# Patent

## Title
Heterocyclic substituted carbonyl derivatives and their use as dopamine D3 receptor ligands

## Abstract

The invention relates to heterocyclic substituted carbonyl derivatives that display selective binding to dopamine D3 receptors. In another aspect, the invention relates to a method for treating central nervous system disorders associated with the dopamine D3 receptor activity in a patient in need of such treatment comprising administering to the subject a therapeutically effective amount of said compounds for alleviation of such disorder. The central nervous system disorders that may be treated with these compounds include Psychotic Disorders, Substance Dependence, Substance Abuse, Dyskinetic Disorders (e.g. Parkinson's Disease, Parkinsonism, Neuroleptic-Induced Tardive Dyskinesia, Gilles de la Tourette Syndrome and Huntington's Disease), Dementia, Anxiety Disorders, Sleep Disorders, Circadian Rhythm Disorders and Mood Disorders. The subject invention is also directed towards processes for the preparation of the compounds described herein as well as methods for making and using the compounds as imaging agents for dopamine D3 receptors.

## Background
BACKGROUND OF THE INVENTION
The subject invention relates to novel heterocyclic derivatives that selectively bind to the dopamine D3 receptor. The therapeutic effect of currently available antipsychotic agents (neuroleptics) is generally believed to be exerted via blockade of D2 receptors; however this mechanism is also thought to be responsible for undesirable extrapyramidal side effects (eps) associated with many neuroleptic agents. Without wishing to be bound by theory, it has been suggested that blockade of the dopamine D3 receptor may give rise to beneficial antipsychotic activity without significant eps. (see for example Sokoloff et al, Nature, 1990; 347: 146-151; and Schwartz et al, Clinical Neuropharmacology, Vol 16, No. 4, 295-314, 1993). This receptor is shown in high abundance in brain regions associated with emotional and cognitive functions. Compounds that selectively bind to the dopamine D3 receptor are useful in treating certain central nervous system disorders. These central nervous system disorders include the following indications:

    1) Psychoses (including schizophrenia)—See, for example, Biochem Pharmacol, 1992, 3(4), 659-66; Clin Neuropharmacol, 1993, 16(4), 295-314; Neuropsychopharmacology, 1997, 16(6), 375-84; Am J Psychiatry, 1999, 156(4), 610-616; Psychopharmacology (Berl), 1995, 120(1), 67-74.
    2) Substance dependence and substance abuse—See, for example, Neuroreport, 1997, 8(9-10), 2373-2377; J Phammacol Exp Ther, 1996, 278(3), 1128-37; Brain Res Mol Brain Res, 1997, 45(2), 335-9.
    3) Mood Disorders (including mania, depressive disorders and bipolar disorders)—See, for example, Clin Neuropharmacol, 1998, 21(3), 176-80; Am J Med Genet, 1998, 81(2), 192-4; J Clin Psychiatry, 1995, 56(11), 514-518; J Clin Psychiatry, 1995, 56(9), 423-429; Am J Med Genet, 1995, 60(3), 234-237; Pharmacopsychiatry, 1999, 32(4), 127-135; J Affect Disord, 1999, 52(1-3), 275-290; Am J Psychiatry, 1999, 156(4), 610-616.
    4) dyskinetic disorders (including Parkinson's Disease, Parkinsonism, Neuroleptic-Induced Tardive Dyskinesia and Gilles de la Tourette Syndrome)—See, for example, Clin Neuropharmacol, 2000, 23(1), 34-44; Eur J Pharmacol, 1999, 385(11, 39-46.
    5) sleep disorders (including narcolepsy)—The D3 agonist pramipexole causes narcolepsy. A D3 antagonist would be useful for reversing this undesirable side effect. See Aust Fam Physician, 1999, 28(7), 737; Neurology, 1999, 52(9), 1908-1910.
    6) anxiety disorders (including obsessive compulsive disorders)—See, for example, Physiol Behav, 1997, 63(1), 137-141; J Clin Psychiatry, 1995, 56(9), 423-429; J Psychiatry Neurosci, 2000, 25(2), 185; J Affect Disord, 1999, 56(2-3), 219-226.
    7) nausea—Dopamine antagonists are used alone and in combination with 5HT3 antagonists. See, for example, Support Care Cancer, 1998, 6(1), 8-12; Support Care Cancer, 2000, 8(3), 233-237; Eur J Anaesthesiol, 1999, 16(5), 304-307.
    8) dementia—See, for example, Behav Brain Res, 2000, 109(1), 99-111; Neuroscience, 1999, 89(3), 743-749.


Dopamine D3 receptor ligands are also useful for the treatment of renal dysfunction. See, for example, WO 200067847.
SUMMARY OF THE INVENTION
This invention relates to a class of compounds and pharmaceutically acceptable salts thereof which are selective modulators of dopamine D3 receptors. The compounds may act as agonists, partial agonists, antagonists or allosteric modulators of dopamine D3 receptors, and are useful for a variety of therapeutic applications.
In another aspect, the invention relates to a method for treating central nervous system disorders associated with the dopamine D3 receptor activity in a patient in need of such treatment comprising administering to the subject a therapeutically effective amount of a compound described herein for alleviation of such disorder. The central nervous system conditions or disorders that may be treated with these compounds include Psychotic Disorders, Substance Dependence, Substance Abuse, Dyskinetic Disorders (e.g. Parkinson's Disease, Parkinsonism, Neuroleptic-Induced Tardive Dyskinesia, Gilles de la Tourette Syndrome and Huntington's Disease), Nausea, Dementia, Anxiety Disorders, Sleep Disorders, Circadian Rhythm Disorders and Mood Disorders.
In yet another aspect, the subject invention is directed toward a pharmaceutical composition comprising an effective amount of a compound described herein with a pharmaceutically-acceptable carrier or diluent optionally in conjunction with one or more dopamine D1, D2, D4, D5 or 5HT receptor antagonists.
In yet another aspect, the subject invention is directed towards processes for the preparation of the class of compounds described herein.
Also within the scope of this invention are methods for using these novel compounds as imaging agents for dopamine D3 receptors. Methods of using these compounds as imaging agents are presented, as are intermediates and methods for making the imaging agents.


DETAILED DESCRIPTION OF THE INVENTION
In accordance with the present invention, there are provided compounds of formula I
A compound of the formula (I):




    
    
        wherein
        
            A is CH or N;
            n is 1 or 2;
            when n is 1, y is 0 or 2;
            when n is 2, y is 0;
            g is 1 or 2;
            each R3 is independently hydrogen, C1-C6alkyl, or
        
        
    
    






    
    
        
        
            
            
                wherein w is 1, 2, or 3;
            
            
            R is selected from the group consisting of (a)-(w):
        
        
    
    









    
    
        
        
            wherein
            
                each R4, R5, R6, R7, R5, R9, R10, R11, R12 and R18 is independently hydrogen, C1-C6alkyl, halogen, trifluoromethyl, —CO2C1-C6alkyl or —CH2OC1-C6alkyl;
                each R71, R72, R74 and R80 is independently hydrogen, C1-C6alkyl, C1-C6alkoxy, halogen, trifluoromethyl, —CO2C1-C6alkyl or —CH2OC1-C6alkyl;
                R73 is hydrogen, alkyl, pyridyl, benzyl, —CH2CF3, —CO2C1-C6alkyl, phenyl optionally substituted with halogen, trifluoromethyl, trifluoromethoxy or R73 is
            
            
        
        
    
    






    
    
        
        
            
            
                 wherein w is 1, 2 or 3 as hereinbefore defined;
                each R75 is hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                each R76 is hydrogen, halogen, —CN or C1-C6alkyl;
                each R77 is hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                each R78 hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                each R79 hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                p, s and x are 0, 1, or 2;
                each R13 is independently hydrogen, C1-C6alkyl, halogen, benzyl, trifluoromethyl, C1-C6alkoxy, nitro, —CN, or —COC1-C6alkyl;
                R16 is C1-C6alkyl;
                each R14 and R1s is independently hydrogen or C1-C6alkyl;
                R17 is hydrogen, C1-C6alkyl, Ar, —COAr, —CONHAr or —SO2—Ar wherein
                
                    Ar is a phenyl group which is optionally mono- or di-substituted with substituents independently selected from C1-C6alkyl, halogen, trifluoromethyl, C1-C6alkoxy, nitro, CN and COC1-C6alkyl; and
                
                
                m is 0, 1, or 2;
            
            
        
        
    
    






    
    
        
        
            
            
                 represents a group selected from (a)-(f):
            
            
        
        
    
    






    
    
        
        
            
            
                wherein
                
                    each R19 and R20 is independently hydrogen, hydroxy or C1-C6alkyl;
                    R21, R22, and R23 are each independently hydrogen or C1-C3 linear alkyl; and
                    d is 3 or 4;
                
                
            
            
            R1 is
            
                a) hydrogen;
                b) C1-C6alkyl optionally mono- or di-substituted with hydroxy; or
                c)
            
            
        
        
    
    






    
    
        
        
            
            
                wherein
                
                    each R24 is independently hydrogen or C1-C6alkyl;
                    each R25, and R26 is independently hydrogen or C1-C6alkyl;
                    t is 0 or 1; and
                    q is 0 or 1;
                
                
            
            
        
        
        R2 is a group selected from (a)-(jj):
    
    












    
    
        
        
            wherein
            
                each R27 and R28 is independently selected from:
                
                    (1) hydrogen;
                    (2) C1-C6alkyl;
                    (3) C1-C6alkoxy;
                    (4) —CO2—R43 wherein R43 is hydrogen or C1-C6alkyl;
                    (5) hydroxy;
                    (6) (CH2)a—OR44 wherein a is 1, 2 or 3 and R44 is hydrogen or C1-C6alkyl;
                    (7) —CO)—NR45R46 
                     wherein R45 and R46 are each independently hydrogen, C1-C2alkyl, or R45 and R46 taken together form a 5-membered monocyclic ring;
                
                
                z is 0 or 1;
                e is 2, 3, 4, 5, 6 or 7;
                h is 0, 1, 2 or 3;
                u is 0, 1, 2, 3 or 4;
                o is 0 or 1;
                l is 0 or 1;
                j is 0, 1, 2 or 3;
                v is 0, 1, 2, 3 or 4;
                w is 1, 2 or 3 as hereinbefore defined;
                f is 1, 2, 3 or 4;
                t is 0 or 1 as hereinbefore defined;
                b is 0, 1 or 2;
                q is 0 or 1 as hereinbefore defined;
                aa is 0 or 2;
                X is O, S or NR90 wherein R90 is hydrogen, C1-C6alkyl, or
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein R143 is hydrogen or alkyl,
                
                
                each M and V is a group independently selected from hydrogen, halogen, C1-C6alkyl, C1-C6alkoxy, trifluoromethyl, hydroxy, phenyl, phenoxy, —SO2NH2 or
            
            
        
        
    
    






    
    
        
        
            
            
                —NR48R49 wherein R48 and R49 are each independently hydrogen or C1-C2alkyl;
                each R31, R32, R33, R34, R35, R36, R37, R38, R39, R40, R68, and R69 is independently hydrogen or C1-C6alkyl;
                each R29, R30 is independently hydrogen, phenyl or C1-C6alkyl;
                each R83, R84, R86, R87, R88, R89, R92, R93, R98, R99, R94, R95, R100, R101, R103, R104, R105, R106, R108, R109, R110, R111, R113, R114, R115, R116, R117, R118, R119, R120, R122, R123, R124, R125, R127, R128, R130, R131, R133, R134, R135, R136, R137, R138, R139 and R140 is independently hydrogen or C1-C6alkyl;
                each R63, R64 and R65 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                each R66 is independently hydrogen, hydroxy, C1-C6alkyl or C1-C6alkoxy;
                Q is CH2, CHOH or C═O;
                X5 is O or S;
                each R57 is independently hydrogen or C1-C6alkyl;
                R70 is hydrogen, C1-C6alkyl, halogen, nitro or a phenyl group optionally mono-substituted with C1-C6alkyl, halogen or trifluoromethyl;
                R81 is hydrogen, C1-C6alkyl, or —CO2C1-C6alkyl;
                R91 is hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                R96 is hydrogen, C1-C6alkyl or
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein R145 and R146 are each independently hydrogen or C1-C6alkyl and b is 0, 1 or 2 as hereinbefore defined;
                
                
                R97 is hydrogen or C1-C6alkyl;
                each R102 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                R107 is hydrogen or C1-C6alkyl;
                each R121 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
                R127 is hydrogen or C1-C6alkyl;
                R126 is C1-C6alkyl or benzyl;
                R129 is hydrogen or C1-C6alkyl;
                R132 is hydrogen, C1-C6alkyl, halogen or C1-C6alkoxy;
                X3 is 0 or —NR127 wherein R127 is hydrogen or C1-C6alkyl;
                X4 is O, S or —NR143 wherein R143 is hydrogen or —C1-C6alkyl;
                R141 is hydrogen, C1-C6alkyl or amino;
                R142 is benzyl or phenyl each of which may be optionally substituted with C1-C6alkyl, halogen or C1-C6alkoxy;
                R144 is hydrogen or C1-C6alkyl;
                R85 is hydrogen, C1-C6alkoxy, C1-C6alkyl, —CO2C1-C6alkyl, C(O)C1-C6alkyl or a group selected from the following:
            
            
        
        
    
    






    
    
        
        
            wherein
            j is 0, 1, 2 or 3 as hereinbefore defined;
            w is 1, 2 or 3 as hereinbefore defined;
            m is 0, 1 or 2 as hereinbefore defined;
            e is 2, 3, 4, 5, 6 or 7 as hereinbefore defined;
            each R147, R148, R150, R151, R152, R153 R156, R157, R159, R160 R162 and R163 is independently hydrogen or C1-C6alkyl;
            R149 is hydrogen, halogen, C1-C6alkyl, phenoxy, trifluoromethyl or trifluoromethoxy;
            R155 is hydrogen, halogen or C1-C6alkyl;
            R158 is hydrogen or C1-C6alkyl;
            R161 is hydrogen or C1-C6alkyl;
            R164 is hydrogen, halogen, C1-C6alkyl or trifluoromethyl;
            R165 is hydrogen, C1-C6alkyl or halogen;
            X7 is O or S or —NR167 wherein R167 is hydrogen or C1-C6alkyl;
            R166 is hydrogen or C1-C6alkyl;
            or R1 and R2 are joined together to form a 5-, 6-, or 7-membered monocyclic saturated ring, and in which the ring is optionally mono- or di-substituted, the substituents independently selected from:
            (1) C1-C6alkyl;
            (2) —CO2—(C1-C6alkyl);
            (3) —NR50R51 wherein R50 and R51 are each independently hydrogen, C1-C6alkyl, or a phenyl group which is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl;
            (4) —C(O)phenyl wherein the phenyl group is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl;
            (5) —(CH2)mOR52 wherein R52 is hydrogen or C1-C2alkyl or a phenyl group which is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl, and m is 0, 1 or 2 as hereinbefore defined;
            (6) —NR54—COR53 wherein R54 is hydrogen or C1-C6alkyl and R53 is hydrogen or C1-C2alkyl;
            (7) ═O;
            (8) CN;
            (9)
        
        
    
    






    
    
        
        
            
            
                wherein
                
                    b is 0, 1 or 2 as hereinbefore defined;
                    w is 1, 2 or 3 as hereinbefore defined;
                    t is 0 or 1 as hereinbefore defined;
                    i is 0, 1 or 2;
                    v is 0, 1, 2, 3 or 4 as hereinbefore defined;
                    k is 0 or 1 as hereinbefore defined;
                    c are 0, 1 or 2;
                    R167 is hydrogen or C1-C6alkyl;
                    each R55, R56, R58, R59, R169 and R170 is independently hydrogen or C1-C6alkyl;
                    each R57 is independently hydrogen, halogen or C1-C6alkyl;
                    each R60 is independently hydrogen, halogen or C1-C6alkyl;
                    R61 and R62 are each independently hydrogen or C1-C6alkyl;
                    R168 is hydrogen, thienyl or furanyl;
                    R171 is hydrogen, C1-C6alkyl, halogen, trifluoromethyl or trifluoromethoxy;
                
                
            
            
            or R1 and R2 are joined together to form a group of formula X;
        
        
    
    






    
    
        
        
            or R1 and R2 are joined together to form the group of formula (Y)
        
        
    
    






    
    
        
        
            or R1 and R2 are joined together to form any of the following groups:
        
        
    
    






    
    
        
        
            
            
                wherein
                g is 1 or 2 as hereinbefore defined;
                p is 0, 1 or 2 as hereinbefore defined;
                R172 is hydrogen, C1-C6alkyl or C1-C6alkoxy;
                R173 is hydrogen, C1-C6alkyl or phenyl optionally mono- or disubstituted with C1-C6alkyl or halogen; and
                R82 is a substituent selected from the following groups:
                (a) C1-C6alkyl optionally substituted with hydroxy;
                (b) C1-C6alkenyl;
                (c) C1-C6alkoxy;
                (d) —(CH2)OC1-C6alkyl;
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein X8 is —(CR175R176)h— or —(CR177═CR188)—
                     wherein each R174 is independently hydrogen, C1-C6alkyl, halogen, trifluoromethyl, C1-C6alkoxy or benzyloxy;
                     h is 0, 1, 2 or 3 as hereinbefore defined;
                     each R175, R176, R177 and R178 is independently hydrogen or C1-C6alkyl; and
                     j is 0, 1, 2 or 3 as hereinbefore defined;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein X9 is —(CR180R181)j— or

—(CR184R185CR186═CR187)— or

—(CR182═CR183)—

                     wherein
                     aa is 0 or 2 as hereinbefore defined;
                     R179 is hydrogen, C1-C6alkyl, halogen, trifluoromethyl,
                     C1-C6alkoxy, benzyloxy or phenyl;
                     each R180, R181, R182, R183, R184, R185, R186 and R187 is independently hydrogen or C1-C6alkyl;
                     j is 0, 1, 2, or 3 as hereinbefore defined;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein w is 1, 2 or 3 as hereinbefore defined;
                     each R188 and R189 is independently hydrogen or C1-C6alkyl;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein
                     i is 0, 1 or 2 as hereinbefore defined;
                     each R190 is independently hydrogen, alkyl or halogen;
                     b is 0, 1, or 2 as hereinbefore defined;
                     each R191 and R192 is independently hydrogen or C1-C6alkyl;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein
                     a is 1, 2 or 3 as hereinbefore defined;
                     each R193 and R194 is independently hydrogen or C1-C6alkyl;
                     R195 is hydrogen, halogen or C1-C6alkyl;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein
                     e is 2, 3, 4, 5 or 6 as hereinbefore defined;
                     b is 0, 1 or 2 as hereinbefore defined;
                     each R196 and R197 is independently hydrogen or C1-C6alkyl;
                     each R198 and R199 is independently hydrogen or C1-C6alkyl;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein
                     each R200 and R201 is independently hydrogen or C1-C6alkyl;
                     w is 1, 2 or 3 as hereinbefore defined;
                
                
            
            
        
        
    
    






    
    
        
        
            
            
                
                
                    wherein
                    each R202, R203, R204 and R205 is independently hydrogen or C0-C6alkyl; and
                    w is 1, 2 or 3 is as hereinbefore defined;

—(CR206R207)w—OC1-C6alkyl  (n)

                    wherein
                    C1-C6alkyl is optionally substituted with hydroxy;
                    each R206 and R207 is independently hydrogen or C1-C6alkyl; and
                    w is 1, 2 or 3 as hereinbefore defined;

—(CR208R209)w—NR210R211  (o)

                    wherein
                    each R208, R209, R210 and R211 is independently hydrogen or C1-C6alkyl;
                    w is 1, 2 or 3 as hereinbefore defined;
                
                
            
            
        
        
        (p)
    
    






with the proviso that when n is 1; and y is 0; and R3 is hydrogen or C1-C6alkyl; and





is group (a);

and R is group:

    
    
        (a) wherein R4 is hydrogen, halogen or C1-C6alkyl, and R1 is hydrogen or unsubstituted C1-C6alkyl,
        
            then R2 cannot be a group of the following formula:
            (a) wherein z is 0,
            (b) wherein u is 0 and M is hydrogen, halogen, C1-C6alkyl, or trifluoromethyl,
            (c) wherein o is 0,
            (d) wherein l is 0,
            (e) wherein j is 0,
            (g) wherein v is 0, or
            (i);
            and also when R is the group of formula (a), R1 and R2 cannot be joined together to form the group of formula Y or a 5-, 6-, or 7-membered monocyclic ring wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl;
        
        
        (b) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (c) wherein o is 0,
            (d) wherein l is 0,
            (i),
            (k),
            (l), or
            (m) wherein 0 is CH2;
            and also when R is the group of formula (b), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (c) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (c) wherein o is 0,
            (d) wherein l is 0, or
            (i);
        
        
        (d) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b) wherein u is 1,
            (c) wherein o is 0,
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (d), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (e) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (c) wherein o is 0,
            (d),
            (i),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (e), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (f) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (c) wherein o is 0,
            (d),
            (i),
            (k),
            (l), or
            (m) wherein 0 is CH2;
            and also when R is the group of formula (f), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (g) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b) wherein u is 1,
            (c) wherein o is 0,
            (d),
            (i),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (g), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (h) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (c) wherein o is 0,
            (d),
            (i),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (h), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (j), then R1 and R2 cannot be joined together to form a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
        
            wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl.
        
        
    
    


The subject invention is directed toward compounds or pharmaceutically acceptable salts of Formula I as depicted above in either racemic or pure stereoisomeric forms.
Terms used herein have the following meanings:

    a) “Pharmaceutically acceptable salts” means either an acid addition salt or a basic addition salt which is compatible with the treatment of patients for the intended use.


“Pharmaceutically acceptable acid addition salt” is any non-toxic organic or inorganic acid addition salt of the base compounds represented by Formula I or any of its intermediates. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acid and acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include the mono-, di- and tri-carboxylic acids. Illustrative of such acids are, for example, acetic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, salicyclic, 2-phenoxybenzoic, p-toluenesulfonic acid and sulfonic acids such as methanesulfonic acid and 2-hydroxyethanesultonic acid. Either the mono- or di-acid salts can be formed, and such salts can exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of these compounds are more soluble in water and various hydrophilic organic solvents and which in comparison to their free base forms, generally demonstrate higher melting points.
“Pharmaceutically acceptable basic addition salts” means non-toxic organic or inorganic basic addition salts of the compounds of Formula (I) or any of its intermediates. Examples are alkali metal or alkaline-earth metal hydroxides such as sodium, potassium, calcium, magnesium or barium hydroxides; ammonia, and aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline. The selection criteria for the appropriate salt will be known to one skilled in the art.

    b) “Stereoisomers” is a general term for all isomers of the individual molecules that differ only in the orientation of their atoms in space. It includes mirror image isomers (enantiomers), geometric (cis/trans) isomers, and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereoisomers).
    c) “Alkyl” means a branched or straight chain alkyl or alkylene group, as is appropriate to the formula, specified by the amount of carbons in the alkyl, e.g., C1-C6 alkyl means a one, two, three, four, five or six carbon branched or straight chain alkyl or alkylene, as the case may be, or any ranges thereof, for example, but not limited to, C1-2, C1-3, C1-4, C1-5, C2-3, C2-4, C2-5, C2-C6, C3-C4, C3-5, C3-6, C4-5, C4-6, C5-6, etc.
    d) “Patient” means a warm blooded animal, such as for example rat, mice, dogs, cats, guinea pigs, and primates such as humans.
    e) “Treat” or “treating” means to alleviate symptoms, eliminate the causation of the symptoms either on a temporary or permanent basis, or to prevent or slow the appearance of symptoms of the named disorder or condition.
    f) “Therapeutically effective amount” means a quantity of the compound which is effective in treating the named disorder or condition.
    g) “Pharmaceutically acceptable carrier” is a non-toxic solvent, dispersant, excipient, adjuvant or other material which is mixed with the active ingredient in order to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to the patient. One example of such a carrier is a pharmaceutically acceptable oil typically used for parenteral administration.
    h) “Psychoses” or “Psychotic Disorders” means conditions wherein the patient experiences a major mental disorder of organic and/or emotional origin characterized by derangement of the personality and loss of contact with reality, often with delusions, hallucinations or illusions. Included under the term psychoses are the disorders schizophrenia, schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, shared psychotic disorder, psychotic disorder not otherwise specified, and substance-induced psychotic disorder, as defined by the Diagnostic and Statistical Manual of Mental Disorders, 4th ed., published 1994 by the American Psychiatric Association, Washington D.C. USA, incorporated herein by reference.
    i) “Substance Dependence” means a condition wherein the patient exhibits a maladaptive pattern of substance use, leading to clinically significant impairment or distress. There is a pattern of repeated self-administration that usually results in tolerance, withdrawal, and compulsive drug-taking.
    j) “Substance Abuse” means a condition wherein the patient exhibits a maladaptive pattern of substance use manifested by recurrent and significant adverse consequences related to the repeated use of substances. There may be repeated failure to fulfill major role obligations, repeated use in situations in which it is physically hazardous, multiple legal problems, and recurrent social and interpersonal problems. Unlike the criteria for Substance Dependence, the criteria for Substance Abuse do not include tolerance, withdrawal, or a pattern of compulsive use and instead only include the harmful consequences of repeated use.
    k) “Parkinson's Disease” means a slowly progressive neurological condition, characterized by tremor, rigidity, bradykinesia, and postural instability. Other manifestations include depression and dementia.
    l) “Parkinsonism” means a condition where the patient exhibits Parkinsonian signs or symptoms (i.e. tremor, muscular rigidity, or akinesia) that develop in association with the use of neuroleptic medication.
    m) “Neuroleptic-Induced Tardive Dyskinesia” means a disorder characterized by involuntary movements of the tongue, jaw, trunk, or extremities which have developed in association with the use of neuroleptic medication. The involuntary movements may be choreiform, athetoid or rhythmic.
    n) “Gilles de la Tourette Syndrome” means a condition manifested by motor and vocal tics. (A tic is a sudden, rapid, recurrent, nonrhythmic, stereotyped motor movement or vocalization.) The disturbance causes marked distress or significant impairment in social, occupational, or other important areas of functioning. The onset is before age eighteen years and the disturbance is not due to the physiological effects of a substance or general medical condition.
    o) “Dementia” means disorders characterized by the development of multiple cognitive deficits that include memory impairment and are due to the direct physiological effects of a general medical condition, to the persisting effects of a substance, or to multiple etiologies (e.g., the combined effects of cerebrovascular disease and Alzheimer's disease). Memory impairment is required to make the diagnosis of a dementia and is a prominent early symptom. Dementia disorders share a common symptom presentation but are differentiated based on etiology. See Diagnostic and Statistical Manual of Mental Disorders, 4th ed., American Psychiatric Association, for diagnostic criteria.
    p) “Anxiety Disorders” means disorders that include Panic Disorder Without Agoraphobia, Panic Disorder with Agoraphobia, Agoraphobia Without History of Panic Disorder, Specific Phobia, Social Phobia, Obsessive-Compulsive Disorder, Post-traumatic Stress Disorder, Acute Stress Disorder, Generalized Anxiety Disorder, Anxiety Disorder Due to a General Medical Condition, Substance-Induced Anxiety Disorder, and Anxiety Disorder Not Otherwise Specified, as defined by the Diagnostic and Statistical Manual of Mental Disorders, 4th ed.
    q) “Sleep Disorders” means disorders that include Primary Sleep Disorders, Sleep Disorder Related to Another Mental Disorder, Sleep Disorder Due to a General Medical Condition, and Substance-Induced Sleep Disorder as defined by the Diagnostic and Statistical Manual of Mental Disorders, 4th ed. Primary Sleep Disorders are those In which none of the etiologies listed below (i.e., another mental disorder, a general medical condition, or a substance) is responsible. Primary Sleep Disorders are presumed to arise from endogenous abnormalities in sleep-wake generating or timing mechanisms, often complicated by conditioning factors. Primary Sleep Disorders in turn are subdivided into Dyssomnias (characterized by abnormalities in the amount, quality, or timing of sleep) and Parasomnias (characterized by abnormal behavioral or physiological events occurring in association with sleep, specific sleep stages, or sleep-wake transitions). A representative example of a Primary Sleep Disorder is Narcolepsy. Narcolepsy is characterized by repeated irresistible attacks of refreshing sleep, cataplexy, and recurrent intrusions of elements of rapid eye movement (REM) sleep into the transition period between sleep and wakefulness.
    r) “Mood Disorders” are disorders that have a disturbance in mood as the predominant feature. As defined by the Diagnostic and Statistical Manual of Mental Disorders, 4th ed., Mood Disorders are divided into the Depressive Disorders (“unipolar depression”), the Bipolar Disorders, and two disorders based on etiology—Mood Disorder Due to a General Medical Condition and Substance-Induced Mood Disorder. The Depressive Disorders (i.e., Major Depressive Disorder, Dysthymic Disorder, and Depressive Disorder Not Otherwise Specified) are distinguished from the Bipolar Disorders by the fact that there is no history of ever having had a Manic, Mixed, or Hypomanic Episode. The Bipolar Disorders (i.e., Bipolar I Disorder, Bipolar II Disorder, Cyclothymic Disorder, and Bipolar Disorder Not Otherwise Specified) involve the presence (or history) of Manic Episodes, Mixed Episodes, or Hypomanic Episodes, usually accompanied by the presence (or history) of Major Depressive Episodes.
    s) “Circadian Rhythm Disorder” means a persistent or recurrent pattern of sleep disruption leading to excessive sleepiness or insomnia that is due to a mismatch between the sleep-wake schedule required by a person's environment and his or her circadian sleep-wake pattern. The sleep disturbance causes clinically significant distress or impairment in social, occupational, or other important areas of functioning. The disturbance does not occur exclusively during the course of another Sleep Disorder or other mental disorder. The disturbance is not due to the direct physiological effects of a substance (e.g., a drug of abuse, a medication) or a general medical condition.


Preferred compounds are those wherein R is group (a) or group (k). When R is group (a), R4 is further preferred to be halogen or CF3. When R is group (k), R12 is further preferred to be hydrogen, C1-C6alkyl, or —CH2OC1-C6alkyl. R2 is preferred to be group (a), (b) or (n). When R2 is group (a), z is further preferred to be 0 or 1; e is further preferred to be 5; and each R27 and R28 is further preferred to be independently selected from hydrogen or C1-C6alkyl. When to R2 is group (b), M is further preferred to be hydrogen, C1-C6alkoxy or C1-C6alkyl and u is further preferred to be 0 or 1. When R2 is group (n), R70 is further preferred to be hydrogen and t is further preferred to be 3.
Specific embodiments of the invention include the compounds set forth in the tables herein.
Preferred embodiments of the invention are those compounds of Formula I set forth in Table 1 that exhibit enhanced D3 potency. Particularly preferred compounds include the following.

