
0.01



26-177
3



26-241
0.2



26-293
0.5



26-417
1.4



26-444
0.03



38-3
1.8










For preparing pharmaceutical compositions from the compounds described by this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powders and tablets may be comprised of from about 5 to about 70 percent active ingredient. Suitable solid carriers are known in the art, e.g. magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration.
For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredient is dispersed homogeneously therein as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.
Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection.
Liquid form preparations may also include solutions for intranasal administration.
Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.
The compounds of the invention may also be deliverable transdermally. The transdermal compositions can take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.
Preferably the compound is administered orally or via inhalation.
Preferably, the pharmaceutical preparation is in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component, e.g., an effective amount to achieve the desired purpose.
The quantity of active compound of formula I in a unit dose of preparation may be varied or adjusted from about 0.1 mg to 1000 mg, more preferably from about 1 mg to 300 mg, according to the particular application.
The actual dosage employed may be varied depending upon the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired.
The amount and frequency of administration of the compounds of the invention and the pharmaceutically acceptable salts thereof will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the patient as well as severity of the symptoms being treated. A typical recommended dosage regimen for compounds of formula I is oral administration of from 10 mg to 2000 mg/day preferably 10 to 1000 mg/day, in two to four divided doses to provide relief from allergic and inflammatory diseases or the other disease or conditions listed above.
The doses and dosage regimen of the additional agents administered in the combinations of the invention will be determined by the attending clinician in view of the approved doses and dosage regimen in the literature, e.g., the package insert, taking into consideration the age, sex and condition of the patient and the severity of the disease.
While the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present invention.

## Claims

1. A compound represented by the structural formula




or a pharmaceutically acceptable salt thereof, wherein



R is H or alkyl;
X is O or S;
R1 is H, alkyl, cycloalkyl, cycloalkyl(C1-C4)alkyl-, —CH2F, —CHF2, —CF3, —C(O)alkyl or —C(O)NR18R19;
R3 and R4 are independently selected from the group consisting of H, alkyl, hydroxyalkyl and —C(O)Oalkyl;
R5 and R6 are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH2F, —CHF2, —CF3, —C(O)OH, —C(O)Oalkyl and —C(O)NR43R44;
t is 1 or 2;
R7 is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R17-phenyl)alkyl or —CH2—C(O)—O-alkyl;
R8 is H, alkyl, alkenyl, alkoxy, alkoxyalkyl, hydroxyalkyl, dihydroxyalkyl, alkyl-NR18R19, cyanoalkyl, haloalkyl, R23-heteroaryl, R23-heteroarylalkyl, R36-heterocycloalkyl, (R36-heterocycloalkyl)alkyl, R17-phenyl, (R17-phenyl)alkyl, R17-naphthyl, (R17-naphthyl)alkyl, R17-benzyloxy, -alkyl-C(O)—NR18R19, -alkyl-C(O)—N(R30)—(R23-heteroaryl), -alkyl-C(O)—(R17-phenyl), -alkyl-C(O)—(R36-heterocycloalkyl); -alkyl-N(R30)—C(O)Oalkyl, -alkyl-N(R30)—C(O)—NR18R19, -alkyl-N(R30)—C(O)alkyl, -alkyl-N(R30)—C(O)-(fluoroalkyl), -alkyl-N(R30)—C(O)—(R39-cycloalkyl), -alkyl-N(R30)—C(O)—(R17-phenyl), -alkyl-N(R30)—C(O)—(R23-heteroaryl), -alkyl-N(R30)—C(O)-alkylene-(R23-heteroaryl), -alkyl-NH—SO2—NR18R19, -alkyl-N(R30)—(R17-phenyl), -alkyl-N(R30)—(R23-heteroaryl), -alkyl-O—(R17-phenyl), -alkyl-O—(R23-heteroaryl), -alkyl-N(R30)—SO2-alkyl, alkylthioalkyl-, alkyl-SO2-alkyl-, (R35-phenylalkyl)-S-alkyl-, (hydroxyalkyl)-S-alkyl-, (alkoxyalkyl)-S-alkyl-, -alkyl-CO2-alkyl, R45-hydroxyalkyl, dihydroxyalkyl substituted by R17-benzyloxy, dihydroxyalkyl substituted by R17-phenyl, alkoxyalkyl substituted by R17-phenyl, (R17-phenyl)alkyl substituted by —CO2alkyl, (R17-phenyl)alkyl substituted by —C(O)N(R30)2, alkyl substituted by (R23-heteroaryl) and —C(O)NR37R38, haloalkyl substituted by CO2alkyl, R12-cycloalkyl, (R12-cycloalkyl)alkyl,







or R7 and R8 and the nitrogen to which they are attached together form a ring system selected from the group consisting of








