WO2010133569A1

WO2010133569A1 - Azabicyclo[4.1.0]heptane derivatives

Info

Publication number: WO2010133569A1
Application number: PCT/EP2010/056774
Authority: WO
Inventors: Giorgio Bonanomi; Fabrizio Micheli
Original assignee: Glaxo Group Ltd
Current assignee: Glaxo Group Ltd
Priority date: 2009-05-20
Filing date: 2010-05-18
Publication date: 2010-11-25
Anticipated expiration: 2011-11-20

Abstract

The invention relates to compounds of formula (I), processes for their preparation, intermediates used in these processes, pharmaceutical compositions containing them and their use in therapy, as serotonin (5-HT), dopamine (DA) and norepinephrine (NE), re-uptake inhibitors.

Description

TITLE

Azabicyclo[4.1.0]heptane Derivatives

The present invention relates to novel compounds, processes for their preparation, intermediates used in these processes, pharmaceutical compositions containing the compounds and their use in therapy as inhibitors of serotonin (5-HT), dopamine (DA) and norepinephrine (NE) reuptake.

BACKGROUND International patent publication WO 2008031772 discloses 1 ,6-disubstituted azabicyclo[4.1.0]heptane derivatives for use in therapy as inhibitors of 5-HT, DA and NE reuptake.

It is an object of the present invention to provide further inhibitors of 5-HT, DA and NE re-uptake.

SUMMARY OF THE INVENTION

According to a first aspect, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof

(I) wherein

R1 is H, halo, C-_|_4alkoxy or C-_|_4alkoxymethyl and R^ is H; or R^ and R^ are both fluoro; or R-I and R^ together form an oxo group;

R3 is 3,4-dichlorophenyl or naphthyl; R4 is C-_|.galkyl, haloC-_|.galkyl, C2-4alkenyl, C3_gcycloalkyl or C3_gcycloalkylC-_|_

4alkyl; and

R5 is C-_|_4alkyl; or

R4 and R^, together with the atoms to which they are attached, form a tetrahydrofuran ring. DESCRIPTION OF THE EMBODIMENTS

(I) wherein

R1 is H, halo, C-_|_4alkoxy or C-_|_4alkoxymethyl and R^ is H; or R-I and R^ are both fluoro; or R^ and R^ together form an oxo group; R3 is 3,4-dichlorophenyl or naphthyl;

R4 is C-_|.galkyl, haloC-_|.galkyl, C2-4alkenyl, C3_gcycloalkyl or C3_gcycloalkylC-_|. 4alkyl; and

R^ is C-|_4alkyl; or

R^ and R^, together with the atoms to which they are attached, form a tetrahydrofuran ring.

As used herein, alkyl is a univalent radical derived by removal of a hydrogen atom from an acyclic alkane of formula C_nH2n+2- For example, a C-|.galkyl is alkyl comprising from 1 to 6 carbon atoms. Alkyl may be straight chain or branched chain. Examples of alkyl are methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, iso-propyl, iso-butyl, sec-butyl, tert-butyl, iso-pentyl and neo-pentyl.

As used herein, halo is a fluorine, chlorine, bromine or iodine radical, i.e. fluoro, chloro, bromo or iodo.

As used herein, haloalkyl is alkyl (as defined hereinbefore) substituted by one or more halo substituents (as defined hereinbefore), which halo substituents may be the same or different. For example, haloC-_|.galkyl is haloalkyl consisting of 1 to 6 carbon atoms. Examples of haloalkyl are monofluoromethyl, difluoromethyl, trifluoromethyl and 1-chloro-2-fluoroethyl. As used herein, alkoxy is a group of formula "R-O-" where R is alkyl (as defined hereinbefore). For example, C-|.galkoxy is alkoxy consisting of 1 to 6 carbon atoms. Examples of alkoxy are methoxy, ethoxy, n-propoxy, n-butoxy, n-pentoxy, n- hexyloxy, iso-propoxy, iso-butoxy, sec-butoxy, tert-butoxy, iso-pentoxy and neo- pentoxy.

As used herein, alkenyl is a univalent radical derived by removal of a hydrogen atom from an acyclic alkene. For example, C2-6alkenyl is alkenyl consisting of 2 to 6 carbon atoms. Alkenyl contains one or more double bonds and may be straight or branched chain. Examples of alkenyl are vinyl, 1-propenyl, allyl, 1 ,3-butadienyl, isopropenyl and 3-methylbut-2-enyl).

As used herein, cycloalkyl is a univalent radical derived by removal of a hydrogen atom from a monocyclic cycloalkane. For example, C3_gcycloalkyl is cycloalkyl consisting of 3 to 6 ring-carbon atoms. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term oxo is a bivalent radical of formula =0.

The skilled chemist will appreciate from formula (I) that R³ and -CH(R⁴ )(OR⁵) (i.e. the radicals attached to the two fused carbons) possess a cis relationship, i.e. both R3 and -CH(R⁴ )(OR5) are on the same face of the bicyclic ring system. The skilled chemist will also appreciate from formula (I) that when R^ and R^ are different, the carbon to which they are attached is stereogenic. In addition the skilled chemist will appreciate that the carbon attached to R⁴ is also stereogenic. It will be appreciated therefore that compounds of formula (I) may exist in a variety of stereoisomers.

Mixtures of enantiomers, may be separated by appropriate optical resolution techniques (for example chiral HPLC).

In an embodiment, R^ and R^ are H.

In an embodiment, R^ is 3,4-dichlorophenyl. In an embodiment, R^ is C-_|_4alkyl or C2-4alkenyl.

In an embodiment, the compound defined in the first aspect is selected from the list:

(1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)1-(methyloxy)ethyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 1 );

(1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)propyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 2);

(1 R,6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)propyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 3); (1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)butyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 4);

(1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)butyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 5);

(1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[3-methyl-(R or S)-1-(methyloxy)butyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 6);

6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 7);

(1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(1 R or 1S)-methyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 8); (1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(2R or 2S)-tetrahydro-2-furanyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 9);

(1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(1 R or 1S)-1-(ethyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 10); or a pharmaceutically acceptable salt thereof.

The compounds of formula (I) as defined in the first aspect contain a basic centre and may form non-toxic acid addition salts formed with inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, with carboxylic acids or with organo-sulfonic acids. Examples include the HCI, HBr, HI, sulfate or bisulfate, nitrate, phosphate or hydrogen phosphate, acetate, benzoate, succinate, saccharate, fumarate, maleate, lactate, citrate, tartrate, gluconate, camsylate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts. For reviews on suitable pharmaceutical salts see Berge et al, J.

Pharm, ScL, 66, 1-19, 1977; P L Gould, International Journal of Pharmaceutics, 33 (1986), 201-217; and Bighley et al, Encyclopedia of Pharmaceutical Technology, Marcel Dekker Inc, New York 1996, Volume 13, page 453-497. In an embodiment, the salt is pharmaceutically acceptable.

It will be appreciated by those skilled in the art that certain protected derivatives of the compounds of formula (I) as defined in the first aspect, which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds defined in the first aspect which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". All protected derivatives and prodrugs of compounds defined in the first aspect are included within the scope of the invention. Examples of suitable pro-drugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31 , pp 306 - 316 and in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as "pro- moieties", for example as described by H. Bundgaard in "Design of Prodrugs" (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within the compounds defined in the first aspect. Therefore, in a further aspect, the invention provides a prodrug of a compound defined in the first aspect.

The compounds defined in the first aspect, their salts or prodrugs, may exist in solvated or hydrated form. Therefore, in a further aspect, the invention provides a solvate or hydrate of a compound defined in the first aspect or a salt thereof.

The compounds of formula (I) and their salts, as defined in the first aspect or solvates or hydrates of either, may exist in one or more polymorphic form. Therefore, in a further aspect, the invention provides a polymorph of a compound of formula (I) defined in the first aspect or their salts, or a polymorph of a solvate or hydrate of a compound of formula (I) defined in the first aspect, or a salt thereof.

Hereinafter, compounds of formula (I) as defined in the first aspect, their salts and prodrugs; any solvates or hydrates of any salt or prodrug; and any polymorph of any compound, salt, solvate or hydrate are referred to as "compounds of the invention". The term "compounds of the invention" also includes all embodiments of the first aspect.

The invention also includes all suitable isotopic variations of a compound of the invention. An isotopic variation of a compound of the invention is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸0, ³⁵S, ¹⁸F and ³⁶CI, respectively. Certain isotopic variations of the invention, for example, those in which a radioactive isotope such as ³H or ¹⁴C is incorporated, are useful in drug and/or substrate tissue distribution studies. Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with isotopes such as deuterium, i.e., ²H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements and hence may be preferred in some circumstances. Isotopic variations of the compounds of the invention can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the Experimental section hereafter using appropriate isotopic variations of suitable reagents.

Compounds of the invention may be prepared in a variety of ways. In the following reaction schemes and hereafter, unless otherwise stated R^ to R^ are as defined in the first aspect.

Throughout the specification, general formulae are designated by Roman numerals (I), (II), (III), (IV) etc. Subsets of these general formulae are defined as (Ia), (Ib), (Ic) etc .... (IVa)₁ (IVb), (IVc) etc.

Compounds of formula (I) may be obtained according to Scheme 1 , by deprotecting compounds of formula (II), wherein Pg is a suitable N-protecting group (typically Boc). When Pg is Boc, typical deprotection conditions comprise treatment with TFA in DCM at a temperature between 0 ⁰C and room temperature. Scheme 1

(H) (I)

Compounds of formula (II) may be obtained according to Scheme 2, by reacting compounds of formula (III) with NaH in DMF at 0 ⁰C and subsequently with an appropriate alkylating agent (e.g. methyl iodide in DMF) between 0 ⁰C and room temperature.

Compounds of formula (III) may be obtained according to Scheme 3, by reacting compounds of formula (IV) in THF at -2O⁰C with the appropriate Grignard reagent.

Compounds of formula (Na), i.e. compounds of formula (II) where R^ and R^, together with the atoms to which they are attached, form a tetrahydrofuran ring may be prepared according to Scheme 4, by reacting compounds of formula (II) where R^ is vinyl and R^ is allyl, via ring closing metathesis reaction using an appropriate catalyst (e.g. [1 ,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](dichloro)- (phenylmethylidene)-ruthenium/tricyclohexylphosphane (1 :1 ) ), followed by reduction of the resulting double bond under standard techniques (e.g. PtO₂ and H₂ in EtOH at room temperature)

Compounds of formula (IV) may be prepared according to reaction scheme 5 by oxidation of compounds of formula (V).

Compounds of formula (V) where R^ and R^ are H may be prepared according to procedures described in International patent publication WO2008/031772 (see Preparation 5 therein).

Compounds of formula (V) may also be prepared according to reaction scheme 6 from compounds of formula (Vl) by treatment with UAIH4 or DIBAL-H in THF between 0 ⁰C and room temperature.

Scheme 6

Compounds of formula (Via), i.e. compounds of formula (Vl) wherein R¹ is alkoxy, may be prepared according to reaction scheme 7 by akylating compounds of formula (VII) under standard conditions, e.g. using NaH in DMF between O ⁰C and room temperature,

Scheme 7

(VII) (Via)

Compounds of formula (VII) may be obtained according to Scheme 8 from compounds of formula (VIII) using standard double bond oxidation procedures, such as treatment with Borane-DMS followed by treatment with sodium hydroxide and hydrogen peroxide.

Scheme 8

(VIM)

Compounds of formula (VIb) may be obtained according to reaction scheme 9 from compounds of formula (VII) by an oxidation process. Typical conditions comprise treatment with Jones reagent at 0⁰C in DCM or Dess-Martin periodinane.

Scheme 9

(VII) (VIb)

Compounds of formula (VIc) may be obtained from compounds of formula (IX) according to Scheme 10, by treatment with a reducing agent such as Et₃SiH in CF₃COOH.

Scheme 10

(IX) (VIc)

Compounds of formula (VId) may be prepared according to reaction scheme 1 1 from compounds from compounds of formula (IX), by standard treatment with Selectfluor (N-fluoro-N'-chloromethyl triethylenediamine bis tetrafluoroborate).