    2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-trans-cyclopropanecarboxylic acid (trans-4-ethyl-cyclohexyl)-amide
    2-[4-(2,4-Dimethyl-phenyl)-piperazin-1-ylmethyl]-(2R,3R)-cyclopropanecarboxylic acid (trans-4-ethyl-cyclohexyl)-amide
    2-[4-(Chloro-trifluoromethyl-pyridin-2-yl)-piperazin-1-ylmethyl]-(2R,3R)-cyclopropanecarboxylic acid (trans-4-ethyl-cyclohexyl)-amide
    2-[4-(2,5-Dimethyl-phenyl)-piperazin-1-ylmethyl]-(2R,3R)-cyclopropanecarboxylic acid (trans-4-ethyl-cyclohexyl)-amide
    2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-(2R,3R)-cyclopropanecarboxylic acid (trans-4-methylcyclohexyl)-amide
    2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-(2R,3R)-cyclopropanecarboxylic acid (trans-4-methyl-cyclohexyl)-amide
    2-[4-o-Tolyl-piperazin-1-ylmethyl]-(2R,3R)-cyclopropanecarboxylic acid (trans-4-ethyl-cyclohexyl)-amide
    4-[4-(6-Fluoro-benzo[b]thiophen-3-yl)-piperazin-1-yl]-N-(trans-4-methyl-cyclohexyl)-butyramide
    2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-(2R,3R)-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide
    2-(4-Thieno[2,3d]isoxazol-3-yl-piperazin-1-ylmethyl)-(2R,3R)-yclopropanecarboxylic acid (trans-4-methyl-cyclohexyl)-amide
    2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperazin-1-ylmethyl]-cyclopropane-1R-carboxylic trans-(4-methyl-cyclohexyl)-amide (AVE1734)
    2R-[4-(5-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-cyclopropane-1R-carboxylic acid trans-(4-methyl-cyclohexyl)-amide. MDL 834012.
    (3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(7-methoxy-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002437359)
    2R-[4-(1-Methyl-7-trifluoromethyl-1H-indazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 832654)
    (3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(7-trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002437360)
    2R-[4-(7-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropanecarboxylic acid (trans-4-methyl-yclohexyl)-amide (MDL 833690)
    (3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(1-methyl-6-trifluoromethyl-1H-indazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002437353)
    2R-[4-(6-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (AD02287765)
    (3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002609935)
    2R-[4-(6-Fluoro-7-methoxy-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 831361)
    2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (A002436291)
    2R-{4-[1-(2,2,20Trifluoro-ethyl)-1H-thieno[3,2-c]pyrazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (A002287767)
    2R-(4-Thieno[2,3-d]isoxazol-3-yl-piperazin-1-ylmethyl)-1R-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 831493)
    2R-(4-Benzo[b]thiophen-2-yl-piperidin-1-ylmethyl)-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 831148)
    2R-[4-(5,6-Dihydro-4H-imidazo[4,5,1-ij]quinolin-2-yl)-piperazin-1-ylmethyl]-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 833699)
    2R-(4-Thieno[2,3-b]pyridin-3-yl-piperazin-1-ylmethyl)-1R-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 833821)
    1-{2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl}-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 832231)


The compounds of the present invention may be prepared by various methods. Schemes I through VI show the different ways of preparing the compounds of Formula I.
The compounds of formula (I) of this invention can be synthesized by following or combining one or more of the steps described below, not necessarily in the order presented. Throughout the description of the synthetic steps, the definitions of R. R1, R2, R3, g, n, y, B and A are as given above unless otherwise stated or indicated, and other nomenclatures appearing below shall have the same meanings defined in their respective first appearances unless otherwise stated or indicated.
Compounds of the present invention may be prepared according to a process which comprises

    (a) reacting a compound of formula (II):






    
    
        wherein R, A, n, y, R3 and 9 are as defined in formula I with a compound of formula (III)
    
    






    
    
         is as defined in formula I and “LG” is a suitable leaving group selected from chlorine, bromine, iodine, mesyl, tosyl, brosyl, triflyl, nosyl, nonaflyl or tresyl:

to provide a compound of formula (IV):

    
    






wherein R3, g, R, A, y, n and





are defined as in formula I;

    (b) hydrolyzing a compound of formula (IV) to provide a compound of formula (V):






    (c) reacting a compound of formula (V) with a compound of formula (VI):






    
    
        wherein R1 and R2 are as defined In formula I;

to provide a compound of formula (I).

    
    


Typically, the reaction in step (a) is carried out in polar solvent such as, for example, acetonitrile and an amine base such as, for example, triethylamine. The reaction is typically conducted at a temperature of about 80° C. to about 85° C. for about 1 to 3 hours.
The reaction in step (b) is typically carried out in a water miscible solvent such as, for example, methanol or 1,4-dioxane in the presence of an aqueous hydroxide base such as, for example, 5 N sodium-hydroxide. The reaction is typically carried out at a temperature of about 55° C. to about 65° C. for about 2 to 3 hours.
The reaction in step (c) is typically carried out in a polar solvent such as, for example, N,N′-dimethylformamide, in the presence of a weak base such as, for example, N-methylmorpholine with a suitable coupling reagent. A suitable coupling reagent is, for example, DCC (1,3-dicyclohexylcarbodiimide), EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2 dihydroquinoline) or TOTU {O-[(ethoxycarbonyl)cyanomethyleneamino]-N,N,N′,N′-tetramethyluronium tetrafluoroborate}. Typically, the reaction takes place at a temperature of about 20° C. to about 25° C. for about 16 to 20 hours.
Alternatively, compounds of the present invention may be prepared according to a process which comprises:

    (a) reacting a compound of formula (VII) wherein R1 and R2 are as hereinbefore defined and “LG” is a suitable leaving group selected from chlorine, bromine, iodine, mesyl, tosyl, brosyl, triflyl, nosyl, nonaflyl or tresyl:






with a compound of formula (II) to provide a compound of formula (I). This reaction is typically carried out in a polar solvent and an amine base such as acetonitrile and triethylamine, at a temperature of about 55° C. to about 65° C. for about 16 to 20 hours.

Compounds of formula (II) are either commercially available or may be prepared via synthetic methods well known in the art. For example, Scheme I describes the coupling of a benzthiophene with a commercially-available substituted piperazine. The synthesis is analogous for the un-substituted piperazine analogs. The less sterically hindered piperazine nitrogen is more reactive and cleanly gives a single product in the benzo[b]thiophene coupling. The more sterically hindered nitrogen can then be alkylated as before in the un-substituted piperazines.




Piperidine-substituted compounds may be prepared via syntheses analogous to those shown in the following reaction Schemes I and III.








The preparation of various substituted aza- and diazacycloheptanes is described by Treiber et al. in WO 9725324.
The synthesis of compounds of formula (I) wherein the variable designated as




contains a carbocycle is shown in general reaction Scheme IV. For the sake of simplification, the description of synthetic schemes is presented below for compounds which contain this carbocyclic group, but it will be apparent that compounds which do not contain a carbocyclic group can be prepared by utilizing the synthetic schemes and making necessary modifications.





    
    
        wherein R, A, R3, y, g and n are as hereinbefore defined.
    
    


Many of the dicarboxylates or more advanced intermediates that are generically described in scheme IV are commercially available. Several of these are shown in Table 1. This table is used for illustrative purposes only and is not intended to limit the scope of the present invention in any way.
Starting Materials:










TABLE I





Structure
Name
CAS #
Supplier











Dimethyl cis-1,2-cyclopropane dicarboxylate
826-34-6
Acros


 






Dimethyl trans-1,2-cyclopropane dicarboxylate
826-35-7
Acros


 






Dimethyl 1-methyl-trans-1,2- cyclopropane dicarboxylate
702-92-1
Acros


 






Dimethyl 3-methyl-trans-1,2- cyclopropane dicarboxylate
28363-79-3
Acros


 






trans-Cyclobutane-1,2-dicarboxylic acid dimethylester

Syntec


 






trans-D,L-1,2-Cyclopentane- dicarboxylic acid
1461-97-8
Aldrich


 






trans-2-Carbomethoxy cyclopentane-1-carboxylic acid

Rieke


 






trans-1,2-Cyclohexane dicarboxylic acid
2305-32-0
Aldrich Acros


 






trans-2-Carbomethoxy cyclohexane-1-carboxylic acid

Rieke


 






cis-1,2-Cyclohexane dicarboxylate acid
610-09-3
Acros


 






cis-2-Carbomethoxy cyclohexane- 1-carboxylic acid

Rieke









When not commercially available, the appropriate starting material may be obtained via standard synthetic methods.
When a compound of formula (I) is obtained as a mixture of enantiomers these may be separated by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, for example using a chiral HPLC column.
Compounds of formula (I) have been found to exhibit affinity for dopamine receptors, in particular D3 receptors, and are expected to be useful in the treatment of disease states which require modulation of such receptors, such as psychotic conditions. Preferred compounds of the present invention are those which have higher affinity for dopamine D3 than dopamine D2 receptors.
A major challenge in antipsychotic research is to produce agents with reduced side effects. Orthostatic hypotension is a common side effect in antipsychotics that is associated with the high potency that these agents have at the alpha-1 receptor (hereinafter referred to as “α-1”). A major goal of this work was to find agents with reduced α-1 potency.
The 6-trifluoromethyl benzo[b]thiophenes described herein have a clear and somewhat surprising advantage over the 8-fluoro benzo[b]thiophenes as is shown in the following table. The 6-fluoro benzo[b]thiophenes are clearly more potent at the alpha-receptor than are the 6-trifluoromethyl benzo[b]thiophenes. This is shown by comparing pairs of analogs that only differ in substitution at the 6-position of the benzo[b]thiophene. In every case, the 6-fluoro benzo[b]thiophene is more potent than the corresponding 6-trifluoromethyl analog. In some cases this small structural difference in substitution at the 6-position produces a dramatic change in α-1 potency. There are several examples wherein 6-fluoro analogs exhibited nanomolar potency while the corresponding 6-trifluoromethyl analog exhibited micromolar potency.















halpha1Ki





(nM)
halpha1 % I


NUM
MOLSTRUCTURE
h = human
h = human


















826844




23.4


 


826804





32.8% Inh @ 1 uM


 


826845




5.44


 


826805





27.8% Inh @ 1 uM


 


826846




93.9


 


826806





10.3% Inh @ 1 uM


 


826849




35.2


 


826809





30.5% Inh @ 1 uM


 


826857




9.82


 


826817





44.4% Inh @ 1 uM


 


826861




9.55


 


826821




1280
24% Inh @ 0.1 nM


 


826847




0.513


 


826807




166000


 


826848




12.3


 


826808




126


 


826850




28.4


 


826810




200


 


826851




18.9


 


826811




223


 


826852




6.16


 


826812




658


 


826853




0.258


 


826813




57


 


826854




0.0713


 


826814




89.3


 


826855




0.889


 


826815




80.5


 


826856




15.3


 


826816




249


 


826858




9.42


 


826818




124


 


826859




4


 


826819




49


 


826860




0.516


 


826820




129


 


826862




0.637


 


826822




32.2


 


826863




32.2


 


826823




148









Especially preferred compounds of the instant invention are those with a reduced liability for α-1 receptor binding while at the same time having a higher affinity for dopamine D3 than dopamine D2 receptors.
Receptor affinity can be measured using standard methodology such as is described below.
[N-Methyl-3H]Spiroperidol Binding to Cloned Human Dopamine D3 Receptors
Purpose
This assay measures the in vitro activity of compounds on cloned human dopamine (D3) receptors and predicts the direct dopamine-blocking properties of putative neuropsychiatric agents at human dopamine D3 receptors.
Methods
A. Cloning
The D3 gene was isolated from a human striatal cDNA library (Stratagene). The gene was sequenced and sub-cloned into the expression vector RC/RSV (Invitrogen). CHO (Chinese Hamster Ovary) cells were stably transfected with 10 μg of the D3/RSV plasmid using the DOTAP method from Boehringer Mannheim and 72 clones that were G418 resistant were isolated. Using mRNA and binding displacement data a single high expressing clone was identified. This clone was then grown in large batches for the purpose of developing a 96 well format assay.
B. Cell Culture


    1. One plate (10 cm) with approximately 2-3 million D3 cells per plate is incubated with 1 ml of Trypsin-EDTA at room temperature for  ̃2 min or until cells have lifted off plates. Four ml of Ham's F12+10% Fetal Bovine Serum+1% Penicillin/Streptomycin+G418 (400 μg/ml) medium are added to resuspend cells and 1 ml of this is added to each large plate (15 cm) containing 19 ml of the same medium as mentioned above.
    2. The 5 large plates are incubated at 37° C.+5% CO2 for  ̃3 days or until the cells are confluent.
    3. After these plates are confluent, they are split into 10 large plates. Medium is aspirated off, 2 ml of Trypsin-EDTA are added to each plate and plates are incubated at RT for 2 min or until cells have lifted off the plate. Eight ml of the F12 medium (same medium as #1 above) are added to each plate (10 ml total) to resuspend the cells and 5 ml are transferred to the 2 new plates containing 15 ml of the F12 media.
    4. The 10 large plates are incubated at 37° C.+5% CO2 for  ̃2 days or until the cells are confluent.
    5. The 10 large plates are split into 60 large plates (using Trypsin-EDTA as #3 except 4 ml of F12 medium are added to resuspend cells and 1 ml is aliquoted to 6 new plates containing 19 ml of F12 medium each).
    6. Plates are incubated at 37° C.+5% CO2 for  ̃3 days or until cell are confluent.
    7. The 60 large plates are then split into 60 roller bottles (100-150 million cells/bottle). Medium is aspirated off, 2 ml of Trypsin-EDTA are added to each plate and incubated at RT for  ̃2 minutes or until cells have lifted off plates. Eight ml of F12 medium are added to each plate to resuspend cells and the entire 10 ml are added to 1 roller bottle containing 90 ml of the F12 medium.
    8. The 60 roller bottles are immediately placed on their sides and transferred to the roller bottle incubator. They are incubated at 37° C.+5% CO2 for  ̃3-5 days. Cells are spun at 30-40% motor speed in the Form a incubator.
    9. Medium is poured off and cells are washed 2× in PBS.
    10. Cells are then scraped off in 20 ml of PBS and the bottles are rinsed again with 5 ml of PBS to remove any remaining cells. Cells are stored on ice before membrane preparation.
    11. The yield for 60 D3 roller bottles has varied from  ̃260-500 mg.
    
        Note: All tissue culture reagents are from Gibco-BRL.

C. Membrane Preparation

    
    


The cells are harvested into 250 ml centrifuge tubes with 100 volumes of cold phosphate buffered saline (PBS) and spun down (1200×G for 10 min at 4° C.). The medium is removed and 100 ml PBS are added to each centrifuge tube, cells are resuspened and spun down again. The PBS is removed and the final pellet is homogenized in an appropriate volume of 10% DMSO with a polytron on ice at a medium setting.
D. Lowry Protein Assay
A 200 μl sample membrane homogenate is added to 200 μl of 1% SDS, vortexed and allowed to stand for 5 min. Aliquots (25, 50 and 100 μl) of this mixture are assayed in duplicate following the standard Bio-Rad DC protein assay protocol (kit catalog number 500-0112) and using reagent S. Absorbance readings are made at 750 nm (note: the most accurate protein OD readings are between 0.1-0.5 units). The protein concentration is calculated using a standard curve generated concurrently with bovine serum albumin as standard.
E. Storage/Freezing conditions
Following the determination of the protein concentration and Scatchard analysis, the protein is diluted into distilled water with 10% DMSO to the appropriate volume based on expression levels (Bmax). The concentrated protein is then aliquoted into 1.5 ml screw top cap Eppendorf tubes and placed into a −80° C. freezer.
F. Binding Assay Reagents


    1. 0.5M Tris Buffer, pH 7.7
    
        a) 44.4 g Tris HCl
        
            26.5 g Tris Base
            q.s. to 1 Liter (0.5 M Tris buffer, pH 7.7 at 37° C.)
        
        
        b) make a 1:10 dilution in distilled H2O (0.05 M. Tris buffer, pH 7.7)
    
    
    2. Tris Buffer containing physiological salts
    
        a) Stock buffer
        
            NaCl 7.014 g
            KCl 0.372 g
            CaCl2 0.222 g
            MgCl2 0.204 g
            q.s. To 100 ml with 0.5 M. Tris Buffer
        
        
        b) Dilute 1:10 in distilled H2O
    
    


This yields 0.05 M. Tris HCl, pH 7.7, containing NaCl (120 mM), KCl (5 mM), CaCl2 (2 mM) and MgCl2 (1 mM)

    
    
        Optional: add 0.1% ascorbic acid and check pH (in assays with compounds that may oxidize.
    
    
    3. a) 1.0% polyethyleneimine stock in 0.5M Tris (reagent 1.a)
    
        b) Dilute 1:10 in distilled H2O
    
    
    4. [N-methyl-3H]-Spiroperidol (60-90 Ci/mmol) is obtained from New England Nuclear; catalog #NET-856.


For Ki determinations: [3H]NMSP is made up to a concentration of 2.7 nM in buffer 2b, such that when 150 μl is added to each tube a final concentration of 0.4 nM is attained in the 1 ml assay. Samples of total CPM added are taken for each experiment to calculate the total ligand concentration.

    5. S(−)-Eticlopride is obtained from Research Biochemicals International (RBI catalog number E-101). A refrigerated stock (good for up to a month) solution of S(−)-eticlopride is made at a concentration of 30 μM in buffer 2b. One hundred microliters are added to 3 wells for the determination of nonspecific binding (this yields a final concentration of 3 μM in the 1 ml assay).
    6. Test Compounds


For most assays, a 100 μM stock solution of the test compound is made up in a suitable solvent (usually <0.1% acetic acid) and serially diluted with buffer 2b, such that when 100 μl of drug is combined with the total 1 ml assay, final concentrations ranging from 10−5-10−8 M are attained. Characteristically eight concentrations are studied for each assay; however, higher or lower concentrations may be used, depending on the potency of the drug.
G. Binding Assay


    
    
        750 μl Tissue
        150 μl [3H]NMSP
        100 μl vehicle (for total binding) or 30 μM (−)eticlopride (for nonspecific binding) or appropriate drug concentration.
    
    


The 96-Well Packard Unifilters GF/B are incubated for >1 h at 25° C. in 0.1% polyethylamine (from 3,b). The cold tissue is added last and mixed on a orbital shaker for a few seconds and is then incubated at 37° C. for 30 min in a shaking water bath. The assay is stopped by rapid filtration through Packard Unifilter plates. The filter membranes are then washed with 15 ml of ice-cold 0.05 M Tris buffer. The filters are then dried ( ̃15 min under a heat lamp or incubated for 15 min in a 60° C. oven) and a bottom seal is applied. Then 40 μl of Packard Microscint 20 scintillation cocktail is added and a permanent topseal (Type P) is applied and heat sealed. The plates are then shaken on an orbital shaker for 1 h and placed in the Packard Topcount and counted for at least 5 minutes for each point.
Specific binding is defined as the difference between total binding and the binding in the presence of 3 μM S-(−)-eticlopride. Total binding is approximately 10% of the total added ligand. Cheng-Prusoff determination (Ki's) are performed using Prism software using a one-site competition curve analysis where the top and the bottom of the non-linear regression are held constant at 0% and 100% percent inhibition. The percent inhibition at each drug concentration is the mean of duplicate determinations.
[N-Methyl-3H]Spiroperidol Binding to Cloned Human Dopamine D2Long Receptors
Purpose:
This assay measures the in vitro activity of drugs on cloned human dopamine D2Long (D2L) receptors and predicts the direct dopamine-displacing properties of neuropsychiatric, cardiovascular and renal agents at human dopamine D2 receptors.
Methods:
A. Cloning
The D2L gene was isolated from a human striatal (caudate/putamen) cDNA library. The gene was sequenced and sub-cloned into the expression vector pRC/RSV (Invitrogen). CHO (Chinese Hamster Ovary) cells were stably transfected and 72 clones that were geneticin (G418) resistant were isolated, Using mRNA and binding data a single high expressing cell line was identified (#44). This cell line was then grown in suspension culture for the purpose of developing a 96 well format assay.
B. Cell Culture Conditions


    1. Medium for adherent CHO cultures:

Ham's F12+10% fetal bovine serum (FBS)+400 g/mil geneticin (G418)+10 ml/L penicillin-streptomycin (pen-strep)

    2. Cells are transferred to suspension culture when at least 1.5 million cells are available (this allows for 300,000 cells/ml in a 50 ml spinner flask; this is the ideal suspension density). Cell are removed from flasks with trypsin, spun down (1000×G) and resuspended in fresh medium:

50% CHO—SFM II+50% Ham's F12 w/10% FBS (final FBS conc. 5%)+400 μg/ml G418+pen-strep (10 ml/L)

    3. After the transfer to suspension culture, growth is monitored and cell viability is assessed using trypan blue exclusion. Total and viable cell count on 5 sectors of the hemocytometer are recorded. When the viable cell density reaches 600,000 cell/ml, the volume is doubled.
    4. After 1 week of growth in the 50/50 medium, the cells are spun down and transferred to a new spinner flask and replaced with 75% CHO—SFM II 125% Ham's F12+10% FBS plus the pen-strep and G418. Thereafter every 3 days, the medium is replaced with new medium containing a decreasing amount of FBS as follows:
















ml of CHO SFM:ml of Ham'S F12
Final % FBS conc.



















87.50:12.5
1.25



93.75:6.25
0.625



99.00:1.00
0.1










    5. The final maintenance culturing medium is made up as follows:


A stock mixture of 10 ml of pen-strep, 0.5 ml of 400 mg/ml (active; final concentration: 200 mg/ml) G418 and 1 ml of FBS are mixed and filtered and refrigerated. A volume (11.5 ml) of this mixture is added to a freshly opened 1 L bottle of CHO—SFM II.
C. Membrane Preparation
The cells are harvested into 250 ml centrifuge tubes with 100 volumes of cold phosphate buffered saline (PBS) and spun down (1200×G for 10 min at 4° C.). The medium is removed and 100 ml PBS are added to each centrifuge tube, cells are resuspened and spun down again. The PBS is removed and the final pellet is homogenized in an appropriate volume of PBS with a polytron on ice at a medium setting.
D. Lowry Protein Assay
A 200 μl sample membrane homogenate is added to 200 μl of 1% SDS, vortexed and allowed to stand for 5 min. Aliquots (25, 50 and 100 μl) of this mixture are assayed In duplicate following the standard Bio-Rad DC protein assay protocol (kit catalog number 500-0112) and using reagent S. Absorbance readings are made at 750 nm (note: the most accurate protein OD readings are between 0.1-0.5 units). The protein concentration is calculated using a standard curve generated concurrently with bovine serum albumin as standard.
E. Storage/Freezing Conditions
Following the determination of the protein concentration, the protein is diluted into distilled water with 10% DMSO to the appropriate volume based on expression levels (Bmax). The concentrated protein is aliquoted into 1.5 ml screw top eppendorf tubes and placed into a −80° C. freezer.
F. Binding Assay Reagents


    1. 0.5M Tris Buffer, pH 7.7
    
        a) 44.4 g Tris HCl
        
            26.5 g Tris Base
            q.s. to 1 Liter (0.5 M Tris buffer, pH 7.7 at 37° C.)
        
        
        b) make a 1:10 dilution in distilled H2O (0.05 M. Tris buffer, pH 7.7)
    
    
    2. Tris Buffer containing physiological salts
    
        a) Stock buffer
        
            NaCl 7.014 g
            KCl 0.372 g
            CaCl2 0.222 g
            MgCl2 0.204 g
            q.s. To 100 ml with 0.5 M. Tris Buffer
        
        
        b) Dilute 1:10 in distilled H2O
    
    


This yields 0.05 M. Tris HCl, pH 7.7, containing NaCl (120 mM), KCl (5 mM), CaCl2 (2 mM) and MgCl2 (1 mM)
Optional: add 0.1% ascorbic acid and check pH (in assays with compounds that may oxidize.


    3.
    
        a) 1.0% polyethyleneimine stock in 0.5M Tris (reagent 1.a)
        b) Dilute 1:10 in distilled H2O
    
    
    4. [N-methyl-3H]-Spiroperidol (60-90 Ci/mmol) is obtained from New England Nuclear; catalog #NET-856.


For Ki determinations: [3H]NMSP is made up to a concentration of 2.7 nM in buffer 2b, such that when 150 μl is added to each tube a final concentration of 0.4 nM is attained in the 1 ml assay. Samples of total CPM added are taken for each experiment to calculate the total ligand concentration.

    5. S(−)-Eticlopride is obtained from Research Biochemicals International (RBI catalog number E-101). A refrigerated stock (good for up to a month) solution of S(−)-eticlopride is made at a concentration of 30 μM in buffer 2b. One hundred microliters are added to 3 wells for the determination of nonspecific binding (this yields a final concentration of 3 μM in the 1 ml assay).
    6. Test Compounds


For most assays, a 100 μM stock solution of the test compound is made up in a suitable solvent (usually <0.1% acetic acid) and serially diluted with buffer 2b, such that when 100 μl of drug is combined with the total 1 ml assay, final concentrations ranging from 10−5-10−8 M are attained. Characteristically eight concentrations are studied for each assay; however, higher or lower concentrations may be used, depending on the potency of the drug.
G. Binding Assay


    
    
        750 μl Tissue
        150 μl [3H]NMSP
        100 μl vehicle (for total binding) or 30 μM (−)eticlopride (for nonspecific binding) or is appropriate drug concentration.
    
    


The 96-Well Packard Unifilters GF/B are incubated for >1 h at 25° C. in 0.1% polyethylamine (from 3,b). The cold tissue is added last and mixed on a orbital shaker for a few seconds and is then incubated at 37° C. for 30 min in a shaking water bath. The assay is stopped by rapid filtration through Packard Unifilter plates. The filter membranes are then washed with 15 ml of ice-cold 0.05 M Tris buffer. The filters are then dried ( ̃15 min under a heat lamp or incubated for 15 min in a 60° C. oven) and a bottom seal is applied. Then 40 μl of Packard Microscint 20 scintillation cocktail is added and a permanent topseal (Type P) is applied and heat sealed. The plates are then shaken on an orbital shaker for 1 h and placed in the Packard Topcount and counted for at least 5 minutes for each point.
Specific binding is defined as the difference between total binding and the binding In the presence of 3 μM S-(−)-eticlopride. Total binding is approximately 10% of the total added ligand. Cheng-Prusoff determination (Ki's) are performed using Prism software using a one-site competition curve analysis where the top and the bottom of the non-linear regression are held constant at 0% and 100% percent inhibition. The percent inhibition at each drug concentration is the mean of duplicate determinations.
3H]Prazosin Binding to Cloned Human Alpha-1A Adrenergic Receptors (α1a) Expressed in Chinese Hamster Ovary Cells (CHO)


    Purpose: This in vitro assay is designed as a screen to identify compounds displaying a affinity for the human α1a adrenoceptor subtype expressed in the membrane fragments of CHO cells. The assay measures the ability of the test compounds to displace [3H] prazosin from α1a sites.
     The identification of multiple vascular α1-addrenoceptors (α1a, α1b, α1d) in vitro has provided impetus to define the role(s) of these subtypes in cardiovascular regulation in vivo (Vargas and Gorman, 1995). Hemodynamic studies in the unanesthetized rat suggest that vascular α1a receptors are the major subtype involved in the sympathetic regulation of peripheral resistance and systemic arterial pressure (Piascik et al., 1989). Additional evidence for an involvement of peripheral α1a receptors in the maintenance of arterial pressure was demonstrated by the findings that the selective α1a antagonist 5-MU dose dependently lowered resting arterial pressure in awake conscious dogs (Piascik et al., 1989). A demonstrated inability of the irreversible antagonist, chloroethylclonidine, to reduce arterial pressure in rats when administered intravenously, is strong evidence against the role of α1b and α1d receptors in the acute regulation of arterial pressure (Vargas et al., 1993).
     Therefore, the binding of compounds to α1a adrenergic receptors is believed to be a good measure of a compound's potential to cause orthostatic hypotension and sedation as side effects. Prazosin is a potent antagonist of the human α1a-adrenoceptor subtype, which has been cloned and is expressed in the membrane fragments of CHO cells.
    hα1a receptor: The cloning of the human α1a cDNA was accomplished first by screening a human prostate cDNA library (Clontech), from which a portion of the coding region was obtained. This DNA fragment was then used to screen a human leukocyte genomic library (Clontech), and the rest of the coding sequence was obtained. Later these two fragments were spliced together. The entire coding sequence was then fully sequenced including matching PCR sequence with original genomic coding sequence, thus ensuring splice sites were joined correctly (Schwinn et al., 1995). Once sequenced, the gene was subcloned into the expression vector pcDNA3 (Invitrogen). Plasmid DNA was then used for transfection into CHO cells and G418 resistant clones were isolated. A clone expressing high levels of the hα1a receptor (as determined by mRNA and receptor binding data) was chosen and pharmacologically characterized.
    Culture Media Ingredients for Adherent α1a expressing CHO Culture:
     A. HAM's F-12 (Celigro)
     B. 10% 0.2 micron filtered Fetal Bovine Serum (FBS)(Celigro)
     C. 1% 0.2 micron filtered Penicillin-Streptomycin (Celigro)
     D. G418 0.2 micron filtered (Geneticin 400 μg/ml)(Celigro)
     Cells are cultured using established methods and procedures in either 150×25 mm culture plates (scale up to 100 plates) or a combination of these plates and 70 roller bottles. One culturing/harvest cycle typically requires 2 weeks and yields between 100-400 mg protein. Plates or bottles are incubated at 37° C.+5% CO2.
    Storage: Cells are harvested by mechanical scraping, washed using PBS, collected in 250 ml Corning polypropylene centrifuge tubes, spun down (1500 RPM) and resuspended in dH2O 10% DMSO (final volume per harvest is approximately 50 ml). Protein determination is made using the Biorad DC Assay Kit. Finally, the appropriate volume is aliquoted into a 2 ml Corning Cryovial (10 mg/1-1.5 ml) which is stored at −80° C.












Current Lot Data:


α1a (clone # 7)


Batch Jan. 14, 1998


















Receptor Concentration
2418 fmoles/mg protein



Kd
0.18 nM



Volume
1.5 ml/cryovial



Protein Concentration
approx. 10 mg/1.5 ml










    Assay Requirement: 0.5 cryovials per 96 well plate (assay volume=200 μl/well)
    [3H]-Ligand: [7-methoxy-3H]-Prazosin: 1.0 nM (NEN, NET-823)
     70-87 Ci/mmol
    Materials: Phentolamine mesylate (Research Biochemicals Int. #P-131)
     96 well flat bottom plates (Beckman)
     Unifilter GF/B Plate (Packard)
     Polyethylenimine (Sigma #P-3134)
     TomTec or Packard Filtermate 196 Cell Harvesters
     Packard TopCount Scintillation Counter
    Buffers: A: 50 mM Tris HCl; 0.1% ascorbate, pH 7.7 (incubation buffer)
     B: 50 mM Tris HCl; pH 7.7 (wash buffer)
    Procedure: Assay additions are as follows (in the order indicated):
     Total Binding=50 μl buffer A+50 μl [3H]prazosin+100 μl membrane
     Nonsp. Bd.=50 μl 10 μM phentolamine+50 μl [3H]prazosin+100 μl membrane
     Test Cpd.=50 μl compound+50 μl [3H]prazosin+100 μl membrane
     Compounds to be evaluated are weighed out to yield a 10 mM stock solution in DMSO in a 24 well polystyrene plate. This is diluted to a 0.5 mM stock in dH2O, Serial dilutions in Buffer A are made from which 50 μl additions to the plate are made in duplicate in order to achieve the final concentrations desired. Typically, one 96 well plate is used to evaluate 11 compounds at 4 concentrations (10−6-10−9 M) in duplicate. Total binding and nonspecific binding are determined in quadruplicate. Usually one standard is run with each assay.
     [3H]Prazosin is made up in Buffer A such that when 50 μl are added per well the final concentration is 1.0 nM in a final assay volume of 200 μl. The final concentration should be verified by running a sample in a scintillation counter prior to adding the [3H]prazosin to the 96 well plate. Note: The radioactivity should be prepared just before the additions are made so that it is not allowed to sit on the bench for very long.
     Packard GFIB Plate Pretreatment: The filter plates are presoaked for at least 30 min in ice cold Buffer B containing 0.05% polyethyleneimine (200 μl/200 ml) to maximize filtration efficiency and minimize filter blank binding.
     Incubation & Filtration: Once buffer, compounds, [3H]prazosin and membrane have been added (and mixed), the 96 well plates are incubated for 40 min at 37° C. and spaced 3-5 min apart. At 40 min, the plates are filtered using a Tomtec Automated Cell Harvester. Filtration is immediately followed by washes of ice cold Buffer. B (total vol.  ̃7 ml).
     Drying and Counting: Each filter plate is dried under a heat lamp for 15 min. The back of the plate is sealed and 40 μl of Packard microscint fluid are added per well. Using Packard film, each plate is heat sealed prior to being counted in a Packard Topcount Scintillation counter. A program has been written that counts each plate twice sending DPM, CPM and TSIS data to disk and printer.
     Analysis of Results: Raw DPM and CPM data are captured on disk and are imported into one of several software packages (Graphpad Prism Ver 2.0, Excel) residing on the LAN. Specific binding is defined as the difference between total binding and the binding in the presence of 10 μM phentolamine. Total binding is less than 10% of the total added ligand. Software using one-site competition curve analysis is employed in the calculation of IC50 and Ki (Cheng-Prusoff equation, 1973). The top and bottom of the non-linear regression are held constant at 0% and 100% inhibition. The percent Inhibition at each drug concentration is the mean of duplicate determinations.
    References:
    
        Vargas, H. M and A. J. Gorman. Life Sciences. Vol. 57, No. 25, pp. 2291-2308, 1995.
        Morrow, A. L. and I. Creese. Mol. Pharmacol. 29: 321-330, 1986.
        Piasdik, M. T., J. W. Kusiak, and K. W. Barron. Eur. J. Pharmacol. 11:101-107, 1989.
        Vargas, H. M., D. Cunningham, L. Zhou, H. B. Hartman and A. J. Gornan. J. Pharmacol. Exp. Ther. 267:264-272, 1993.
    