comprises an R35-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl or pyrrolidinyl ring;
p is 0 or 1;
q is 0 or 1;
the dotted line represents an optional double bond;
R9 is H, halo, alkyl, cycloalkyl, —CH2F, —CHF2 or CF3;
R10, R11, and R13 are independently selected from the group consisting of H and halo;
R12 is 1-3 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl, —(CH2)n—N(R30)—C(O)-cycloalkyl, —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)C(O)Oalkyl, —(CH2)n—N(R30)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)—NR18R19, —(CH2)n—C(O)—NR18R19, R17-phenyl, R35-heteroarylalkyl, R35-heteroaryloxy, —C(O)-heterocycloalkyl, —O—C(O)-heterocycloalkyl, —O—C(O)—NR18R19, —NH—SO2-alkyl, —NH—C(═NH)NH2, and





or two R12 substituents on the same carbon form ═O, ═NOR30 or ═CH2;
R14 is 1 or 2 substituents independently selected from the group consisting of H, OH, halo, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, —CF3, CN, R17-phenyl, (R17-phenyl)alkyl, —NR18R19, alkyl-NR18R19, —(CH2)n—C(O)OH, —(CH2)n—C(O)Oalkyl, —(CH2)n—C(O)alkyl, —(CH2)n—C(O)(R35-phenyl), —(CH2)n—C(O)(R23-heteroaryl), —(CH2)n—C(O)NR18R19, (CH2)n—C(O)N(R30)—(CH2)n—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)—C(O)-(fluoroalkyl), —(CH2)n—N(R30)—C(O)-(cycloalkyl), —(CH2)n—N(R30)—C(O)(R35-phenyl), —(CH2)n—N(R30)—C(O)(R23-heteroaryl), —(CH2)n—N(R30)C(O)NR18R19, —(CH2)n—N(R30)—C(O)Oalkyl, —(CH2)n—N(R30)cycloalkyl, —(CH2)n—N(R30)(R17-phenyl), —(CH2)n—N(R30)(R23-heteroaryl), —(CH2)n—N(R18)SO2alkyl, —(CH2)n—N(R20)SO2—(R17-phenyl), —(CH2)n—N(R30)SO2—CF3, —CH2S(O)0-2(R35-phenyl), —(CH2)n—OC(O)N(R30)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, (R23-heteroaryl)oxy, (R23-heteroaryl)amino, —CH(OH)—(R17-phenyl), —CH(OH)—(R23-heteroaryl), —C(═NOR30)—(R17-phenyl), —C(═NOR30)—(R23-heteroaryl), morpholinyl, thiomorpholinyl,




w is 0 or 1;
or two R14 substituents and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
each n is independently 0, 1, 2 or 3;
R15 is H, alkyl, cycloalkyl, (cycloalkyl)alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, —C(O)Oalkyl, —C(O)O(R30-cycloalkyl), -alkyl-C(O)O-alkyl, —C(O)O-alkylene-(R35-phenyl), R17-phenyl, (R17-phenyl)alkyl, —CH—(R17-phenyl)2, R23-heteroaryl, —(CH2)n—C(O)NR18R19, —SO2-alkyl, —SO2-cycloalkyl, —SO2—CF3, —SO2—(R35-phenyl), —SO2—NR18R19, —C(O)alkyl, —C(O)-(fluoroalkyl), —C(O)—C(CH3)(CF3)2, —C(O)—(R17-phenyl), —C(O)—(R23-heteroaryl), —C(O)-hydroxyalkyl, —C(O)alkoxyalkyl, —C(O)—(R39-cycloalkyl), —C(O)-alkylene-(R17-phenyl), —C(O)-alkylene-R23-heteroaryl), —C(O)-alkylene-S—C(O)alkyl, —C(═S)—(R17-phenyl), hydroxyalkyl substituted by R17-phenyl, hydroxyalkyl substituted by R23-heteroaryl, alkoxyalkyl substituted by R17-phenyl, alkoxyalkyl substituted by R23-heteroaryl,