Scheme 1 1

(IX) (VId)

Compounds of formula (IX) may be obtained from compounds of formula (VIII), according to reaction scheme 12 by treatment with Selectfluor (N-fluoro-N'- chloromethyl triethylenediamine bis tetrafluoroborate).

Scheme 12

Compounds of formula (VIII) may be prepared using procedures similar to those described in International patent publication WO2008/031772, Preparation 57.

A specific enantiomer or diastereoisomer of a compound of the invention may be obtained for example by optical resolution of a mixture of enantiomers or diastereoisomers using conventional methods, such as chiral chromatography.

The compounds of the invention may be used to treat diseases or conditions for which inhibition of 5-HT, DA and NE re-uptake is beneficial, i.e. inhibition of their respective monoamine transporter proteins referred to as SERT for 5-HT, DAT for DA and NET for NE. Therefore according to a further aspect, the invention provides a compound of the invention for use in treating a disease or condition. In an embodiment the disease or condition is a human disease or condition. In an embodiment, the invention provides a compound of the invention for use in treating a disease or condition for which inhibition of monoamine neurotransmitter re-uptake is beneficial, i.e. inhibition of one or more of SERT, NET and DAT.

In an embodiment the disease or condition which may benefit from inhibition of monoamine neurotransmitter re-uptake, i.e. inhibition of one or more of 5-HT, DA and NE re-uptake, is selected from the list consisting of: Parkinsonism, depression, eating disorders, sleep disorders, substance related disorders, attention-deficit hyperactivity disorders, anxiety disorders, cognition impairment, sexual dysfunctions, obsessive compulsive spectrum disorders, Gilles de Ia Tourettes disease and senile dementia, as well as other disorders sensitive to the monoamine neurotransmitter re- uptake-inhibiting activity of the compounds.

The terms describing some indications used hereinabove are classified in the

Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, published by the American Psychiatric Association (DSM-IV) and/or the International Classification of Diseases, 10th Edition (ICD-10). The various subtypes of the disorders mentioned hereinabove are contemplated as part of the present invention. Numbers in brackets after the listed diseases below refer to the classification code in DSM-IV.

The term "depression" includes:

Depression and mood disorders including Major Depressive Episode, Manic Episode, Mixed Episode and Hypomanic Episode; Depressive Disorders including Major Depressive Disorder, Dysthymic Disorder (300.4), Depressive Disorder Not Otherwise Specified (311 ); Other Mood Disorders including Mood Disorder Due to a General Medical Condition (293.83) which includes the subtypes With Depressive Features, With Major Depressive-like Episode, With Manic Features and With Mixed Features), Substance-Induced Mood Disorder (including the subtypes With

Depressive Features, With Manic Features and With Mixed Features) and Mood Disorder Not Otherwise Specified (296.90). Bipolar Disorders including Bipolar I Disorder, Bipolar Il Disorder (Recurrent Major Depressive Episodes with Hypomanic Episodes) (296.89), Cyclothymic Disorder (301.13) and Bipolar Disorder Not Otherwise Specified (296.80); The term "anxiety disorders" includes:

Anxiety disorders including Panic Attack; Panic Disorder including Panic Disorder without Agoraphobia (300.01 ) and Panic Disorder with Agoraphobia (300.21 ); Agoraphobia; Agoraphobia Without History of Panic Disorder (300.22), Specific Phobia (300.29, formerly Simple Phobia) including the subtypes Animal Type, Natural Environment Type, Blood-lnjection-lnjury Type, Situational Type and Other Type), Social Phobia (Social Anxiety Disorder, 300.23), Obsessive-Compulsive Disorder (300.3), Posttraumatic Stress Disorder (309.81 ), Acute Stress Disorder (308.3), Generalized Anxiety Disorder (300.02), Anxiety Disorder Due to a General Medical Condition (293.84), Substance-Induced Anxiety Disorder, Separation Anxiety Disorder (309.21 ), Adjustment Disorders with Anxiety (309.24) and Anxiety Disorder Not Otherwise Specified (300.00):

The term "substance related disorder" includes:

Substance-related disorders including Substance Use Disorders such as Substance Dependence, Substance Craving and Substance Abuse; Substance-Induced Disorders such as Substance Intoxication, Substance Withdrawal, Substance- Induced Delirium, Substance-Induced Persisting Dementia, Substance-Induced Persisting Amnestic Disorder, Substance-Induced Psychotic Disorder, Substance- Induced Mood Disorder, Substance-Induced Anxiety Disorder, Substance-Induced Sexual Dysfunction, Substance-Induced Sleep Disorder and Hallucinogen Persisting Perception Disorder (Flashbacks); Alcohol-Related Disorders such as Alcohol

Dependence (303.90), Alcohol Abuse (305.00), Alcohol Intoxication (303.00), Alcohol Withdrawal (291.81 ), Alcohol Intoxication Delirium, Alcohol Withdrawal Delirium, Alcohol-Induced Persisting Dementia, Alcohol-Induced Persisting Amnestic Disorder, Alcohol-Induced Psychotic Disorder, Alcohol-Induced Mood Disorder, Alcohol- Induced Anxiety Disorder, Alcohol-Induced Sexual Dysfunction, Alcohol-Induced Sleep Disorder and Alcohol-Related Disorder Not Otherwise Specified (291.9); Amphetamine (or Amphetamine-I_ike)-Related Disorders such as Amphetamine Dependence (304.40), Amphetamine Abuse (305.70), Amphetamine Intoxication (292.89), Amphetamine Withdrawal (292.0), Amphetamine Intoxication Delirium, Amphetamine Induced Psychotic Disorder, Amphetamine-Induced Mood Disorder, Amphetamine-Induced Anxiety Disorder, Amphetamine-Induced Sexual Dysfunction, Amphetamine-Induced Sleep Disorder and Amphetamine-Related Disorder Not Otherwise Specified (292.9); Caffeine Related Disorders such as Caffeine Intoxication (305.90), Caffeine-Induced Anxiety Disorder, Caffeine-Induced Sleep Disorder and Caffeine-Related Disorder Not Otherwise Specified (292.9); Cannabis- Related Disorders such as Cannabis Dependence (304.30), Cannabis Abuse (305.20), Cannabis Intoxication (292.89), Cannabis Intoxication Delirium, Cannabis- lnduced Psychotic Disorder, Cannabis-lnduced Anxiety Disorder and Cannabis- Related Disorder Not Otherwise Specified (292.9); Cocaine-Related Disorders such as Cocaine Dependence (304.20), Cocaine Abuse (305.60), Cocaine Intoxication (292.89), Cocaine Withdrawal (292.0), Cocaine Intoxication Delirium, Cocaine- Induced Psychotic Disorder, Cocaine-Induced Mood Disorder, Cocaine-Induced Anxiety Disorder, Cocaine-Induced Sexual Dysfunction, Cocaine-Induced Sleep Disorder and Cocaine-Related Disorder Not Otherwise Specified (292.9); Hallucinogen-Related Disorders such as Hallucinogen Dependence (304.50), Hallucinogen Abuse (305.30), Hallucinogen Intoxication (292.89), Hallucinogen Persisting Perception Disorder (Flashbacks) (292.89), Hallucinogen Intoxication Delirium, Hallucinogen-Induced Psychotic Disorder, Hallucinogen-Induced Mood Disorder, Hallucinogen-Induced Anxiety Disorder and Hallucinogen-Related Disorder Not Otherwise Specified (292.9); Inhalant-Related Disorders such as Inhalant Dependence (304.60), Inhalant Abuse (305.90), Inhalant Intoxication (292.89), Inhalant Intoxication Delirium, Inhalant-Induced Persisting Dementia, Inhalant- Induced Psychotic Disorder, Inhalant-Induced Mood Disorder, Inhalant-Induced Anxiety Disorder and Inhalant-Related Disorder Not Otherwise Specified (292.9); Nicotine-Related Disorders such as Nicotine Dependence (305.1 ), Nicotine

Withdrawal (292.0) and Nicotine-Related Disorder Not Otherwise Specified (292.9); Opioid-Related Disorders such as Opioid Dependence (304.00), Opioid Abuse (305.50), Opioid Intoxication (292.89), Opioid Withdrawal (292.0), Opioid Intoxication Delirium, Opioid-lnduced Psychotic Disorder, Opioid-lnduced Mood Disorder, Opioid- Induced Sexual Dysfunction, Opioid-lnduced Sleep Disorder and Opioid-Related Disorder Not Otherwise Specified (292.9); Phencyclidine (or Phencyclidine-Like)- Related Disorders such as Phencyclidine Dependence (304.60), Phencyclidine Abuse (305.90), Phencyclidine Intoxication (292.89), Phencyclidine Intoxication Delirium, Phencyclidine-lnduced Psychotic Disorder, Phencyclidine-lnduced Mood Disorder, Phencyclidine-lnduced Anxiety Disorder and Phencyclidine-Related Disorder Not Otherwise Specified (292.9); Sedative-, Hypnotic-, or Anxiolytic-Related Disorders such as Sedative, Hypnotic, or Anxiolytic Dependence (304.10), Sedative, Hypnotic, or Anxiolytic Abuse (305.40), Sedative, Hypnotic, or Anxiolytic Intoxication (292.89), Sedative, Hypnotic, or Anxiolytic Withdrawal (292.0), Sedative, Hypnotic, or Anxiolytic Intoxication Delirium, Sedative, Hypnotic, or Anxiolytic Withdrawal Delirium, Sedative-, Hypnotic-, or Anxiolytic-Persisting Dementia, Sedative-, Hypnotic-, or Anxiolytic- Persisting Amnestic Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Psychotic Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Mood Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Anxiety Disorder Sedative-, Hypnotic-, or Anxiolytic-lnduced Sexual Dysfunction, Sedative-, Hypnotic-, or Anxiolytic-lnduced Sleep Disorder and Sedative-, Hypnotic-, or Anxiolytic-Related Disorder Not Otherwise Specified (292.9); Polysubstance-Related Disorder such as Polysubstance Dependence (304.80); and Other (or Unknown) Substance-Related Disorders such as Anabolic Steroids, Nitrate Inhalants and Nitrous Oxide;

The term "Sleep disorder" includes:

Sleep disorders including primary sleep disorders such as Dyssomnias such as Primary Insomnia (307.42), Primary Hypersomnia (307.44), Narcolepsy (347), Breathing-Related Sleep Disorders (780.59), Circadian Rhythm Sleep Disorder

(307.45) and Dyssomnia Not Otherwise Specified (307.47); primary sleep disorders such as Parasomnias such as Nightmare Disorder (307.47), Sleep Terror Disorder (307.46), Sleepwalking Disorder (307.46) and Parasomnia Not Otherwise Specified (307.47); Sleep Disorders Related to Another Mental Disorder such as Insomnia Related to Another Mental Disorder (307.42) and Hypersomnia Related to Another Mental Disorder (307.44); Sleep Disorder Due to a General Medical Condition; and Substance-Induced Sleep Disorder including the subtypes Insomnia Type, Hypersomnia Type, Parasomnia Type and Mixed Type;

The term "eating disorder" includes:

Eating disorders such as Anorexia Nervosa (307.1 ) including the subtypes Restricting Type and Binge-Eating/Purging Type; Bulimia Nervosa (307.51 ) including the subtypes Purging Type and Nonpurging Type; Obesity; Compulsive Eating Disorder; Binge Eating Disorder; and Eating Disorder Not Otherwise Specified (307.50):

The term "Attention-Deficit/Hyperactivity Disorder" includes:

Attention-Deficit/Hyperactivity Disorder including the subtypes Attention-Deficit /Hyperactivity Disorder Combined Type (314.01 ), Attention-Deficit /Hyperactivity Disorder Predominantly Inattentive Type (314.00), Attention-Deficit /Hyperactivity Disorder Hyperactive-Impulse Type (314.01 ) and Attention-Deficit /Hyperactivity Disorder Not Otherwise Specified (314.9); Hyperkinetic Disorder; Disruptive