    


The functional activity of compounds of the invention (i.e. whether they are antagonists, agonists or partial agonists) can readily be determined using the microphysiometer test method that follows:
Chinese Hamster Ovary (CHO) cells, expressing the human dopamine D3 receptor, were grown on the surface of a capsule cup. Cups are assembled and placed on the microphysiometer, and buffer (Dulbecco's Modified Eagle's Medium without sodium bicarbonate and without serum) is perfused through the cup assembly until a stable baseline is achieved (4 hours). Buffer perfusion rate and solution changes are controlled by a computer. Intracellular acidification rate is measured in each of the 8 cup assemblies and recorded by the computer. Buffer containing test compound (10 nM, 100 nM, and 1 uM) was perfused-through the cup assembly for 20 min, then buffer containing quinpirole (a D3 agonist) (10 nM) and test compound (same concentrations) is perfused for an additional 1 min. This is followed by a recovery period of 10-60 min where buffer alone is perfused through the cups. Quinpirole increases acidification rate. A D3 antagonist will inhibit this acidification rate in a concentration dependent manner.
D3 antagonists are of potential use as antipsychotic agents for example in the treatment of schizophrenia, schizo-affective disorders, psychotic depression and mania. Conditions which may be treated by D3 agonists include dyskinetic disorders such as Parkinson's disease, neuroleptic-induced parkinsonism and tardive dyskinesias; depression; anxiety; dementia; circadian rhythm disorders, and drug (e.g. cocaine) dependency.
D3 receptor ligands are also useful for treating renal dysfunction.
In accordance with yet another embodiment of the present invention, there is provided a method of modulating the activity of dopamine D3 receptors, said method comprising: contacting cell-associated dopamine D3 receptors with a concentration of a compound of formula IB, or a physiologically acceptable salt thereof, sufficient to modulate the activity of said dopamine D3 receptor. As employed herein, a compound of formula IB shall refer to the compound of formula I except that the proviso therein is deleted therefrom.
As employed herein, the phrase “modulating the activity of dopamine D3 receptors” refers to a variety of therapeutic applications. Said therapeutic applications refer to the treatment of conditions or disorders which include dyskinetic disorders, psychoses, anxiety disorders, mood disorders, dementia, sleep disorders, nausea substance dependence, substance abuse and nausea.
The instant invention also provides a method of treating conditions or disorders of the central nervous system comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I, IA, or IB or a pharmaceutically acceptable salt thereof. The compounds of formula IA are preferred for this method. As employed herein, a “compound of formula IA” shall refer to the compound of formula I except that the proviso therein i.e. “Proviso A” is deleted therefrom and inserted therefor is the following proviso (hereinafter referred to as “Proviso B”):
“when n is 1; and y is 0; and R3 is hydrogen or C1-C6alkyl; and




is a group of formula (a); and R is group:

    
    
        (a) wherein R4 is hydrogen, halogen or C1-C6alkyl, and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a) wherein z is 0,
            (b) wherein u is 0 and M is hydrogen, halogen, C1-C6alkyl, or trifluoromethyl,
            (c) wherein o is 0,
            (d) wherein l is 0,
            (e) wherein j is 0,
            (g) wherein v is 0, or
            (i);
            and also when R is the group of formula (a), R1 and R2 cannot be joined together to form the group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl;
            
            
        
        
        (b) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (d) wherein l is 0,
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (b), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (d) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b) wherein u is 1,
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (d), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (e) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (e), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (f) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (f), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (g) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b) wherein u is 1,
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (g), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubsfituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (h) and R1 is hydrogen or unsubstituted C1-C6alkyl, then R2 cannot be a group of the following formula:
        
            (a),
            (b),
            (d),
            (k),
            (l), or
            (m) wherein Q is CH2;
            and also when R is the group of formula (h), R1 and R2 cannot be joined together to form a group of formula X or a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
            
                wherein said ring is unsubsubtituted or mono- or di-substituted with C1-C6alkyl or
            
            
        
        
    
    






    
    
        (j) then R1 and R2 cannot be joined together to form a group of formula Y or a 5-, 6-, or 7-membered monocyclic ring
        
            wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl”.
        
        
    
    


The instant invention further provides a method of treating conditions or disorders of the central nervous system comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I, IA or IB or a pharmaceutically acceptable salt thereof, in conjunction with one or more D1, D2, D4, Ds or 5HT receptor antagonists. Compounds of formula IA are preferred for this method.
Also provided herein is a method for treating renal dysfunction comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I, IA or IB.
In treating a patient afflicted with a condition or disorder described above, a compound of formula I, IA, or IB can be administered in any form or mode which makes the compound bioavailable in therapeutically effective amounts, including orally, sublingually, buccally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, rectally, topically, and the like. One skilled in the art of preparing formulations can determine the proper form and mode of administration depending upon the particular characteristics of the compound selected for the condition or disease to be treated, the stage of the disease, the condition of the patient and other relevant circumstances. For example, see Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co. (1990), incorporated herein by reference.
The compounds of Formula I, IA, or IB can be administered alone or in the form of a pharmaceutical composition in combination with pharmaceutically acceptable carriers, the proportion and nature of which are determined by the solubility and chemical properties of the compound selected, the chosen route of administration, standard pharmaceutical practice and other relevant criteria.
The compounds of formula I, IA, or IB may be administered orally, for example, in the form of tablets, troches, capsules, elixirs, suspensions, solutions, syrups, wafers, chewing gums and the like and may contain one or more of the following adjuvants: binders such as microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; and sweetening agents such as sucrose or saccharin may be added or a flavoring agent such as peppermint, methyl salicylate or orange flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or a fatty oil. Other dosage unit forms may contain other various materials which modify the physical form of the dosage unit, for example, as coatings. Thus, tablets or pills may be coated with sugar, shellac, or other enteric coating agents. A syrup may contain, in addition to the present compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.
The compounds of Formula I, IA, or IB may also be administered topically, and when done so the carrier may suitably comprise a solution, ointment or gel base. The base, for example, may comprise one or more of petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers.
The solutions or suspensions may also include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylene diaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials.
The highly lipophilic esters, amides and carbamates of compounds I, IA, or IB are capable of sustained release in mammals for a period of several days or from about one to four weeks when formulated and administered as depot preparations, as for example, when injected in a properly selected pharmaceutically acceptable oil. The preferred oils are of vegetable origin such as sesame oil, cottonseed oil, corn oil, coconut oil, soybean oil, olive oil and the like, or they are synthetic esters of fatty acids and polyfunctional alcohols such as glycerol or propyleneglycol.
The depot compositions of formula I, IA, or IB are prepared by dissolving a highly lipophilic ester, amide or carbamate of the instant invention in a pharmaceutically acceptable oil under sterile conditions. The oil is selected so as to obtain a release of the active ingredient over a desired period of time. The appropriate oil may easily be determined by consulting the prior art, or without undue experimentation by one skilled in the art.
The dosage range at which the compounds of formula I, IA or IB exhibit their ability to act therapeutically can vary depending upon the particular disease or condition being treated and its severity, the patient, the formulation, other underlying disease states that the patient is suffering from, and other medications that may be concurrently administered to the patient. Generally, the compounds of formula I, IA or IB will exhibit their therapeutic activities at dosages of between about 0.001 mg/kg of patient body weight/day to about 100 mg/kg of patient body weight/day.
In a further aspect, the present invention provides novel radiolabeled imaging agents of formula I, IA, or IB useful, inter alia, for imaging dopamine D3 receptors in the CNS to diagnose CNS abnormalities.
In a further aspect, the present invention provides novel radiolabeled imaging agents of formula I, IA, or IB useful, inter alia, for imaging dopamine D3 receptors in the CNS to diagnose CNS abnormalities.
The radiolabeled (tritiated and C-14 labeled) forms compounds of formula I, IA, or IB are useful as radioligands to determine the binding of compounds to the dopamine D3 receptor. They are also useful as labeled parent compounds to determine the metabolism of the compound in animals. Preferred for this purpose are compounds of formula I and IA wherein R is group (a) with a radiolabeled 14C in the 3-position of the benzo[b]thiophene ring, R4 is trifluoromethyl, s is 1, R3 is hydrogen, n is 1, y is 0, and A is N. Particularly preferred for this purpose are compounds of formula IC. As employed herein, a “compound of formula IC” shall refer to the compound of formula I wherein R is group (a) with a radiolabeled 14C in the 3-position of the benzo[b]thiophene ring, R4 is trifluoromethyl in the 6-position of the benzo[b]thiophene ring, s is 1, R3 is hydrogen, n is 1, y is 0, and A is N. Compounds of formula IC may be prepared in a manner analogous to that set forth in Example 33.
Imbalances in dopamine production have been implicated in a variety of mental and physical disorders, such as Parkinson's disease (PD). It is thus desirable to diagnose and monitor such imbalances and to monitor the effectiveness of drugs and substances that affect brain chemistry. New and powerful imaging methods that enable one to assess the living brain in vivo and thereby monitor brain chemistry and the effectiveness of drugs and substances that affect brain chemistry have been developed. Methods such as positron emission tomography (PET) and single photon emission computed tomography (SPECT) involve administering to a patient a radioactive tracer substance comprising a ligand that binds to the presynaptic or postsynaptic neuroreceptors in the patient's brain. Emissions (primarily gamma rays are emitted from the positrons or photons from the radioactive tracer) are measured. These emissions are indicative of the number and degree of occupancy of blocking of the neuroreceptors. The number of neuroreceptors and the degree of occupancy or blocking is calculated utilizing a mathematical model, and compared with an intra-person or inter-person control to determine the degree of drug response. Further treatment of the patient with drugs is based on the comparisons made. For these methods to be useful, however, a ligand that has a high specificity and affinity for the desired receptor is required.
It is believed that certain radioactive ligands may be selective for dopamine, transporters and are thus potentially useful in evaluating changes in dopamine function in vivo to and in vitro, especially for patients with Parkinson's disease (PD), which is characterized by a selective loss of dopamine neurons in the basal ganglia and substantia nigra.
Another aspect of this invention relates to methods for utilizing the compounds of the invention as CNS imaging agents. Imaging techniques are non-invasive diagnostic techniques that generally involve administering a compound with marker atoms that can be detected externally to the mammal. Generally, these methods comprise administering to a mammal a compound of the invention, dissolved or dispersed in a suitable pharmaceutical carrier or diluent. The compound of the invention selectively binds to dopamine D3, thus permitting the imaging of CNS receptors and the ability to, inter alia, evaluate brain chemistry, the effectiveness of drugs, and neuronal functions. Imaging techniques suitable for practicing the present invention include, but are not limited to, single photon emission computed tomography (SPECT) and positron emission tomography (PET).
Radionuclides that are widely used in diagnostic nuclear medicine include technetium [99Tc], iodine [123I], carbon [11C], and fluorine [18F]. The radiolabeled imaging agent specifically exemplified herein contains the radionuclide 11C. It should be noted that the same or similar radiochemistry reactions can be carried out using radionuclides other than 11C.
The invention is further illustrated by the following non-limiting examples and tabulated information. These examples are understood to be illustrative only and are not intended to limit the scope of the present invention in any way. As used herein, the following terms have the indicated meanings: “g” refers to grams; “mmol” refers to millimoles; “ml” refers to milliliters; “° C.” refers to degrees Celsius; “TLC” refers to thin layer chromatography; “LC/MS” refers to liquid chromatography mass spectrometry; “APCI” refers to atmospheric pressure chemical ionization; “mp” refers to melting point.
EXAMPLES
Example 1
Synthesis of Intermediate Substituted Piperazines




Example 1(a)
Preparation of Intermediate 3-benzyl-piperazine
To a suspension of 3-benzyl-piperazine-2,5dione (14.98 g, 73 mmol, prepared following generally the procedure of Halpern and Westley, J. Org. Chem. 1968, 33, 864) in dry diethyl ether (500 mL) is added dropwise to a solution of lithium aluminum hydride (400 mL of a 1 M solution in diethyl ether, 400 mmol, 5.4 eq). The suspension is heated at reflux for 23 hours and then cooled to 0° C. Water (70 mL) is then cautiously added and the resulting suspension is warmed to room temperature. After 3 hours the suspension is filtered and the solid washed with diethyl ether (1 L). The filtrate is concentrated under vacuum to provide crude title compound (11.40 g, 88%) as a yellow, crystalline solid. A sample (2 g) is recrystallized from cyclohexane and then from toluene to provide the purified title compound (0.83 g) as a fine, white crystals: mp 80-81° C.
Anal. Calcd. For C11H16N2: C, 74.96; H, 9.15; N, 15.89.
Found: C, 74.84; H, 9.01; N, 16.15.




Example 1(b)
To a solution of LDA (295 mL, 0.59 mol, 2 M in heptane/THF/ethylbenzene) in anhydrous THF (300 mL) cooled to 40° C. was added 2-methylpyrazine (48.5 mL, 0.531 mol) dropwise via an addition funnel. The reaction was allowed to warm to −20° C. and was stirred for 90 minutes when a solution of benzaldehyde (54 mL, 0.531 mol) in anhydrous THF (200 mL) was added dropwise via an addition funnel. After complete addition, the reaction was allowed to warm to room temperature and was stirred for 20 hours. The reaction was then cooled in an ice bath and saturated NH4Cl (500 mL) was added. The resulting mixture was extracted with EtOAc (500 mL, 250 mL). The combined extracts were dried (Na2SO4), filtered and concentrated to a damp, beige solid. The product was triturated with Et2O and collected then dried overnight to yield 56.0 g (53%) of a light brown solid, mp 81-84° C.
A solution of the above-obtained solid (56.0 g, 0.28 mol) in MeOH (1.1 L) and conc. HCl (290 mL) was stirred at reflux for 24 hours. The reaction was cooled to room temperature and concentrated to a dark liquid. The dark liquid was cooled in an ice bath and water (1 L) was added. The resulting solution was neutralized with a saturated solution of Na2CO3 and the product was extracted with EtOAc (1 L, 2×500 mL). The combined extracts were dried (Na2SO4), filtered and concentrated to yield 46 g of a dark brown solid. The solid was purified via flash column chromatography (40% EtOAc in heptane) yielding 22.7 g of the olefin as a brown foam.
A 1 L Parr shaker bottle was flushed with nitrogen and charged with 10% Pd/C (4.5 g, Degussa type) and the above-obtained olefin (20.0 g, 0.110 mol) in EtOH (450 mL). The reaction was hydrogenated at 50 psi for 3.5 hours when the reaction was filtered through a celite plug and rinsed with ethanol. The bottle was recharged with fresh 10% Pd/C (4.5 g, Degussa type), the filtrate and conc. HCl (15 mL). The reaction was hydrogenated at 50 psi for 18 hours when the reaction was diluted with warm MeOH and filtered through a plug of celite. The solid was thoroughly washed with hot MeOH and the filtrate was concentrated to yield 11.2 g (39%) of the final product as the di-HCl salt, mp 297-300.
See: Tetrahedron, 30, 1974 pp 667-673 and Tet. Lett. 1979, pp 4483-4486




Example 1(c)
DBU (14.0 g, 92 mmol) was added to a solution of the piperazine diacetate (18.2 g, 92 mmol) and aldehyde (12.3 g, 92 mmol) in 92 mL of DMF at ambient temperature. The resulting mixture was stirred at room temperature for 5 h. The precipitated product was collected by filtration, providing 17.1 g of product.




The monoacetate (17.0 g, 62.8 mmol) and hydrazine hydrate (9.4 g, 188.6 mmol) in 125 mL of DMF were stirred at room temperature for 20 h. The precipitated solid was collected by filtration, and washed with water and ethanol, leaving 13.7 g of product.




The olefin (13.6 g, 59.1 mmol) and palladium on carbon (2.7 g, 10% Pd/C, Degussa type, 50% H2O) in 1.2 l of methanol were shaken on a Parr hydrogenation apparatus at 40 psi of hydrogen, until hydrogen uptake ceased. The mixture was diluted with dichloromethane and filtered through celite. Concentration of the filtrate provided 12.1 g of product.




A solution of LAH (156 mL, 156 mmol, 1 M in THF) was added dropwise to a 0° C. solution of the piperazine dione (12.1 g, 52.1 mmol) in 100 ml of THF. The mixture was heated to reflux and stirred overnight. The mixture was cooled to 0° C. and 38 mL of water in 200 mL of THF was carefully added. The resulting mixture stirred for 1 h, then it was filtered, the filter cake was washed with THF, and the filtrate was concentrated in vacuo to give 7.4 g of product.
Example 2




1-(6-(trifluoromethyl)-benzo[b]thien-3-yl)-piperazine hydrochloride
2a: 2-Carbomethoxy-3-amino-6-trifluoromethylbenzo[b]thiophene
Equip a 22-L, 3-necked, round-bottom flask with a mechanical stirrer, nitrogen bubbler, and a thermocouple probe, charge with 1.20 kg (5.55 mole) of 2-nitro-4-trifluoromethylbenzonitrile, 589.3 g (496 mL, 5.55 mole) of methyl thioglycolate, and 4.3 L of NMP. Cool the resulting yellow solution to 2° C., and add slowly, over a period of 78 min a solution prepared from 466.0 g (11.11 mole, 2.0 eq) of lithium hydroxide monohydrate in 3.38 L of water while maintaining the temperature between 2-20° C. Allow the brown slurry to warm to 21° C. over a 2 h period, and then dilute with 8.0 L of water (observe exotherm->27° C.). Stir for 40 min and cool to 18° C., collect the product by filtration, rinsing with 10 L of water, then air-drying at ambient temperature to give 1.295 kg (84.7% yield) of 2-carbomethoxy-3-amino-6-trifluoromethylbenzo[b]thiophene, as a light-yellow solid, 99.8% pure by HPLC assay.
2b: 1-(6-(trifluoromethyl)-benzo[b]thien-3-yl)-piperazine hydrochloride
Equip a 12-L, 3-necked, round-bottom flask with a mechanical stirrer, nitrogen bubbler, and a thermocouple probe, and charge with 1.14 kg (4.14 mole) of 2-carbomethoxy-3-amino-6-trifluoromethylbenzo-[b]thiophene Example 2a, 196.0 g (2.28 mole, 0.55 eq) of piperazine, 4.0 L of NMP, and 570 mL of xylene. Heat the solution, and hold at 170-180° C. for 4 h, at which time the reaction is ca. 98% complete as determined by HPLC assay. Cool the brown solution to 168° C., and then add 1.605 kg (18.63 mole, 4.5 eq) of piperazine (temp->109° C.) following with 1.575 kg (28.28 mole, 2.0 eq) of p-toluenesulfonic acid monohydrate (observe exotherm, 109->130° C.). Connect a Dean-Stark trap to the condenser, and heat the reaction to collect an azeotrope. Remove a total of 410 mL of an aqueous distillate, while allowing the pot temperature to increase from 145 to 165° C. Monitor the progress of the reaction by GC/MS and HPLC assays. After 14 h at ca. 165° C. (>99% conversion by HPLC and GC/MS assay), cool the reaction to 30-35° C., and then quench into an extractor that contains 5 kg of ice, 12 L of water, and 8.5 L of toluene. Separate the phases, wash the organic extract with 11 L of 0.5 N NaOH, 2 L of saturated aq. NaCl., and then extract with 8 L of 1 N HCl. Dilute the acidic aqueous extract with 1 kg of ice, and basify to pH 11.2 by adding 624 g of 50% NaOH. Extract the resulting mixture with 9.5 L of toluene. Wash the toluene extract with 2 L of saturated aqueous NaCl, dry (Na2SO4), and filter. Charge the filtrate into a 22 L 3-necked, round-bottomed flask (N2, mechanical stirring, temperature control probe), and add a total of 3.7 L of 1 N ethereal HCl at 20-27° C. so that the mixture is positive to Congo Red Indicator paper. During the HCl addition, add a total of 2.5 L of toluene to improve the stirring of the thick slurry that results. Stir at ambient temperature for 40 min, filter the slurry and wash with 4.5 L of toluene. After air drying, obtain 1.165 kg (87% yield) of 3-piperazinyl-6-trifluoromethyl-benzo[b]thiophene hydrochloride as a light pink-beige solid, 99.1% pure by GC/MS assay.
Example 3




3a: trans-Cyclopropane-1,2-dicarboxylic acid monomethyl ester
Suspend trans-cyclopropane-1,2-dicarboxylic acid dimethylester (59.8 g, 0.378 mol) is suspended in 1.0N phosphate buffer (1.5 L, pH=7) add pig liver esterase (2.25 mL, 7500 units), and monitor NaOH consumption with a pH meter to control the reaction. After 3 h the consumption of 189 mL of 2N NaOH indicates the complete hydrolysis of the diester to the monomethylester. Acidified the clear solution by the addition of SN HCl to a pH=1. Separate the enzyme by addition of dichloromethane (500 mL) and diatomaceous earth (25 g). Stir for 5 min, and then filter the mixture. Saturate the filtrate with NaCl, and extract with ethyl acetate (5 times). Combine the extracts, dry (Na2SO4) and evaporate to obtain 50.8 g (93%) of solid, mp 46-47° C., m/z=45 (M+H)+
3b: (S,S)-(+)-Cyclopropane-1,2-dicarboxylic acid monomethyl ester
Add trans-cyclopropane-1,2-dicarboxylic acid monomethyl ester, Example 3a, (19.46 g) in acetone to quinine (43.8 g) In one portion. Heat the reaction to reflux, and then add methylcyclohexane (150 mL). After crystallization (5 times) from acetone/methylcyclohexane, collect 6.2 g of the diastereomeric salt (═D:+173, c:7.3 CHCl3)
3c: (R,R)-(−)-Cyclopropane-1,2-dicarboxylic acid monomethyl ester
Concentrate the filtrate from 3b above and treat the residue with 1N KHSO4 solution to yield 12.0 g of the crude (R,R) enantiomer. Dissolve this material in acetone and add 1 equivalent of quinidine in one portion. Heat the reaction to reflux, and then add methylcyclohexane. After crystallization overnight, collect 10.3 g of the diastereomeric salt (αD: −235, c: 8.5 CHCl3)
Example 4




4a: trans-2-Hydroxymethyl-cyclopropanecarboxylic acid methyl ester
Add boranemethyl sulfide complex (177 mL, 0.354 mol), slowly, by means of a dropping funnel, to a stirring solution of trans-cyclopropane-1,2-dicarboxylic acid monomethyl ester (Example 3a) (25.5 g, 0.177 mol), trimethyl borate (60.3 mL, 0.531 mol) and tetrahydrofuran (150 mL) at 0° C. After complete addition, allow the reaction to come to ambient temperature and stir for 2 h more. Pour the reaction mixture into a stirring solution of 50% aqueous sodium chloride solution (1.5 L)-concentrated HCl (10 mL). Extract the mixture with ethyl acetate (EtOAc) (3 times), combine the extracts, dry (Na2SO4) and concentrate the solvent to obtain a colorless oil: 22.6 g.
4b: (S,S)-(+)-2-Hydroxymethyl-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 4a, and substitute (S,S)-(+)-cyclopropane-1,2-dicarboxylic acid monomethyl ester (Example 3b) therein to obtain the title compound, αD: +54, c: 1.5 CHCl3 (Tetrahedron Asymmetry Vol. 6, No. 3, pp. 683-684, 1995)
4-c: (R,R)-(−)-2-Hydroxymethyl-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 4a, and substitute (R,R)-(−)-cyclopropane-1,2-dicarboxylic acid monomethyl ester (Example 3c) therein to obtain the title compound (αD: −78.6, c: 4.3 CHCl3)
Example 5




5a: trans-2-Methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester
Add, dropwise, triethylamine (7.74 mL, 56 mmol) and 4-dimethylaminopyridine (0.013 g, 0.106 mmol) in dichloromethane (30 mL) to a stirred solution of trans-2-hydroxymethyl-cyclopropanecarboxylic acid methyl ester (Example 4a) (2.4 g, 18.64 mmol), at 0-5° C. After 0.5 h, pour the reaction mixture into water and extract the mixture with dichloromethane (3 times). Wash the combined extracts with 1N KHSO4, dry (Na2SO4) and concentrate to yield 4.29 g of a pale yellow oil, which solidifies when stored at 0° C., m/z=209 (M+H)+
5b: (S,S)-(+)-2-Methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 5a, and substitute (S,S)-(+)-2-hydroxymethyl-cyclopropanecarboxylic acid methyl ester (Example 4b) therein to obtain the title compound (αD: +75, c: 4.7 CHCl3)
5c: (R,R)-(−)-2-Methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 5a, and substitute (R,R)-(−)-2-hydroxymethyl-cyclopropanecarboxylic acid methyl ester (Example 4c) therein to obtain the title compound (αD: −74.4, c: 5.9 CHCl3).
Example 6




6a: trans-2-[4-(6-Trfluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester
Heat at reflux for 16 h, a mixture of 1-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazine, free base of Example 2b, (23.0 g, 71.3 mmol), trans-2-methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester (Example 5a) (15.3 g, 73.5 mmol), and triethylamine (40 mL, 288 mmol) in acetonitrile (600 mL). Concentrate the reaction mixture under reduced pressure and dilute the resultant oil with EtOAc (30 mL). Filter the resulting precipitate (unreacted starting piperazine) away and purify the filtrate by column chromatography over silica gel (EtOAc/heptane/MeOH/triethylamine, 20:20:1). Concentration of the appropriate fractions gives 18.0 g of colorless oil, m/z—413 (M+H)+.
6b: (S,S)-(+)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 6a, and substitute (S,S)-(+)-2-methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester (Example 5b) therein to obtain the title compound (αD: +48, c: 2.8 EtOH).
6c: (R,R)-(−)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 5, and substitute (R,R)-(−)-2-methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester, Example 5c, therein, to obtain the title compound (αD: −49.3, c: 3.5 CHCl3).
Example 7




7a: trans-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid
Add 5N NaOH solution (425 mL, 226 mmol) to a solution of trans-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester, Example 6a, (18.0 g, 45.1 mmol) in dioxane/methanol (400 mL, 3:1) and heat the reaction at 60° C. for 6 h. Selectively remove most of the methanol and acidify the remaining dioxane solution to pH 5-6 with 2N acetic acid. Collect the desired compound, which precipitates from solution, to obtain 14.08 g (81%) of colorless crystals, m/z=385 (M+H)+
7b: (S,S)-(+)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid
Follow the procedure of Example 7a, and substitute (S,S)-(+)-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester (Example 6b) therein to obtain the title compound (αD: +55, c: 1.05 EtOH).
7c: (R,R)-(−)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid
Follow the procedure of Example 7a, and substitute (R,R)-(−)-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid methyl ester (Example 5c) therein to obtain the title compound (αD: −40.5, c: 0.79 CHCl3).
Example 8




8a: trans-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Stir a solution of trans-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid (Example 7a) (1.99 g, 5.0 mmol), and O-[(ethoxycarbonyl)-cyanomethylenamino]-N,N,N′,N′-tetramethyluronium-tetrafluoroborate (TOTU, 2.05 g, 6.3 mmol) in DMF (50 mL) at 20° C. for 0.5 h. Add N-methylmorpholine (0.58 mL, 5.3 mmol), after which add trans-4-methylcyclohexylamine (1.13 g, 10 mmol), and stir the reaction for 5 h. Concentrate the reaction under reduced pressure to obtain an oil. Purify by column chromatography over silica gel (EtOAc(heptane/methanol/triethylamine, 20:20:1:1) to obtain 1.86, (76%) of colorless crystals LC/MS, m/z=480 (M+H)+.
8b: (S,S)-(+)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Follow the procedure of Example 8a, and substitute (S,S)-(+)-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid (Example 7b) therein to obtain the title compound, LC/MS, m/z=480 (M+H)+.
8c: (R,R)-(−)-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methytcyclohexyl)-amide
Follow the procedure of Example 8a, and substitute (R,R)-(−)-2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid (Example 7c) therein to obtain the title compound LC/MS, m/z=480 (M+H)+.
Example 9




trans-4-([2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-ylmethyl]-cyclopropanecarbonyl}-amino)-trans-cyclohexanecarboxylic acid ethyl ester
Follow the procedure of Example 8a, and substitute trans-4-amino-cyclohexane carboxylic acid ethyl ester (J. Med. Chem. (1971), 14(7), 600-614) for trans-4-methylcyclohexylamine therein to obtain the title compound LC/MS, m/z=538 (M+H)+.
Example 10




trans-2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 8a, and substitute trans-4-ethylcyclohexylamine for trans-4-methylcyclohexylamine therein to obtain the title compound LC/MS, m/z=494 (M+H)+.
Example 11




trans-2-[4-(3-Chloro-phenyl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide




11a: trans-2-(4-Methyl-cyclohexylcarbamoyl)-cyclopropanecarboxylic acid methyl ester
Stir a solution of trans-cyclopropane-1,2-dicarboxylic acid monomethyl ester (Example 3a) (3.0 g, 20.8 mmol) and TOTU (8.5 g, 26 mmol) in DMF (300 mL) at ambient temperature for 20 min at which time add N-methylmorpholine (2.4 mL 22.9 mmol) and trans 4-methylcyclohexylamine 3.06 g, 27.0 mmol). After 3 h, remove the solvent and dissolve the residue in H2O/EtOAc. Extract the aqueous layer 3 more times with EtOAc and combine the extracts. Wash the extract with saturated NaHCO3 solution, brine and dry over MgSO4. Concentrate the solvent to afford the crude product, and purify by column chromatography over silica gel to give 4.76 g (95%) of colorless crystals, m/z=240 (M+H)+.




11b: trans-2-(trans-2-Methylcyclohexylcarbamoyl)-cyclopropanecarboxylic acid
Stir a solution of trans-2-(4-methyl-cyclohexylcarbamoyl)-cyclopropane-carboxylic acid monomethyl ester (Example 11a) (4.5 g, 19.8 mmol), 5N NaOH (40 mL) and MeOH/dioxane overnight. Concentrate the reaction mixture under vacuum, cool the resulting solution and acidify to pH 5. Collect the precipitate, wash (H2O) and dry under vacuum at 40° C. to obtain 3.7 g (87%) of solid.




11c: trans-2-Hydroxymethyl-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Stir a solution, under argon, at 0° C., of trans-2-(trans-2-methylcyclohexylcarbamoyl)-cyclopropanecarboxylic acid (Example 11b) (3.69 g, 16.4 mmol), trimethyl borate (5.6 mL, 50 mmol), and add slowly 2N borane dimethylsulfide complex in THF (16.4 mL, 32.8 mmol). Continue to stir the reaction at 0° C. for 1 h, and then at ambient temperature for 2 h. Carefully pour the reaction mixture into ice-H2O and acidify with 2N HCl. Extract the mixture with EtOAc (3 times), wash with brine, dry over MgSO4 and concentrate to obtain 2.85 g (83%) of product as a solid.




11d: Methane sulfonic acid 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester
Add, dropwise, to a stirring suspension at −20° C. of trans-2-hydroxymethyl-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide (Example 11c) (2.85 g, 13.5 mmol), triethylamine (3.74 mL, 27.0 mmol) and 4-(dimethylamino)pyridine (0.1 g, 0.82 mmol) in dichloromethane (100 ml) a solution of methanesulfonic acid (1.59 mL, 20.3 mmol) in DCM (10 mL). When the reaction is complete, pour the mixture into ice water and extract the mixture with DCM. Combine the extracts, wash with 0.1N KHSO4, brine, dry over MgSO4 and concentrate under vacuum. Treat the residue with a mixture of isopropanol/heptane and collect the precipitate by filtration to obtain 2.91 g (74%) of solid, MS m/z=290 (M+H)+.
11e: trans-2-[4-(3-Chloro-phenyl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Reflux a solution of methane sulfonic acid 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 11d) (30.0 mg, 0.1 mmol), 1-(3-chloro-phenyl)-piperazine hydrochloride (29.0 mg, 0.13 mmol) triethylamine (0.05 mL, 0.4 mmol) in acetonitrile (5 ml) for 15 h. Concentrate the reaction mixture under vacuum and chromatograph the residue over silica gel to obtain 11.3 mg (28%) of solid, LC/MS, m/z=390 (M+H)+.
Example 12




trans-2-[4-(pyrazin-2-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute 1-(pyrazin-2-yl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound LC/MS, m/z=358 (M+H)+.
Example 13




trans-2-[4(3-Chloro-5-trifluoromethyl-pyridin-2-yl)-piperazin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide




13a: trans-2-(4-Ethyl-cyclohexylcarbamoyl)-cyclopropanecarboxylic acid methyl ester
Follow the procedure of Example 11a, and substitute trans-(4-ethylcyclohexylamine) for trans-(4-methylcyclohexylamine) therein to obtain the title compound, m/z=254 (M+H)+.