wherein z is 0, 1 or 2;
R16 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, R17-phenyl, (R17-phenyl)alkyl, (R23-heteroaryl)alkyl, hydroxyalkyl, alkoxyalkyl and —(O)Oalkyl, or two R16 groups and the carbon to which they are both attached form —C(O)—;
R17 is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, cycloalkyl, —OH, hydroxyalkyl, alkoxy, alkoxyalkyl, —CN, —CF3, —OCF3, —OCHF2, —OCH2F, —C(O)OH, —C(O)Oalkyl, —C(O)O—(R36-phenyl), —C(O)alkyl, —C(O)—(R35-phenyl), —SOalkyl, —SO2alkyl, —O2—CF3, alkylthio, —NR43R44, -alkyl-NR43R44, R35-phenyl, R35-phenoxy, R35-heteroaryl, R35-heteroaryloxy, R36-heterocycloalkyl, —C(O)—(R36-heterocycloalkyl), hydroxyalkyl-NH—, —C(O)N(R30)2, —N(R43)—(R35-cycloalkyl) and —C(═NOR30); or two R17 substituents on adjacent carbon atoms together form —O—CH2—O—, —O—(CH2)2—O—, —(CH2)2—O— or —O—CH2—O—CH2—;
R18 and R19 are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, R17-phenyl, (R17-phenyl)alkyl, naphthyl and cycloalkyl;
R20 is H, alkyl, or cycloalkyl;
R22 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF3, —NH2 and R35-phenyl;
R23 is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF3, —NR18R19, —CN, —C(O)Oalkyl, —SO2-alkyl, —NHSO2-alkyl, R35-phenyl, R35-heteroaryl, morpholinyl, and —(CH2)n—C(O)—N(R30)2;
R24 is H, OH or alkoxy; or when the optional double bond is present, R24 and the adjacent carbon atom form the double bond;
R25 is H or R35-phenyl;
R27 is 1 to 3 substituents independently selected from the group consisting of H, halo, OH, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, haloalkyl, —CN, —C(O)OH, —C(O)Oalkyl, —C(O)N(R30)(R18), —C(O)—(R36-hetercycloalkyl), R17-phenyl, (R17-phenyl)-alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, (R23-heteroaryl)oxy, (R23-heteroaryl)amino NR18R19, NR18R19-alkyl, —(CH2)n—N(R30)—C(O)alkyl, —(CH2)n—N(R30)—C(O)-(fluoroalkyl), —(CH2)n—N(R30)—C(O)alkoxyalkyl, —(CH2)n—N(R30)—C(O)(cycloalkyl), —(CH2)n—N(R30)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)—(R23-heteroaryl), —(CH2)n—N(R30)—C(O)O-alkyl, —(CH2)n—N(R30)—C(O)O—(CF3alkyl), —(CH2)n—N(R30)—C(O)O—(R39-cycloalkyl), —(CH2)n—N(R30)—C(O)O-alkylene-cycloalkyl, —(CH2)n—N(R30)—C(O)—N(R30)(R20), —(CH2)n—N(R30)—SO2-alkyl, —(CH2)n—N(R30)—SO2—CF3, —(CH2)n—N(R30)—SO2—N(R30)2 and




or two R27 groups and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
R28 is H, alkyl, R35-benzyl or -alkyl-C(O)O-alkyl;
R29 is alkyl, haloalkyl, —C(O)Oalkyl, —C(O)alkyl, —C(O)CF3, —C(O)—(R12-cycloalkyl), —C(O)—(R17-phenyl), —C(O)—(R23-heteroaryl), —C(O)—(R36-hetercycloalkyl), —SO2-alkyl, —SO2—(R35-phenyl), —C(O)NR18R19, R35-phenyl, (R35-phenyl)alkyl or R23-heteroaryl;
R30 is independently selected from the group consisting of H, alkyl, R35-benzyl and R35-phenyl;
R31 is H, alkyl, R36-benzyl or phenoxyalkyl;
R33 is H, OH or alkoxy;
R34 is H, alkyl, hydroxyalkyl, alkoxyalkyl or —C(O)Oalkyl;
R35 is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, OH, —CF3, alkoxy, —CO2alkyl and —N(R43)(R44);
R36 is 1 or 2 substituents independently selected from the group consisting of H, alkyl, R17-phenyl, —OH, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl and —NR18R19; or two R36 groups and the carbon to which they are both attached form —C(═NOR30)— or —C(O)—;
R37 and R38 are independently selected from the group consisting of H and alkyl, or R37 and R38 together are —(CH2)3— or —(CH2)4—, and together with the nitrogen to which they are attached, form a ring;
R39 is H, OH, alkyl, alkoxy, or CF3;
R40 is —OR30 or —NHC(O)alkyl;
R41 is H or —SO2alkyl;
R42 is —(CH2)n—(R35-phenyl), —(CH2)n—(R23-heteroaryl), —C(O)Oalkyl or —C(O)alkyl;
R43 and R44 are independently selected from the group consisting of H and alkyl; and
R45 is 1 or 2 substituents independently selected from the group consisting of halo, alkoxyalkyl, —CO2alkyl, R17-phenyl, R23-heteroaryl and cycloalkyl.

2. A compound of claim 1 wherein X is O.

3. A compound of claim 2 wherein

4. A compound of claim 3 wherein R10, R11 and R13 are each H; R1 is H, alkyl, cycloalkyl or —CF3; and R9 is H, alkyl or —CF3.