Behaviour Disorders such as Conduct Disorder including the subtypes childhood- onset type (321.81 ), Adolescent-Onset Type (312.82) and Unspecified Onset (312.89), Oppositional Defiant Disorder (313.81 ) and Disruptive Behaviour Disorder Not Otherwise Specified; and Tic Disorders such as Tourette's Disorder (307.23);

The term "Cognition impairment" includes:

Cognition impairment including cognition impairment in other diseases such as schizophrenia, bipolar disorder, depression, other psychiatric disorders and psychotic conditions associated with cognitive impairment, e.g. Alzheimer's disease;

The term "Sexual dysfunctions" includes:

Sexual dysfunctions including Sexual Desire Disorders such as Hypoactive Sexual Desire Disorder (302.71 ), and Sexual Aversion Disorder (302.79); sexual arousal disorders such as Female Sexual Arousal Disorder (302.72) and Male Erectile Disorder (302.72); orgasmic disorders such as Female Orgasmic Disorder (302.73), Male Orgasmic Disorder (302.74) and Premature Ejaculation (302.75); sexual pain disorder such as Dyspareunia (302.76) and Vaginismus (306.51 ); Sexual Dysfunction Not Otherwise Specified (302.70); paraphilias such as Exhibitionism (302.4), Fetishism (302.81 ), Frotteurism (302.89), Pedophilia (302.2), Sexual Masochism (302.83), Sexual Sadism (302.84), Transvestic Fetishism (302.3), Voyeurism (302.82) and Paraphilia Not Otherwise Specified (302.9); gender identity disorders such as Gender Identity Disorder in Children (302.6) and Gender Identity Disorder in Adolescents or Adults (302.85); and Sexual Disorder Not Otherwise Specified (302.9);

The term Obsessive compulsive spectrum disorder" includes:

Obsessive compulsive spectrum disorder including Obsessive compulsive disorders (300.3), somatoform disorders including body dysmorphic disorder (300.7) and hyperchondriasis (300.7), bulimia nervosa (307.51 ), anorexia nervosa (307.1 ), eating disorders not elsewhere classified (307.50) such as binge eating, impulse control disorders not elsewhere classified (including intermitted explosive disorder (312.34), compulsive buying or shopping, repetitive self-mutilation, onychophagia, psychogenic excoriation, kleptomania (312.32), pathological gambling (312.31 ), trichotillomania (312.39) and internet addiction), paraphilia (302.70) and nonparaphilic sexual addictions, Sydeham's chorea, torticollis, autistic disorders (299.0), compulsive hoarding, and movement disorders, including Tourette's syndrome (307.23).

All of the various forms and sub-forms of the disorders mentioned herein are contemplated as part of the present invention.

In an embodiment, compounds of the invention may be useful as analgesics. For example they may be useful in the treatment of chronic inflammatory pain (e.g. pain associated with rheumatoid arthritis, osteoarthritis, rheumatoid spondylitis, gouty arthritis and juvenile arthritis); musculoskeletal pain; lower back and neck pain; sprains and strains; neuropathic pain; sympathetically maintained pain; myositis; pain associated with cancer and fibromyalgia; pain associated with migraine; pain associated with influenza or other viral infections, such as the common cold; rheumatic fever; pain associated with functional bowel disorders such as non-ulcer dyspepsia, non-cardiac chest pain and irritable bowel syndrome; pain associated with myocardial ischemia; post operative pain; headache; toothache; and dysmenorrhea.

Compounds of the invention may be useful in the treatment of neuropathic pain. Neuropathic pain syndromes can develop following neuronal injury and the resulting pain may persist for months or years, even after the original injury has healed. Neuronal injury may occur in the peripheral nerves, dorsal roots, spinal cord or certain regions in the brain. Neuropathic pain syndromes are traditionally classified according to the disease or event that precipitated them. Neuropathic pain syndromes include: diabetic neuropathy; sciatica; non-specific lower back pain; multiple sclerosis pain; fibromyalgia; HIV-related neuropathy; post-herpetic neuralgia; trigeminal neuralgia; and pain resulting from physical trauma, amputation, cancer, toxins or chronic inflammatory conditions. These conditions are difficult to treat and although several drugs are known to have limited efficacy, complete pain control is rarely achieved. The symptoms of neuropathic pain are incredibly heterogeneous and are often described as spontaneous shooting and lancinating pain, or ongoing, burning pain. In addition, there is pain associated with normally non-painful sensations such as "pins and needles" (paraesthesias and dysesthesias), increased sensitivity to touch (hyperesthesia), painful sensation following innocuous stimulation (dynamic, static or thermal allodynia), increased sensitivity to noxious stimuli (thermal, cold, mechanical hyperalgesia), continuing pain sensation after removal of the stimulation (hyperpathia) or an absence of or deficit in selective sensory pathways (hypoalgesia).

Compounds of the invention may also be useful in the amelioration of inflammatory disorders, for example in the treatment of skin conditions (e.g. sunburn, burns, eczema, dermatitis, psoriasis); ophthalmic diseases such as glaucoma, retinitis, retinopathies, uveitis and of acute injury to the eye tissue (e.g. conjunctivitis); lung disorders (e.g. asthma, bronchitis, emphysema, allergic rhinitis, respiratory distress syndrome, pigeon fancier's disease, farmer's lung, chronic obstructive pulmonary disease, (COPD); gastrointestinal tract disorders (e.g. aphthous ulcer, Crohn's disease, atopic gastritis, gastritis varialoforme, ulcerative colitis, coeliac disease, regional ileitis, irritable bowel syndrome, inflammatory bowel disease, gastroesophageal reflux disease); other conditions with an inflammatory component such as migraine, multiple sclerosis, myocardial ischemia.

In one embodiment, compounds of the invention are useful in the treatment of depression and anxiety disorders.

In another embodiment, compounds of the invention are useful in the treatment of depression. "Treatment" includes prophylaxis, where this is appropriate for the relevant condition(s).

The compounds of the invention may also be used in combination with other therapeutic agents. The invention thus provides, in a further aspect, a combination comprising a compound of the invention together with a further therapeutic agent.

The compounds of the invention may be used in combination with the following agents to treat or prevent psychotic disorders: i) antipsychotics; ii) drugs for extrapyramidal side effects, for example anticholinergics (such as benztropine, biperiden, procyclidine and trihexyphenidyl), antihistamines (such as diphenhydramine) and dopaminergics (such as amantadine); iii) antidepressants; iv) anxiolytics; and v) cognitive enhancers for example cholinesterase inhibitors (such as tacrine, donepezil, rivastigmine and galantamine).

The compounds of the invention may be used in combination with antidepressants to treat or prevent depression and mood disorders.

The compounds of the invention may be used in combination with the following agents to treat or prevent bipolar disease: i) mood stabilisers; ii) antipsychotics; and iii) antidepressants.

The compounds of the invention may be used in combination with the following agents to treat or prevent anxiety disorders: i) anxiolytics; and ii) antidepressants.

The compounds of the invention may be used in combination with the following agents to improve nicotine withdrawal and reduce nicotine craving: i) nicotine replacement therapy for example a sublingual formulation of nicotine beta- cyclodextrin and nicotine patches; and ii) bupropion.

The compounds of the invention may be used in combination with the following agents to improve alcohol withdrawal and reduce alcohol craving: i) NMDA receptor antagonists for example acamprosate; ii) GABA receptor agonists for example tetrabamate; and iii) Opioid receptor antagonists for example naltrexone. The compounds of the invention may be used in combination with the following agents to improve opiate withdrawal and reduce opiate craving: i) opioid mu receptor agonist/opioid kappa receptor antagonist for example buprenorphine; ii) opioid receptor antagonists for example naltrexone; and iii) vasodilatory antihypertensives for example lofexidine.

The compounds of the invention may be used in combination with the following agents to treat or prevent sleeping disorders: i) benzodiazepines for example temazepam, lormetazepam, estazolam and triazolam; ii) non-benzodiazepine hypnotics for example Zolpidem, zopiclone, zaleplon and indiplon; iii) barbiturates for example aprobarbital, butabarbital, pentobarbital, secobarbita and phenobarbital; iv) antidepressants; v) other sedative-hypnotics for example chloral hydrate and chlormethiazole.

The compounds of the invention may be used in combination with the following agents to treat anorexia: i) appetite stimulants for example cyproheptidine; ii) antidepressants; iii) antipsychotics; iv) zinc; and v) premenstral agents for example pyridoxine and progesterones.

The compounds of the invention may be used in combination with the following agents to treat or prevent bulimia: i) antidepressants; ii) opioid receptor antagonists; iii) antiemetics for example ondansetron; iv) testosterone receptor antagonists for example flutamide; v) mood stabilisers; vi) zinc; and vii) premenstral agents.

The compounds of the invention may be used in combination with the following agents to treat or prevent autism: i) antipsychotics; ii) antidepressants; iii) anxiolytics; and iv) stimulants for example methylphenidate, amphetamine formulations and pemoline.

The compounds of the invention may be used in combination with the following agents to treat or prevent ADHD: i) stimulants for example methylphenidate, amphetamine formulations and pemoline; and ii) non-stimulants for example norepinephrine reuptake inhibitors (such as atomoxetine), alpha 2 adrenoceptor agonists (such as clonidine), antidepressants, modafinil, and cholinesterase inhibitors (such as galantamine and donezepil). The compounds of the invention may be used in combination with the following agents to treat personality disorders: i) antipsychotics; ii) antidepressants; iii) mood stabilisers; and iv) anxiolytics.

The compounds of the invention may be used in combination with the following agents to treat or prevent male sexual dysfunction: i) phosphodiesterase V inhibitors, for example vardenafil and sildenafil; ii) dopamine agonists/dopamine transport inhibitors for example apomorphine and buproprion; iii) alpha adrenoceptor antagonists for example phentolamine; iv) prostaglandin agonists for example alprostadil; v) testosterone agonists such as testosterone; vi) serotonin transport inhibitors for example serotonin reuptake inhibitors; vii) noradrenaline transport inhibitors for example reboxetine and viii) 5-HT1A agonists, for example flibanserine.

The compounds of the invention may be used in combination with the same agents specified for male sexual dysfunction to treat or prevent female sexual dysfunction, and in addition an estrogen agonist such as estradiol.

Antipsychotic drugs include Typical Antipsychotics (for example chlorpromazine, thioridazine, mesoridazine, fluphenazine, perphenazine, prochlorperazine, trifluoperazine, thiothixine, haloperidol, molindone and loxapine); and Atypical Antipsychotics (for example clozapine, olanzapine, risperidone, quetiapine, aripirazole, ziprasidone and amisulpride).

Antidepressant drugs include serotonin reuptake inhibitors (such as citalopram, escitalopram, fluoxetine, paroxetine and sertraline); dual serotonin/noradrenaline reuptake inhibitors (such as venlafaxine, duloxetine and milnacipran); Noradrenaline reuptake inhibitors (such as reboxetine); tricyclic antidepressants (such as amitriptyline, clomipramine, imipramine, maprotiline, nortriptyline and trimipramine); monoamine oxidase inhibitors (such as isocarboxazide, moclobemide, phenelzine and tranylcypromine); and others (such as bupropion, mianserin, mirtazapine, nefazodone and trazodone).

Mood stabiliser drugs include lithium, sodium valproate/valproic acid/divalproex, carbamazepine, lamotrigine, gabapentin, topiramate and tiagabine. Anxiolytics include benzodiazepines such as alprazolam and lorazepam.

The compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient by an appropriate route. Accordingly, in another aspect, the invention provides pharmaceutical compositions comprising a compound of the invention and one or more pharmaceutically-acceptable excipients.

As used herein, "pharmaceutically acceptable excipient" means any pharmaceutically acceptable material present in the pharmaceutical composition or dosage form other than the compound or compounds of the invention. Typically the material gives form, consistency and performance to the pharmaceutical composition.

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

Such pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be dispensed and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged as dosage forms wherein each physically discrete dosage form contains a safe and effective amount of a compound of the invention. Accordingly, in another aspect, the invention provides dosage forms comprising pharmaceutical compositions of the invention. Each discrete dosage form typically contains from 1 mg to 500 mg of a compound of the invention.