13b: trans-2-trans-2-Ethylcyclohexylcarbamoyl)-cyclopropanecarboxylic acid
Follow the procedure of Example 11b, and substitute trans-2-(4-ethyl-cyclohexylcarbamoyl)-cyclopropanecarboxylic acid monomethyl ester (Example 13a) for trans-2-(4-methyl-cyclohexylcarbamoyl)-cyclopropanecarboxylic acid monomethyl ester therein to obtain the title compound.




13c: trans-2-Hydroxymethyl-cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11c, and substitute trans-2-(4-ethyl-cyclohexylcarbamoyl)-cyclopropanecarboxylic acid (Example 13b) for trans-2-(4-methyl-cyclohexylcarbamoyl)-cyclopropane carboxylic acid therein to obtain the title compound.




13d: Methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester
Follow the procedure of Example 11d, and substitute trans-2-hydroxymethyl-cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide, Example 13c for methane sulfonic acid 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester therein, to obtain the title compound, m/z=304 (M+H)+.
13e: trans-2-[4-(3-Chloro-5-trifluoromethyl-pyridin-2-yl)-piperazin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(3-chloro-5-trifluoromethyl-pyridin-2-yl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound LC/MS m/z=473 (M+H)+.
Example 14




trans-2-[4-(4-Trifluoromethyl-phenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-methylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute 1-(4-trifluoromethyl-phenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound MS, m/z=424 (M+H)+, 85.2% pure by HPLC assay.
Example 15




trans-2-[4-(4-Trifluoromethyl-phenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for methane sulfonic acid 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(4-trifluoromethyl-phenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound MS m/z=438 (M+H)+, 78.5% pure by HPLC assay.
Example 16




trans-2-[4-(4-Chlorophenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethyl cyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester, Example 13d for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(4-chlorophenyl)-piperazine hydrochloride for 1-(3-chlorophenyl)-piperazine hydrochloride therein, to obtain the title compound LC/MS, m/z=404 (M+H)+.
Example 17




trans-2-[4-(3,4-Dichlorophenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(3,4-dichlorophenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound LC/MS, m/z=438 (M+H)+.
Example 18




trans-2-[4-(2,5-Dimethylphenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(2,5-dimethylphenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound LC/MS, m/z=398 (M+H)+.
Example 19




trans-2-[4-(2,5-Dimethylphenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 1e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(2,5-dimethylphenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein, to obtain the title compound LC/MS, m/z=398 (M+H)+.
Example 20




trans-2-[4-(2-Methylphenyl)-piperazin-1-ylmethyl]cyclopropanecarboxylic acid trans-(4-ethylcyclohexyl)-amide
Follow the procedure of Example 11e, and substitute methane sulfonic acid 2-(trans-4-ethyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester (Example 13d) for 2-(trans-4-methyl-cyclohexylcarbamoyl)-trans-cyclopropylmethyl ester and 4-(2-methylphenyl)-piperazine hydrochloride for 1-(3-chloro-phenyl)-piperazine hydrochloride therein to obtain the title compound LC/MS, m/z=384 (M+H)+.
Example 21




3-Piperidinyl-4-yl-thieno[2,3-d]isoxazole hydrochloride




21a: 4-(3-Bromo-thiophene-2-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester
Stir a solution, under nitrogen, of 3-bromothiophene (21.0 mL, 0.224 mol) in tetrahydrofuran (1.0 L) at −78° C., and add a 2.0M solution of lithium disopropylamide in heptane/tetrahydrofuran/ethylbenzene (112 mL, 0.224 mol) for 45 min. Add, dropwise, over 2 h, a solution of 4-(N-methoxy-N-methylcarboxamido)-1-piperidinecarboxylic acid 1,1-dimethylethyl ester (prepared according to U.S. Pat. No. 5,134,139) (79.4 g, 0.291 mol) in tetrahydrofuran (800 mL). Stir for 2 h, add a saturated ammonium chloride solution, and stir for an additional 0.5 h. Filter the resulting solid, and pour the filtrate into water (800 mL). Extract the aqueous mixture with ether and concentrate to obtain a dark liquid. Pour the liquid into water (400 mL), add NaCl and extract the aqueous mixture with ether. Wash the extract with water, brine, and dry over Na2SO4. Filter and concentrate to obtain the crude product. Chromatograph the product over silica gel (pet.ether/ether, 4:1) to obtain 41.5 g (50%) of white solid.




21b: 4-[(3-Bromo-thiophen-2-yl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester
Stir a mixture of 4-(3-bromo-thiophene-2-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester (Example 21a) (41.5 g, 0.11 mol), hydroxylamine hydrochloride (15.4 g, 0.23 mol) and pyridine (190 mL) at ambient temperature overnight. Pour the reaction into water (500 mL) and extract with dichloromethane (3 times). Wash the combined extracts with saturated CuSO4 solution (2 times), dry (MgSO4) and concentrate to a green solid. Dissolve the solid in toluene (175 mL) and let stand at ambient temperature for 3 h. Collect the resulting crystals that form and wash with toluene (60 mL). Concentrate the filtrate and again dissolve the residue in toluene and proceed to collect additional crystals to obtain a total yield of 25 g (58%) of the title compound as a light, green solid.




21c: 4-Thieno[2,3-d]isoxazol-3-yl-piperidine-1-carboxylic acid tert-butyl ester
Add to a stirring solution of 4-[(3-bromo-thiophen-2-yl)-hydroxyimino-methyl]piperidine-1-carboxylic acid tert-butyl ester (Example 21b)(25 g., 64.2 mmol) in 2-methoxyethanol (200 mL), a solution of potassium hydroxide (7.2 g, 128.4 mmol) in water (20 mL). Heat the reaction to 60° C. and then add copper powder (1.25 g). Stir at 60-70° C. for 6 h and then at ambient temperature overnight. Pour the reaction mixture into water (500 mL) and extract with EtOAc (3 times). Concentrate to a dark residue and purify by column chromatography over silica gel (heptane/EtOAc, 4:1) to provide 9.8 g (50%) of a white solid.
21d: 3-Piperidinyl-4-yl-thieno[2,3-d]isoxazole hydrochloride
Add ethereal HCl (10 mL) to 4-thieno[2,3-d]isoxazol-3-yl-piperidine-1-carboxylic acid-tert-butyl ester (Example 21c)(1.0 g, 3.2 mmol) and then methanol (1 mL) to effect solution. Permit to stand at ambient temperature for 1 h and then collect 0.34 g of white solid, mp 240-241° C. From the filtrate collect 0.25 g of additional white solid, mp 263-265° C. Both samples: MS, m/z=209 (M+H)+.
Analysis (sample mp 263-265° C.):















Calc. For: C10H12N2OS•HCl:
49.08% C
5.35% H
11.45% N


Found:
49.03% C
5.29% H
11.25% N









Example 22




trans-2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid methyl ester




22a: trans-2-(4-Thieno[2,3-d]isoxazol-3-1-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid methyl ester
Reflux a solution of 3-piperidinyl-4-yl-thieno[2,3-d]isoxazole trifluoroacetate, (trifluoroacetic acid salt of Example 21d)(6.0 g, 18.6 mmol), trans-2-methanesulfonyloxymethyl-cyclopropanecarboxylic acid methyl ester (Example 5a)(3.99 g, 19.2 mmol) and triethylamine (7.6 g, 75 mmol) in acetonitrile (100 mL) for 15 h. Concentrate and purify the residue by column chromatography over silica gel (dichloromethane/EtOAc/diethylamine (8:2:1). Further purify by another chromatography dichloromethane/MeOH (95:5) to obtain 1.4 g (24%) of product, MS, m/z=321 (M+H)+.
22b: trans-2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid methyl ester
Add 5N NaOH (4.4 mL, 21.9 mmol) to a solution of trans-2-(4-thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid methyl ester, Example 22a (1.4 g, 4.37 mmol) in dioxane/methanol (24 mL, 3:1), and heat to 60° C. for 3 h. Concentrate the reaction and treat the residue with acetic acid. After 36 h dilute the acetic acid solution with dichloromethane and wash the organic phase with 5% aqueous HCl, water and brine. Dry the organic phase and concentrate to a yellow oil. Evacuate the oil at high vacuum and 60° C. for 20 h to obtain 0.191 g of oil.
Treat the aqueous washings with NaCl and collect an additional 1.0 g of the product as a solid. MS, m/z 307 (M+H)+.
Example 23




Racemic-trans-2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethyl-cyclohexyl)-amide
Add TOTU (415 mg, 1.06 mmol) to a solution of trans-2-(4-thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid methyl ester (Example 22b)(300 mg, 1.0 mmol), in dimethylformamide (approximately 10 mL), and allow the reaction to stir at ambient temperature for 0.5 h. Add N-methylmorpholine (107 mg, 1.26 mmol) and trans-4-ethylcyclohexylamine (254 mg, 2.0 mmol), and stir for 6 h. Add additional equivalents of N-methylmorpholine and TOTU and continue to stir for overnight. I think again I ddin't write it down Concentrate the reaction and purify the residue by column chromatography over silica gel (dichloromethane/MeOH, gradient 0→30%) to obtain 270 mg (65%), LC/MS, m/z=416 (M+H)+.
Example 24




Chiral-trans-2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethyl-cyclohexyl)-amide
Dissolve racemic-trans-2-(4-thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethyl-cyclohexyl)-amide (Example 23)(14 mg) in heptane/ethanol (1.5 mL,  ̃1:1) and make 20 separate injections of 50 μL each on to a preparative HPLC apparatus (Chiralpak AD 10μ 250×4.6 mm, heptane/ethanol, 85:15). Collect and combine fractions that elute at tR=14.2 min and obtain 3.0 mg of enantiomer with an enantiomeric excess>99%.
Example 25




Chiral-trans-2-(4-Thieno[2,3-d]isoxazol-3-yl-piperidin-1-ylmethyl)-cyclopropanecarboxylic acid trans-(4-ethyl-cyclohexyl)-amide
Follow the procedure of Example 24, but collect and combine the fractions that elute at tR=23.6 min to obtain 6.0 mg of enantiomer with opposite chirality and an enantiomeric excess>99%.
Example 26




The cyclobutane carboxylic acid (26-1)(1.43 g, 9.05 mmol) and TOTU (3.7 g, 9.5 mmol) were combined in 90 mL of DMF and stirred at ambient temperature for 0.5 h. N-Methylmorpholine (0.96 g, 9.5 mmol) and the amine (26-2)(1.53 g, 13.5 mmol) were added and the mixture stirred at room temperature for 18 h. The mixture was diluted with ethyl acetate and the organic phase was sequentially washed with saturated sodium bicarbonate solution (3×), 10% HCl solution and brine (2×), then dried over magnesium sulfate, filtered and concentrated. The residue was dissolved in ethyl acetate and the mixture filtered. Concentration of the filtrate provided 1.5 g of the product (263).
Lithium borohydride 93.0 mL, 5.93 mmol, 2M in THF) was added dropwise to a solution of the ester (26-3)(1.5 g, 5.93 mmol) in 20 mL of THF at ambient temperature. The solution was stirred at room temperature for 20 h, and tehn lit was quenched by the careful addition of water. The product was extracted into ethyl acetate, and the combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered and concentrated to leave 1.1 g of product. Chromatography on silica gel (40 g Biotage column) eluting with ethyl acetate and dichloromethane gave 618 mg of the alcohol (26-4).
Methanesulfonyl chloride (0.276 g, 2.41 mmol) was added dropwise to a solution of the alcohol (26-4)(0.493 g, 2.19 mmol) and triethylamine (0.332 g, 3.29 mmol) in 11 mL of dichloromethane at 0° C. The mixture was stirred at 0° C. for 1 h, and then it was stirred at ambient temperature for 2 h. The resulting solution was diluted with dichloromethane and washed with saturated sodium bicarbonate solution, water, and brine and then filtered and the filtrate was concentrated to provide 627 mg of the product (26-5).




In Scheme V, R is as hereinbefore defined. For example, the mesylate (26-5)(75 mg, 0.025 mmol), potassium carbonate (52 mg, 0.038 mmol) and the amine (Example 2)(72 mg, 0.025 mmol) in 2 mL of acetonitrile were heated at 80° C. in a JKEM reaction block for 20 h. The mixture was cooled to room temperature, diluted with 6 mL of dichloromethane and 400 mg of polymer supported isocyanate resin was added (Argonaut Technologies, resin loading 1.49 mmol/gram) and the mixture shaken at ambient temperature for 20 h. The reaction mixture was deposited on a 2 g Varian strong cation exchange (SCX) column. The resin was washed twice with 4 mL ethyl acetate and thrice with 4 mL of dichloromethane. The column was further washed with 15 mL of ethyl acetate. The product was next eluted using a 4:1 mixture of 2% triethyl amine in ethyl acetate and methanol. The volatiles were removed in vacuo and the residue was chromatographed on silica gel, eluting with methanol/ethyl acetate. Concentration of the fractions containing the product provided 67 mg of product.
The following compounds were prepared using this method.












CPD


Compound
#











829997


 






829998


 






829999


 






830000


 






829996









Characterizing data for the compounds is as follows:

















MOL
Amt


R.T.
MS %


CPD #
WT
obt
yield
basepeak
(min.)
area





















829997
457.65
0.051
45
458.3
1.6
100


829998
493.63
0.067
54
494.3
1.64
100


829999
443.63
0.062
56
444.3
1.57
100


830000
430.61
0.040
37
431.3
1.44
100


829996
416.59
0.052
50
417.3
1.43
100









Example 27




1-(6-Fluoro-benzo[b]thiophen-3-yl)-[1,4]diazepane
27a. 3-Amino-6-fluoro-benzo[b]thiophene-2-carboxylic acid
At 50° C., add to a stirring solution of 2-carbomethoxy-3-amino-6-fluoro-benzo[b]thiophene (prepared according to U.S. Pat. No. 5,143,923), (90.1 g, 0.4 mol) in H2O (450 mL), a 50% aqueous solution of NaOH (64 g, 0.8 mol) over 2-3 min. Heat the reaction to 70-73° C. and continue to stir for 3 h. Add 10% aqueous isopropanol (45 mL) and bring to reflux. Remove the isopropanol under N2 and add H2O (300 mL). Cool the reaction mixture to between 7-10° C. and add concentrated HCl (80 mL). Add H2O (650 mL), cool to 5-70° C., filter the resulting solid, and wash the filter cake with H2O (2×150 mL). Dry the solid under vacuum at 35° C. to obtain 80.6 g (94.7%) of solid mp 160-163° C., TLC on silica gel (dichloromethane/methanol, 3:1), Rf=0.69.
27b. 1-(6-Fluoro-benzo[b]thiophen-3-yl)-[1,4]diazepane
Heat a solution of 3-amino-6-fluoro-benzo[b]thiophene-2-carboxylic acid (5.0 g, 24 mmol) in 1-methyl-2-pyrrolidinone (5 ml) to 100° C. for 2 h., and then, introduce a stream of nitrogen, to cool the solution to room temperature. Add homopiperazine (9.5 g, 95 mmol) and p-toluene sulfonic acid monohydrate (9.0 g, 47 mmol) and heat the mixture to 145° C. for 4 h. After that time, cool the reaction mixture to room temperature, dilute with ethyl acetate (30 mL) and wash with brine (3×15 mL). Separate the organic layer and dry over Mg SO4. Evaporate the solvent and purify the crude product by column chromatography (SiO2, 100 g CH2Cl2MeOH 9:2, then CH2Cl2/MeOH/NH4OH 9:2:0.15) to give 3.9 g (65%) of yellowish oil LC/MS (LiChrospher 5μ, RP-18, 250 mm CH3CN/Water-radient 20%→100% (25 min), Flow: 1.5 mL/min) tR=10.74 min, m/z=250.3.
Example 28




1-(2,6-Difluoro-benzo[b]thien-3-yl)-piperazine trifluoroacetate




28a: 4-(6-Fluoro-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester
Add a solution of di-tert-butyl dicarbonate (5.15 g, 23.6 mmol) in CHCl3 (15 mL), dropwise, over 45 min to a solution at −65° C. of 1-(6-fluorobenzo[b]thiophen-3-yl)-piperazine (prepared according to U.S. Pat. No. 5,143,923), (2.8 g, 11.8 mmol), 4-(dimethyl-amino)pyridine (0.16, 1.3 mmol), and diisopropylethylamine (4.3 mL, 3.2 g, 24.8 mmol) in CHCl3 (50 mL). Following complete addition, stir the reaction at ambient temperature for 20 h, and then pour the reaction into a mixture of cold (5° C.) 5% aqueous NaOH/EtOAc (150/150 mL). Extract the product into EtOAc, wash the extract with H2O, brine and concentrate to a red oil. Purify the crude oil over silica gel (EtOAc), to obtain 3.6 g, of red oil, LC/MS m/z=337 (M+H)+.




28b: 4-(2-Bromo-6-fluoro-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester
Add N-bromosuccinimide (0.59 g, 3.3 mmol) to a stirring solution of 4-(6-fluoro-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester (Example 28a)(1.00 g, 2.97 mmol) in CHCl3 (32.8 mL) and reflux for 30 min. Allow cooling to room temperature and filter. Evaporate the solvent and purify the residue by chromatography over silica gel (EtOAc/heptane, 9:1) to obtain 0.53 g (43%) of oil, MS, m/z=416 (M+H)+.
In an alternative procedure, add N-bromosuccinimide (1.319 g, 6.62 mmol) to a stirring solution of 4-(6-fluoro-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester (Example 46a)(2.226 g, 6.62 mmol) in CCl4 and reflux for 2 h. Allow cooling to room temperature and filter. Evaporate the solvent and purify the residue by chromatography over silica gel (EtOAc/heptane, 9:1) to obtain 2.34 g (94%) of oil.




28c: 4-(2-Fluoro-6-fluoro-benzo[b]thiophen-3-yl)-piperazine-1 carboxylic acid tert-butyl ester
At a temperature of −65° C. stir, under nitrogen, a solution of the 4-(2-bromo-6-fluo-o-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester (Example 28b)(15.59 g, 37.55 mmol) in anhydrous THF (247 mL) and add, dropwise, n-butyllithium in hexane (2.5M, 19.53 mL, 48.82 mmol). Stir for 30 min and then add, dropwise, N-fluorobenzenesulfonimide (17.76 g, 56.33 mmol) dissolved in anhydrous THF. Stir overnight at ambient temperature, cool the reaction to 0° C., add saturated NaCl solution and then water. Extract the mixture with EtOAc (3x's), combine the extracts and wash with water and brine. Dry the extract (MgSO4), and concentrate to obtain 11.0 g of oil. Chromatograph the oil over silica gel (ether/pet. ether, 9:1) and obtain 6.28 g (52%) of red oil, MS, m/z, 354 (M+H)+.
28d:1-(2,6-Difluoro-benzo[b]thien-3-yl)-piperazine trifluoroacetate
Stir a solution of 4-(2-fluoro-6-fluoro-benzo[b]thiophen-3-yl)-piperazine-1-carboxylic acid tert-butyl ester (Example 28c)(250 mg, 0.70 mmol) in trifluoroacetic acid (2.2 mL) at ambient temperature for 30 min. Evaporate the trifluoroacetic acid and treat the residue with ether. Stir the suspension at ambient temperature for 2 h, and filter the resulting white solid to obtain 191 mg (56%) of the trifluoroacetate salt. MS, m/z=255 (M+H)+.
Examples 29-31
The following HPLC conditions are referred to in examples 29-31:
HPLC Condition I:
A) 95/5/0.1% Water/Acetonitrile/Formic Acid,
B) 5/95/0.1% Water/Acetonitrile/Formic Acid.
Column: YMC ODS-A 4×50 mm, Flow rate: 2 mL/minute.
The initial HPLC conditions consisted of 100% (A) flowing at 2 mL/minute. After the initial injection a linear gradient was performed so that at 2 minutes the HPLC conditions were 100% B. These conditions were then held for 3.4 minutes at which time the system switched back to initial conditions and equilibrated for the next analysis.
HPLC Condition II:
A) 95/5/0.1% Water/Acetonitrile/Formic Acid,
B) 5/95/0.1% Water/Acetonitrile/Formic Acid.
Column: YMC ODS-A 2×50 mm, Flow rate=1 mL/minute.
The initial HPLC conditions consisted of 100% (A) flowing at 0.1 mL/minute. After the initial injection a linear gradient was performed so that at 2 minutes the HPLC conditions were 100% B. These conditions were then held for 3.5 minutes at which time the system switched back to initial conditions and equilibrated for the next analysis.
Example 29




6-Methyl-3-piperidin-4-yl-thieno[2,3-d]isoxazole hydrochloride




29a: Preparation of 4-[1-(3-bromo-4-methyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester
Under inert conditions, add a 2.0 M solution (in tetrahydrofuran/n-heptane) of lithium diisopropylamide (29.65 mmol, 14.83 mL, 1.05 equivalents) to a cold (−78° C.) solution of 3-bromo-4-methylthiophene (28.24 mmol, 5.00 g, 1.00 equivalents) in dry tetrahydrofuran (27.33 mL). Stir at −78° C. for 1 hour and add a solution of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid tert-butyl ester (28.24 mmol, 7.69 g, 1.00 equivalents), dropwise. Continue stirring at −78° C. for 3 hours. Quench the reaction mixture with saturated ammonium chloride (aqueous, 55 mL) and allow to warm to room temperature. Extract the reaction mixture with a mixture of ethyl acetate:diethyl ether (1:1, 3 times 40 mL). Combine the extracts and dry over magnesium sulfate, filter and evaporate. Purify the residue via flash column chromatography using a mixture of n-heptane:ethyl acetate (4:1) to yield a yellow, crystalline solid (9.84 g).
MS (CI, methane) m/e 388 (MH+), LC/MS (APCI), m/e 288 (M-100), retention time 2 min. 43 sec. Condition I




29b: Preparation of 4-[1-(3-bromo-4-methyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester
Add ammonium hydroxide hydrochloride (50.68 mmol, 3.52 g, 2.00 equivalents) to a stirred solution of 4-[1-(3-bromo-4-methyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester (25.54 mol, 9.84 g, 1.00 equivalents) in pyridine (47.5 mL). Stir at room temperature overnight and at 70° C. for 4 hours. Cool the reaction mixture and add hydrochloric acid (3 M solution, 115 mL). Extract the reaction mixture with dichloromethane (115 mL), filter the organic layer, wash with water (100 mL), dry over magnesium sulfate, filter and evaporate. Recrystallize the resulting residue from toluene to yield a white solid (4.84 g).
LC/MS (APCI), m/e 403 (MH+), retention time 2 min. 32 sec. Condition I.




29c: Preparation of 4-(6-methyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester
Add cesium carbonate (3.72 mmol, 1.21 g, 1.50 equivalents) and copper iodide (0.25 mmol, 47 mg, 0.10 equivalents) to a stirred solution of 4-[1-(3-bromo-4-methyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (2.48 mmol, 1.00 g, 1.00 equivalents) in 2-methoxy ethanol (25 mL). Stir the resulting mixture at room temperature overnight and filter to remove the inorganic material. Concentrate the filtrate and partition the resulting oil between ethyl acetate (75 mL) and water (25 mL). Extract the aqueous layer with ethyl acetate (2×75 mL) and wash the combined organic layers with saturated sodium chloride (aqueous, 25 mL), dry over magnesium sulfate, filter and evaporate. Purify the residue via flash column chromatography eluting with n-heptane:ethyl acetate (4:1) to yield a white solid (588 mg). MS (CI, methane) m/e 323 (MH+), LC/MS (ESI), m/e 345 (MNa+), retention time 2.05 minutes. Condition II.
29d: Preparation of 6-methyl-3-piperidin-4-yl-thieno[2,3-d]isoxazole hydrochloride




Stir a solution of 4-(6-methyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (8.84 mmol, 2.85 g, 1.00 equivalents) in hydrochloric acid (48.75 mL, 1 M solution in diethyl ether) and methanol (2.00 mL) at room temperature for 3.5 hours. Filter the suspension, collect the white solid and dry to yield the desired product (659 mg). Allow the mother liquor to age overnight, filter, collect the white solid and dry to yield additional desired product (1.252 g). LC/MS (ESI), m/e 223 (MH+), retention time 1.14 minutes. Condition II.
Example 30




4-(5-Methyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid




30a: Preparation of 4-[1-(3-bromo-5-methyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester
Prepared essentially as in 29a except that 2-bromo-5-methyl thiophene is used as the starting material. In addition, 1.20 equivalents of lithium diisopropylamide and 1.24 equivalents of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid. tert-butyl ester are used for the reaction. Accordingly, stirring time of the reaction mixture may vary. Purification of the residue via flash column chromatography uses a gradient with a mixture of ethyl acetate: n-heptane (1:9) to ethyl acetate: n-heptane (2:8) to yield a yellow oil. LC/MS (ESI), m/e 332 (M-56) and 388 (MH+), retention time 2.15 minutes. Condition II.




30b: Preparation of 4-[1-(3-bromo-5-methyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester
Prepared essentially as 29b except that 4-[1-(3-Bromo-5-methyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester is used as the starting material and the reaction mixture was stirred at 70° C. for 6 hours. LC/MS (ESI), m/e 347 (M-56) and 403 (MH+), retention time 2.03 minutes. Condition II.




30c: Preparation of 4-(5-methyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid
Prepared essentially as in 29c except that 4-[1-(3-bromo-5-methyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester is used as the starting material. Two other differences are: 1) 0.05 equivalents of copper iodide is used, and 2) no partition between ethyl acetate and water accompanied by subsequent extraction with ethyl acetate is required. Purification of the residue via flash column chromatography uses a mixture of ethyl acetate: n-heptane (1:4) to yield a white solid. LC/MS (ESI), m/e 345 (MNa+), retention time 2.12 minutes. Condition II.
Example 31




5-Methoxymethyl-3-piperidin-4-yl-thieno[2,3-d]isoxazole hydrochloride




31a: Preparation of (4-bromo-thiophen-2-yl)-methanol
Under inert conditions, add sodium borohydride (13.82 mmol, 0.523 g, 2.08 equivalents) in absolute ethanol (16 mL) dropwise over a period of 15 minutes to a stirred mixture of 4-bromothiophene-2-carboxaldehyde (26.58 mmol, 5.08 g, 1.00 equivalents) in cold (0° C.) absolute ethanol (32 mL). Stir the resulting mixture at room temperature for 2.5 hours and add glacial acetic acid dropwise until the effervescence ceases. Evaporate the resulting solution, take the residue up in diethyl ether (75 mL), wash with water (15 mL) and brine (15 mL) and dry over magnesium sulfate. Filter and evaporate to yield the product as a colorless oil (5.13 g).




31b: Preparation of 4-bromo-2-methoxymethyl-thiophene
Add sodium hydride (737 mg, 29.23 mmol, 1.10 equivalents, 95%) to a solution containing methyl iodide (1.65 mL, 26.57 mmol, 1.00 equivalents) and (4-bromo-thiophen-2-yl)-methanol (5.13 g, 26.57 mmol, 1.00 equivalents) in tetrahydrofuran (dry, 25 mL). Stir the resulting mixture at room temperature overnight and evaporate. Partition the residue between water (100 mL) and dichloromethane (100 mL). Extract the aqueous layer with dichloromethane (100 mL), combine the organic layers, dry over magnesium sulfate, filter and evaporate to yield the desired product as a yellow oil.




31c: Preparation of 4-[1-(3-bromo-5-methoxymethyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester
Add lithium diisopropyl amide (13.20 mL, 26.37 mmol, 1.05 equivalents) to a stirred, cold (−78° C.) solution of 4-bromo-2-methoxymethyl-thiophene (5.20 g, 25.11 mmol, 1.00 equivalents) in tetrahydrofuran (dry, 24.30 mL). Stir at −78° C. for 1 hour and add a solution of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid tert-butyl ester (6.84 g, 25.11 mmol, 1.00 equivalents) in tetrahydrofuran (dry, 16.40 mL), dropwise. Stir the resulting solution at −78° C. for 3 hours. Quench the reaction mixture with saturated sodium chloride (aqueous, 50 mL). Allow the resulting mixture to warm to room temperature and extract with a mixture of ethyl acetate: diethyl ether (1:1, 3×35 mL). Combine the extracts, dry over magnesium sulfate, filter and evaporate. Purify the residue via flash column chromatography eluting with a mixture of n-heptane:ethyl acetate (4:1) to yield the desired product as a yellow oil (9.47 g). LC/MS (ESI), m/e 362 (M-56) and 418 (MH+), retention time 2.08 minutes. Condition II.




31d: Preparation of 4-[1-(3-bromo-5-methoxymethyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester
Add hydroxylamine hydrochloride (2.29 g, 45.27 mmol, 2.00 equivalents) to a stirred solution of 4-[1-(3-bromo-5-methoxymethyl-thiophen-2-yl)-methanoyl]-piperidine-1-carboxylic acid tert-butyl ester (9.47 g, 22.64 mmol, 1.00 equivalents) in pyridine (42.40 mL). Stir the resulting solution at room temperature overnight and then at 70° C. for 4 hours. Cool the reaction mixture slightly, add hydrochloric acid (3N, 100 mL) and extract the resulting mixture with dichloromethane (100 mL). Wash the extract with water (100 mL), dry over magnesium sulfate, filter and evaporate to yield the desired product as a yellow oil (9.48 g).




31e: Preparation of 4-(5-methoxymethyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester
Add cesium carbonate (1.13 g, 3.46 mmol, 1.50 equivalents) and copper iodide (44 mg, 0.23 mmol, 0.10 equivalents) to a stirred solution of 4-[1-(3-bromo-5-methoxymethyl-thiophen-2-yl)-1-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (1.00 g, 2.31 mmol, 1.00 equivalents) in 2-methoxy ethanol (23.30 mL). Stir the resulting mixture at room temperature overnight or up to 3 days and filter through celite. Evaporate the filtrate, partition the residue between ethyl acetate (70 mL) and water (23 mL) and separate. Extract the aqueous layer with ethyl acetate (3×70 mL), combine the organic layers, dry over magnesium sulfate, filter and evaporate. Purify the residue via flash column chromatography eluting with a mixture of hexane:ethyl acetate (4:1) to yield the desired product as a yellow oil. LC/MS (ESI), m/e 375 (MNa+), retention time 1.98 minutes. Condition II.