5. A compound of claim 4 wherein R10, R11 and R13 are each H, R1 is alkyl, and R9 is —CF3.

6. A compound of claim 5 wherein t is 1, R5 is H, R6 is H, alkyl or hydroxyalkyl, and R3 and R4 are each H or alkyl.

7. A compound of claim 6 wherein R7 is H, alkyl, cycloalkyl, hydroxyalkyl or alkoxyalkyl, and R8 is R12-cycloalkyl, (R12-cycloalkyl)alkyl, R45-hydroxyalkyl, R17-phenyl, (R17-phenyl)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, -alkyl-N(R30)—C(O)—NR18R19, -alkyl-N(R30)—C(O)alkyl, -alkyl-N(R30)—C(O)—(R17-phenyl), -alkyl-N(R30)—C(O)—(R23-heteroaryl), -alkyl-N(R30)—(R23-heteroaryl),

8. A compound of claim 7 wherein R8 is R12-cycloalkyl, R45-hydroxyalkyl, (R17-phenyl)alkyl, R23-heteroaryl, (R23-heteroaryl)alkyl, -alkyl-N(R30)—(R23-heteroaryl), -alkyl-N(R30)—C(O)alkyl,




wherein p is 0, or




wherein p is 1.

9. A compound of claim 8 wherein R12 is OH, —(CH2)n—N(R30)—C(O)-cycloalkyl Or —(CH2)n—N(R30)—(R23-heteroaryl), R45 is R17-phenyl, or R29 is heteroaryl, —C(O)alkyl or —C(O)cycloalkyl.

10. A compound of claim 6 wherein R7 and R8 and the nitrogen to which they are attached form

11. A compound of claim 10 wherein R7 and R8 form




the optional double bond is not present, and R14 is selected from the group consisting of H, OH, alkoxy, —(CH2)n—N(R30)(R23-heteroaryl), R23-heteroaryl and (R23-heteroaryl)-alkyl; or wherein R7 and R8 form




R15 is selected from the group consisting of alkyl, R17-phenyl, R23-heteroaryl, —C(O)alkyl, —C(O)(fluoroalkyl), —C(O)—(R23-heteroaryl), —C(O)-alkoxyalkyl, —C(O)—(R38-cycloalkyl), —SO2-alkyl, —SO2—NR18R19 and




and R16 is selected from the group consisting of H and alkyl, or two R16 groups and the carbon to which they are attached form —C(O)—.

12. A compound of claim 10 wherein R7 and R8 form




q is 1, and R27 is 1, 2 or 3 substituents independently selected from the group consisting of H, OH, alkyl, alkoxy, alkoxyalkyl, R17-phenyl, —C(O)OH, —C(O)Oalkyl, R23-heteroaryl, (R23-heteroaryl)amino and —(CH2)n—N(R30)—C(O)(cycloalkyl) wherein n is 0.

13. A compound of claim 10 wherein R7 and R8 form




wherein p is 0, R34 is H, and R35 is 1 or 2 substituents independently selected from the group consisting of H, OH, halo and alkyl.

14. A compound of claim 10 wherein R7 and R8 form




wherein p is 0, B is a pyrazolyl or thiazolyl ring, and R35 is 1 or 2 substituents independently selected from the group consisting of H and alkyl.

15. A compound of claim 7 wherein R7 is H or alkyl and R8 is (R17-phenyl)alkyl, R45-hydroxyalkyl or -alkyl-N(R30)—(R23-heteroaryl), wherein R45 is R17-phenyl; heteroaryl is pyridinyl, pyrimidinyl, pyrazinyl, indolyl, benzothienyl or benzofuranyl; R17 is 1 to 3 substituents independently selected from the group consisting of halogen, OH, alkoxy and alkyl; and R23 is 1 or 2 substituents independently selected from the group consisting of H, alkyl, alkoxy and halogen.

16. A compound of claim 1 selected from the group consisting of

17. The compound of claim 1 having the formula

18. The compound of claim 1 having the formula

19. The compound of claim 1 having the formula

20. The compound of claim 1 having the formula

21. The compound of claim 1 having the formula

22. The compound of claim 1 having the formula

23. The compound of claim 1 having the formula

24. The compound of claim 1 having the formula

25. The compound of claim 1 having the formula

26. A pharmaceutical composition comprising an effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier.

27. A method of treating a PDE4 mediated disease, where the disease is selected from COPD asthma, depression or multiple sclerosis comprising administering an effective amount of a compound of claim 1 to a patient in need of such treatment.

28. The method of claim 27 for treating dermatitis in dogs.

29. The method of claim 27 further comprising administering to said patient the compound of formula I in combination with at least one other medicament selected from the group consisting of disease modifying antirheumatic drugs, nonsteroidal anitinflammatory drugs, COX-2 selective inhibitors, COX-1 inhibitors, Immunosuppressives, steroids, biological response modifiers and other anti-inflammatory agents or therapeutics useful for the treatment of PDE4 mediated diseases.