The compositions of the invention will typically be formulated into dosage forms which are adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets and cachets; (2) parenteral administration such as sterile solutions, suspensions, implants and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal and vaginal administration such as suppositories, pessaries and foams; (5) inhalation and intranasal such as dry powders, aerosols, suspensions and solutions (sprays and drops); (6) topical administration such as creams, ointments, lotions, solutions, pastes, drops, sprays, foams and gels; (7) ocular administration such as drops, ointment, sprays, suspensions and inserts; (8) buccal and sublingual administration such as lozenges, patches, sprays, drops, chewing gums and tablets.

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

Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, colouring agents, anticaking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, rate modifying agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents. The skilled artisan will appreciate that certain pharmaceutically acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation. Skilled artisans possess the knowledge and skill in the art to enable them to determine suitable pharmaceutically acceptable excipients in appropriate amounts for use with the compounds of the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically acceptable excipients and may be useful in selecting suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press). The pharmaceutical compositions of the invention may be prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

It will be appreciated that the compounds of the combination or composition may be administered simultaneously (either in the same or different pharmaceutical formulations), separately or sequentially.

All publications, including, but not limited to, patents and patent applications cited in this specification, are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

It will be appreciated that the invention includes the following further aspects. The diseases and conditions described above extend, where appropriate, to these further aspects.

i) The use of a compound of the invention in the manufacture of a medicament in treating a disease or condition for which inhibition of monoamine neurotransmitter re-uptake is beneficial, i.e. inhibition of one or more of SERT, NET and DAT. In an embodiment the disease or condition is depression or an anxiety disorder. In a further embodiment the disease or condition is an eating disorder

ii) A method of treating a disease or condition for which inhibition of monoamine neurotransmitter re-uptake is beneficial, i.e. inhibition of one or more of SERT, NET and DAT in a mammal (preferably a human) comprising administering an effective amount of a compound of the invention. In an embodiment the disease or condition is a depression or an anxiety disorder. In a further embodiment the disease or condition is an eating disorder.

Supporting Compounds

The preparation of a number of the compounds of the invention are described below.

In the procedures that follow, after each starting material, reference to an intermediate is typically provided. This is provided merely for assistance to the skilled chemist. The starting material may not necessarily have been prepared from the batch referred to.

Compounds of the invention and intermediates are named using ACD/Name PRO 6.02 chemical naming software (Advanced Chemistry Development Inc., Toronto, Ontario, M5H2L3, Canada).

Where reference is made to the use of a "similar" or "analogous" procedure, as will be appreciated by those skilled in the art, such a procedure may involve minor variation, for example reaction temperature, reagent/solvent amount, reaction time, work-up conditions or chromatographic purification conditions.

All temperatures refer to ⁰C.

Proton Magnetic Resonance (^ H NMR) spectra are typically recorded either on a Varian instrument at 300, 400 or 500 MHz or on a Bruker instrument at 300 and 400 MHz. Chemical shifts are reported in ppm (δ) using the residual solvent line as internal standard. Splitting patterns are designed as s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; b, broad. The NMR spectra were recorded at a temperature ranging from 25 to 9O⁰C. When more than one conformer is detected the chemical shifts for the most abundant one is reported.

Mass spectra (MS) are typically taken on a 4 Il triple quadrupole Mass Spectrometer (Micromass UK) or on a Agilent MSD 1100 Mass Spectrometer, operating in ES (+) and ES (-) ionization mode or on an Agilent LC/MSD 1100 Mass Spectrometer, operating in ES (+) and ES (-) ionization mode coupled with HPLC instrument Agilent 1100 Series [LC/MS - ES (+):analysis performed on a Supelcosil ABZ +Plus (33x4.6 mm, 3μm) (mobile phase: 100% [water +0.1% HCO₂H] for 1 min, then from 100% [water +0.1% HCO₂H] to 5% [water +0.1% HCO₂H] and 95% [CH₃CN ] in 5 min, finally under these conditions for 2 min; T=40°C; flux= 1 mL/min; LC/MS - ES

(-):analysis performed on a Supelcosil ABZ +Plus (33x4.6 mm, 3μm) (mobile phase: 100% [water +0.05% NH₃] for 1 min, then from 100% [water +0.05% NH₃ to 5% [water +0.05% NH₃] and 95% [CH₃CN ] in 5 min, finally under these conditions for 2 min; T=40°C; flux= 1 mL/min] ; in the mass spectra only one peak in the molecular ion cluster is reported.

DAD chromatographic traces, mass chromatograms and mass spectrums may be taken on a on a UPLC/MS AcquityTM system coupled with a Micromass ZQTM mass spectrometer operating in ESI positive or negative. The phases used are: A) H₂O/ACN 95/5 + 0,1 % TFA; B) H₂O/ACN 5/95 + 0,1% TFA. The gradient is: t=0min) 95%A 5%B, t=0,25) 95%A 5%B, t=3,30) 100%B, t=4,0) 100%B, followed by 1 min of reconditioning.

Flash silica gel chromatography was carried out on silica gel 230-400 mesh (supplied by Merck AG Darmstadt, Germany) or over Varian Mega Be-Si pre-packed cartridges or over pre-packed Biotage silica cartridges.

SPE-SCX cartridges are ion exchange solid phase extraction columns supplied by Varian. The eluent used with SPE-SCX cartridges is methanol followed by 2N ammonia solution in methanol.

In a number of preparations, purification was performed using either Biotage manual flash chromatography (Flash+) or automatic flash chromatography (Horizon, SP1 ) systems. All these instruments work with Biotage Silica cartridges.

SPE-Si cartridges are silica solid phase extraction columns supplied by Varian.

A number of the supporting compounds have been prepared as racemic mixtures and a number have been prepared as single enantiomers. The absolute stereochemistry of those compounds prepared as single enantiomers have not been assigned, but may be assigned using ab initio vibrational circular dichroism (VCD).

Chiral molecules exhibit VCD. VCD is the differential interaction of a chiral molecule with left and right circularly polarized infrared radiation during vibrational excitation.

The VCD spectrum of a chiral molecule is dependent on its three-dimensional structure. Most importantly, the VCD spectrum of a chiral molecule is a function of its absolute configuration and, in the case of flexible molecules, of its conformation. In principle, therefore, VCD permits the determination of the structure of a chiral molecule. VCD spectra were first measured in the 1970s. Subsequently, VCD instrumentation has developed enormously in spectral range and in sensitivity. Currently, VCD spectra of liquids and solutions can be measured over the majority of the fundamental infrared (IR) spectral range (v≥ 650 cm-1 ) with high sensitivity at acceptable resolution (1-5 cm-1 ) using both dispersive and Fourier Transform (FT) VCD instrumentation. Very recently, commercial FT VCD instrumentation has become available, greatly enhancing the accessibility of VCD spectra.

The use of VCD as a reliable method for the determination of absolute configuration of chiral molecules is now well established (see for example Shah RD, et a!., Curr

Opin Drug Disc Dev 2001 ;4:764-774; Freedman TB, et al., HeIv Chim Acta 2002;

85:1160-1 165; Dyatkin AB, et al. Chirality 2002;14:215-219; Solladie^'-Cavallo A,

Balaz Met al., Tetrahedron Assym 2001 ;12:2605-2611 ; Nafie LA, et al. Circular dichroism, principles and applications, 2nd ed. New York: John Wiley & Sons; 2000. p 97-131 ; Nafie LA, et al. in: Yan B, Gremlish H-U, editors. Infrared and Raman spectroscopy of biological materials. New York: Marcel Dekker; 2001. p 15-54;

Polavarapu PL, et al., J Anal Chem 2000;366:727-734; Stephens PJ, et al., Chirality

2000;12:172-179; Solladie^' -Cavallo A, et al., Eur J Org Chem 2002: 1788-1796).

The method entails comparison of observed IR and VCD spectra with calculations of the spectra for a specific configuration and provides information both on the absolute configuration and on the solution conformation.

Given an experimental spectrum of a chiral molecule whose absolute configuration and/or conformation are unknown and to be determined, the general procedure is as follows: 1 ) all possible structures are defined; 2) the spectra of these structures are predicted; and 3) predicted spectra are compared to the experimental spectrum. The correct structure will give a spectrum in agreement with experiment; incorrect structures will give spectra in disagreement with experiment.

VCD spectra are always measured simultaneously with vibrational unpolarized absorption spectra ("infrared (IR) spectra") and the two vibrational spectra together provide more information than does the VCD spectrum alone. In addition, vibrational unpolarized absorption spectra are automatically predicted simultaneously with VCD spectra.

For ab initio assignments, VCD and unpolarized IR spectra were calculated using the Gaussian 98 software package.

Abreviations

Intermediate 1 : 1 ,1-dimethylethyl (1S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-formyl-3- azabicyclo[4.1.01heptane-3-carboxylate (single enantiomer)

In a round bottom flask, 1 ,1-dimethylethyl (1S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1- (hydroxymethyl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (500 mg, 1.343 mmol) (Intermediate 18) was dissolved in dry DCM (15 ml.) and stirred under a nitrogen atmosphere at 0 ⁰C. After 10 minutes, Dess-Martin periodinane (684 mg, 1.612 mmol) was added portionwise and the mixture slowly warmed to room temperature and stirred for 1 h. It was then quenched by the addition of NaHCC>3 saturated solution (50 ml) and an aqueous solution of Na2S2U3 δh^O (9.22 g dissolved in 40 ml of water). The two phases were stirred for 30 minutes and then the organic layer was separated, dried on IS^SC^ and concentrated under reduced pressure to give the title compound as a yellow oil (505 mg); ¹H NMR (400 MHz, CHLOROFORM-c/) δ ppm 8.46 - 8.66 (1 H, m), 7.36 - 7.45 (2 H, m), 7.12 (1 H, dd), 4.25 - 4.42 (1 H, m), 3.16 - 3.93 (3 H, m), 2.04 - 2.20 (2 H, m), 2.00 (1 H, d), 1.47 - 1.64 (10 H, m); 314 [MH-56]⁺. Intermediate 2: 1 ,1-Dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-((R or SVI-hvdroxy^-propen-i-vD-S-azabicvcW.I .OIheptane-S-carboxylate (single enantiomer)

1 ,1-Dimethylethyl (1S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-formyl-3- azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (505 mg, 1.364 mmol) was dissolved in dry THF (20 ml) and cooled to -20 ⁰C under a nitrogen atmosphere. Vinylmagnesium bromide (4092 μl, 4.09 mmol) was added dropwise whilst keeping the internal temperature below -20 ⁰C. The mixture was stirred at this temperature for 1 h. Additional vinylmagnesium bromide (4092 μl, 4.09 mmol) was required to drive the reaction to completion. The solution was cooled to -78⁰C and then slowly warmed to rt and stirred overnight. The reaction mixture was quenched with NH4CI saturated solution (20 ml), the organic phase was extracted with EtOAc (3X25 ml), dried on Na2SC>4 and concentrated under vacuum. The residue was purified by chromatography on silica gel eluting with a gradient from 10% to 50% ethyl acetate in cyclohexane. One diastereomer was isolated (353 mg); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.48 - 7.56 (1 H, m), 7.34 - 7.43 (1 H, m), 7.27 (3 H, s), 5.84 - 5.98 (1 H, m), 5.11 - 5.35 (2 H, m), 3.85 - 4.01 (1 H, m), 3.43 - 3.54 (1 H, m), 3.06 - 3.42 (2 H, m), 1.85 - 2.34 (2 H, m), 1.50 (9 H, s), 0.95 - 1.10 (2 H, m); LCMS: Rt 0.95 min (Acquity™ UPLC BEH C18 column (50 x 21 mm, 1.7 μm particle size), column temperature 40 ⁰C. (LC/MS - ES (+): mobile phase: A-water + 0.1% HCOOH / B - CH₃CN + 0.06% HCOOH, Flow rate: 1.0 mL/min, Gradient: t = 0 min 3% B, t = 0.05 min 6% B, t = 0.57 min 70% B, t = 1.449 min 99% B, t = 1.45 min 3% B). Intermediate 3: 1 ,1-dimethylethyl (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-(1- hydroxyethyl)-3-azabicvclo[4.1.OIheptane-3-carboxylate (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-formyl- 3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (0.100 g, 0.270 mmol) in dry THF (3.5 ml.) at -2O⁰C was added bromo(methyl)magnesium (3.2M solution in 2- methyltetrahydrofuran; 0.169 ml_; 0.540 mmol) and the reaction mixture was stirred at -2O⁰C for 30 minutes and then at room temperature overnight. The reaction mixture was quenched with NH4CI sat. solution (4 ml), extracted with ethyl acetate (4 ml) and concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 8/2) to obtain one diastereomer of the title compound as colourless oil (0.050 g); ¹H NMR (500 MHz, DMSO-c/₆) δ ppm 7.62 (d, 1 H) 7.53 (d, 1 H) 7.35 (dd, 1 H) 4.38 (d, 1 H) 3.52 - 3.90 (m, 2 H) 3.09 - 3.32 (m, 2 H) 2.63 - 2.71 (m, 1 H) 1.76 - 2.05 (m, 2 H) 1.41 (s, 9 H)1.03 (d, 3 H) 0.89 (d, 1 H)0.73 - 0.84 (m, 1 H); MS (m/z) = 386 [MH].