31f: Preparation of 5-methoxymethyl-3-piperidin-4-yl-thieno[2,3-d]isoxazole hydrochloride
Stir a solution of 4-(5-methoxymethyl-thieno[2,3-d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (2.21 g, 6.68 mmol, 1.00 equivalents) and hydrochloric acid (1.0 M in diethly ether, 35 mL) overnight to form a suspension. Add additional hydrochloric acid (1.0 M in diethyl ether, 10 mL). Stir the suspension overnight, filter and wash the solid with ether. Collect the solid and dry to yield the desired product as a dark blue solid. LC/MS (ESI), m/e 253 (MH+), retention time 1.17 minutes. Condition II.
Example 32




Synthesis of BOC protected piperazine-thienylisoxazole
3-Bromothiophene-2-carbaldehyde oxime
3-Bromothiophene-2-carbaldehyde (Maybridge)(28.7 gm, 0.15 mol) in ethanol (50 ml) was added in one portion to a solution of hydroylamine hydrochloride (13.8 gm, 0.2 mole), sodium hydroxide (8 gm, 0.2 mol) in water (30 ml) and ethanol (100 ml). The mixture was stirred at 0° C. for 2 hours and was kept at 0° C. overnight. The reaction mixture was diluted with cold water (600 ml), and the precipitated solids were collected by filtration to provide 20.5 gm, (67%) of product. The aqueous layer was further extracted with ethyl acetate and, the combined organic layers were washed with brine, dried with magnesium sulfate filtered and concentrated in vacuo to leave an additional 6.9 g of product.
3-bromothiophene-2-hydroximidoyl chloride
To a solution of 3-bromothiophene-2-carbaldehyde oxime (10.8 gm, 52.4 mmol), hydrogen chloride (14.5 ml, 4M in dioxane) in DMF (100 ml) was added oxone (16.9 gm, 1.05 eqiv) in one portion at room temperature. The mixture was stirred at ambient temperature overnight.
At the end of the reaction, DMF solution was poured into water and product was extracted into ethyl acetate. The organic solution was washed with brine, dried over magnesium sulfate, filtered and concentrated in vacuo to 12.68 gm of product which was used in the next reaction without furthur purification.
(4-t-Butoxycarbonylpiperazinyl)-3-bromo-2-thienyl methanone oxime
3-bromothiophene-2-hydroximidoyl chloride (16.4 gm, 68 mmol) in tetrahydrofuran (THF, 70 ml) was added dropwise to a solution of N-(t-butoxycarbonyl)piperazine (14 gm, 1.1 equiv.), DABCO (9.5 gm, 1.25 eqiv.) in DMF (100 ml) at 0° C. over 25 minutes. The mixture was stirred for 3.5 hrs. At the end, the mixture was poured into water and was extracted with ethyl acetate. The organic was washed with brine and dried over magnesium sulfate. The solvent was removed on a rotary evaporator. The crude product (30.5 gm) was purified by chromatography on a Biotage cartridge (400 gm of silica gel), eluting with methanol in dichloromethane (0-5% of MeOH). The product thus obtained weighed 24.6 gm (85%).
(t-BOC-piperazine)-3-thienylbenzisoxazole
A mixture of (4-t-Butoxycarbonylpiperazinyl)-3-bromo-2-thienyl methanone oxime (10.3 gm, 26.4 mmol), cesium carbonate (10.7 gm, 32.7 mmol), and copper iodide (500 mg) in methoxyethanol (200 ml) was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate, the washed with water. The aqueous solution was extracted three times with ethyl acetate. The organic solution (total 600 ml) was washed with brine and was dried over magnesium sulfate then concentrated to an oil ( ̃10 gm). This material was purified by chromatography using a Biotage cartridge (120 gm of silica gel, eluting with 0-8% Methanol in dichloromethane). The product thus obtained as light oil (5.1 gm, 62%).
Example 33
Synthesis of radiolabeled C14 intermediate useful for the preparation of certain compounds within the scope of the present invention.




General: Analytical thin layer chromatography (TLC) was performed on E. Merck TLC plates with silica gel 60 F254 (0.25 mm). TLC plates used in the analysis of radioactive samples were scanned on a BIOSCAN system 2000 Imaging Scanner using P-10 gas (10% methane, 90% argon). Identity of the intermediates was established by co-migration in radio-TLC and/or radio-H PLC with the standard samples of unlabeled analogues. Flash chromatography was performed using silica gel with a particle size of 40-63 μm. Specific activity was determined on a Packard Minaxi Tri-Carb Uquid Scintillation Analyzer (Model 1600 TR) using Bio-Safe II as scintillation cocktail.
Purification of compounds VI-2, VI-3, VI-4, VI-5, and VI-6 was monitored by HPLC (conditions:A) which was carried out on Waters 600 Controller, Waters 996 Photodiode Array Detector, Millennium Chromatography Manager and Beta-Ram Radioactive Flow Through Monitor System, Model 2 (IN/US Systems Inc.). Final purity determination of VI-7 by HPLC (conditions:B) was performed on Waters Model 510 Pumps, Waters 680 Gradient Controller, Waters 715 Ultra Wisp Autosampler, Waters 484 Tunable Absorbance Detector and Beta-Ram Radioactive Flow-Through Monitor System, Model 2 (IN/US Systems Inc.).
Conditions A: YMC Basic 5 μm, C18, 4.6×250 mm, mobile phase A: (v/v) 50/50 acetonitrile/0.1N ammonium formate, mobile-phase B: (v/v) 75(25 acetonitrile/0.1N ammonium formate, flow rate 1.0 mL/min, uv detection at 254 nm.















Time (minutes)
% MP:A
% MP:B





















Gradient:
0
100
0




15
100
0




25
0
100




30
0
100




35
100
0










Conditions B: Ultremex 5 μm, C8, 4.6×150 mm, mobile phase (v/v/v) 50/50/0.25 acetonitrile/0.05 M potassium phosphate buffer, pH 3.0/triethylamine, flow rate 1.0 mL/min, uv detection at 210 nm.
[14C] Copper (I) Cyanide (VI-1):
A Solution of Copper (II) Sulfate Pentahydrate
(4.16 g, 16.67 mmol) in water (13.3 mL) was heated to 70° C. and a solution of sodium metabisulfite (1.94 g, 6.28 mmol) in water (3.3 mL) at 70° C. was added in one minute. Immediately a solution of [14C] potassium cyanide (245.5 mg, 200 mCi, 3.77 mmol, S.A. 53.0 mCi/mmol) and unlabeled potassium cyanide (0.84 g, 12.9 mmol) in water (3.3 mL) at 70° C. was added in one minute. A white solid precipitated out of solution and blue color of the solution was discharged. After stirring for 10 min at 70° C., the mixture was filtered hot and the solid was washed with hot water (15 mL) and ethanol (15 mL). The white solid was dried under vacuum (0.1 mm Hg) for 27 h 45 min to prove VI-1 (1.393 g, 186.6 mCi) in 93.3% yield.
2-Nitro-4-(trifluoromethyl)[7-14C]benzonitrile (VI-2)
To a suspension of [14C]copper (I) cyanide (VI-1)(1.393 g, 15.55 mmol, 186.6 mCi) in 1-methyl-2-pyrrolidinone (NMP, 10 mL) was added 4-bromo-3-nitrobenzotrifluoride (6.33 g, 23.45 mmol) and the mixture was heated at 190-195° C. for 1 h. Ethyl acetate (25 mL) and water (20 mL) were added at room temperature and the mixture was filtered through celite. To the filtrate more water (20 mL) and ethyl acetate (25 mL) were added and the aqueous layer was extracted with ethyl acetate (90 mL). The organic extract was washed with iron (III) chloride solution (50 mL) prepared by dissolving iron (III) chloride (7.468 g, 46.04 mmol) in water (50 mL). The organic extract was further washed with water (30 mL), sat. sodium chloride (15 mL), dried (Na2 SO4) and the solvent was removed in vacuo. The residue was purified by flash chromatography on silica gel (hexane/ethyl acetate, 9/1-7/3) to provide an oil which was dissolved in hexane (70 mL). The solvent was removed under reduced pressure and residue was dried under vacuum for 15 h 40 min to provide VI-2 (3.01 g, 167.13 mCi, 89.6% yield) as a yellow solid. Radio-TLC (hexane/ethyl acetate, 9/1), Rf=0.21; HPLC (System A), RCP 99.86% (ret. time, 9.2 min).
[3-14C]-3-Amino-2-carbomethoxy-6-trifluoromethylbenzo[b]thiophene (VI-3)
Nitrile (VI-2)(3.01 g, 13.9 mmol, 167.13 mCi) was dissolved in DMF (14 mL) and methyl thioglycolate (1.78 g, 15.94 mmol, 95%) was added in one minute. The mixture was cooled to 0-5° C. and a solution of lithium hydroxide (0.689 g, 28.77 mmol) in water (9.2 mL) was added dropwise in 12 minutes. After the addition, cooling bath was removed and the mixture was stirred at room temperature for 4 hours. Water (70 mL) was added at 0-5° C. and the mixture was stirred for 15 min at 0-5° C. the solid was collected on a filter, washed with water (20 mL) and dried under vacuum (0.1 mm Hg) for 40 h 15 min to provide VI-3 (3.469 g, 151.24 mCi, 90.49% yield). Radio-TLC(CH2Cl2), Rf=0.372; HPLC (system A), RCP 99.92% (ret. time, 16.722 min).
[3-14C]-3-Amino-6-trifluoromethylbenzo[b]thiophene (VI-4)
To a solution of benzo[b]thiophene (VI-3)(3.469 g, 12.6 mmol, 151.2 mCi) in NMP (14 mL) was added 1-methylpiperazine (6.699, 66.79 mmol) and the mixture was heated at 140-145° C. for 5 h. The mixture was allowed to cool to room temperature, poured into water (60 mL) and extracted with ethyl acetate (140 mL). The organic extract was washed with water (30 mL), sat. sodium chloride (10 mL), dried (Na2SO4) and the solvent was removed in vacuo.
The residue was purified by flash chromatography on silica gel (hexane/ethyl acetate, 1/1) to yield a greenish solid which was dried under vacuum (0.1 mm HG) for 14 h to provide VI-4 (w.66 g, 146.95 mCi, 97.16% yield).). Radio-TLC (hexane/ethyl acetate, 1/5), Rf=0.407; HPLC (system A), RCP 99.44% (ret. time, 10.552 min).
1-[6-trifluoromethyl)benzo[b]thien-3-yl-[3-14C]piperazine (VI-5)
To a solution of benzo[b]thiophene (VI-4)(2.66 g, 12.24 mmol, 146.95 mCi) in NMP (17 mL) was added piperazine (4.309 g, 50.02 mmol) and p-toluenesulfonic acid (4.769, 25.02 mmol) at room temperature. The mixture was heated at 170° C. for 20 m h 24 min, allowed to cool to room temperature and poured into a solution of sodium carbonate (4.70 g, 44.3 mmol) in water (60 mL). The mixture was extracted with ethyl acetate (20 mL), dried (Na2SO4) and the solvent was removed in vacuo. The residue was purified by flash chromatography on silica gel (CH2Cl2/MeOH/NH4OH, 9/1/0.2) and product was dried under vacuum (0.1 mm Hg) for 11 h 50 min. Ethanol (absolute, 30 mL) was added to the product and solvent was removed under reduced pressure. The residue was dried under vacuum (0.1 mm Hg) for 24 h 55 min to provide VI-5 (3.44 g, 144.18 mCi, 98.1% yield) as an oil. Radio-TLC(CH2Cl2/MeOH/NH4OH, 9/110.2), Rf=0.46; HPLC (system A), RCP 99.88% (ret. time, 5.807 min).
Example 34







3-Bromo-thiophene-2-carboxylic acid. To a solution of 3-bromothiophene (600.0 g, 3.68 mol) in THF (3 L) cooled to −72° C. was added LDA (1.93 L, 3.86 mol, 2 N) slowly over 2 hours. The rate of LDA addition is such that the reaction temperature never exceeded 48° C. After complete addition, the solution is stirred for an additional 40 minutes. Diethyl ether (3 L) is then added via an addition funnel such that the temperature is maintained below −65° C. The addition funnel is then replaced with a dispersion tube and CO2 gas is bubbled through the solution for 3 hours. Dry ice (500 g) is then added and the mixture is stirred overnight. The reaction flask is then placed in an ice bath and 6 N HCl is added slowly to prevent excessive bubbling until the pH of the solution is adjusted to 1-2. The resulting mixture is then extracted with EtOAc. The extract is washed with brine then dried over MgSO4, filtered and evaporated. The product is dried under vacuum at room temperature yielding 585.15 g (77%) as an off-white solid.




1-(3-Bromo-thiophene-2-carboxylic acid)-2-(4-toluenesulfonyl)-hydrazine. To a stirred suspension of the acid (285.53 g, 1.38 mol) in DCM (1.5 L) was added a catalytic amount of NMP (2 mL). Thionyl chloride (105.8 mL, 1.45 mol) is then added and the solution is refluxed until the solids have completely dissolved. The solution is further refluxed for 1 hour, cooled to room temperature and evaporated to afford a light, brown solid. The crude material is dried under vacuum overnight. The brown solid is taken up in toluene (3.5 L) and p-toluenesulfonhydrazine (402.25 g, 2.16 mol) is added. The mixture is stirred at 100° C. for 8 hours then at room temperature overnight. The resulting mixture was cooled with an ice bath and the resulting solids were collected by filtration and washed with toluene. The solids were then stirred as a slurry in 1 N HCl for 1 hour. The solids were collected by filtration and washed with copious ambunts of water. The solid were dried under vacuum at 40° C. then recrystallized from toluene/isoproyl alcohol yielding 484.28 g (93%) of the desired product.




N-((4-Methylphenyl)-sulfonyl)-3-bromo-thiophene-2-carbohydrazonyl chloride. 1-(3-Bromo-thiophene-2-carboxylic acid)-2-(4-toluenesulfonyl)-hydrazine (60.80 g, 0.161 mol) was added to thionyl chloride (70.5 mL, 0.966 mol). The resulting mixture was stirred at 80° C. until the mixture becomes homogenous. The solution is then stirred at 70° C. for 30 minutes and heptane (300 mL) is added over a period of 20 minutes. The solution was cooled slowly to room temperature then cooled further to 5° C. The solids are collected by filtration, washed with heptane (3×100 mL) and dried under vacuum yielding 62.1 g (98%) of the desired product as an off-white solid.




3-(4-Benzyl-piperazin-1-yl)-1-(toluene-4-sulfonyl)-1H-thieno[3,2-c]pyrazole. To a stirred solution of DABCO (14.18 g, 112.18 mol) and benzylpiperazine (35.35 g, 0.200 mol) in DMF (200 mL) cooled to −30° C. was added via cannula a solution of N-((4-Methylphenyl)-sulfonyl)-3-bromo-thiophene-2-carbohydrazonyl chloride (62.1 g, 0.158 mol) in TH F (100 mL). The addition is controlled to prevent the reaction temperature from exceeding −30° C. After complete addition precipitation occurs and the mixture is then allowed to stir at room temperature overnight when K2CO3 (65.41 g, 0.473 mol) and CuCl (1.0 g, 0.010 mol) was added. The resulting mixture is heated to 110° C. and the THF is removed by distillation at this point. The temperature is then increased to 140° C. and the mixture is stirred for 6 hours, cooled to room temperature and stirred overnight. The mixture was then poured overwater (100 mL) and EtOAc (100 mL). The EtOAC layer is then separated and the aqueous layer is extracted with EtOAC (3×500 mL). The combined EtOAC layers were washed with water (500 mL) and then filtered through celite and concentrated. The solids were collected by filtration and washed with cold water then EtOAc/heptane (1:4) and dried under vacuum yielding 66.05 g (95%) of the desired product as an off-white solid.




3-(4-Benzyl-piperazin-1-yl)-1H-thieno[3,2-c]pyrazole. To a stirred mixture of KOH(S) (56.09 g, 2.66 mol) in methly alcohol (1.33 L) is added 3-(4-benzyl-piperazin-1-yl)-1-(toluene-4-sulfonyl)-1H-thieno[3,2-c]pyrazole (241 g, 0.532 mol). The mixture is heated at reflux for 1.25 hours, cooled to room temperature and evaporated. The residue is taked up in EtOAc (1 L) washed with water (2 L), dried (MgSO4) filtered and evaporated. The residue was recrystallized from EtOAc/Heptane yielding 129 g (81%).




3-(4-Benzyl-piperazin-1-yl)-1-methyl-1H-thieno[3,2-c]pyrazole. To a stirred solution of 3-(4-benzyl-piperazin-1-yl)-1H-thieno[3,2-c]pyrazole (318.0 g, 1.07 mol) in THF (2.5 L) was added a mixture of potassium t-butoxide (134.4 g, 1.2 mol) in THF (1.5 L) dropwise over a period of 1 hour while keeping the reaction temperature below 25° C. After complete addition, the mixture was cooled to −30° C. and Mel (65.4 mL, 1.05 mol) was added dropwise over a period of 30 minutes. The mixture is then slowly warmed to room temperature overnight. To the reaction mixture is slowly added saturated NaHCO3 (1 L). The solution is then evaporated to remove the THF and the resulting aqueous mixture is taken up in EtOAc and washed with water and brine. The EtOAc extract is dried (Na2SO4), filtered and evaporated. The viscous concentrate is filtered through a silica gel plug with 1:1 EtOAc/heptane and evaporated yielding a viscous oil that is then dried under vacuum where it solidifies and yields 326.03 g (98%) as a 12:1 ratio of regioisomers in favor of the desired product.




1-Methyl-3-piperazin-1-yl-1 H-thieno[3,2-c]pyrazole. To a solution of a mixture of 3-(4-Benzyl-piperazin-1-yl)-1-methyl-1H-thieno[3,2-c]pyrazole and the 2-methyl analog (189.0 g, 0.60 mol) is dissolved in DCM (1.25 L) is added 1-chloroethylchloroformate (78.6 mL, 0.72 mol). The solution is heated at reflux for 1 hour when the mixture is cooled and the solvent is removed by evaporation. The residue is taken up in methanol (1 L) and heated at reflux for 30 minutes. After cooling, the solution is treated with 1 N HCl in ether (200 mL) and an additional 1 L of ether to afford the precipitation of the product. The solid is collected via filtration and washed with cold ether. The solid is recrystallized from methanol (1 L) and the HCl salt is collected by filtration, washed with ether and dried under vacuum yielding 123.04 g (80%) of the desired product as an 80:1 mixture of regioisomers in favor of the desired regioisomer as seen by NMR.




2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperazin-1-ylmethyl]-cyclopropane-1R-carboxylic acid methyl ester. A mixture of 1-methyl-3-piperazin-1-yl-1H-thieno[3,2-c]pyrazole (2.75 g, 12.37 mmol), 2R-bromomethyl-cyclopropane-1R-carboxylic acid methyl ester (2.41 g, 12.5 mmol) and K2CO3 (3.46 g, 25.0 mmol) in acetonitrile (75 mL) was heated at reflux for 1 hour and then at room temperature overnight. The mixture was diluted with EtOAc (100 mL) and then poured into water (100 mL). The layers were separated and the aqueous layer was extracted with EtOAc (50 mL). The combined organic layers were washed with brine (30 mL), dried (MgSO4), filtered and evaporated. The residue was separated via column chromatography (9:1; EtOAc/MeOH) yielding 3.65 g (88%) of the desired product.




2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperazin-1-ylmethyl]-cyclopropane-1R-carboxylic trans(4-methyl-cyclohexyl)-amide. To a solution of 2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperazin-1-ylmethyl]-cyclopropane-1R-carboxylic acid methyl ester (10.0 g, 29.9 mmol) and trans-4-methylcyclohexylamine (6.77 g, 60.0 mmol) in toluene (100 mL) was added sodium methoxide (9.72 g, 45.0 mmol, 25% in MeOH). The resulting mixture was heated to 80° C. for 21 hours. Upon cooling to room temperature, the mixture was partitioned between water (800 mL) and EtOAc (200 mL) and separated. The aqueous layer is salted out with the addition of NaCl then extracted with EtOAc (200 mL). The combined organic layers are washed with brine (50 mL), dried (Na2SO4), filtered and evaporated. The residue was recrystallized from toluene (100 mL) and EtOAc (20 mL) to yield 9.30 g (75%) of the desired product as a white solid, MP 185-186° C. Anal. Calcd. for C22H33N5OS: C, 63.58; H, 8.00; N, 16.85. Found: C, 63.68; H, 8.24; N, 16.62.
Example 35




Trityloxymethyl-(1R, 2R)-cyclopropanecarboxylic acid ethyl ester. To a suspension of sodium hydride (15.20 g, 380 mmol, 60% oil dispersion) in xylenes (300 mL) was added triethylphosphonoacetate (85.07 g, 379 mmol) in a controlled manner to avoid the excessive evolution of gas and to maintain the internal temperature less than 55° C. After the complete addition, the mixture was stirred for 20 minutes when the yellow solution was added via cannula to a solution of (R)-trityl glycidyl ether (100.0 g, 316 mmol) in xylenes (300 mL). The resulting solution was heated to 125° C. for 2 hours. The resulting solution was cooled to room temperature, acidified with the addition of 10% HCl (320 mL) and extracted with EtOAc (2×300 mL). The combined extracts were washed with brine (100 mL), dried (MgSO4), filtered and evaporated yielding a 175 g of a crude product as an oil. The material was carried on crude.




2R-bromomethyl-cyclopropane-1R-carboxylic acid methyl ester. A solution of triphenylphosphine (124.7 g, 1.34 mol) in CH2Cl2 (260 mL) was cooled to 5° C. when a solution of bromine (24.4 mL, 1.34 mol) in CH2Cl2 (65 mL) was added over 20 minutes while the temperature was maintained below 12° C. The mixture was stirred at 5° C. for 1 hour when 2 M HCl/Et2O (16 mL, 32 mmol) was added followed by the addition of crude trityloxymethyl-(1R,2R)-cyclopropane carboxylic acid ethyl ester (124 g, 0.32 mol). The resulting mixture was stirred at room temperature overnight when saturated NaHCO3 (600 mL) was added. The mixture was separated and the aqueous layer was extracted with CH2Cl2 (200 mL). The combined organic layers were washed with water (400 mL), dried (MgSO4), filtered and evaporated. The residue was diluted with heptane (200 mL) and evaporated two times to remove excess CH2Cl2. The residue was allowed to stand for 30 minutes when the solid impurities were removed by filtration. The filter cake was washed with heptane (2×400 mL). The combined organic layers were evaporated to provide 92.68 g of a crude yellow liquid. The crude liquid was distilled (BP=80-85° C./1.5 torr) to provide 55.19 g (84% yield for the two steps) of a colorless liquid.
Example 36




4-(2-Fluoro-5-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester. A solution of 4-fluorobenzotrifluoride (25 g, 0.152M) in anhydrous THF (300 ml) was cooled to 60° C. (IPA/CO2 bath) and treated with n-butyl lithium (84 mL of a 2.0M solution in Hexane, 0.168M-1.1 eq) with a maximum rate so not to exceed −60° C. The reaction was stirred for 3 hours (temperature maintained) and then treated with a solution of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid tert-butyl ester (51.86 g, 0.190M-1.25 eq, in 130 mL of anhydrous THF) with a maximum rate so as not to exceed −55° C. The mixture was stirred for a further two hours before allowing to warm to room temperature and stirred for 0.5 hours. The reaction was quenched with saturated ammonium chloride solution (75 mL) and the THF removed under reduced pressure. The residue was dissolved in ethylacetate (800 mL), washed with. 1N Hydrochloric acid (400 ml), 5% aq NaHCO3 (400 mL), water (400 mL) and brine (400 mL) successively. The organics were dried over MgSO4, filtered and concentrated to give a brown oil, which on triturating in ethyl acetate gave a white solid 27.6 g (48%).




4-[(2-Fluoro-5-trifluoromethyl-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester. A solution of 4-(2-fluoro-5-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester (59, 0.013M) in pyridine (25 mL) was treated with hydroxylamine hydrochloride (1.11 g, 0.015 M-1.2 eq). The reaction was stirred under N2 at room temperature for 14 hours and then poured onto ice water (250 mL). The mixture was stirred at 0° C. for 1 hour, the product was then filtered off, washed with cold water (3×15 mL) and dried in a vacuum oven at 50° C. A white solid was obtained (5.03 g, 97%).




4-(5-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester. A solution of 4-[(2-Fluoro-5-trifluoromethyl-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (4.969 g, 0.013 M) in anhydrous THF (59 mL) was treated with Potassium tert-butoxide (13.4 mL of a 1M solution in THF, 0.0133 M-1.05 eq). The mixture was stirred at ambient temperature for 1 hour and then heated to 65° C. for 2 hours. The THF was removed under reduced pressure. The residue was dissolved in ethyl acetate (100 mL), washed with H2O (50 mL) and brine (50 mL) respectively. It was then dried over MgSO4, filtered and concentrated to give a solid (5 g) which was purified on silica  ̃120 g, (eluting with ethylacetate heptane (30:70) to give the product as a white solid (2.69 g, 57%).




3-Piperidin-4-yl-5-trifluoromethyl-benzo[d]isoxazole. 4-(5-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (2.69 g, 0.007M) was suspended in a 50:50 mixture of DCM/Trifluoroacetic acid (4 mL). The mixture was heated for 30 minutes at 50° C. and then concentrated to give the product as the TFA salt. This was dissolved in dichloromethane (10 mL), washed with saturated Na2CO3 solution (3×3 mL), dried over MgSO4, filtered and concentrated to give the product as an oil (0.91 g, 46%)
Example 37




2R-[4-(5-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-cyclopropane-1R-carboxylic acid trans-(4-methyl-cyclohexyl)-amide. A solution 3-Piperidin-4-yl-5-trifluoromethyl-benzo[d]isoxazole (0.1 g, 0.37 mM) was dissolved in acetonitrile (4.5 mL) and treated with potassium carbonate (0.06 g, 0.46 mM) dissolved in water (0.5 mL). The reaction was stirred at ambient temperature for 10 minutes and then treated with methane sulfonic acid 2-(4-methyl-cyclohexyl carbonyl)-trans-cyclopropyl methyl ester (0.11 g, 0.38 mM-1.04 eq). Heated mixture at 56° C. for 14 hours. Product filtered off and washed with hexanes (2 mL). Dried product under vacuum to obtain the product as a white solid (0.118 g, 91%).
Example 38
7-Methoxy benzisoxazolyl piperidine




4-(2-Fluoro-3-methoxy-benzoyl)-piperidine-1 carboxylic acid tert-butyl ester. To a stirred solution of 2-fluoroanisole (6.00 g, 47.6 mmol) and anhydrous THF (125 mL) at −78° C. under nitrogen was added butyllithium (35 ml of a 1.6 M solution in hexanes, 56.0 mmol). After stirring for 13 min, N,N,N′,N′,N′′-Pentamethyldiethylenetriamine (12.9 mL, 61.8 mmol) was added dropwise and the reaction stirred at −78° C. After 168 min, a solution of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid tert-butyl ester (16.8 g, 61.7 mmol) in anhydrous THF (40 mL) was added dropwise over 25 min. The reaction was stirred at −78° C. for 35 min and at room temperature for 65 min. The reaction was diluted with ethyl acetate (400 mL) and washed with cold 0.5 N aqueous HCl (2×200 mL), 5% aqueous potassium carbonate (200 mL), water (200 mL), and brine (200 mL) successively. The organic phase was dried over magnesium sulfate, filtered, and the solvent removed to give 20.1 g of a yellow oil. The product was chromatographed on silica gel (350 g), using a step gradient elution of 20% ethyl acetate/heptane to 30% ethyl acetate/heptane, to afford 12.0 g (75%) of the desired product as a white solid.




4-[(2-Fluoro-3-methoxy-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester. A mixture of 4-(2-Fluoro-3-methoxy-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester (11.6 g, 34.4 mmol), hydroxylamine hydrochloride (2.87 g, 41.3 mmol) and pyridine (50 mL) was stirred at room temperature under nitrogen overnight. The yellow reaction solution was poured into cold water (500 mL) and the mixture aged at 0° C. for 15 min. The product was collected by filtration, washed with water, and dried under vacuum at 50° C. to afford 11.6 g (96%) of the desired product as a white powder. Proton NMR showed product to be a 2:1 mixture of Z- to E-isomers.




4-(7-Methoxy-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 831478). To a room temperature mixture of 4-[(2-Fluoro-3-methoxy-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (5.00 g, 14.2 mmol) in THF (50 mL) under nitrogen was added potassium tert-butoxide (15.0 mL of a 1M THF solution, 15.0 mmol) rapidly and the reaction refluxed for 4 h. After cooling to room temperature, the reaction was diluted with ethyl acetate (250 mL) and washed with water (100 mL) and brine (100 mL) successively. The organics were dried over magnesium sulfate, filtered, and concentrated to give a waxy solid. Recrystallization of the solid did not remove impurities so the crude product was chromatographed on silica using a step gradient elution of 10% ethyl acetate/dichloromethane to 40% ethyl acetate/dichloromethane to afford 3.04 g (64%) of the desired product as a white powder, mp: 130-132° C.




7-Methoxy-3-piperidin-4-yl-benzo[d]isoxazole hydrochloride (MDL 831587A). A mixture of 4-(7-Methoxy-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (3.00 g, 9.03 mmol), HCl (35 mL of a 1 M ether solution, 35.0 mmol), and methanol (25 mL) was stirred at room temperature under nitrogen for 18 h. Ether (75 mL) was added, the mixture stirred at room temperature for 15 min, and the product collected by filtration to afford 2.37 g (98%) of the desired product as a white powder, mp: >250° C.
Example 39




(3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(7-methoxy-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002437359). The target was synthesized via the method analogous to the one described previously. 3R-Imidazol-1-ylmethyl-piperidine was obtained via the method describe by Guzi, et. al. WO 0037458.
Example 40
7-trifluoromethyl-N-methyl-indazole piperidine




4-(2-Fluoro-3-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester. To a stirred solution of 2-fluorobenzotrifluoride (19.4 g, 118 mmol) and anhydrous THF (300 mL) at −78° C. under nitrogen was added was added LDA (65.0 mL of a 2.0 M solution in heptane/tetrahydrofuran/ethylbenzene, 130 mmol) dropwise over 25 min. After 165 min, a solution of 4-(methoxy-methyl-carbamoyl)-piperidine-1-carboxylic acid tert-butyl ester (40.1 g, 147 mmol) in anhydrous THF (100 mL) was added dropwise over 50 min and the reaction stirred at −78° C. for 20 min and at room temperature for 85 min. The reaction was quenched with saturated ammonium chloride (40 mL) and the mixture concentrated. The residue was taken up in ethyl acetate (400 mL) and washed successively with 1 N HCl (200 mL), 5% aqueous potassium carbonate, water (200 mL), and brine (200 mL). The organic phase was dried over magnesium sulfate, filtered, and the solvent removed to give an amber oil. The product was chromatographed on silica gel (300 g), using a step gradient elution of 20% ethyl acetate/heptane to 60% ethyl acetate/heptane, to afford 23.2 g (52%) of the desired product as an off-white solid.




4-(1-Methyl-7-trifluoromethyl-1H-indazol-3-yl)-piperidine-1-carboxylic acid tert-butyl esters (MDL 832782). A solution of 4-(2-Fluoro-3-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester (7.00 g, 18.6 mmol), methylhydrazine (1.13 g, 24.4 mmol), and n-butanol (50 mL) was heated at reflux for 135 min. The reaction was cooled to room temperature and diluted with ethyl acetate (250 mL). The oganics were washed with 5% aqueous potassium carbonate (150 mL), water (150 mL), and brine (150 mL) successively, dried over magnesium sulfate, filtered, and the solvent partially removed to give a viscid liquid. The remaining solvent was removed under high vacuum at 65° C. to provide an off-white solid. The product was chromatographed on silica, eluting with 30% ethyl acetate/heptane, to afford 6.63 g (93%) of the desired product as an off-white solid, mp: 104.7° C.




1-Methyl-3-piperidin-4-yl-7-trifluoromethyl-1 indazole hydrochloride (832641A). Added 1N HCl (100 mL) to a solution of 4-(1-methyl-7-trifluoromethyl-1H-indazol-3-yl)-piperidine-1-carboxylic acid tert-butyl esters (6.50 g, 16.9 mmol) in ethanol (25 mL) and the reaction stirred at room temperature overnight. The mixture was diluted with ether (350 mL) and cooled 0° C. for 20 min. The solid was collected, washed with ether, and dried at 45° C. under high vacuum to afford 4.66 g (86%) of the desired product as a white powder.
Example 41




2R-[4-(1-Methyl-7-trifluoromethyl-1H-indazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 832654). The target was synthesized via the method analogous to the one described previously.
Example 42
7-trifluoromethyl benzisoxazol piperidine




4-[(2-Fluoro-3-trifluoromethyl-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (MDL 832163). A mixture of 4-(2-Fluoro-3-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester (9.00 g, 24.0 mmol), hydroxylamine hydrochloride (2.00 g, 28.8 mmol) and pyridine (50 mL) was stirred at room temperature under nitrogen overnight. The yellow reaction solution was poured into cold water (500 mL) and the mixture aged at 0° C. for 1 h. The product was collected by filtration, washed with water, and dried under vacuum at 50° C. to afford 9.54 g of a while solid. Trituration of the solid with hot 25% ethyl acetate/heptane afforded 8.50 g (91%) of the desired product as a white solid. Proton NMR showed product to be a 3.8 to 1 mixture of isomers.