Intermediate 4 and Intermediate 5: 1 ,1-dimethylethyl (1 R.6S or 1S,6R)-6-(3,4- dichlorophenyl)-1-((R or S)-1-hvdroxypropyl)-3-azabicvclor4.1.Olheptane-3- carboxylate (each a single diastereoisomer and a single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-formyl- 3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (0.285 g, 0.770 mmol) in dry THF (8 ml.) at -2O⁰C was added bromo(ethyl)magnesium (1.539 mL, 1.539 mmol) and the reaction mixture was stirred at -2O⁰C for 30 minutes and then at RT overnight. The reaction mixture was quenched with NH4CI sat. solution (4 ml), extracted with ethyl acetate (4 ml) and concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 8/2) to give two diastereomers each as a single enantiomer:

Intermediate 4 (first eluting compound) as a yellow oil (0.084 g); ¹H NMR (400 MHz, DMSOd₆) δ ppm 7.64 (d, 1 H) 7.52 (d, 1 H) 7.36 (dd, 1 H) 4.34 (d, 1 H) 3.48 - 3.84 (m, 1 H 3.09 - 3.45 (m, 3 H)) 2.24 - 2.44 (m, 1 H) 1.89 - 2.07 (m, 1 H) 1.74 - 1.91 (m, 1 H) 1.28 - 1.53 (m, 2 H) 1.41 (s, 9 H) 0.98 (d, 1 H) 0.80 (d, 1 H) 0.77 (t, 3 H); MS (m/z) = 400 [MH]; and

Intermediate 5 (second eluting compound) as yellow oil (0.019 g); ¹H NMR (400 MHz, DMSOd₆) δ ppm 7.47 - 7.63 (m, 2 H) 7.29 (dd, 1 H) 4.39 - 4.58 (m, 1 H) 3.05 - 3.95 (m, 4 H 2.09 - 2.30 (m, 2 H)) 1.71 - 1.90 (m, 1 H) 1.41 (s, 9 H) 1.19 - 1.40 (m, 3 H)0.78 - 0.88 (m, 1 H) 0.54 (t, 3 H); MS (m/z) = 400 [MH].

Intermediate 6 and Intermediate 7: 1.1-Dimethylethyl (1 R.6S or 1 S.6R)-6-(3.4- dichlorophenyl)-1-(R or S)-1-hvdroxybutyl)-3-azabicvclo[4.1.OIheptane-3-carboxylate (each a single diastereoisomer and a single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-formyl- 3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (0.350 g, 0.945 mmol) in dry THF (8 ml.) at -2O⁰C was added bromo(propyl)magnesium (0.945 mL, 1.890 mmol) and the reaction mixture was stirred at -2O⁰C for 30 minutes and then at RT overnight. The temperature was decreased to -2O⁰C and further bromo(propyl) magnesium (0.47 mmol, 0.236 ml) was added and the reaction mixture was stirred at -2O⁰C for 30 minutes and at room temperature for 3h. The reaction mixture was quenched with NH4CI sat. solution (4 ml), extracted with ethyl acetate (4 ml) and concentrated under reduced pressure. The residue was purified by chromatography eluting with Cy/EtOAc 9/1 10 CV, to 8/2 4 CV, 8/2 1 1 CV) to give two diastereomers each as a single enantiomer:

Intermediate 6 (first eluting compound) as a white foam (0.208 g); ¹H NMR (500 MHz, CHLOROFORM-d) δ ppm 7.47 (br. s., 1 H) 7.38 (d, 1 H) 7.23 - 7.27 (m, 1 H) 3.14 - 4.03 (m, 4 H) 2.64 (br. s., 1 H) 2.09 (br. s., 1 H) 1.94 (br. s., 1 H) 1.50 (s, 9 1-1)1.13 - 1.44 (m, 4 H) 0.78 - 1.01 (m, 5 H); MS (m/z) = 414 [MH]; and Intermediate 7 (second eluting compound) as a colourless oil (0.064 g); ¹H NMR (500 MHz, CHLOROFORM-d) δ ppm 7.35 - 7.40 (m, 2 H) 7.12 (dd, 1 H) 3.09 - 4.03 (m, 4 H) 2.61 - 2.79 (m, 1 H) 2.13 - 2.23 (m, 1 H) 1.87 - 2.03 (m, 1 H) 1.64 - 1.76 (m, 1 H) 1.48 (s, 9 H) 0.82 - 1.43 (m, 5 H) 0.69 (t, 3 H); MS (m/z) = 414 [MH].

Intermediate 8 and Intermediate 9: 1.1-dimethylethyl (1 R.6S or 1S.6RV6-(3.4- dichlorophenvD-i-d-hvdroxy-S-methylbutvD-S-azabicvcloK.I .OIheptane-S- carboxylate (each a single diastereoisomer and a single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-formyl- 3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (0.35 g, 0.945 mmol) in dry THF (8 mL) at -2O⁰C was added bromo(2-methylpropyl)magnesium (0.473 ml_, 0.945 mmol) and the reaction mixture was stirred at -2O⁰C for 30 minutes and then at room temperature over night. Further bromo(2-methylpropyl)magnesium (0.473 mL, 0.945 mmol) was added and the reaction stirred at room temperature for 1.5 hours. Still further bromo(2-methylpropyl)magnesium (0.473 mL, 0.945 mmol) was added and the reaction stirred at room temperature for 3h. The reaction was quenched with NH4CI saturated solution (4 ml), extracted with ethyl acetate (4 ml) and concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 85/15 15 CV, to 70/30 0.5 CV, 70/30 5 CV) to give two diastereomers:

Intermediate 8 (first eluting compound) as a colourless oil (0.102 g); ¹H NMR (500 MHz, CHLOROFORM-d) δ ppm 7.48 (s, 1 H) 7.39 (d, 1 H) 7.24 (br. s., 1 H) 3.47 - 4.07 (m, 2 H) 3.16 - 3.48 (m, 2 H) 1.83 - 2.24 (m, 2 H) 2.60 - 2.84 (m, 1 H) 1.53 -

1.69 (m, 1 H) 1.43 - 1.58 (m, 1 H) 1.50 (s, 9 H) 1.21 - 1.34 (m, 1 H) 0.82 - 0.98 (m, 5 H) 0.74 (br. s., 3 H); MS (m/z) = 428 [MH]; and

Intermediate 9 (second eluting compound) which was further purified by Liquid Chromatography (0.0067 g of product was obtained); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.34 - 7.41 (m, 2 H) 7.12 (dd, 1 H) 3.24 - 4.04 (m, 4 H)

2.70 - 2.84 (m, 1 H) 2.14 - 2.26 (m, 1 H) 1.89 - 2.03 (m, 1 H) 1.44 - 1.64 (m, 1 1 H) 1.25 - 1.37 (m, 1 H)1.23 (d, 1 H) 1.10 - 1.21 (m, 1 H) 1.04 (d, 1 H) 0.85 (d, 2 H) 0.39 (d, 3 H); MS (m/z) = 428 [MH]. Intermediate 10 and Intermediate 1 1 : 1 ,1-dimethylethyl (1S,6R or 1 R,6S)-6-(3,4- dichlorophenyl)-1-(1-hydroxy-3-buten-1-yl)-3-azabicvclo[4.1.01heptane-3-carboxylate (each a single diasteroisomer and a single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-formyl- 3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (390 mg, 1.053 mmol) in THF (15 ml.) at 20 ⁰C was added bromo(2-propen-1-yl)magnesium (6.32 mL, 6.32 mmol). After 1.5 hrs NH4CI (sat sol)/water (1/1 ) was added, the mixture was stirred for 30 min and then extracted twice with DCM. The organic phase was dried and the solvent removed under reduced pressure. The residue was purified by chromatography (c-Hex/EtOAc 0 to 30%; 100 g silica column) to give two diastereomers:

Intermediate 10 (first eluting compound) (210 mg) as colorless oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.5 (s, 1 H), 7.4 (d, 1 H), 7.3 (d, 1 H), 5.65 (m, 1 H), 5. 1 (m, 2H), 3.95 (m, 1 H), 3.65 (bs, 1 H), 3.4 (bs, 1 H), 3.3 (m, 1 H), 2.65 (bs, 1 H), 2.3 (bs, 2H), 2.05 (bs, 1 H), 1.95 (bs, 1 H), 1.6 (m, 1 H), 1.5 (s, 9H), 0.95 (s, 2H); MS (m/z) = 412 [MH⁺]; and

Intermediate 11 (second eluting compound) (50 mg) as colorless oil: ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.4 (d, 2 H), 7.1 (d, 1 H), 5.5 (m, 1 H), 5.0 (m, 2H), 3.8 (m, 2H), 3.4 (bs, 1 H), 3.25 (m, 1 H), 2.8 (bs, 1 H), 2.25 (m, 2H), 2.20 (m, 1 H), 1.95 (bs, 1 H), 1.6 (bs, 1 H), 1.5 (s, 9H), 1.3 (d, 1 H), 1.1 (d, 2H); MS (m/z) = 412 [MH⁺].

Intermediate 2 and Intermediate 12: 1 ,1-dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4- dichlorophenvO-i-π-hvdroxy^-propen-i-ylKS-azabicvcW.I .OIheptane-S- carboxylate (each a single diastereoisomer and a single enantiomer)

1 ,1-Dimethylethyl (1S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-formyl-3- azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 1 ) (505 mg, 1.364 mmol) was dissolved in dry THF (20 ml) and cooled to -20 ⁰C under a nitrogen atmosphere. Vinyl magnesium bromide (4092 μl, 4.09 mmol) was added dropwise keeping the internal temperature below -20 ⁰C. The mixture was stirred at this temperature for 1 h. Additional vinyl magnesium bromide (4092 μl, 4.09 mmol) was required to drive the reaction to completion. The solution was cooled to -78⁰C and then slowly warmed to room temperature and stirred overnight. The reaction mixture was quenched with NH4CI saturated solution (20 ml), extracted with ethyl acetate (3 x 25 ml), dried over Na2SC>4 and concentrated under reduced pressure. The residue was purified by chromatography on silica gel eluting with a gradient from 10% to 50% ethyl acetate in cyclohexane to give two diastereomers: Intermediate 2 (first eluting compound) (353 mg) as white foam, ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.48 - 7.56 (1 H, m), 7.34 - 7.43 (1 H, m), 7.27 (3 H, s), 5.84 - 5.98 (1 H, m), 5.11 - 5.35 (2 H, m), 3.85 - 4.01 (1 H, m), 3.43 - 3.54 (1 H, m), 3.06 - 3.42 (2 H, m), 1.85 - 2.34 (2 H, m), 1.50 (9 H, s), 0.95 - 1.10 (2 H, m); MS (m/s) = 397 [MH⁺] LCMS: Rt 0.95 min (Acquity™ UPLC BEH C18 column (50 x 21 mm, 1.7 μm particle size), column temperature 40 ⁰C. (LC/MS - ES (+): mobile phase: A-water + 0.1% HCOOH / B - CH₃CN + 0.06% HCOOH, Flow rate: 1.0 mL/min, Gradient: t = 0 min 3% B, t = 0.05 min 6% B, t = 0.57 min 70% B, t = 1.449 min 99% B, t = 1.45 min 3% B); and

Intermediate 12 (second eluting compound) (240 mg) as white foam; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.4 2- 7.39 (m, 2 H) 7.25 (m, 1 H) 5.85 (m, 1 H) 5.05 (d, 1 H) - 5.15 (m, 1 H) 3.93-3.1 (br. m., 6 H) 2.32 (m, 1 H) 2.04 - 2.14 (m, 2 H) 1.96 (br. s., 1 H) 1.65 (m, 2 H), 1.6 (s, 9 H) 1.35 -0.8 (m, 4 H); MS (m/s) = 397 [MH⁺].