4-(7-Trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 832159). To a room temperature mixture of 4-[(2-fluoro-3-trifluoromethyl-phenyl)-hydroxyimino-methyl]-piperidine-1-carboxylic acid tert-butyl ester (1.40 g, 3.59 mmol) in THF (20 mL) under nitrogen was added potassium tert-butoxide (3.60 mL of a 1M THF solution, 3.60 mmol) in one portion and the reaction heated at 60° C. for 1.5 h. After standing at room temperature overnight, the solvent was removed and the residue diluted with ethyl acetate (60 mL). The organics were washed with water (30 mL) and brine (30 mL) successively, dried over magnesium sulfate, filtered, and concentrated to give an amber solid. The crude product was chromatographed on silica using 40% ethyl acetate/heptane as eluent to afford 0.97 g (73%) of the desired product as a white solid, mp:111-113° C.




3-Piperidin-4-yl-7-trifluoromethyl-benzo[d]isoxazole (MDL 832106A). A mixture of 4-(7-trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (8.00 g, 21.6 mmol), HCl (100 mL of a 1 M ether solution, 100 mmol), and methanol (50 mL) was stirred at room temperature under nitrogen overnight. The reaction was concentrated and the solid triturated with methanol/ether to afford 5.84 g (88%) of the desired product as a white powder, mp: 242-243° C.
Example 43




(3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(7-trifluoromethyl-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002437360). The target was synthesized via the method analogous to the one described previously. 3R-Imidazol-1-ylmethyl-piperidine was obtained via the method describe by Guzi, et. al. WO 0037458.
Example 44
7-Trifluoromethyl benzo[b]thienyl piperidine




4-(3-Hydroxy-2-methoxycarbonyl-7-trifluoromethyl-2,3-dihydro-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 832712). To a room temperature solution of 4-(2-fluoro-3-trifluoromethyl-benzoyl)-piperidine-1-carboxylic acid tert-butyl ester (9.00 g, 24.0 mmol), methyl thioglycolate (2.40 mL, 26.8 mmol), and anhydrous THF (200 mL) under nitrogen was added NaH (1.15 g of a 60% oil dispersion, 28.7 mmol) in one portion. After the gas evolution ceased, the reaction was stirred at 55° C. After 100 min, the reaction was cooled to room temperature and diluted with ethyl acetate (500 mL). The mixture was washed with water (300 mL) and brine (300 mL) successively, dried over magnesium sulfate, filtered, and the solvent removed to afford a sticky white solid. Trituration with 20% ethyl acetate/heptane afforded 6.20 g (56%) of the desired product as a white powder.




3-Piperidin-4-yl-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester. To a room temperature solution of 4-(3-hydroxy-2-methoxycarbonyl-7-trifluoromethyl-2,3-dihydro-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (6.00 g, 13.0 mmol) in DCM (30 mL) was added TFA (30 mL) causing rapid gas evolution. After 5 min, the reaction was stirred at 40° C. for 5.5 h. After cooling to room temperature, the reaction was poured into 20% aqueous potassium carbonate (400 mL) and extracted with DCM (2×200 mL). The combined extracts were dried over magnesium sulfate, filtered, and the solvent removed to give a thick oil. After drying under high vacuum 4.37 g (98%) of the desired product was obtained as a white foam.




3-(1-Acetyl-piperidin-4-yl)-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester. To a room temperature solution of 3-piperidin-4-yl-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester (4.37 g, 12.7 mmol), triethylamine (2.70 mL. 19.4 mmol), and anhydrous THF (80 mL) under nitrogen was added acetyl chloride (1.10 mL, 15.5 mmol) in one portion and the reaction stirred at room temperature overnight. The reaction was diluted with ethyl acetate (300 mL) and washed with water (150 mL) and brine (150 mL) successively. The organic layer was dried over magnesium sulfate, filtered, and the solvent removed. The residue was chromatographed on silica, eluting with 10% methanol/ethyl acetate, to afford 4.28 g (88%) of the desired product as a white solid, mp: 155.2° C.




3-(1-Acetyl-piperidin-4-yl)-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid. To a solution of 3-(1-acetyl-piperidin-4-yl)-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester (4.10 g, 10.6 mmol) in THF (25 mL) was added 0.5 N aqueous sodium hydroxide (23.4 mL, 11.7 mmol) and the reaction stirred at room temperature. After 18 h, the reaction was acidified with 1 N HCl (200 mL) and the mixture extracted with DCM (2×100 mL). The organics were washed with water (100 mL), dried over magnesium sulfate, filtered, and concentrated to give 4.13 g of the desired product as a white foam.




1-[4-(7-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-yl]-ethanone (MDL 832823). A mixture of 3-(1-acetyl-piperidin-4-yl)-7-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid (4.13 g, 1.1 mmol), Cu powder (0.706 g, 11.1 mmol), and quinoline (20 mL) was heated to 200° C. under nitrogen. After 10 min, no gas evolution was observed and the reaction cooled at room temperature. The mixture was diluted with ethyl acetate (100 mL), filtered through a Celite bed and the filtrate washed with 1 N HCl (2×100 mL), 5% aqueous potassium carbonate (100 mL), water (100 mL), and brine (100 mL) successively. The organics were dried over magnesium sulfate, filtered, and concentrated to give an amber oil. The oil was chromatographed on silica, eluting with 10% methanol/ethyl acetate to afford 2.69 g (74%) of the desired product as a tan solid.




4-(7-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine. A mixture of 1-[4-(7-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-yl]-ethanone (2.95 g, 9.01 mmol), concentrated HCl (30 mL), and ethanol was heated at 80° C. for 18 h. After cooling to room temperature, the reaction was basified with 20% aqueous potassium carbonate (150 mL) and the mixture extracted with DCM (2×100 mL). The organics were washed with water (100 mL), dried over potassium carbonate, filtered, and concentrated to give 2.42 g (94%) the desired product as an amber waxy solid.
Example 45




2R-[4-(7-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropanecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 833690). The target was synthesized via the method analogous to the one described above.
Example 46




1-Dimethoxymethyl-2-fluoro-4-trifluoromethyl-benzene. A solution of 2-fluoro-4-trifluoromethylbenzaldehyde (10.0 g, 52.0 mmol), trimethyl orthoformate (55.0 g, 520 mmol) and a catalytic amount of p-toluene sulfonic acid (5 mol %) was stirred at room temperature for 24 hours. The resulting solution was saturated with the addition of solid Na2CO3 and stirred for 15 minutes. The mixture was then filtered and the solution was evaporated. The residue was diluted with ether and evaporated yielding 12.1 g (98%) of the desired product as an oil.




[(2-Fluoro-4-trifluoromethyl-phenyl)-methoxy-methyl]-phosphinic acid diethyl ester. To a solution of the 1-dimethoxymethyl-2-fluoro-4-trifluoromethyl-benzene (5.0 g, 21.0 mmol) and P(OEt)3 (4.18 g, 25.2 mmol) in CH2Cl2 (42 mL) cooled to −78° C. was added titanium tetrachloride (42 mL, 42 mmol, 1 M in DCM) portion-wise. The resulting solution was stirred at −78° C. for 2 hours when the reaction is quenched with the addition of water (10 mL). The mixture was warmed to room temperature and separated. The aqueous layer was extracted with DCM (3×10 mL). The combined organic layers were washed with brine (25 mL), dried (MgSO4), filtered and evaporated. The residue is separated via chromatography (1:1; EtOAc/DCM) yielding 6.1 g (85%) of the desired product.




1-Benzyl-4-[(2-fluoro-4-trifluoromethyl-phenyl)-methoxy-methylene]-piperidine. To a solution of [(2-fluoro-4-trifluoromethyl-phenyl)-methoxy-methyl]-phosphinic acid diethyl ester (16.1 g, 46.5 mmol) in THF (200 mL) cooled to −78° C. was added n-Butyl lithium (32.1 mL, 51.5 mmol) drop-wise. To the resulting solution was added a solution of 1-benzyl-4-piperidone (9.71 g, 51.5 mmol) in THF (20 mL) drop-wise. The mixture was stirred at −78° C. for 3 hours and then was allowed to slowly warm to room temperature overnight. The reaction mixture was partitioned between water (50 mL) and EtOAc (200 mL) and separated. The aqueous layer was extracted with EtOAc (100 mL) and the combined organic layers were washed with brine (50 mL), dried (MgSO4), filtered and evaporated. The residue was purified via chromatography (gradient elution, heptane to 40% EtOAc in heptane) to obtain 9.9 g (56%) of the desired product as an oil.




(1-Benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone. A solution of the 1-benzyl-4-[(2-fluoro-4-trifluoromethyl-phenyl)-methoxy-methylene]-piperidine (9.9 g, 26.1 mmol) in acetone (100 mL) and conc. HCl (10 mL) in a sealed vessel was heated at 60° C. for 2 hours. Upon cooling to room temperature the solution was basified with the addition of 3 N NaOH. The resulting mixture was extracted with EtOAc (2×200 mL). The combined extracts were washed with brine (50 mL), dried (MgSO4), filtered and evaporated yielding 8.8 g (92%) of the desired product as a solid.




3-(1-Benzyl-piperidin-4-yl)-1-methyl-6-trifluoromethyl-1 Hindazole (MDL 833796). A solution of (1-benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone (2.0 g, 5.46 mmol), methylhydrazine (327 mg, 7.10 mmol), and n-butanol (22 mL) was heated at 120° C. in a seal vessel for 18 hours. The reaction was cooled to room temperature and basified with NaHCO3 and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over magnesium sulfate, filtered, and evaporated. The product was chromatographed on silica, eluting with 1:1 ethyl acetate/heptane, to afford 1.68 g (82%) of the desired product as an oil.




1-Methyl-3-piperidin-4-yl-6-trifluoromethyl-1 H-indazole hydrochloride (MDL 833799A). To the 3-(1-benzyl-piperidin-4-yl)-1-methyl-6-trifluoromethyl-1H-indazole (1.41 g, 3.78.mmol) in DCM (20 mL) was added 1-chloroethyl chloroformate (0.49 mL, 4.50 mmol). The resulting solution was stirred at room temperature overnight when the volitiles were removed in vacuo. The residue was taken un in methanol (20 mL) and the resulting solution was heated at reflux for 1 hour. The mixture was cooled to room temperature and the solution was evaporated. The residue was taken up in EtOAc and the solid product was collected by filtration yielding 1.03 g (86%) of the HCl salt as a white solid.
Example 47




(3S-Imidazol-1-ylmethyl-piperidin-1-yl)-[2R-[4-(1-methyl-6-trifluoromethyl-1H-indazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl]-methanone (A002437353). The target was synthesized via the method analogous to the one described previously. 3R-Imidazol-1-ylmethyl-piperidine starting material was obtained via the method describe by Guzi, et. al. WO 0037458.
Example 48




(1-Benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone oxime. A mixture of (1-benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone (5.0 g, 13.66 mmol), hydroxylamine hydrochloride (1.1 g, 16.39 mmol) and pyridine (50 mL) was stirred at room temperature overnight when the mixture was distilled to remove pyridine (35 mL). The solid residue was washed with heptane then ether. The resulting solid was partitioned between a saturated solution of NaHCO3 and EtOAc. The organic layer was dried (MgSO4) filtered and evaporated. The solid residue was washed with 3:1 heptane/EtOAc and dried under vacuum to obtain 2.1 g (40%) of the desired product as a white solid.




3-(1-Benzyl-piperidin-4-yl)-6-trifluoromethyl-benzo[b]isoxazole. To a room temperature mixture of (1-benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone oxime (2.1 g, 5.51 mmol) in THF (20 mL) under nitrogen was added potassium tert-butoxide (5.78 mL of a 1M THF solution, 5.78 mmol) in one portion. The resulting solution was stirred at room temperature for 6 hours when the mixture was partitioned between water (60 mL) and ethyl acetate (60 mL). The aqueous layer was extracted with EtOAc (60 mL). The combined organic layers were washed with water (30 mL) and brine (30 mL), dried over magnesium sulfate, filtered, and concentrated to give 1.9 g (96%) as the desired product.




3-Piperidin-4-yl-6-trifluoromethyl-benzo[d]isoxazole hydrochloride. To the 3-(1-Benzyl-piperidin-4-yl)-6-trifluoromethyl-benzo[b]isoxazole (1.9 g, 5.27 mmol) in DCM (26 mL) was added 1-chloroethyl chloroformate (0.69 mL, 6.33 mmol). The resulting solution was stirred at room temperature overnight when the volitiles were removed in vacuo. The residue was taken un in methanol (25 mL) and the resulting solution was heated at reflux for 1 hour. The mixture was cooled to room temperature and the solution was evaporated. The residue was taken up in EtOAc and the solid product was collected by filtration yielding 1.2 g (74%) of the HCl salt as a white solid.
Example 49




2R-[4-(6-Trifluoromethyl-benzo[d]soxazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (A002287765). The target was synthesized via the method analogous to the one described previously.
Example 50




3-(1-Benzyl-piperidin-4-yl)-6-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester (MDL 833803). To a room temperature solution of (1-benzyl-piperidin-4-yl)-(2-fluoro-4-trifluoromethyl-phenyl)-methanone (7.5 g, 20.5 mmol), methyl thioglycolate (2.0 mL, 22.5 mmol), and DMF (100 mL) was added K2CO3 (5.65 g, 41.0 mmol). The reaction was stirred at 60° C. for 24 hours, cooled to room temperature and diluted with ethyl acetate (500 mL). The mixture was washed with water (2×300 mL) and brine (300 mL) successively, dried over magnesium sulfate, filtered, and the solvent removed to afford an oil. The oil was purified via chromatography (30% EtOAc in heptane) yielding 5.91 g (67%) as a solid.




4-(2-Methoxycarbonyl-6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester. To a solution of 3-(1-benzyl-piperidin-4-yl)-6-trifluoromethyl-benzo[b]thiophene-2-carboxylic acid methyl ester (5.9 g, 13.6 mmol) in DCM (50 mL) was added methyl chloroformate (1.26 mL, 16.3 mmol) drop-wise. The resulting solution was stirred overnight when the volatiles were removed in vacuo. The residue was washed with heptane to yield 4.2 g (77%) of the desired product as a white solid.




4-(2-Carboxy-6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester. To a stirred solution of 4-(2-Methoxycarbonyl-6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester (1.1 g, 2.7 mmol) in THF (7.0 mL) was added 1 N NaOH (2.97 mL). The resulting mixture was stirred at room temperature overnight when the mixture was diluted with water (50 mL) and washed with ether (100 mL). The aqueous layer was acidified with the addition of 3 N HCl and the product was extracted with EtOAc (2×150 mL). The combined organic layers were washed with brine (50 mL), dried (MgSO4), filtered and evaporated yielding 960 mg (92%) of the desired product as a white solid.




4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester. A mixture of 4-(2-carboxy-6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester (4.3 g, 11.1 mmol) and copper (705 mg, 11.1 mmol) in quinoline (28 mL) was heated at 200° C. for 45 minutes, Upon cooling to room temperature the mixture was diluted with EtOAc (50 mL) and filtered. The filtrate was washed with 5% HCl (2×20 mL), water (20 mL) and brine (20 mL), dried (MgSO4), filtered and evaporated. The residue was separated via chromatography (30% EtOAc in heptane) yielding 3.14 g (82%) of the desired product as a white solid.




4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine hydrobromide. A mixture of 4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidine-1-carboxylic acid methyl ester (3.1 g, 9.0 mmol) in HBr (45 mL, 30% in acetic acid) was stirred at room temperature for 20 hours when the volatiles were removed in vacuo. The residue was washed with EtOAc and the product was collected by filtration yielding 3.09 g (94%) of the desired product as a white solid.
Example 51




(3S-Imidazol-1-ylmethyl-piperidin-1-yl)-{2R-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperidin-1-ylmethyl]-1R-cyclopropyl}-methanone (A002609935). The target was synthesized via the method analogous to the one described previously. 3R-Imidazol-1-ylmethyl-piperidine starting material was obtained via the method describe by Guzi, et. al. WO 0037458.
Example 52




4-(6-Fluoro-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 811778). To a stirred suspension of 4-(6-fluoro-benzo[d]isoxazol-3-yl)-piperidine (1.00 g, 454 mmol) in dry dichloromethane (10.0 mL) was added triethylamine (0.95 mL, 6.82 mmoles), 4-dimethylaminopyridine (55 mg, 0.454 mmoles) and di-tert-butyl dicarbonate (1.98 g, 9.09 mmoles). Gas spontaneously evolved for several minutes upon the addition of di-tert-butyl dicarbonate. The resulting solution was stirred at room temperature for 1 hour when the solution was diluted with CH2Cl2 (50 mL) and washed with water (10 mL), 10% HClaq (10 mL), water (10 mL), saturated NaHCO3 (10 mL), water (10 mL) and brine (10 mL) and dried (MgSO4), filtered and evaporated. The residue was recrystallized from diethyl ether yielding 1.31 g (90%) as a white, crystalline solid, MP 117-188° C. Analysis calculated for C17H21N2FO3: 63.74% C, 6.61% H, 8.74% N. Found: 63.66% C, 6.64% H, 8.73% N.




4-(6-Fluoro-7-hydroxy-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 811820). To a stirred solution of 4-(6-fluoro-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid ter-butyl ester (1.00 g, 3.13 mmol) in dry tetrahydrofuran (31.3 mL) cooled to −78° C. was added lithium diisopropylamide (1.72 mL, 3.35 mmoles). The resulting solution was stirred at −78° C. for 2 hours when trimethylborate (0.44 mL, 3.84 mmoles) was added. The resulting solution was stirred at −78° C. for 1 hour then was allowed to warm to room temperature over 3 hours when hydrogen peroxide (2.00 mL) and acetic acid (1.00 mL) were added. The resulting mixture was stirred at room temperature overnight when the mixture was quenched with saturated NH4Claq (20 mL) and 10% HClaq (20 mL). The resulting mixture was extracted with CH-2C12 (4×50 mL). The combined extracts were washed with brine (50 mL), dried (MgSO4), filtered and evaporated. The residue was separated via column:chromatography (1:1; Et2O/Pet. ether) yielding 0.619 g (59%) of the phenol as a white, crystalline solid, mp 169-170° C. Analysis calculated for C17H21N2FO4; 60.70% C, 6.29% H, 8.33% N. Found: 60.72% C, 6.15% H, 8.22% N.




4-(6-Fluoro-7-methoxy-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (MDL 811841). To a stirred solution of 4-(6-fluoro-7-hydroxy-benzo[d]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (1.28 g, 3.80 mmol) in N-methyl-2-pyrrolidone (33 mL) was added potassium tert-butoxide (2.09 g, 17.12 mmoles). To the resulting deep red solution was added iodomethane (1.20 mL, 19.02 mmoles). The resulting yellow solution was stirred at room temperature for 6 hours when the reaction was quenched with water (55 mL) and acidified with HClaq. The resulting mixture was extracted with. Et2O (4×110 mL). The combined extracts were washed with brine (110 mL), dried (MgSO4), filtered and evaporated. The residue was separated via column chromatography (1:1; Et2O/Pet. ether) yielding 1.2 g of the methyl ether. The white, solid product was further purified via recrystallization from 1:1; Et2O/Pet. ether yielding 963 mg (72%) as a white, crystalline solid, mp 94-96° C. Analysis calculated for C18H23N2FO4: 61.70% C, 6.62% H, 7.99% N. Found: 61.75% C, 6.73% H, 7.94% N.




6-Fluoro-7-methoxy-3-piperidin-4-yl-benzo[d]isoxazole hydrochloride (MDL 811998). To a stirred solution of 4-(6-fluoro-7-methoxy-benzo[o]isoxazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (4.00 g, 11.43 mmol) in dry hydrochloric acid in diethyl ether (100 mL) was added methanol (7.62 mL). The resulting solution was stirred at room temperature for 5 hours when a white solid precipitate formed. The resulting suspension was filtered and the white solid was wash thoroughly with ether yielding 1.76 g of the desired product as a white solid. The mother liquor precipitated yielding an additional 0.94 g of product providing a total of 2.70 g (83%) of the desired product as a pure, white solid, mp 246-248° C.
Example 53




2R-[4-(6-Fluoro-7-methoxy-benzo[d]isoxazol-3-yl)-piperidin-1-ylmethyl]-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 831361). The target was synthesized via the method analogous to the one described above.
Example 54




4-[(3-Bromo-thiophen-2-yl)-(methyl-hydrazono)-methyl]-piperidine-1-carboxylic acid tert-butyl ester. A mixture of 4-(thiophene-2-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester (1.96 g, 5.2 mmol) in methylhydrazine (2 mL) was heated at 75° C. overnight. The excess methyl hydrazine was then removed with a vacuum pump. The residue was purified by chromatography (eluted with 0-8% of MeOH in DCM) yielding 0.95 g (45%) of the desired product.




4(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester. 4-[(3-Bromo-thiophen-2-yl)-(methyl-hydrazono)-methyl]-piperidine-1-carboxylic acid tert-butyl ester (700 mg, 1.74 mmol) was mixed with CuI (20 mg), CsCO3 (650 mg, 1.15 eq) in methoxyethanol (10 mL). The mixture was heated to 70° C. for 2 hr. then stirred overnight at room temperature. The solvent was stripped on rotary evaporator. The residue was extracted into EtOAc then washed with brine and concentrated down to an oil. This oil was purified via chromatography (eluted with 0-10% MeOH in DCM) yielding 520 mg (68%) of the desired product.




1-Methyl-3-piperidin-4-yl-1H-thieno[3,2-c]pyrazole hydrochloride (A002436287A). 4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperidine-1-carboxylic acid tert-butyl ester (520 mg, 1.6 mmol) was stirred at room temperature in a solution of HCl (5 mL, 4N HCl in dioxane) for 4 hours. The volatiles were removed in vacuo and the residue was triturated with ether (twice) to yield off white solids 304 mg (74%) as the desired hydrochloride salt.
Example 55




2R-[4-(1-Methyl-1H-thieno[3,2-c]pyrazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (A002436291). The target was synthesized via the method analogous to the one described previously.
Example 56




4-{(3-Bromo-thiophen-2-yl)-[(2,2,2-trifluoro-ethyl)-hydrazono]-methyl}-piperidine-1-carboxylic acid tert-butyl ester. To a mixture of 4-(thiophene-2-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester (2.34 g, 6.24 mmol) in n-butanol (20 mL) was added trifluoroethylhydrazine (2.43 g, 12.4 mmol). The resulting mixture was heated at 110° C. overnight. The volatiles were then removed in vacuo. The residue was purified by chromatography (eluted with 0-10% MeOH in DCM) yielding 2.41 g (92%) of the desired product.




4-[1-(2,2,2-Trifluoro-ethyl)-1H-thieno[3,2-c]pyrazol-3-yl]-piperidine-1-carboxylic acid tert-butyl ester. 4-{(3-Bromo-thiophen-2-yl)-[(2,2,2-trifluoro-ethyl)-hydrazono]-methyl}-piperidine-1-carboxylic acid tert-butyl ester (2.34 g, 4.98 mmol) was mixed with CuI (50 mg), CsCO3 (1.9 g, 1.2 eq) in methoxyethanol (25 mL). The mixture was heated to 75° C. for 1 hour. The mixture was then diluted with EtOAc and filtered. The filtrate was evaporated and the residue was purified via chromatography (eluted with 0-10% MeOH in DCM) yielding 2.03 g (>95%) of the desired product.




3-Piperidin-4-yl-1-(2,2,2-trifluoro-ethyl)-1H-thieno[3,2-c]pyrazole hydrochloride (833906). 4-[1-(2,2,2-Trifluoro-ethyl)-1H-thieno[3,2-c]pyrazol-3-yl]-piperidine-1-carboxylic acid tert-butyl ester (1.9 g, 4.87 mmol) was stirred at room temperature in a solution of HCl (6 mL, 4N HCl in dioxane) for 4 hours. The volatiles were removed in vacuo and the residue was triturated with ether (twice) to yield off white solids 2.1 g (74%) as the desired hydrochloride salt.
Example 57




2R-{4-[1-(2,2,20Trifluoro-ethyl)-1H-thieno[3,2-c]pyrazol-3-yl)-piperidin-1-ylmethyl]-1R-cycloproanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (A002287767). The target was synthesized via the method analogous to the one described previously.
Example 58




3-Bromo-thiophene-2-carbaldehyde oxime. 3-Bromothiophene-2-carbaldehyde (28.7 g, 0.15 mol) in ethanol (50 mL) was added in one portion to a solution of hydroxylamine hydrochloride (13.8 g, 0.2 mol), sodium hydroxide (8 g, 0.2 mol) in water (30 mL) and ethanol (100 mL). The mixture was stirred at 0° C. for 2 hours and was kept at 0° C. overnight when a precipitate formed. The mixture was diluted with cold water (600 ml) and the solid was collected by filtration yielding 20.5 g, (67%). The aqueous solution was further extracted with ethyl acetate. The organic solution was washed with brine, dried with magnesium sulfate, filtered and evaporated yielding 6.9 g of additional product as a light yellow solid. The total yield was 27.4 g (89%).




3-Bromo-thiophene-2-(chloro-carbaldehyde) oxime. To the solution of 3-bromo-thiophene-2-carbaldehyde oxime (10.8 g, 52.4 mmol), hydrogen chloride (14.5 mL, 4M in dioxane) in DMF (100 mL) was charged with oxone (16.9 g, 1.05 eqiv) in one portion at room temperature. The mixture was stirred at room temp overnight when the solution was poured in to water and extracted with ethyl acetate. The organic solution was washed with brine and dried over magnesium sulfate, filtered and evaporated to dryness to give a yellow solid (12.68 g, quantitative by weight) which was used in the next reaction without further purification.




4-[(3-Bromo-thiophen-2-yl)-hydroxyimino-methyl]-piperazine)-1-carboxylic acid tert-butyl ester. A solution of 3-bromo-thiophene-2-(chloro-carbaldehyde) oxime (16.4 g, 68 mmol) in THF (70 mL) was added drop-wise to a solution of N-(t-butoxycarbonyl)piperazine (14 g, 1.1 equiv.), DABCO (9.5 g, 1.25 eqiv.) in DMF (100 mL) at 0° C. over 25 minutes. The mixture was stirred at 0° C. for 3.5 hours when the mixture was poured into water and was extracted with ethyl acetate. The organic layer was washed with brine and dried over magnesium sulfate, filtered and evaporated. The crude product (30.5 g) was purified via chromatography (eluted with 0-5% of MeOH in DCM) yielding 24.6 g (85%) of the desired product.




4-Thieno[2,3-d]isoxazol-3-yl-piperazine-1-carboxylic acid tert-butyl ester. A mixture of 4-[(3-bromo-thiophen-2-yl)-hydroxyimino-methyl]-piperazine)-1-carboxylic acid tert-butyl ester (10.3 g, 26.4 mmol), cesium carbonate (10.7 g, 32.7 mmol), copper iodide (500 mg) in methoxyethanol (200 mL) was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate and washed with water. The aqueous solution was extracted three times with ethyl acetate. The combined organic layers (total 600 ml) were washed with brine, dried over magnesium sulfate, filtered and evaporated. The residue was purified via chromatography (120 gm of silica gel, eluted with 0-8% Methanol in dichloromethane) yielding 5.1 g (62%) of the desired product as light oil.




3-piperazin-1-yl-thieno[2,3-d]isoxazole. 4-Thieno[2,3-d]isoxazol-3-yl-piperazine-1-carboxylic acid tert-butyl ester (5.0 g, 16.2 mmol) was stirred at room temperature in a solution of HCl (25 mL, 4N HCl in dioxane) for 4 hours. The volatiles were removed in vacuo and the residue was triturated with ether (twice) to yield off white solids 3.3 g (84%) as the desired hydrochloride salt.
Example 59




2R-(4-Thieno[2,3-d]isoxazol-3-yl-piperazin-1-ylmethyl)-1R-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 831493). The target was synthesized via the method analogous to the one described previously.
Example 60




2R-(4-Benzo[b]thiophen-2-yl-piperidin-1-ylmethyl)-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 831148). The target was synthesized via the method analogous to the one described above. 4-Benzo[b]thiophen-2-yl-piperidine was obtained via the method described by Bernasconi, DE 2456246.
Example 61




2R-[4-(5,6-Dihydro-4H-imidazo[4,5,1-ij]quinolin-2-yl)-piperazin-1-ylmethyl]-1R-cyclopranecarboxylic acid (trans-4-methyl-cyclohexyl)-amide (MDL 833699). The target was synthesized via the method analogous to the one described above. 2-Piperazin-1-yl-5,6-dihydro-4H-imidazo[4,5,1-ij]quinoline was obtained via the method described by Glamkowski and Freed, EP 95-113849.
Example 62




2R-(4-Thieno[2,3-b]pyridin-3-yl-piperazin-1-ylmethyl)-1R-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 833821). The target was synthesized via the method analogous to the one described above. 3-piperazin-1-yl-thieno[2,3-b]pyridine was obtained via the method described by Hrib and Jurcak, US 92-942232.
Example 63




Cyclopropanecarboxylic acid tert-butyl ester. To a stirred suspension of 12.09 (107.1 mmol) of potassium t-butoxide in 200 mL ether at 0° C. under nitrogen was added 13.4 g (128.6 mmol) of cyclopropanecarboxylic acid chloride over 5 min. After 30 min at 0° C. the mixture was stirred at ambient temperature for an additional 30 min. The reaction mixture was poured into aqueous saturated sodium bicarbonate and extracted with ether. The organic layer was dried and carefully concentrated to deliver 15.0 g (99%) of a yellow oil as the desired ester product.




1-Allyl-cyclopropanecarboxylic acid tert-butyl ester. Lithium diisopropyl amide was generated from 7.5 g (58.1 mmol) diisopropyl amine and 23.2 mL of 2.5 M n-butyl lithium in 200 mL THF at 0° C. under nitrogen. After stirring for 30 minutes at 0° C. the solution was taken to −78° C. where 7.5 g (52.8 mmol) of cyclopropanecarboxylic acid tert-butyl ester in 30 mL of THF was added dropwise over 5 min. After 4 h 12.8 g (106 mmol) of allyl bromide in 30 mL THF was added drop-wise over 10 min. to the clear golden solution. The reaction was allowed to slowly warm to room temperature. After 19 hours the reaction was poured into aqueous saturated ammonium chloride solution, extracted with ether, dried and concentrated to deliver an oil which was purified via Kugelrohr distillation (approx. 20 mm Hg; 60-75° C. oven) to deliver 5.4 g (56%) of the desired product as a clear colorless oil.




1-(2-Oxo-ethyl)-cyclopropanecarboxylic acid tert-butyl ester. A solution of 5.7 g (31.3 mmol) of 1-allyl-cyclopropanecarboxylic acid tert-butyl ester in 50 mL methanol and 50 mL dichloromethane under nitrogen was taken to −78° C. where ozone was bubbled in for 1 hour. Nitrogen was bubbled in until the familiar blue color dissipated. Three drops of pyridine followed by 2 mL of dimethyl sulfide were added and the cooling bath removed. After 2 hours the reaction was poured into aqueous saturated ammonium chloride solution, extracted with dichloromethane, dried and concentrated to deliver a quantitative yield of the desired aldehyde as an oil.
Example 64




1-{2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl}-cyclopropanecarboxylic acid tert-butyl ester. To a solution of 4.2 g (14.7 mmol) of 1-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazine in 100 mL of dichloromethane under nitrogen was added 3.5 g (19.1 mmol) of 1-(2-Oxo-ethyl)-cyclopropanecarboxylic acid tert-butyl ester. After stirring 10 minutes at room temperature 6.5 g (30.7 mmol) of sodium triacetoxyborohydride was added portion-wise. After 30 minutes 100 mL of 0.5 N aqueous sodium hydroxide was added and the reaction was stirred for 15 minutes. The organic layer was separated, dried and concentrated to deliver a paste. This was taken up in methanolic hydrogen chloride. The salt thus obtained was recrystallized from ethyl acetate/methanol to deliver 6.6 g (13.4 mmol) of the desired product as a white powder mp=187-9° C.




1-{2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl}-cyclopropanecarboxylic acid. 1-[2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl]-cyclopropanecarboxylic acid tert-butyl ester, 0.7 g (1.54 mmol), was stirred in 10 mL of trifluoroacetic acid. After 2 hours the reaction mixture was concentrated to an oil. This was taken up in methanolic hydrogen chloride. The salt thus obtained was recrystallized from ethyl acetate/methanol to deliver 0.4 g (60%) of white powder as the desired carboxylic acid mp=255-7° C.




1-{2-[4-(6-Trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl}-cyclopropanecarboxylic acid (3-imidazol-1-yl-propyl)-amide (MDL 832231). A suspension of 0.17 g (0.39 mmol) of 1-{2-[4-(6-trifluoromethyl-benzo[b]thiophen-3-yl)-piperazin-1-yl]-ethyl}-cyclopropanecarboxylic acid in 10 mL dichloromethane under nitrogen was treated with 1 mL of oxalyl chloride followed by 3 drops of DMF. After 2 hours the reaction was concentrated to a slurry, diluted with 10 mL DMF and treated with 0.10 g (0.78 mmol) of 1-(3-aminopropyl)imidazole followed by 0.12 g (1.2 mmol) of triethylamine. After 12 hours the reaction was poured into aqueous saturated sodium bicarbonate and extracted with ethyl acetate. The organic layer was separated, dried and concentrated to deliver an oil. Chromatographic purification gave 0.075 g (38%) of the desired final product as an oil.







TABLE 2













 















    No.
    R
    n




    R1
    A
    R2
    R3
    d3Ki


















827055 (racemic)




2




H
N




CH2Ph
792


 


827071 (racemic)




2




H
N




(CH2)2Ph
454


 


827074 (racemic)




2




CH3
N




(CH2)2Ph
267


 


827079 (racemic)




2




CH3
N




(CH2)3Ph
126


 


827085 (racemic)




2




H
N




CH2Ph
126


 


827086 (racemic)




2




H
N




CH2Ph
334


 


827087 (racemic)




2




H
N




CH2Ph
178


 


827088 (racemic)




2




H
N




CH2Ph
284


 


827089 (racemic)




2




CH3
N




CH2Ph
307


 


827090 (racemic)




2




CH3CH2
N




CH2Ph
346


 


827101 (racemic)




2




H
N




CH2Ph
148


 


827102 (racemic)




2




H
N




CH2Ph
218


 


827105 (racemic)




2




H
N




(CH2)3Ph
117


 


827106 (racemic)




2




H
N




(CH2)3Ph
317


 


827107 (racemic)




2




H
N




(CH2)3Ph
489


 


827108 (racemic)




2




H
N




(CH2)3Ph
302


 


827109 (racemic)




2




CH3 
N




(CH2)3Ph
78.6


 


827110 (racemic)




2




CH3CH2
N




(CH2)3Ph
208


 


827111 (racemic)




2




HO(CH2)2
N




(CH2)3Ph
14.3


 


817047 (racemic)




2




H
N




H
2.86


 


817048 (racemic)




2




H
N




H
2.85


 


817049 (racemic)




2




H
N




H
7.02


 


817050 (−)-R,R




2




H
N




H
2.15


 


817051 (−)-R,R




2




H
N




H
3.47


 


817244 (racemic)




2




HO(CH2)2
N




H


 


817569A (−)-R,R




2




H
N




H
5.38


 


818499 (racemic)




2




H
N




H
22.1


 


818497 (racemic)




2




H
N




H
5.52


 


818673 (Isomer 1)




2




CH2Ph
N




H


 


818674 (Isomer 2)




2




CH2Ph
N




H


 


827196 (racemic)




2




H
N




H
2.35


 


817037 (racemic)




2




H
N




H
0.98


 


817038 (racemic)




2




H
N




H
0.952


 


817039 (racemic)




2




H
N




H
14.8


 


817040 (racemic)




2




H
N




H
0.804


 


817045 (racemic)




2




H
N




H
5.36


 


817041 (racemic)




2




H
N




H
0.776


 


817042 (racemic)




2




H
N




H
0.615


 


817043 (racemic)




2




H
N




H
7.2


 


817044 (racemic)




2




H
N




H
1.1


 


817046 (racemic)




2




H
N




H
1.92


 


817090 (−)-R,R




2




H
N




H
2.17


 


817091 (−)-R,R




2




H
N




H
3.98


 


817092 (−)-R,R




2




H
N




H
3.39


 


817094




2




H
N




H
8.89


 


817095




2




H
N




H
0.671


 


817096




2




H
N




H
9.09


 


817097




2




H
N




H
0.636


 


817098




2




H
N




H
1.06


 


817099




2




H
N




H
1.1


 


817100




2




H
N




H
0.75


 


817101




2




H
N




H
0.547


 


817102 (−)-R,R




2




H
N




H
7.75


 


827253




2




H
N




H
15.8


 


827254




2




H
N




H
8.66


 


817039A (racemic)




2




H
N




H
35


 


817090A (racemic)




2




H
N




H
5.04


 


817571A (racemic)




2




H
N




H
11.5


 


817092A (racemic)




2




H
N




H
22.3


 


817570A (racemic)




2




H
N




H
5.04


 


817478A (racemic)




2




H
N




H
1.99


 


817043A (racemic)




2




H
N




H
18.6


 


817091A (−)-R,R




2




H
N




H
4.04


 


817102A (−)-R,R




2




H
N




H
7.57


 


818675DA (−)-R,R




2




H
N




H
5.69


 


827201 (racemic)




2




H
N




H
126


 


817093 (−)-R,R




2




H
N




H
5.97


 


817480A (racemic)




2




H
N




H
5.33


 


817566




2




H
N




H
2.46


 


817565




2




H
N




H
1.73


 


827198




2




H
N




H
161


 


818427 (racemic)




2




H
N




H
6.07


 


817892 (−)-R,R




2




H
N




H
5.47


 


817893 (+)-S,S




2




H
N




H
245


 


817892A (−)-R,R




2




H
N




H
3.02


 


818574 (racemic)




2




H
N




H
9.6


 


817787 (−)-R,R




2




H
N




H
5.81


 


817786 (+)-S,S




2




H
N




H
125


 


817787A (−)-R,R




2




H
N




H
1.76


 


817272 (racemic)




2




H
N




H
1.72


 


817273 (racemic)




3




H
N




H
3.05


 


817260 (racemic)




3




H
N




H
12.5


 


817261 (racemic)




3




H
N




H
9.23


 


827731 (racemic)




3




H
N




H
19.5


 


827732 (racemic)




3




H
N




H
10.5


 


817567 (racemic)




3




H
N




H
80.6


 


817568 (racemic)




3




H
N




H
36.3


 


827733 (racemic)




3




H
N




H
1.19


 


818498 (racemic)




3




H
N




H
25.6


 


827800 (racemic)




3




H
N




H
19.7


 


814648




2




CH3(CH2)2
N




H
39.6


 


814649




2








N




H
38.9


 


814650A




2




CH3
N




H
18


 


813384




2




H
N




H
74


 


822129




2




CH3
N




H
38


 


822130




2




CH3CH2
N




H
51


 


822131




2




CH3
N




H
61


 


822132




2




CH3CH2
N




H
18


 


822133




2




CH3
N




H
17


 


822134




2




H
N




H
82


 


822135




2




H
N




H
3.6


 


822136




2




H
N




H
207


 


822137




2




H
N




H
36


 


822138




2




CH3
N




H
81


 


822139




2




CH3
N




H
21.2


 


822140




2




CH3
N




H
112.8


 


822142




2




CH3
N




H
39.1


 


822143




2








N




H
338


 


826804




2




H
N




H
67.4


 


826805




2




H
N




H
14.2


 


826806




2




H
N




H
40


 


826807 (racemic)




2




H
N




H
43


 


826808




2




CH3
N




H
17.4


 


826809




2




CH3CH2
N




H
71


 


826810




2




HO(CH2)2
N




H
86.7


 


826820




2




H
N




H
52.8


 


826821




2




H
N




H
70.1


 


826824




2




H
N




H
34.2


 


826825




3




H
N




H
74.9


 


826826




3




H
N




H
27.4


 


826827 (racemic)




3




H
N




H
39.5


 


826828




3




CH3
N




H
14.4


 


826829




3




CH3CH2
N




H
1.5


 


826830




3




HO(CH2)2
N




H
25.1


 


826840




3




H
N




H
11


 


826841




3




H
N




H
22


 


822144




2




CH3
N




H
144


 


822145




2




H
N




H
176


 


822147




2




H
N




H
217


 


822148




2




H
N




H
41.4


 


826844




2




H
N




H
0.591


 


826845




2




H
N




H
4.42


 


826846




2




H
N




H
17.6


 


826847




2




H
N




H
1.01


 


826848




2




CH3
N




H
4.35


 


826849




2




CH3CH2
N




H
54.2


 


826850




2




HO(CH2)2
N




H
18.8


 


826860




2




H
N




H
3.48


 


826861




2




H
N




H
2.89


 


826857 Racemic




2




H
N




H
137


 


826860 Racemic




2




H
N




H
130


 


825870 Racemic




2




H
N




H
100


 


825871 Racemic




2




H
N




H
137


 


825872 Racemic




2




H
N




H
104


 


825881




2




H
N




H
189


 


825882




2




H
N




H
186


 


825883




2




H
N




H
140


 


825886




2




H
N




H
40.2


 


825887




2




H
N




H
90.4


 


825888




2




H
N




H
89.4


 


825893




2




H
N




H
73.8


 


825894




2




H
N




H
59


 


825895




2




H
N




H
31.7


 


825896




2




H
N




H
53.3


 


825897




2




H
N




H
140


 


825898




2




H
N




H
49


 


825899




2




H
N




H
42.9


 


826065




2




H
N




H
175


 


822202




2




H
N




H
8.1


 


826066




2




H
N




H
60.3


 


826067




2




H
N




H
19.3


 


826068




2




H
N




H
54.6


 


826069




2




H
N




H
27.2


 


826070




2




H
N




H
12.6


 


826071




2




H
N




H
165


 


822203




2




H
N




H
30


 


826072




2




H
N




H
62.6


 


826073




2




H
N




H
32.3


 


826074




2




H
N




H
72.6


 


826075




2




H
N




H
55.6


 


826076




2




H
N




H
15


 


826077




2




H
N




H
88.9


 


826078




2




H
N




H
162


 


822204




2




H
N




H
12.6


 


826079




2




H
N




H
106


 


826080




2




H
N




H
14.8


 


815916




2




H
N




H
5


 


(+)-816589 (S,S)




2




H
N




H
1070


 


(−)-817569 (R,R)




2




H
N




H
2.62


 


815917




2




H
N




H
18


 


815918




2




H
N




H
15


 


826655




2




H
N




H
31


 


826656




2




H
N




H
45


 


826658




2




H
N




H
17


 


826884




2




H
N




H
15.2


 


826885




2




CH3
N




H
20.3


 


826886




2








N




H
136


 


826887




2




H
N




H
83.8


 


826888




2




H
N




H
37.2


 


826889




2




H
N




H
36.2


 


826890




2




H
N




H
38


 


826891




2




H
N




H
263


 


826892




2




H
N




H
25.9


 


826893




2




H
N




H
20.4


 


826894




2




CH3CH2
N




H
35


 


826895




2




HO(CH2)2
N




H
48.8


 


826897




2




H
N




H
128


 


826898




2




H
N




H
50.5


 


816078A HCl salt




2




H
N




H
89.7


 


826678




3




H
N




H
15


 


826679




3




H
N




H
9.1


 


826680




3




H
N




H
58


 


826681




3




H
N




H
48


 


826682




3




H
N




H
103


 


826684




3




H
N




H
7.9


 


826665




2




H
N




H
1.9


 


826666




2




H
N




H
6.3


 


826667




2




H
N




H
2.5


 


826668




2




H
N




H
3.9


 


826670




2




H
N




H
4.2


 


826677




2




H
N




H
2.3


 


826864




3




H
N




H
2.16


 


826865




3




H
N




H
3.68


 


826866




3




H
N




H
1.24


 


826867




3




H
N




H
1.89


 


826868




3




CH3
N




H
3.49


 


826869




3




CH3CH2
N




H
6.75


 


826870




3




HO(CH2)2
N




H
7.4


 


826880




3




H
N




H
2.86


 


826881




3




H
N




H
2.38


 


825825




2




H
N




H
152


 


825826G (Maleate Salt)




2




H
N




H
32.5


 


825827




2




H
N




H
54.8


 


825828




2




H
N




H
21.8


 


825829G (Maleate Salt)




2




H
N




H
40.1


 


825830




2




H
N




H
64.6


 


825831




2




H
N




H
48.1


 


825832




2




H
N




H
60.3


 


825834G (Maleate Salt)




2




H
N




H
6.9


 


825900




2




H
N




H
22.4


 


826057




2




H
N




H
90.9


 


826058




2




H
N




H
45.2


 


826059




2




H
N




H
27.1


 


826060




2




H
N




H
40.2


 


826061




2




H
N




H


 


826062




2




H
N




H


 


826063




2




H
N




H


 


826064




2




H
N




H


 


826086




2




H
N




H
103


 


826092




2




H
N




H
293


 


826098




2




H
N




H
451


 


826100




2




H
N




H
193


 


826102




2




H
N




H
210


 


826104




2




H
N




H
16.8


 


826105




2




H
N




H
690


 


826106




2




H
N




H
230


 


826108




2




H
N




H
100


 


826113




2




H
N




H
200


 


826114




2




H
N




H
133


 


826115




2




H
N




H
72.3


 


826116




2




H
N




H
59.9


 


826117




2




H
N




H
241


 


826118




2




H
N




H
568


 


826120




2




H
N




H
487


 


826121




2




H
N




H
502


 


826122




2




H
N




H
130


 


816592




2




H
N




H
11


 


816593




2




H
N




H
7.47


 


827173




2




H
N




H
2.78


 


827174




2




H
N




H
8.7


 


827176




2




H
N




H
1.18


 


827177




2




H
N




H
1


 


827178




2




H
N




H
12.4


 


827179




2




H
N




H
1.04


 


827180




2




H
N




H
2.47


 


827181




2




H
N




H
97.2


 


827182




2




H
N




H
73


 


827183




2




H
N




H
3.29


 


827185




2




H
N




H
4.89


 


827186




2




H
N




H
8.21


 


827187




2




H
N




H
10.6


 


827189




2




H
N




H
126


 


827190




2




H
N




H
2.42


 


827191




2




H
N




H
0.724


 


827193




2




H
N




H
40.6


 


827194




2




H
N




H
24.4


 


827195




2




H
N




H
8.18


 


827175




2




H
N




H
4.0


 


827197




2




H
N




H
27.6


 


827199




2




H
N




H
4.34


 


827200




2




H
N




H
119


 


827205




2




H
N




H
200


 


827206




2




H
N




H
59.7


 


827212




2




H
N




H
1.2


 


827213




2




H
N




H
11.1


 


827215




2




H
N




H
0.229


 


827216




2




H
N




H
2.16


 


827218




2




H
N




H
1.22


 


827219




2




H
N




H
2.63


 


817480 (Racemic)




2




H
N




H
3.07


 


827221




2




H
N




H
99.2


 


827222




2




H
N




H
2.09


 


828393




2




H
N




H
3.45


 


817570 (Racemic)




2




H
N




H
6.24


 


827226




2




H
N




H
16.1


 


827228




2




H
N




H
62


 


827229




2




H
N




H
1.71


 


827230




2




H
N




H
2.09


 


817571 (racemic)




2




H
N




H
2.85


 


827233




2




H
N




H
27.7


 


827234




2




H
N




H
3.15


 


827235




2




H
N




H
4.49


 


827214




2




H
N




H
6.13


 


827236




2




H
N




H
12.9


 


827238




2




H
N




H
2.48


 


827239




2




H
N




H
90.8


 


827244




2




H
N




H
197


 


827245




2




H
N




H
43


 


818675 (Racemic)




2




H
N




H
0.711


 


817478 (Racemic)




2




H
N




H
1.46

















  CMPD NUMBER
    R
  n




    A
    R1R2
    R3
    d3Ki

















826811




2




N




H
40.4


 


826816




2




N




H
26.3


 


826817




2




N




H
20


 


826818




2




N




H
29.3


 


826819




2




N




H
11.6


 


826822




2




N




H
2.79


 


826823




2




N




H
19


 


826831




3




N




H
2.55


 


826832




3




N




H
16.1


 


826833




3




N




H
0.872


 


826834




3




N




H
8.01


 


826835




3




N




H
2.12


 


826836




3




N




H
2.23


 


826837




3




N




H
8.8


 


826838




3




N




H
14.6


 


826839




3




N




H
9.37


 


826842




3




N




H
3.66


 


826843




3




N




H
126


 


826851




2




N




H
2.77


 


826852




2




N




H
3.54


 


826853




2




N




H
0.128


 


826854




2




N




H
0.116


 


826855




2




N




H
0.58


 


826856




2




N




H
1.8


 


826857




2




N




H
0.817


 


826858




2




N




H
3.71


 


826859




2




N




H
3.53


 


826862




2




N




H
0.951


 


825868




2




N




H
13


 


825891




2




N




H
118


 


826081




2




N




H
18.6


 


826657




2




N




H
14


 


826659




2




N




H
140


 


826660




2




N




H
53


 


826661




2




N




H
25


 


826662




2




N




H
69


 


826664




2




N




H
9.4


 


826896




2




N




H
89.5


 


826899




2




N




H
45.5


 


826900




2




N




H
181


 


826901




2




N




H
42.4


 


826902




2




N




H
13.1


 


826903




2




N




H
7.47


 


826904




2




N




H
39.4


 


826905




2




N




H
14.5


 


826906




2




N




H
39.5


 


826907




2




N




H
18.8


 


826908




2




N




H
7.06


 


826909




2




N




H
28.9


 


826910




2




N




H
17.5


 


826911




2




N




H
93


 


826912




2




N




H
58.5


 


826913




2




N




H
86.1


 


826914




2




N




H
101


 


826916




2




N




H
157


 


826917




2




N




H
196


 


826918




2




N




H
30.3


 


826919




2




N




H
329


 


826920




2




N




H
67.6


 


826921




2




N




H
183


 


826922




2




N




H
183


 


826923




2




N




H
141


 


826924




2




N




H
256


 


826926




2




N




H
86.5


 


826928




2




N




H
11.2


 


826683




3




N




H
3.7


 


826685




3




N




H
154


 


826686




3




N




H
26


 


826687




3




N




H
132


 


826688




3




N




H
217


 


826689




3




N




H
674


 


826691




3




N




H
35


 


826669




2




N




H
0.24


 


826671




2




N




H
6.8


 


826672




2




N




H
0.64


 


826673




2




N




H
7.9


 


826674




2




N




H
48


 


826675




2




N




H
8


 


826676




2




N




H
2


 


826871




3




N




H
10.7


 


826872




3




N




H
3.66


 


826873




3




N




H
1.31


 


826874




3




N




H
1.72


 


826875




3




N




H
2.79


 


826876




3




N




H
2.72


 


826877




3




N




H
4.72


 


826878




3




N




H
7.95


 


826879




3




N




H
1.59


 


826882




3




N




H
2.93


 


826883




3




N




H
13.4


 


826056




2




N




H
5.4


 


826119




2




N




H
219


 


826812




2




N




H
7.29


 


826813




2




N




H
5.16


 


826814




2




N




H
10.3


 


826815




2




N




H
31.7


 


826863




2




N




H
26.6


 


827064




2




N




PhCH2
245


 


827065




2




N




PhCH2
168


 


827066




2




N




PhCH2
209


 


827067




2




N




PhCH2
268


 


827068




2




N




PhCH2
197


 


827063




2




N




PhCH2
373


 


827082




2




N




Ph(CH2)3
303


 


827084




2




N




Ph(CH2)3
266


 


827092




2




N




PhCH2
316


 


827093




2




N




PhCH2
84


 


827094




2




N




PhCH2
34.6


 


827095




2




N




PhCH2
37.8


 


827096




2




N




PhCH2
44.7


 


827097




2




N




PhCH2
51.5


 


827098




2




N




PhCH2
82.9


 


827099




2




N




PhCH2
144


 


827100




2




N




PhCH2
63.5


 


827103




2




N




PhCH2
81.7


 


827104




2




N




PhCH2
141


 


827112




2




N




Ph(CH2)3
126


 


827113




2




N




Ph(CH2)3
95


 


827114




2




N




Ph(CH2)3
120


 














826690 (racemic)




3




N




H
83% Inh @ 0.1 nM

























 















D3 Ki








MDL #
(nM)
Chirality
n
X
R1
R2
R3

















817274
4.49
Racemic
1
N




H






 


817275
3.34
Racemic
1
N




H






 


827184
237
Racemic
1
N
t-Boc
H






 


827192
150
Racemic
1
N




Me






 


827202
177
Racemic
1
N
—CH2Ph
H






 


827209
30.4
Racemic
1
CH
—CH2Ph
H






 


827211
353
Racemic
1
CH




H






 


827223
159
Racemic
1
N
t-Boc
H






 


827226
16.1
Racemic
1
N




H






 


827229
1.71
Racemic
1
N




H






 


827231
60.6
Racemic
1
N




CH3






 


827237
153
Racemic
1
N
—CH2CH2Ph
H






 


827240
152
Racemic
1
N




H






 


827241
89.5
Racemic
1
N
—CH2Ph
H






 


827248
19.4
Racemic
1
CH
—CH2Ph
H






 


827250
45.2
Racemic
1
CH




H






 


828716
2.38
R, R
1
N




H






 


829006DA
12.4
R, R
1
N




H






 


829363
63.9
Racemic
1
CH




H






 


830046
16.5
Racemic
1
N




H






 


830112
54
Racemic
1
CH




H






 


830114
5.42
Racemic
1
N




H






 


830116
13.1
Racemic
1
N




H






 


830121
47.1
Racemic
1
N




H






 


830122
67.6
Racemic
2
N




H






 


830123
73.8
Racemic
2
N




H






 


830124
1.84
Racemic
1
N




H






 


830128
69.7
Racemic
2
N




H






 


830129
4.16
Racemic
1
N




H






 


830131
34.1
Racemic
2
N




H






 


830132
9.26
Racemic
1
CH




H






 


830133
176
Racemic
1
CH




H






 


830134
0.602
Racemic
1
CH




H






 


830439
17.6
Racemic
1
N




H






 


831147
40.8
S, S
1
CH




H






 


831148
0.454
R, R
1
CH




H






 


831226
26.7
R, R
1
N




H






 


831269
4.08
R, R
1
N




H






 


831270
17.8
R, R
1
N




H
—NH—(CH2)3Ph


 


831271
6.57
R, R
1
N




H






 


831309
40.6
R, R
1
CH




H






 


831310
103
R, R
1
CH




H
—NH—(CH2)3-Ph


 


831311
38.7
R, R
1
CH




H






 


831312
214
R, R
1
CH




H
—NH—(CH2)3-Ph


 


831313
134
R, R
1
N




H






 


831314
133
R, R
1
N




H
—NH—(CH2)3-Ph


 


831360
17.1
S, S
1
CH




H






 


831361
1.75
R, R
1
CH




H






 


831488
0.725
R, R
1
CH




H






 


831489
16.9
R, R
1
CH




H
—NH—(CH2)3-Ph


 


831491
0.187
R, R
1
N




H






 


831492
0.432
R, R
1
N




H






 


831768
0.904
R, R
1
N




H






 


831808
3.84
R, R
1
N




H






 


831809
0.0284
R, R
1
CH




H






 


831810
124
R, R
1
CH




H






 


831878
189
R, R
1
CH




H






 


831880
100
R, R
1
CH




H






 


831881
109
R, R
1
N




H






 


831882
57.3
R, R
1
N




H






 


831883
48.1
R, R
1
N




H






 


831884
1.94
R, R, (R, S)
1
N




H






 


831885
35.3
R, R
1
N




H






 


831886
16.8
R, R
1
N




H






 


831887
6.5
R, R
1
N




H






 


831888
17.5
R, R
1
N




H






 


831949
29
R, R
1
N




H






 


831950
5.48
R, R
1
CH




H






 


831952
42.8
R, R
1
CH




H






 


831953
37.4
R, R
1
CH




H






 


831958
13
R, R
1
N




H






 


832102
81.5
R, R
1
CH




H
—NH—(CH2)3-Ph


 


832104
619.7
R, R
1
CH




H






 


832108
178
R, R
1
CH




H






 


832203
4.76
R, R
1
CH




H






 


832216
76.5
R, R, R
1
N




H






 


832217
44.1
R, R, S
1
N




H






 


832268
2.62
R, R, (R, S)
1
N




H






 


832271
112.29
R, R
1
CH




H






 


832272
144.72
R, R
1
CH




H






 


832273
205
R, R
1
CH




H






 


832274
52.4
R, R
1
CH




H






 


832494
25
R, R
1
N




H






 


832495
63.6
R, R
1
N




H






 


832496
3.66
R, R
1
N




H






 


832497
38
R, R
1
N




H






 


832498
70.3
R, R
1
N




H






 


832499
19.7
R, R
1
N




H






 


832500
161
R, R (R, S)
1
N




H






 


832501
41.4
R, R (R, S)
1
N




H






 


832502
2.67
R, R
1
N




H






 


832503
11.11
R, R (R, S)
1
N




H






 


832504
5.77
R, R (R, S)
1
N




H






 


832510
25
R, R
1
N




H






 


832511
12.3
R, R
1
N




H






 


832512
15.7
R, R
1
N




H






 


832577
1.85
R, R
1
N




H






 


832578
25.8
R, R, (R, S)
1
N




H






 


832579
6.29
R, R
1
N




H






 


832580
28.83
R, R
1
N




H






 


832581
67.35
R, R
1
N




H






 


832582
25
R, R
1
N




H






 


832583
1.61
R, R
1
N




H






 


832654
4.72
R, R
1
CH




H






 


832714
12.4
R, R
1
CH




H






 


832715
166.46
R, R
1
CH




H






 


832716
7.54
R, R
1
CH




H






 


832734
9.85
R, R
1
N




H






 


832735
36.8
R, R
1
N




H






 


832736
2.74
R, R
1
N




H






 


832737
5.42
R, R
1
N




H






 


832738
3.26
R, R
1
N




H






 


832739
2.89
R, R
1
N




H






 


832740
30.2
R, R
1
N




H






 


832741
6.03
R, R
1
N




H






 


832742
5.75
R, R
1
N




H






 


832743
2.36
R, R
1
N




H






 


832744
1.37
R, R
1
N




H






 


832745
15.04
R, R
1
N




H






 


832747ES
1.05
R, R
1
N




H






 


832748FH
79.07
R, R
1
N




H






 


832789
3.05
R, R
1
N




H






 


832833
18.2
R, R
1
N




H






 


832834
12.2
R, R
1
N




H






 


832835
18.1
R, R
1
N




H






 


832859
14.3
R, R
1
N




H






 


833062
55.4
R, R
1
N




H






 


833064
28.8
R, R, (R, S)
1
N




H






 


833065
692.37
R, R
1
N




H






 


833080
22.9
R, R
1
CH




H






 


833090
41.85
R, R
1
CH




H






 


833091
117.7
R, R
1
CH




H






 


833092
33.47
R, R
1
CH




H






 


833097
0.9
R, R, S
1
N




H






 


833098
8.84
R, R, R
1
N




H






 


833120
138
R, R
1
N




H






 


833121
26.5
R, R
1
N




H






 


833122
52.89
R, R
1
N




H






 


833123
26.26
R, R
1
N




H






 


833124
26.83
R, R
1
N




H






 


833125
70.76
R, R
1
N




H






 


833136
14.3
R, R
1
N




H






 


833151
11.2
R, R
1
CH




H






 


833157
43.88
R, R
1
CH




H






 


833197
45.38
R, R
1
N




H






 


833277
16.7
R, R
1
N




H






 


833278
7.04
R, R
1
N




H






 


833305
8.81
R, R
1
CH




H






 


833322
12.8
R, R
1
N




H






 


833323
3.46
R, R
1
N




H






 


833512
2.17
R, R
1
N




H






 


833513
43.1
R, R
1
N




H






 


833514
0.36
R, R
1
N




H






 


833515
48.3
R, R
1
N




H






 


833516
21.7
R, R
1
N




H






 


833585ES
5.86
R, R
1
CH




H






 


833625
22.2
R, R, R
1
N




H






 


833626
2.17
R, R
1
N




H






 


833627
1.63
R, R
1
N




H






 


833628
95.82
R, R
1
N




H






 


833629
2.69
R, R
1
N




H






 


833630
52.09
R, R, R
1
N




H






 


833631
31.3
R, R
1
N




H






 


833632
4.06
R, R, R
1
N




H






 


833686
7.22
R, R
1
CH




H






 


833690
7.97
R, R
1
CH




H






 


833692
72.9
R, R
1
CH




H






 


833693
18.4
R, R
1
CH




H






 


833694
46.9
R, R
1
CH




H






 


833695
176
R, R
1
CH




H






 


833696
268
R, R
1
CH




H






 


833697
105
R, R
1
CH




H






 


833698
134
R, R
1
CH




H






 


833699
7.2
R, R
1
N




H






 


833700
3.24
R, R
1
N




H






 


833701
1.28
R, R
1
N




H






 


833702
0.495
R, R
1
N




H






 


833703
1.92
R ,R
1
CH




H






 


833704
2.3
R, R
1
N




H






 


833769
1.63
R, R
1
CH




H






 


833821
6.91
R, R
1
N




H






 


833822
0.64
R, R
1
N




H






 


833823
0.637
R, R
1
N




H






 


833834
0.333
R, R
1
N




H






 


833835
0.499
R, R
1
N




H






 


833836
1.36
R, R
1
N




H






 


833910
1.09
R, R, (R, S)
1
CH




H






 


833911
1.26
R, R (R, S)
1
CH




H






 


833934
0.633
R, R
1
CH




H






 


833941
8.98
R, R
1
CH




H






 


834003
3.51
R, R
1
CH




H






 


834012
0.241
R, R
1
CH




H






 


834015
1.24
R, R
1
CH




H






 


834016
1.63
R, R
1
CH




H






 


834017
1.62
R, R
1
CH




H






 


834054
13.8
R, R
1
CH




H






 


834055
62
R, R
1
CH




H






 


834056
93.2
R, R
1
CH




H






 


834057
89.4
R, R
1
CH




H






 


834058
91.4
R, R
1
CH




H






 


834059
2.76
R, R
1
CH




H






 


834129
6.75
R, R
1
CH




H






 


834171
65.2
R, R
1
CH




H






 


834172
0.837
R, R
1
N




H






 


834173
5.15
R, R (R, S)
1
N




H






 


A002200835
0.4
R, R (R, S)
1
CH




H






 


A002200836
1.51
R, R
1
CH




H






 


A002200837
0.368
R, R
1
CH




H






 


A002200838
0.807
R, R
1
CH




H






 


A002243383
25.09
R, R
1
N




H






 


A002243384
133.36
R, R
1
N




H






 


A002287512
0.647
R, R (R, S)
1
CH




H






 


A002287513
3.57
R, R
1
CH




H






 


A002287764
0.48
R, R
1
CH




H






 


A002287765
0.011
R, R
1
CH




H






 


A002287766
126.3
R, R, S
1
CH




H






 


A002328939
201.16
R, R
1
CH




H






 


A002328940
26.8
R, R
1
CH




H






 


A002328941
23.88
R, R
1
CH




H






 


A002329092
321.2
R, R
1
CH




H






 


A002329093
411.4
R, R
1
CH




H






 


A002329094
7.08
R, R
1
CH




H






 


A002329095
26.47
R, R
1
CH




H






 


A002329097
101.2
R, R
1
CH




H






 


A002329098
345.2
R, R
1
CH




H






 


A002329099
2.06
R, R (R, S)
1
CH




H






 


A002329100
6.32
R, R
1
CH




H






 


A002436288A
105.69
R, R
1
CH




H






 


A002437094
91.93
R, R
1
N




H






 


A002437350
16.2
R, R
1
N




H






 


A002437351
32.68
R, R
1
N




H






 


A002437353
0.937
R, R, S
1
CH




H






 


A002437354
1.03
R, R, S
1
CH




H






 


A002437355
1.87
R, R, S
1
CH




H






 


A002437357
14.03
R, R (R, S)
1
N




H






 


A002437358
8.858
R, R (R, S)
1
N




H






 


A002437359
0.541
R, R, S
1
CH




H






 


A002437360
0.239
R, R, S
1
CH




H






 


A002437517
10.51
R, R
1
CH




H






 


A002437802
0.251
R, R, S
1
N




H






 


A002438636
13.04
R, R, S
1
N




H






 


A002438637
21.18
R, R, R
1
N




H






 