Intermediate 13: 1 ,1-dimethylethyl 6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)-3- buten-1-yl1-3-azabicyclo[4.1.OIheptane-3-carboxylate (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-(1- hydroxy-3-buten-1-yl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 10) (0.058 g, 0.141 mmol) in dry DMF (4 ml.) at 0 ⁰C was added NaH (0.00675 g, 0.169 mmol) and the clear reaction mixture was stirred at RT for 30 min. Methyl iodide

(10.55 μl_, 0.169 mmol) was added and the mixture was stirred at RT. After 1 h30\ 1 eq more of NaH (0.141 mmol, 0.0056 g) and methyl iodide (0.141 mmol, 8.8 μl) were added and the reaction was stirred overnight. A further 2 eq of NaH and methyl iodide were added to the solution and the reaction mixture was left at RT for 2.5 hours. The reaction was quenched with NH4CI sat. solution and diluted with diethyl ether. The organic layer was then dried and concentrated under reduced pressure. The residue was purified by chromatography (10 g silica, eluting mixture Cy/EtOAc from 0% EtOAc to 30%) to obtain the title compound (0.0391 g) as colourless oil; NMR(¹H, CDCI3): δ 7.5 (s, 1 H), 7.38 (d,1 H), 7.22 (m,1 H), 5.76 (m, 1 H), 5.09 (d, 1 H), 5.01 (d, 1 H), 3.93 (d, 1 H), 3.69 (m, 1 H), 3.40 (m, 1 H), 3.29 (m, 1 H), 2.97 (s, 3H), 2.46 (m, 1 H), 2.33 (m, 1 H), 2.23 (m, 1 H), 2.06 (m, 1 H), 1.97 (sb, 1 H), 1.50 (s, 9H), 0.95 (sb, 1 H), 0.89 (d, 1 H); MS (m/z) = 426 [MH]. Intermediate 14: 1 ,1-dimethylethyl (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[3- methyl-1-((R or S)-methyloxy)butyl1-3-azabicvclo[4.1.01heptane-3-carboxylate (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-(1- hydroxy-3-methylbutyl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 8) (0.100 g, 0.233 mmol) in dry THF (8 ml.) was added NaH (0.028 g, 0.700 mmol) and the reaction mixture was stirred for 30 min at RT. Then methyl iodide (0.044 ml_, 0.700 mmol) was added and the yellow reaction suspension was stirred at RT for 2h. Further NaH (0.028 g, 0.700 mmol) and methyl iodide (0.31 mmol, 0.058 ml) were added and the reaction mixture was stirred at RT over night. Still further NaH (0.028 g, 0.700 mmol) and methyl iodide (0.31 mmol, 0.058 ml) were added and the reaction was stirred at 4O⁰C for 6h. It was quenched with NH4CI sat. solution (4 ml) and extracted with ethyl acetate (4 ml x3). The organic phase was concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 9/1 , 15 CV) to obtain the title compound (0.0758 g) as a light yellow oil; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.51 (d, 1 H) 7.39 (d, 1 H) 7.19 - 7.24 (m, 1 H) 3.17 - 4.06 (m, 4 H) 2.90 (s, 3 H) 2.29 (t, 1 H) 1.90 - 2.21 (m, 2 H) 1.63 - 1.77 (m, 1 H) 1.37 - 1.62 (m, 1 1 H) 0.75 - 1.06 (m, 8 H). MS (m/z) = 442 [MH]

Intermediate 15: 1.1-dimethylethyl (1 S.6R or 1 R.6S)-6-(3.4-dichlorophenvn-1-ri-(2- propen-1-yloxy)-2-propen-1-yll-3-azabicvclor4.1.0lheptane-3-carboxylate (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-(1- hydroxy-2-propen-1-yl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 2) (90 mg, 0.226 mmol) in DMF (2 ml.) at 20 ⁰C was added sodium hydride (13.56 mg, 0.339 mmol). After 30 min, allyl iodide (0.052 mL, 0.565 mmol) was added. After 4 hr further sodium hydride (13.56 mg, 0.339 mmol) was added, followed, after 30 min, by further allyl iodide (0.052 mL, 0.565 mmol). After 2 days DCM was added and the mixture washed with NaHCC>3 (sat sol)/Na2S2θ3 until mixture was colourless. The organic phase was dried and the solvent was removed under reduced pressure to give a residue which was purified via chromatography (c-Hex/EtOAc 0 to 50%; 25 g SiO2 column) to give title compound (48 mg) as a colorless oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.57 (d, 1 H) 7.37 (d, 1 H) 7.24 (dd, 1 H), 5.79 (m, 2 H) 5.16 (m, 4 H) 3.8 (m, 3 H) 3.26 (m, 3 H) 2.68 (d, 1 H) 2.10 (d, 2 H) 1.48 (s, 9 H) 0.90 (s, 2 H); MS (m/z) = 437 [MH⁺].

Intermediate 16: 1 ,1-dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(2/? or 2S)-2,5-dihydro-2-furanyl1-3-azabicvclo[4.1.01heptane-3-carboxylate (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 S,6R)-6-(3,4-dichlorophenyl)-1-[(1 R)-1-(2- propen-i-yloxy^-propen-i-yO-S-azabicyclo^.i .Olheptane-S-carboxylate (Intermediate 15) (48 mg, 0.109 mmol) in DCM (2 ml_) at 20 ⁰C were added [1 ,3- bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](dichloro)-(phenylmethylidene)- ruthenium/tricyclohexylphosphane (1 :1 ) (4.65 mg, 5.47 μmol) and 2,5- cyclohexadiene-1 ,4-dione (1.184 mg, 10.95 μmol). After 2 days NaHCC>3 (sat sol) was added and the mixture extracted with DCM. The solvent was removed under reduced pressure from organic phase. The residue was purified via chromatography (c-Hex/EtAc 0 to 30%; Silica column) to give the title compound (35 mg) as a colorless oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.44 - 7.54 (m, 1 H) 7.39 (d, J=8.31 Hz, 1 H) 7.27 (d, 1 H) 5.84 - 6.03 (m, 1 H) 5.59 - 5.73 (m, 1 H) 4.65 - 4.76 (m, 1 H) 4.49 - 4.62 (m, 1 H) 3.86 - 4.05 (m, 1 H) 3.09 - 3.84 (m, 4 H) 1.81 - 2.18 (m, 2 H) 1.47 (s, 9 H) 0.96 (s, 2 H); MS (m/z) = 409 [MH⁺].

Intermediate 17: 1.1-dimethylethyl (1 S.6R or 1 R.6S)-6-(3.4-dichlorophenvn-1-r(2/? or 2S)-tetrahydro-2-furanyll-3-azabicvclo[4.1.Olheptane-3-carboxylate (single enantiomer)

1 ,1-Dimethylethyl 1 ,1-dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(2R or 2S)-2,5-dihydro-2-furanyl]-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 16) (35 mg, 0.085 mmol) and platinum(IV) oxide (0.968 mg, 4.26 μmol) in ethanol (3 ml.) was hydrogenated at 20 ⁰C at 1 atmosphere pressure. After 5 hrs, the catalyst was filtered-off and washed with ethanol and the solvent was removed under reduced pressure. The reside was purified via chromatography (c-Hex/EtOAc 0 to 25%; Silica column)) to give the title compound as a colorless oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.47 (s, 1 H) 7.38 (d, 1 H) 7.27 (s, 1 H), 3.85 (m, 1 H) 3.63 (m, 2 H) 2.96 (m, 1 H) 2.04 (m, 1 H) 1.96 (m, 1 H) 1.87 (m, 1 H) 1.76 (m, 2 H) 1.50 (s, 9 H) 0.94 (d, 1 H) 0.83 (d, 1 H). MS (m/z) = 411 [MH⁺]

Intermediate 18 and Intermediate 19: 1.1-dimethylethyl (1S.6R or 1 R.6SV6-(3.4- dichlorophenyl)-1-(hvdroxymethyl)-3-azabicvclor4.1.0lheptane-3-carboxylate (single diastereoisomers, single enantiomers)

Racemate 1 ,1-dimethylethyl (1S,6R/ 1 R,6S)-6-(3,4-dichlorophenyl)-1- (hydroxymethyO-S-azabicyclo^.i .Olheptane-S-carboxylate (3.2 g) (for preparation see international patent publication WO2008/031772, Preparation 5) were separated by Chiral preparative liquid chromatography. Chromatography conditions: Column: Chiralpak AD-H (25 x 2 cm); Mobil phase: n-Hexane/lsopropanol 90/10% v/v; UV detection: 215 nm; Flow Rate (mL/min): 17; Injection: 100 mg.

Intermediate 18 was obtained as the first eluting compound as white foam (1.51 g); ¹H NMR (400 MHz, DMSOd₆) δ ppm 7.43 (br. s, 1 H) 7.39 (d, 1 H) 7.17 (d, 1 H) 3.75 - 3.90 (q, 2 H) 3.30 - 3.45 (m, 3 H) 3.05 - 3.25 (m, 1 H) 2.10(m, 1 H) 2.00 (br. s, 1 H) 1.50 (s, 9 H) 1.05 (d, 1 H) 0.97 (br. s, 1 H); MS (m/z) = 372 [MH];

Intermediate 19 was obtained as the second eluting compound as white foam (1.65 g); ¹H NMR (400 MHz, DMSOd₆) δ ppm 7.43 (br. s, 1 H) 7.39 (d, 1 H) 7.16 (d, 1 H) 3.75 - 3.90 (q, 2 H) 3.30 - 3.45 (m, 3 H) 3.05 - 3.25 (m, 1 H) 2.07 (m, 1 H) 2.00 (br. s, 1 H) 1.53 (s, 9 H) 1.05 (d, 1 H) 0.97 (br. s, 1 H); MS (m/z) = 372 [MH].

Compound 1 : (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)1- (methyloxy)ethyl1-3-azabicvclo[4.1.Oiheptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-((R or S^I-hydroxyethyO-S-azabicyclo^.i .OJheptane-S-carboxylate (Intermediate 3) (0.048 g, 0.124 mmol) in dry THF (3 ml.) was added NaH (0.015 g, 0.373 mmol) and the reaction mixture was stirred for 30 min at RT. Methyl iodide (0.023 ml_, 0.373 mmol) was added and the yellow solution was stirred at RT overnight. Further NaH (0.015 g, 0.373 mmol) and methyl iodide (0.5 mmol, 30.6 μl_) were added and the reaction mixture was stirred at 4O⁰C for 1 h. The reaction was quenched with NH4CI sat. solution (4 ml), extracted with ethyl acetate (4 ml) and concentrated under reduced pressure. The residue was purified by chromatography (eluting with Cy/EtOAc from 10/0 to 8/2) to give 1 ,1-dimethylethyl (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)ethyl]-3-azabicyclo[4.1.0]heptane-3-carboxylate as colourless oil (0.020 g); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.51 (s, 1 H) 7.38 (d, 1 H) 7.24 (dd, 1 H) 3.15 - 4.09 (m, 4 H) 2.96 (s, 3 H) 2.25 - 2.35 (m, 1 H) 1.89 - 2.14 (m, 2 H) 1.49 (S, 9 H) 1.16 (d, 3 H) 0.75 - 0.96 (m, 2 H). MS (m/z) = 400 [MH]. The colourless oil was dissolved in DCM (0.5 ml.) and the temperature was decreased to O⁰C. TFA (0.024 ml_, 0.31 1 mmol) was added and the reaction mixture was stirred for 3h, allowing the temperature to reach RT. NaHCC>3 sat. solution (2 ml) was added. The reaction mixture was diluted with DCM, the organic phase was separated and concentrated under reduced pressure to give the tile compound as a yellow oil (0.008 g); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.55 (d, 1 H) 7.37 (d, 1 H) 7.25 - 7.31 (m, 1 H) 3.48 (d, 1 H) 2.98 (s, 3 H) 2.93 (d, 1 H) 2.79 - 2.89 (m, 1 H) 2.52 - 2.62 (m, 1 H) 2.20 - 2.27 (m, 1 H) 1.83 - 1.99 (m, 2 H) 1.09 (d, 3 H) 0.97 (d, 1 H) 0.83 (d, 1 H); MS (m/z) = 300 [MH].