A002438638
3.28
R, R, R
1
N




H






 


A002438639
97.32
R, R
1
CH




H






 


A002438663
1.37
R, R, S
1
N




H






 


A002438664
6.49
R, R
1
CH




H






 


A002439099
4.91
R, R
1
N




H






 


A002439100
551.94
R, R
1
N




H






 


A002439258
34.08
R, R
1
CH




H






 


A002439259
39.05
R, R
1
CH




H






 


A002439260
0.303
R, R, S
1
CH




H






 


A002439261
55.3
R, R
1
CH




H






 


A002440433A
180.18
R, R
1
CH




H






 


A002440437A
41.24
R, R
1
CH




H






 


A002609935
0.142
R, R, S
1
CH




H





























 














MDL #
D3 Ki (nM)
Chirality
X
Y
R1
n
R2

















827393
391
Racemic
O
CH
H
1






 


827394
391
Racemic
O
CH
H
1






 


827395
31.4
Racemica
O
CH
H
1






 


827397
111
Racemic
O
CH
H
1






 


829460
158
R,R
O
CH
H
1






 


829461
306
R,R
O
CH
H
1






 


829462
9.65
R,R, (R,S)
O
CH
H
1






 


829463
4.76
Racemic
O
CH
H
1






 


829464
22.4
R,R
O
CH
H
1






 


829465
80.3
R,R
O
CH
H
1






 


829466
24.8
R,R
O
CH
H
1






 


829467
9.01
R,R
O
CH
H
1






 


829590
109
Racemic
O
CH
H
1






 


829591
72.4
Racemic
O
CH
H
1






 


829592
42.3
Racemic
O
CH
H
1






 


829593
350
Racemic
O
CH
H
1






 


829594
364
Racemic
O
CH
H
1






 


829595
201
Racemic
O
CH
H
1






 


829596
4.41
R, R
O
CH
H
1






 


829597
226
Racemic
O
CH
H
1






 


830126
23.8
Racemate
O
N
H
2






 


830127
1.66
Racemate
O
N
H
1






 


830250
48.6
R,R
O
CH
H
1






 


830251
11.4
R,R
O
CH
H
1






 


830252
110
R,R
O
CH
H
1






 


830827
9.85
R,R
O
CH
3-CH3
1






 


830828
8.57
R,R
O
CH
H
1
—N—(CH2)3-Ph


 


830829
5.3
R,R
O
CH
H
1
—N—(CH2)2-Ph


 


830830
2.58
R,R
O
CH
H
1






 


830831
3.19
R,R
O
CH
H
1






 


830832
67.7
R,R
O
CH
H
1






 


831045
335
R,R
O
CH
H
1






 


831046
15.5
R,R
O
CH
H
1






 


831047
2.87
R,R
O
CH
H
1
—N—(CH2)4-Ph


831048
11.3
R,R
O
CH
H
1
—N—(CH2)3—N(CH3)2


 


831082
38.2
S,S
O
N
H
1






 


831083
3.2
R,R
O
N
H
1






 


831192
3,44
R,R
O
CH
H
1






 


831193
23.8
R,R
O
CH
H
1






 


831194
5.06
R,R
O
CH
H
1






 


831195
1.84
R,R
O
CH
H
1






 


831196
7.81
R,R
O
CH
H
1






 


831197
4.17
R,R
O
CH
H
1






 


831198
1.76
R,R
O
CH
H
1






 


831199
2.78
R,R
O
CH
H
1






 


831215
5.92
R,R
O
CH
H
1






 


831267
28.3
R,R
N-Ts
N
H
1






 


831272
6.22
R,R
O
CH
H
1






 


831273
68.6
R,R
O
CH
H
1






 


831274
17.4
R,R
O
CH
H
1






 


831275
5.15
R,R
O
CH
H
1






 


831276
3.85
R,R
O
CH
H
1






 


831277
5.38
R,R
O
CH
H
1






 


831278
18.1
R,R
O
CH
H
1






 


831315
29.3
R,R
NH
N
H
1






 


831343
13.7
R,R
O
CH
H
1






 


831344
22.1
R,R
O
CH
H
1






 


831345
34.4
R,R
O
CH
H
1






 


831374
1.12
R,R
O
CH
CH3
1






 


831375
2.9
R,R
O
CH
CH3
1
—N—(CH2)3-Ph


 


831376
40.3
R,R
O
CH
H
1






 


831377
4.68
R,R
O
CH
H
1






 


831378
35.6
R,R
O
CH
H
1






 


831379
46.7
R,R
O
CH
H
1






 


831385
22.7
Racemic
O
CH
H
1






 


831390
0.861
Racemic
O
CH
H
1






 


831453
18.3
Racemic
O
CH
H
1






 


831454
1.86
Racemic
O
CH
H
1






 


831455
6.79
Racemic
O
CH
H
1






 


831456
6.21
Racemic
O
CH
H
1






 


831457
1.83
Racemic
O
CH
H
1






 


831458
7.43
Racemic
O
CH
H
1






 


831493
4.95
R,R
O
N
H
1






 


831494
28.3
R,R
O
N
H
1
—N—(CH2)3-Ph


 


831599
49.8
Racemic
O
CH
H
1






 


831602
19.6
Racemic
O
CH
H
1






 


831603
20.9
Racemic
O
CH
H
1






 


831621
70.3
Racemic
O
CH
H
1






 


831622
34.6
Racemic
O
CH
H
1






 


831649
173
Racemic
O
CH
H
1






 


831673
23.5
Racemic
N-Me
N
H
1






 


831686
25.3
Racemic
O
CH
H
1






 


831687
277
Racemic
O
CH
H
1






 


831689
16.4
Racemic
O
CH
H
1






 


831701
55.7
Racemic
O
CH
H
1






 


831702
27.9
Racemic
O
CH
H
1






 


831703
45
Racemic
O
CH
H
1






 


831704
47.5
Racemic
O
CH
H
1






 


831735
81.3
Racemic
O
CH
H
1






 


831736
291
Racemic
O
CH
H
1






 


831737
1.25
Racemic
O
CH
H
1






 


831738
223
Racemic
O
CH
H
1






 


831740
2.96
Racemic
O
CH
H
1






 


831741
125
Racemic
O
CH
H
1






 


831794
27.3
Racemic
O
CH
H
1






 


831795
1.82
Racemic
O
CH
H
1






 


831798
46.3
Racemic
O
CH
H
1






 


831799
12.1
Racemic
O
CH
H
1






 


831800
5
Racemic
O
CH
H
1






 


831801
11.2
Racemic
O
CH
H
1






 


831802
13.4
Racemic
O
CH
H
1






 


831803
7.44
Racemic
O
CH
H
1






 


831804
9.89
Racemic
O
CH
H
1






 


831805
10.2
Racemic
O
CH
H
1






 


831818
0.117
Racemic
O
CH
H
1






 


831819
0.1
Racemic
O
CH
H
1






 


831820
6.54
Racemic
O
CH
H
1






 


831821
24.1
Racemic
O
CH
H
1






 


831822
1.38
Racemic
O
CH
H
1






 


831823
2.37
Racemic
O
CH
H
1






 


831830
0.76
R,R
N-Me
N
H
1






 


831831
6.31
Racemic
O
CH
H
1






 


831835
234
Racemic
O
CH
H
1






 


831836
94.3
Racemic
O
CH
H
1






 


831837
491
Racemic
O
CH
H
1






 


831838
141
Racemic
O
CH
H
1






 


831936
34.7
R,R, (R,S)
O
CH
H
1






 


831937
35
R,R
O
CH
H
1






 


831938
33
R,R
O
CH
H
1






 


831951
36.7
R,R
N-Ts
N
H
1






 


831954
15.6
R,R
O
CH
CH3
1






 


831955
47.3
R,R
O
N
H
1






 


8319560A
5.91
R,R
N-Me
N
H
1






 


831957
63.2
R,R
O
CH
H
1






 


831991
0.722
Racemic
O
CH
H
1






 


831992
4.81
Racemic
O
CH
H
1






 


831993
2.51
Racemic
O
CH
H
1






 


831994
2.68
Racemic
O
CH
H
1






 


831995
4.73
Racemic
O
CH
H
1






 


831996
7.97
Racemic
O
CH
H
1






 


831997
3.55
Racemic
O
CH
H
1






 


831998
7.46
Racemic
O
CH
H
1






 


831999
473
Racemic
O
CH
H
1






 


832000
79.6
Racemic
O
CH
H
1






 


832001
157
Racemic
O
CH
H
1






 


832044
657
Racemic
O
CH
H
1






 


832045
22
Racemic
O
CH
H
1






 


832046
35.5
Racemic
O
CH
H
1






 


832047
25.8
Racemic
O
CH
H
1






 


832049
6.54
Racemic
O
CH
H
1






 


832055
112
Racemic
O
CH
H
1






 


832057
21.7
Racemic
O
CH
H
1






 


832058
24.2
Racemic
O
CH
H
1






 


832059
4.13
Racemic
O
CH
H
1






 


832153
30.74
Racemic
O
CH
H
1






 


832154
21.86
Racemic
O
CH
H
1






 


832155
0.434
Racemic
O
CH
H
1






 


832156
0.315
Racemic
O
CH
H
1






 


832157
0.226
Racemic
O
CH
H
1






 


832158
0.37
Racemic
O
CH
H
1






 


832164
6.6
Racemic
O
CH
H
1






 


832165
30.13
Racemic
O
CH
H
1






 


832185
7.87
Racemic
O
CH
H
1






 


832186
16.06
Racemic
O
CH
H
1






 


832187
19.14
Racemic
O
CH
H
1






 


832188
10.1
Racemic
O
CH
H
1






 


832189
58.47
Racemic
O
CH
H
1






 


832190
94.61
Racemic
O
CH
H
1






 


832191
8.06
Racemic
O
CH
H
1






 


832192
7.81
Racemic
O
CH
H
1






 


832193
13.1
Racemic
O
CH
H
1






 


832194
23
Racemic
O
CH
H
1






 


832195
22.8
Racemic
O
CH
H
1






 


832196
22.9
Racemic
O
CH
H
1






 


832197
17.1
Racemic
O
CH
H
1






 


832198
5.81
Racemic
O
CH
H
1






 


832199
18.7
Racemic
O
CH
H
1






 


832207
54.84
R,R
N-Me
N
H
1






 


832269
5.1
R,R, (R, S)
O
CH
H
1






 


832340
11.54
Racemic
O
CH
H
1






 


832341
2.63
Racemic
O
CH
H
1






 


832342
0.077
Racemic
O
CH
H
1






 


832343
0.718
Racemic
O
CH
H
1






 


832344
2.76
Racemic
O
CH
H
1






 


832345
1.46
Racemic
O
CH
H
1






 


832346
12.88
Racemic
O
CH
H
1






 


832347
1.88
Racemic
O
CH
H
1






 


832348
0.465
Racemic
O
CH
H
1






 


832349
0.843
Racemic
O
CH
H
1






 


832391
77.93
Racemic
O
CH
H
1






 


832392
2.29
Racemic
O
CH
H
1






 


832393
8.44
Racemic
O
CH
H
1






 


832394
8.32
Racemic
O
CH
H
1






 


832395
30.2
Racemic
O
CH
H
1






 


832397
51.94
Racemic
O
CH
H
1






 


832464
12.8
Racemic
O
CH
H
1






 


832487
66.1
Racemic
O
CH
H
1






 


832488
39.5
Racemic
O
CH
H
1






 


832489
95
Racemic
O
CH
H
1






 


832505
40.6
Racemic
O
CH
H
1






 


832513
55.4
R,R (Isomer 1)
O
CH
H
1






 


832514
114.6
S,S, (Isomer 2)
O
CH
H
1






 


832527
45.4
R,R
O
CH
H
1






 


832528
48.5
R,R
O
CH
H
1






 


832529
145.6
R,R
O
CH
H
1






 


832530
16
R,R
O
CH
H
1






 


832531
67.67
R,R
O
CH
H
1






 


832532
14.4
R,R
O
CH
H
1






 


832533
4.12
R,R, (R,S)
O
CH
H
1






 


832565
1.45
R,R




N
H
1






 


832566
1.76
R,R




N
H
1






 


832573FH
4.52
Racemic
O
CH
H
1






 


832574FH
57.34
R,R
O
CH
H
1






 


832575ES
4.06
Racemic
O
CH
H
1






 


832576ES
17.5
Racemic
O
CH
H
1






 


832634
7.63
R,R




N
H
1






 


832648
20.8
R,R




N
H
1






 


832592
7.29
R,R
N-Ph
N
H
1






 


832723
54
R,R
O
CH
H
1






 


832724
1.44
R,R
O
CH
H
1






 


832725
4.22
R,R
O
CH
H
1






 


832726
7.05
R,R
O
CH
H
1






 


832727
1.19
R,R
O
CH
H
1






 


832728
3.12
R,R
O
CH
H
1






 


832729
1.67
R,R
O
CH
H
1






 


832730
3.62
R,R
O
CH
H
1






 


832731
0.699
R,R
O
CH
H
1






 


832732
0.74
R,R
O
CH
H
1






 


832733
0.656
R,R
O
CH
H
1






 


832788
1.84
R,R
O
CH
H
1






 


832816
2.57
R,R
N-Ph
N
H
1






 


832849
1.12
R,R
N-Ph
N
H
1






 


832850
0.264
R,R




N
H
1






 


832881
37.86
R,R




N
H
1






 


833333
8.11
R,R




N
H
1






 


833372
1.04
Racemic
O
CH
H
1






 


833373
65.46
R,R,S
O
CH
H
1






 


833374
30.6
R,R (R,S)
O
CH
H
1






 


833375
24.2
R,R
O
CH
H
1






 


833382
2.32
R,R
O
CH
H
1






 


833383
2.57
R,R
O
CH
H
1






 


833384
5.68
R,R,R
O
CH
H
1






 


833385
3.31
R,R (R,S)
O
CH
H
1






 


833386
36.46
R,R
O
CH
H
1






 


833482
0.962
R,R
O
CH
H
1






 


833483
0.93
R,R
O
CH
H
1






 


833485
2.99
R,R
O
CH
H
1






 


833509
41.53
R,R
O
CH
H
1






 


833714
4.16
R,R
O
CH
H
1






 


833715
5.7
R,R
O
CH
H
1






 


833748
1.38
R,R
O
CH
H
1






 


833855
12.4
R,R




N
H
1






 


833856
3.01
R,R




N
H
1






 


833876
50.3
R,R




N
H
1






 


833909
1.86
R,R
N-Ph
CH
H
1






 


834205
90.47
R,R




N
H
1






 


834206
88.82
R,R




N
H
1






 


834207
71.4
R,R




N
H
1






 


A002201942
30.2
R,R




N
H
1






 


A002202401
205.3
R,R
N-(CH2)3— CH3
N
H
1






 


A002202402
47.33
R,R
N-Me
N
H
1






 


A002202403
86.78
R,R
N-Me
N
H
1






 


A002243385
660.86
R,R




N
H
1






 


A002243404
492.34
R,R




N
H
1






 


A002287757
2.41
R,R
N-Me
N
H
1






 


A002287758
139.32
R,R




N
H
1






 


A002287767
3.3
R,R
N—CH2—CF3
CH
H
1






 


A002328969
137.1
R,R
N—CH2—CF3
CH
H
1






 


A002328970
36.2
R,R
N—CH2—CF3
CH
H
1






 


A002329040
159.9
R,R
N-Me
N
H
1






 


A002350369
100.28
R,R
N-Me
N
H
1






 


A002350370
81.25
R,R
N-Me
N
H
1






 


A002436290
520.47
R,R
N-Me
CH
H
1






 


A002436291
0.629
R,R
N-Me
CH
H
1






 


A002436292
128.56
R,R
N-Me
CH
H
1






 


A002436428
2.17
R,R
N-Me
N
H
1






 


A002436429
4.16
R,R
N-Me
N
H
1






 


A002437082
9.23
R,R




N
H
1






 


A002437083
11.03
R,R




N
H
1






 


A002437084
7.105
R,R




N
H
1






 


A002437352
1.22
R,R,S
N-Me
CH
H
1






 


A002438640
104.16
R,R
N-Me
CH
H
1






 


A002438641
62.76
R,R
N-Me
CH
H
1






 


A002439262
28.3
R,R
N-Ph
CH
H
1






 


A002439263
30.6
R,R
N-Ph
CH
H
1






 


A002439265
87.98
R,R
N—CH2— CF3
CH
H
1






 


A002439266
66.04
R,R
N—CH2— CF3
CH
H
1






 


A002440434A
462.13
R,R
N-Me
CH
H
1






 


A002440435A
304.55
R,R
N—CH2— CF3
CH
H
1






 


A002440436A
11.38
R,R
N-Ph
CH
H
1






 


833127
79.63
R,R
O
CH
H
1






 


833128
26.69
R,R
O
CH
H
1






 


833129
95.34
R,R
O
CH
H
1






 


833130
8.85
R,R
O
CH
H
1






 


833131
16.8
R,R
O
CH
H
1






 


833132
107.5
R,R
O
CH
H
1






 


833133
113.7
R,R
O
CH
H
1






 


833160
2.09
R,R
O
CH
H
1






 


833163
17.5
R,R
O
CH
H
1






 


833164
52.55
R,R
O
CH
H
1






 


833165
1.07
R,R
O
CH
H
1






 


833166
1.72
R,R
O
CH
H
1






 


833167
1.57
R,R
O
CH
H
1






 


833168
3.11
R,R
O
CH
H
1






 


834126
4.9
R,R
N-Bn
N
H
1





























(All Compounds Racemic)











MDL #
D3 Ki (nM)
R















831452
115
Bn



 



831811
819







 



831812
453






























 












MDL #
D3 Ki (nM)
Chirality
n
X
Ar















829997
10.4
Racemic
2
N






 


829830
65.3
Racemic
1
CH






 


829996
277
Racemic
1
N






 


830000
286
Racemic
2
N








































MDL #
D3 Ki (nM)
Chirality
X
Y
R1
R2
















830557
97.4
S
CH
O
OH






 


830558
34.2
R
CH
O
OH






 


830573
547
R
CH
O
OH






 


832212
301.8
n.a.
N
O
H






 


A002201941
119
n.a.
N
N—CH3
H






































MDL #
D3 Ki (nM)
X
Ar
R














832171
4.9
CH










 


832180
91
CH




—NH—(CH2)2—Ph


 


832231
6.93
N










 


832239
206.8
CH










 


832240
185.96
CH










 


832241A
51.8
CH










 


832242A
436.2
CH










 


832317
943
CH










 


832517
172.58
N










 


832518
98.2
N










 


832519
97.51
N




—NH—(CH2)2—Ph


 


832520
140.93
N










 


833330
72.69
CH










 


833365
166.4
CH










 


833378
23.7
CH










 


833475
22.45
N










 


833476
131.37
CH

































 










MDL #
D3 Ki (nM)
Chirality
Ar













832831DA
1153.74
Racemic






 


833953A
88.82
Racemic





























 













MDL #
D3 Ki (nM)
Chirality
X
R

















830115
45.3
Racemic
H
Me



830130
6.68
Racemic
F
Et

## Claims

1. A compound of the formula (I):



wherein
A is CH;
n is 1;
g is 1 or 2;
each R3 is independently hydrogen, C1-C6alkyl, or







wherein w is 1, 2, or 3;

R is selected from the group consisting of (a) and (k):






wherein
each R4, R12 is independently hydrogen, C1-C6alkyl, halogen, trifluoromethyl, —CO2C1-C6alkyl or —CH2OC1-C6alkyl;
p and s are 0, 1, or 2;








 represents a group (a):








wherein
each R19 and R20 is independently hydrogen, hydroxy or C1-C6alkyl;
and
d is 3 or 4;


R1 is
a) hydrogen;
b) C1-C6alkyl optionally mono- or di-substituted with hydroxy; or








wherein
each R24 is independently hydrogen or C1-C6alkyl;
each R25, and R26 is independently hydrogen or C1-C6alkyl;
t is 0 or 1; and
q is 0 or 1;



R2 is a group selected from (a)-(jj):











wherein
each R27 and R28 is independently selected from:
(1) hydrogen;
(2) C1-C6alkyl;
(3) C1-C6alkoxy;
(4) —CO2—R43 wherein R43 is hydrogen or C1-C6alkyl;
(5) hydroxy;
(6) —(CH2)a—OR44 wherein a is 1, 2 or 3 and R44 is hydrogen or C1-C6alkyl;
(7) —(CO)—NR45R46 
 wherein R45 and R46 are each independently hydrogen, C1-C2alkyl, or R45 and R46 taken together with the nitrogen atom to which they are attached from









z is 0 or 1;
e is 2, 3, 4, 5, 6 or 7;
h is 0, 1, 2 or 3;
u is 0, 1, 2, 3 or 4;
o is 0 or 1;
l is 0 or 1;
j is 0, 1, 2 or 3;
v is 0, 1, 2, 3 or 4;
w is 1, 2 or 3;
f is 1, 2, 3 or 4;
t is 0 or 1;
b is 0, 1 or 2;
q is 0 or 1;
aa is 0 or 2;
X is O, S or NR90 wherein R90 is hydrogen, C1-C6alkyl, or









wherein R143 is hydrogen or alkyl;

each M and V is a group independently selected from hydrogen, halogen, C1-C6alkyl, C1-C6alkoxy, trifluoromethyl, hydroxy, phenyl, phenoxy, —SO2NH2 or









 or

—NR48R49 wherein R48, and R49 are each independently hydrogen or C1-C2alkyl;
each R31, R32, R33, R34, R35, R36, R37, R38, R39, R40, R68, and R69 is independently hydrogen or C1-C6alkyl;
each R29, R30 is independently hydrogen, phenyl or C1-C6alkyl;
each R83, R84, R86, R87, R88, R89, R92, R93, R98, R99, R94, R95, R100, R101, R103, R104, R105, R106, R108, R109, R110, R111, R113, R114, R115, R116, R117, R118, R119, R120, R122, R123, R124, R125, R127, R128, R130, R131, R133, R134, R135, R136, R137, R138, R139 and R140 is independently hydrogen or C1-C6alkyl;
each R63, R64 and R65 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
each R66 is independently hydrogen, hydroxy, C1-C6alkyl or C1-C6alkoxy;
Q is CH2, CHOH or C═O;
X5 is O or S;
each R67 is independently hydrogen or C1-C6alkyl;
R70 is hydrogen, C1-C6alkyl, halogen, nitro or a phenyl group optionally mono-substituted with C1-C6alkyl, halogen or trifluoromethyl;
R81 is hydrogen, C1-C6alkyl, or —CO2C1-C6alkyl;
R91 is hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
R96 is hydrogen, C1-C6alkyl or









wherein R145 and R146 are each independently hydrogen or C1-C6alkyl and b is 0, 1 or 2;

R97 is hydrogen or C1-C6alkyl;
each R102 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
R107 is hydrogen or C1-C6alkyl;
each R121 is independently hydrogen, halogen, C1-C6alkyl or C1-C6alkoxy;
R127 is hydrogen or C1-C6alkyl;
R126 is C1-C6alkyl or benzyl;
R129 is hydrogen or C1-C6alkyl;
R132 is hydrogen, C1-C6alkyl, halogen or C1-C6alkoxy;
X3 is O or —NR127 wherein R127 is hydrogen or C1-C6alkyl;
X4 is O, S or —NR143 wherein R143 is hydrogen or C1-C6alkyl;
R141 is hydrogen, C1-C6alkyl or amino;
R142 is benzyl or phenyl each of which may be optionally substituted with C1-C6alkyl, halogen or C1-C6alkoxy;
R144 is hydrogen or C1-C6alkyl;
R85 is hydrogen, C1-C6alkoxy, C1-C6alkyl, —CO2C1-C6alkyl, C(O)C1-C6alkyl or a group selected from the following:






wherein
j is 0, 1, 2 or 3;
w is 1, 2 or 3;
m is 0, 1 or 2;
e is 2, 3, 4, 5, 6 or 7;
each R147, R148, R150, R151, R152, R153, R156, R157, R159, R160 R162 and R163 is independently hydrogen or C1-C6alkyl;
R149 is hydrogen, halogen, C1-C6alkyl, phenoxy, trifluoromethyl or trifluoromethoxy;
R155 is hydrogen, halogen or C1-C6alkyl;
R158 is hydrogen or C1-C6alkyl;
R161 is hydrogen or C1-C6alkyl;
R164 is hydrogen, halogen, C1-C6alkyl or trifluoromethyl;
R165 is hydrogen, C1-C6alkyl or halogen;
X7 is O or S or —NR167 wherein R167 is hydrogen or C1-C6alkyl;
R166 is hydrogen or C1-C6alkyl;
or R1 and R2 are joined together to form a pyrrolidinyl or piperidinyl ring, and in which the ring is optionally mono- or di-substituted, the substituents independently selected from:
(1) C1-C6alkyl;
(2) —CO2—(C1-C6alkyl);
(3) —NR50R51 wherein R50 and R51 are each independently hydrogen, C1-C6alkyl, or a phenyl group which is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl;
(4) —C(O)phenyl wherein the phenyl group is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl;
(5) —(CH2)mOR52 wherein R52 is hydrogen or C1-C2alkyl or a phenyl group which is optionally mono- or disubstituted with substituents independently selected from C1-C6alkyl, halogen or trifluoromethyl, and m is 0, 1 or 2;
(6) —NR54—COR53 wherein R54 is hydrogen or C1-C6alkyl and R53 is hydrogen or C1-C2alkyl;
(7) ═O;
(8) —CN;





wherein
b is 0, 1 or 2;
w is 1, 2 or 3;
t is 0 or 1;
i is 0, 1 or 2;
v is 0, 1, 2, 3 or 4;
k is 0 or 1;
c are 0, 1 or 2;
R167 is hydrogen or C1-C6alkyl;
each R55, R56, R58, R59, R169 and R170 is independently hydrogen or C1-C6alkyl;
each R57 is independently hydrogen, halogen or C1-C6alkyl;
each R60 is independently hydrogen, halogen or C1-C6alkyl;
R61 and R62 are each independently hydrogen or C1-C6alkyl;
R168 is hydrogen, thienyl or furanyl;
R171 is hydrogen, C1-C6alkyl, halogen, trifluoromethyl or trifluoromethoxy;


or R1 and R2 are joined together to form a group of formula X;




or R1 and R2 are joined together to form the group of formula (Y)




or R1 and R2 are joined together to form any of the following groups:





wherein
g is 1 or 2;
p is 0, 1 or 2;
R172 is hydrogen, C1-C6alkyl or C1-C6alkoxy;
R173 is hydrogen, C1-C6alkyl or phenyl optionally mono- or disubstituted with C1-C6alkyl or halogen; and
R82 is a substituent selected from the following groups:
(a) C1-C6alkyl optionally substituted with hydroxy;
(b) C1-C6alkenyl;
(c) C1-C6alkoxy;
(d) —(CH2)OC1-C6alkyl;
(e)









wherein X8 is —(CR175R176)h— or —(CR177═CR188)—
 wherein each R174 is independently hydrogen, C1-C6alkyl, halogen, trifluoromethyl, C1-C6alkoxy or benzyloxy;
 h is 0, 1, 2 or 3; each R175, R176, R177 and R178 is independently hydrogen or C1-C6alkyl; and
 j is 0, 1, 2 or 3;










wherein X9 is —(CR180R181)j— or —(CR184R185CR186═CR187)— or —(CR182═CR183)—
 wherein
 aa is 0 or 2;
 R179 is hydrogen, C1-C6alkyl, halogen, trifluoromethyl,
 C1-C6alkoxy, benzyloxy or phenyl;
 each R180, R181, R182, R183, R184, R185, R186 and R187 is independently hydrogen or C1-C6alkyl;
 j is 0, 1, 2, or 3;










wherein w is 1, 2 or 3;
 each R188 and R189 is independently hydrogen or C1-C6alkyl;










wherein
 i is 0, 1 or 2;
 each R190 is independently hydrogen, alkyl or halogen;
 b is 0, 1, or 2 as;
 each R191 and R192 is independently hydrogen or C1-C6alkyl;










wherein
 a is 1, 2 or 3;
 each R193 and R194 is independently hydrogen or C1-C6alkyl;
 R195 is hydrogen, halogen or C1-C6alkyl;










wherein
 e is 2, 3, 4, 5 or 6 as hereinbefore defined;
 b is 0, 1 or 2;
 each R196 and R197 is independently hydrogen or C1-C6alkyl;
 each R198 and R199 is independently hydrogen or C1-C6alkyl;










wherein
 each R200 and R201 is independently hydrogen or C1-C6alkyl;
 w is 1, 2 or 3;










wherein
each R202, R203, R204 and R205 is independently hydrogen or C1-C6alkyl; and
w is 1, 2 or 3;


—(CR206R207)w—OC1-C6alkyl  (n)


wherein
C1-C6alkyl is optionally substituted with hydroxy;
each R206 and R207 is independently hydrogen or C1-C6alkyl; and
w is 1, 2 or 3;


—(CR208R209)w—NR210R211  (o)


wherein
each R208, R209, R210 and R211 is independently hydrogen or C1-C6alkyl;
w is 1, 2 or 3;








with the proviso that when n is 1; and R3 is hydrogen or C1-C6alkyl; and




is group (a);
and R is group:
(a) wherein R4 is hydrogen, halogen or C1-C6alkyl, and R1 is hydrogen or unsubstituted C1-C6alkyl,
then R2 cannot be a group of the following formula:
group (a) wherein z is 0,
group (b) wherein u is 0 and M is hydrogen, halogen, C1-C6alkyl, or trifluoromethyl,
group (c) wherein o is 0,
group (d) wherein l is 0,
group (e) wherein j is 0,
group (g) wherein v is 0, or
the group formula (i);
and also when R is the group of formula (a), R1 and R2 cannot be joined together to form the group of formula Y or a pyrrolidinyl or piperidinyl ring wherein said ring is unsubstituted or mono- or di-substituted with C1-C6alkyl.

2. The compound of claim 1 wherein R is group (a).

3. The compound of claim 2 wherein R4 is halogen or CF3.

4. The compound of claim 3 wherein R2 is group (a).

5. The compound of claim 4 wherein z is 0 or 1;
e is 5 and each R27 and R28 is independently selected from hydrogen or C1-C6alkyl.

6. The compound of claim 3 wherein R2 is group (b).

7. The compound of claim 6 wherein M is hydrogen, C1-C6alkoxy or C1-C6alkyl and u is 0 or 1.

8. The compound of claim 3 wherein R2 is group (n).

9. The compound of claim 8 wherein R70 is hydrogen and f is 3.

10. The compound of claim 1 wherein R is group (k).

11. The compound of claim 10 wherein R12 is hydrogen, C1-C6alkyl, or —CH2OC1-C6alkyl.

12. The compound of claim 11 wherein R2 is group (a).

13. The compound of claim 12 wherein z is 0 or 1;
e is 5 and each R27 and R28 is independently selected from hydrogen or C1-C6alkyl.

14. The compound of claim 11 wherein R2 is group (b).

15. The compound of claim 14 wherein M is hydrogen, C1-C6alkoxy or C1-C6alkyl and u is 0 or 1.

16. The compound of claim 11 wherein R2 is group (n).

17. The compound of claim 16 wherein R70 is hydrogen and f is 3.

18. A pharmaceutical composition comprising an effective amount of a compound of claim 1 with a pharmaceutically-acceptable carrier or diluent.

19. A process for preparing a compound of formula I of claim 1 which comprises:
(a) reacting a compound of formula (II):





wherein R3, g, y, R, A and n are as defined in formula I of claim 1; with a compound of formula (III)






wherein “LG” is a suitable leaving group selected from chlorine, bromine, iodine and mesyl; and






 is as defined in formula I of claim 1;

to provide a compound of formula (IV)




(b) hydrolyzing a compound of formula (IV) to provide a compound of formula (V)




and (c) reacting a compound of formula (V) with a compound of formula (VI)





wherein R1 and R2 are as defined in formula (I) of claim 1;

to provide the compound of formula (I).

20. A process for preparing compounds of formula I of claim 1 which comprises:
(a) reacting a compound of formula (VII)





wherein “LG” is a suitable leaving group selected from chlorine, bromine, iodine and mesyl; and






 R1 and R2 are as defined in formula I of claim 1;

with a compound of formula (II)





wherein R3, g, y, R, A and n are as defined in formula I of claim 1;

to provide the compound of formula (I).