Compound 2: (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1- (methyloxy)propyl1-3-azabicyclo[4.1.Oiheptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-((R or S)-1-hydroxypropyl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 4) (0.050 g, 0.125 mmol) in dry THF (4 ml.) was added NaH (0.015 g, 0.375 mmol) and the reaction mixture was stirred for 30 min at RT. Then methyl iodide (0.023 ml_, 0.375 mmol) was added and the yellow suspension was stirred at RT for 3h. Further NaH (0.015 g, 0.375 mmol) and methyl iodide (0.5 mmol, 30.6 μl) were added and the reaction was stirred at RT overnight. Still further NaH (0.015 g, 0.375 mmol) and methyl iodide (0.5 mmol, 30.6 μl) were added and the reaction mixture was stirred at RT for 1 h. NH4CI saturated solution (4 ml) was added and the mixture was extracted with ethyl acetate (4 x 4 ml). The organic phase was concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc from 10/0 to 8/2 10 CV) to give 1 ,1-dimethylethyl (1 R.6S or 1 S,6R)-6-(3,4- dichlorophenyl)-1-[(R or S)-1-(methyloxy)propyl]-3-azabicyclo[4.1.0]heptane-3- carboxylate as a colourless oil (0.0297 g); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.51 (d, 1 H) 7.38 (d, 1 H) 7.23 (dd, 1 H) 3.17 - 4.07 (m, 4 H) 2.93 (s, 3 H) 2.04 - 2.18 (m, 2 H) 1.89 - 2.04 (m, 1 H) 1.68 - 1.82 (m, 1 H) 1.56 (s, 9 H) 0.83 - 1.05 (m, 6 H); MS (m/z) = 414 [MH]. The colourless oil was dissolved in DCM (0.5 ml_), TFA (0.048 ml_, 0.624 mmol) was added and the reaction mixture was stirred at RT for 3h. NaHCC>3 sat. sol. (2 ml) was added and the mixture was diluted with DCM. The organic phase was separated and concentrated under reduced pressure to give the title compound as a yellow oil (0.023 g); ¹H NMR (500 MHz, CHLOROFORM-d) δ ppm 7.46 (s, 1 H) 7.29 (d, 1 H) 7.15 - 7.18 (m, 1 H) 3.36 (d, 1 H) 2.86 (s, 3 H) 2.81 - 2.85 (m, 1 H) 2.71 - 2.80 (m, 1 H) 2.48 - 2.57 (m, 1 H) 1.91 - 1.99 (m, 1 H) 1.82 - 1.89 (m, 2 H) 1.58 - 1.65 (m, 1 H) 1.35 - 1.47 (m, 1 H) 0.94 (s, 2 H) 0.78 (t, 3 H); MS (m/z) = 314 [MH].

Compound 3: (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1- (methyloxy)propyl1-3-azabicyclo[4.1.Oiheptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-((R or S)-1-hydroxypropyl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 5) (0.0177 g, 0.044 mmol) in dry THF (1.5 ml.) was added NaH (0.00530 g, 0.133 mmol) and the reaction mixture was stirred for 30 min at RT. Methyl iodide (8.29 μl_, 0.133 mmol) was added and the suspension was stirred at RT for 3.5 hours. Further NaH (0.00530 g, 0.133 mmol) and methyl iodide (0.176 mmol, 1 1 μl) were added and the reaction was stirred at 4O⁰C for 3h and then at RT over night. The temperature was brought to 4O⁰C and the reaction stirred for 3h. Still further NaH (0.00530 g, 0.133 mmol) and methyl iodide (0.176 mmol, 1 1 ul) were added and the reaction was stirred at 4O⁰C for 3h and then at 6O⁰C for 2h. Still further NaH (0.00530 g, 0.133 mmol) and methyl iodide (0.176 mmol, 11 μl) were added and the reaction mixture was stirred at RT over night. Still further NaH (0.0106 g, 0.266 mmol) and methyl iodide (0.352 mmol, 22 μl) were added and the reaction stirred at RT for 5.5 hours. Still further NaH (0.0106 g, 0.266 mmol) and methyl iodide (0.352 mmol, 22 μl) were added and the reaction stirred at RT for 2h. NH4CI sat. solution (4 ml) was added and the mixture was extracted with ethyl acetate (4 x 4 ml). The organic phase was concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc from 10/0 to 8/2 10 CV) to given (1 R,6S or 1S,6R)-1 ,1- dimethylethyl 6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)propyl]-3- azabicyclo[4.1.0]heptane-3-carboxylate as a colourless oil (0.0107) g; MS (m/z) = 414 [MH]. The colourless oil was dissolved in DCM (0.5 ml_), TFA (0.020 ml_, 0.265 mmol was added and the reaction mixture was stirred at RT for 4h. Toluene (1 ml.) was added and the reaction mixture was concentrated. The residue was dissolved in methanol and loaded on to 500mg SCX cartridge and eluted with MeOH and 2M ammonia sol. in methanol. The ammonia fraction was concentrated under vacuum to give the title compound (0.0059 g) as a colourless oil; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.39 (d, 1 H) 7.36 (d, 1 H) 7.16 (dd, 1 H) 3.27 (s, 3 H) 3.20 (d, 1 H) 3.00 (d, 1 H) 2.69 - 2.77 (m, 1 H) 2.11 - 2.17 (m, 1 H) 1.46 - 1.52 (m, 4 H) 1.43 (d, 1 H) 1.92 - 1.99 (m, 1 H) 1.15 (d, 1 H) 0.64 (t, 3 H); MS (m/z) = 314 [MH].

Compound 4: (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1- (methyloxy)butyl1-3-azabicvclo[4.1.Oiheptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-((R or S)-1-hydroxybutyl)-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 6) (0.100 g, 0.241 mmol) in dry THF (8 ml.) was added NaH (0.029 g, 0.724 mmol) and the reaction mixture was stirred for 30 min at RT. Methyl iodide (0.045 ml_, 0.724 mmol) was added and the yellow suspension was stirred at RT for 4h. Further NaH (0.029 g, 0.724 mmol) and methyl iodide (0.06 ml, 0.989 mmol) were added and the reaction mixture was stirred at RT overnight. Still further NaH (0.029 g, 0.724 mmol) and methyl iodide (0.06 ml, 0.989 mmol) were added and the reaction mixture was stirred at RT for 2h. The temperature was brought to 4O⁰C and the reaction mixture was stirred for 2h. NH4CI sat. solution (4 ml) was added and the mixture was extracted with ethyl acetate (3 x 4 ml). The organic phase was concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 9/1 , 10 CV) to give 1 ,1 -dimethylethyl (1 R,6S or 1S,6R)-6-(3,4- dichlorophenyl)-1-[(R or S)-1-(methyloxy)butyl]-3-azabicyclo[4.1.0]heptane-3- carboxylate (0.070 g) as a colourless oil; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.82 - 1.03 (m, 5 H) 1.15 - 1.31 (m, 1 H) 1.32 - 1.57 (m, 1 1 H) 1.59 - 1.72 (m, 1 H) 1.89 - 2.23 (m, 3 H) 2.92 (s, 3 H) 3.16 - 4.07 (m, 4 H) 7.22 (dd, J=8.34, 1.77 Hz, 1 H) 7.38 (d, J=8.08 Hz, 1 H) 7.51 (d, J=1.52 Hz, 1 H); MS (m/z) = 428 [MH]. The colourless oil was dissolved in DCM (0.5 ml_), TFA (0.121 ml_, 1.569 mmol) was added and the reaction was stirred at RT for 3h. NaHCC>3 sat. solution (2 ml) was added and the mixture was diluted with DCM. The organic phase was separated and concentrated to give the title compound as colourless oil (0.0607 g); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 0.88 (t, J=7.33 Hz, 3 H) 1.10 - 1.18 (m, 2 H) 1.18 - 1.49 (m, 4 H) 1.55 - 1.66 (m, 1 H) 2.04 - 2.16 (m, 2 H) 2.23 - 2.33 (m, 1 H) 2.74 - 2.83 (m, 1 H) 2.98 (s, 3 H) 3.02 - 3.15 (m, 2 H) 3.71 (d, 1 H) 7.32 - 7.38 (m, 1 H) 7.42 (d, 1 H) 7.52 - 7.59 (m, 1 H); MS (m/z) = 328 [MH].

Compound 5: (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1- (methyloxy)butyl1-3-azabicvclo[4.1.Oiheptane (single enantiomer)

To a solution of 1 ,1 -dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-((R or

S^I-hydroxybutyO-S-azabicyclo^.i .OJheptane-S-carboxylate (Intermediate 7) (0.062 g, 0.150 mmol) in dry THF (4 ml.) was added NaH (0.018 g, 0.449 mmol) and the reaction mixture was stirred for 30 min at RT. Methyl iodide (0.028 ml_, 0.449 mmol) was added and the yellow suspension was stirred at RT for 4h. Further NaH (0.018 g, 0.449 mmol) and methyl iodide (0.037 ml, 0.598 mmol) were added and the reaction mixture was stirred at RT overnight. Still further NaH (0.018 g, 0.449 mmol) and methyl iodide (0.037 ml, 0.598 mmol) were added and the reaction was stirred at RT for 2h. The temperature was brought to 4O⁰C and the reaction was stirred for 3h. Still further NaH (0.018 g, 0.449 mmol) and methyl iodide (0.037 ml, 0.598 mmol) were added at RT and the reaction mixture was stirred at 6O⁰C for 3.5 hours and then at RT overnight. Still further NaH (0.018 g, 0.449 mmol) and of methyl iodide (0.037 ml, 0.598 mmol) were added and the reaction mixture was stirred at 6O⁰C for 3h. NH4CI sat. solution (4 ml) was added and the mixture was extracted with ethyl acetate (3 x 4 ml). The organic phase was concentrated under reduced pressure. The residue was purified by chromatography (eluting mixture Cy/EtOAc 9/1 , 10 CV) to give 1 ,1-dimethylethyl (1 R,6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1- (methyloxy)butyl]-3-azabicyclo[4.1.0]heptane-3-carboxylate as colourless oil (0.0165 g); ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.35 - 7.41 (m, 2 H) 7.15 (d, 1 H) 3.21 - 3.99 (m, 7 H) 2.14 - 2.36 (m, 2 H) 1.89 - 2.07 (m, 1 H) 1.41 - 1.63 (m, 1 1 H) 1.20 - 1.38 (m, 2 H) 1.08 - 1.17 (m, 1 H) 0.79 - 0.94 (m, 1 H) 0.64 (t, 3 H); MS (m/z) = 428 [MH]; The colourless oil was dissolved in DCM (0.5 ml_), TFA (0.031 ml_, 0.404 mmol) was added and the reaction was stirred at RT for 4h. NaHCC>3 sat. solution (2 ml) was added and the reaction mixture was diluted with DCM. The organic phase was separated and concentrated under reduced pressure. The residue was purified by Preparative Liquid Chromatography to obtain the title compound as the TFA salt.

Chromatography conditions:

Column : Phenomenex Luna AXIA C18 (100x21.2mm, 5.0um)

Mobil phase "A": 0.1%v/v solution of TFA in Water

Mobil phase "B": MeCN

UV detection: Diode-array 210-350nm MS detection: Electrospray, +ve, 100-900amu, centroid mode

Preparative gradient A%/B%: from 70/30 to 0/100

Flow Rate(mL/min): 17

To the TFA salt was added NaHCC>3 (1 ml) and the mixture was extracted with DCM (2 ml x 2). The organic phase was concentrated under reduced pressure to obtain the title compound (0.004 g) as a colourless oil; ¹H NMR (500 MHz, CHLOROFORM- d) δ ppm 7.41 (d, 1 H) 7.36 (d, 1 H) 7.16 (dd, 1 H) 3.30 (s, 3 H) 3.23 (d, 1 H) 2.99 (d, 1 H) 2.67 - 2.80 (m, 2 H) 2.22 (d, 1 H) 1.89 - 2.06 (m, 2 H) 1.52 - 1.65 (m, 1 H) 1.44 - 1.47 (m, 1 H) 1.44 - 1.47 (m, 1 H) 1.32 - 1.43 (m, 1 H) 1.20 - 1.33 (m, 1 H) 1.15 - 1.19 (m, 1 H) 0.75 - 0.91 (m, 1 H) 0.63 (t, 3 H); MS (m/z) = 328 [MH].

Compound 6: (1 R.6S or 1S.6R)-6-(3.4-dichlorophenylVl-F3-methyl-(R or SVI- (methyloxy)butyll-3-azabicvclor4.1.Olheptane (single enantiomer)

1 ,1-Dimethylethyl (1 R.6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[3-methyl-(R or S)-1- (methyloxy)butyl]-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 14) (0.075 g, 0.170 mmol) was dissolved in DCM (0.5 ml.) and TFA (0.131 mL, 1.695 mmol) was added. The reaction mixture was stirred at RT for 1.5 hours and NaHCC>3 sat. solution (2 ml) was added. The reaction mixture was diluted with DCM (2ml), the organic phase was separated and concentrated under reduced pressure to obtain the title compound as light yellow oil (0.060 g); ¹H NMR (400 MHz, CHLOROFORM- d) δ ppm 7.57 (s, 1 H) 7.43 (d, 1 H) 7.31 - 7.38 (m, 1 H) 3.72 (d, 1 H) 3.02 - 3.17 (m, 2 H) 2.97 (s, 3 H) 2.75 - 2.85 (m, 1 H) 2.27 - 2.37 (m, 1 H) 2.19 - 2.25 (m, 1 H) 2.06 - 2.15 (m, 1 H) 1.57 - 1.71 (m, 2 H) 1.40 - 1.50 (m, 1 H) 1.26 - 1.40 (m, 1 H) 1.09 - 1.21 (m, 2 H) 0.82 (dd, 6 H); MS (m/z) = 342 [MH].

Compound 7: 6-(3.4-dichlorophenyl VI-F(R or SV1-(methyloxyV3-buten-1-yll-3- azabicvcloF4.1.01heptane (single enantiomer)

1 ,1-Dimethylethyl 6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 13) (0.0212 g, 0.050 mmol) was dissolved in DCM (1 mL) and the temperature was decreased to O⁰C. TFA (0.019 mL, 0.249 mmol) was added and the reaction was stirred at O⁰C for 30 mins. Further TFA (0.498 mmol, 0.038 ml) was added and the reaction mixture was stirred at O⁰C for 3h. Still further TFA (0.498 mmol, 0.038 ml) was added and the reaction mixture was stirred for a further 2h. Toluene (2 mL) was added and the reaction mixture was concentrated under reduced pressure. The residue was dissolved in methanol, charged on to a 1g SCX column and eluted with methanol and 2M ammonia in methanol to give the title compound as yellow oil (0.013 g); NMR(¹H, CDCI3): δ 7.53 (s, 1 H), 7.38 (d, 1 H), 7.27 (m, 1 H), 5.73 (m, 1 H), 5.04 (d, 1 H), 4.99 (d, 1 H), 3.50 (d, 1 H), 2.99 (s, 3H), 2.97 (d, 1 H), 2.85 (m. 1 H), 2.59 (m, 1 H), 2.40 (m, 1 H), 2.25 (m, 1 H), 2.17 (m, 1 H), 1.94 (t, 2H), 1.00 (m, 2H); MS (m/z) = 326 [MH].

Compound 8: (1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(1 R or 1S)-methyloxy)-3- buten-1-yl1-3-azabicyclo[4.1.01heptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 R,6S or 1S, 6R)-6-(3,4-dichlorophenyl)-1-((R or S^I-hydroxy-S-buten-i-yO-S-azabicyclo^.i .OJheptane-S-carboxylate (Intermediate 11 ) (50 mg, 0.121 mmol) in DMF (2 mL) at 20 ⁰C was added sodium hydride (5.82 mg, 0.146 mmol). After 15 min, iodomethane (9.86 μL, 0.158 mmol) was added. After 4 hr, further sodium hydride (5.82 mg, 0.146 mmol) was added followed after 20 min by iodomethane (9.86 μl_, 0.158 mmol). After 20 hr, still further sodium hydride (5.82 mg, 0.146 mmol) was added followed after 20 min by further iodomethane (9.86 μl_, 0.158 mmol). After 4 hr, NaHCC>3 (sat. sol.) was added and the mixture was extracted twice with DCM. The organic phase was dried and the solvent was removed under reduced pressure. The residue was purified via chromatography (c- Hex/EtOAc 0 to 50%; 10 g silica column) to give 1 ,1-dimethylethyl (1 S.6R or 1R,6S)- 6-(3,4-dichlorophenyl)-1-[(1 R or 7S)-1-(methyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane-3-carboxylate (30 mg);¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.4 (d, 2 H), 7.15 (dd, 1 H), 5.55 (m, 1 H), 4.85 (dd, 2H), 3.75 (m, 2H), 3.3 (m, 4H), 2.35 (m, 2H), 2.2 (m, 2H), 1.95 (m, 1 H), 1.7 (m, 1 H), 1.5 (s, 9H), 1.25 (d, 1 H), 1.15 (d, 1 H); MS (m/z) = 426 [MH⁺]. This compound was dissolved in DCM (1 ml.) at 20 ⁰C and TFA (0.093 ml_, 1.213 mmol) was added. After 2 hr, toluene (4 ml.) was added and the solvent was removed in vacuo. The residue was dissolved in DCM and washed with NaHCC>3 (sat sol). The organic phase was dried and the solvent removed under reduced pressure to give title compound (20 mg) as yellow oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.42 (s, 1 H), 7.4 (d, 1 H), 7.15 (d, 1 H), 5.55 (m, 1 H), 4.85 (dd, 2H), 3.25 (s, 3H), 3.1 (dd, 2H), 2.85 (m, 2H), 2.35 (m, 2H), 2.2 (m, 1 H), 2.0 (m, 2H), 1.5 (d, 1 H), 1.2 (d, 1 H); MS (m/z) = 326 [MH⁺].

Compound 9 : (1S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(2R or 2S)-tetrahydro-2- furanyl1-3-azabicyclo[4.1.01heptane (single enantiomer)

The title compound was obtained (10 mg) as colourless oil, starting from 1 ,1- dimethylethyl (1 R,6S or 1S,6R)-6-(3,4-dichlorophenyl)-1-[(2R or 2S)-tetrahydro-2- furanyl]-3-azabicyclo[4.1.0]heptane-3-carboxylate (Intermediate 17) (14 mg, 0.034 mmol) in a similar fashion to the preparation of Compound 7; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.42 (d, 1 H) 7.30 (d, 1 H) 7.25 (dd, 1 H) 3.74 - 3.84 (m, 1 H) 3.47 - 3.61 (m, 1 H) 3.38 (d, 1 H) 2.87 (d, 1 H) 2.71 - 2.88 (m, 2 H) 2.44 - 2.59 (m, 1 H) 1.45 - 2.01 (m, 6 H) 0.94 (d, 1 H) 0.83 (d, 1 H); MS (m/z) = 312 [MH⁺].

Compound 10: tf SSR or 7R6S)-6-(3,4-dichlorophenyl)-1-[(1 /? or 7S)- 1 -(ethyloxy)-3- buten-1-yl1-3-azabicyclo[4.1.01heptane (single enantiomer)

To a solution of 1 ,1-dimethylethyl (1 S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(1 R or IS^I-hydroxy-S-buten-i-ylJ-S-azabicyclo^.i .OJheptane-S-carboxylate (Intermediate 10) (105 mg, 0.255 mmol) in DMF (2 ml.) at 20 ⁰C was added sodium hydride (48.9 mg, 2.037 mmol). After 5 min, iodoethane (0.205 ml_, 2.55 mmol) was added. After 2 hr, NaHCC>3 (sat sol) and water were added and the mixture was extracted with DCM. The combined organic phases were dried and the solvent was removed under reduced pressure to give 1 ,1-dimethylethyl (1S,6R or 1 R,6S)-6-(3,4-dichlorophenyl)- 1-[(1 R or ISH-hydroxy-S-buten-i-ylJ-S-azabicyclo^.i .OJheptane-S-carboxylate (107 mg) as yellow oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.51 (s, 1 H) 7.36 (d, 1 H) 7.20 (dd, 1 H) 5.70 - 5.84 (m, 1 H) 4.96 - 5.11 (m, 2 H) 3.81 (br. s., 1 H) 3.24 - 3.44 (m, 3 H) 2.58 - 2.67 (m, 1 H) 2.40 - 2.50 (m, 1 H) 2.33 (br. s., 2 H) 2.04 - 2.14 (m, 1 H) 1.90 - 2.01 (m, 1 H) 1.50 (s, 9 H) 1.28 (s, 1 H) 1.13 (t, 3 H) 0.84 - 0.98 (m, 3 H); MS (m/z) = 440 [MH⁺]. This yellow oil was dissolved in DCM (3 ml.) at 20 ⁰C and TFA (0.020 ml_, 0.255 mmol) was added. After 1 hr, toluene (4 ml.) was added and the solvent was removed in vacuo. The residue was dissolved in DCM and washed with NaHCC>3 (sat sol). The organic phase was separated, dried and solvent was removed in vacuo to give the title compound (88 mg) as colorless oil; ¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 7.55 (d, 1 H) 7.36 (d, 1 H) 7.25 (dd, 1 H) 5.67 - 5.82 (m, 1 H) 4.93 - 5.07 (m, 2 H) 3.55 (d, 1 H) 3.39 (dq, 1 H) 2.96 (d, 1 H) 2.80 - 2.91 (m, 1 H) 2.53 - 2.71 (m, 2 H) 2.34 - 2.45 (m, 1 H) 2.21 - 2.33 (m, 1 H) 1.87 - 1.99 (m, 3 H) 1.10 - 1.18 (m, 2 H) 0.96 - 1.01 (m, 1 H) 0.83 - 0.94 (m, 2 H); MS (m/z) = 340 [MH⁺]. The affinity of compounds of the invention for SERT, NET and DAT may be tested in one or other of the following affinity assays.

a) Filtration Binding Assay using membranes from hSERT, hNET, and hDAT Lewis

Lung Carcinoma Porcine tubule Kidney (LLCPK) cell lines

- Membrane preparation hSERT-LLCPK, hDAT-LLCPK or hNET-LLCPK cell lines are used for the membrane preparations for radioligand binding assays. Stable cell lines may be generated as follows: i) hSERT - generated by transfecting LLC-PK1 or LLCPK cells with hSERT cloned into the mammalian expression vector pCDNA3.1 Hygro(+); ii) hNET - generated by transfecting LLCPK cells with hNET cloned into the mammalian expression vector pRC/CMV; iii) hDAT- generated by transfecting LLCPK cells with hDAT cloned into the mammalian expression vector pDESTCDNA3.1 (an example of a procedure for transfecting LLCPK cells with hDAT, hSERT and hNET may be found in H. Gu, S. C. Wall and G. Rudnick, J. Biol. Chem. (1994) 269 : 7124-7130.)

Each cell line is cultured independently in Dulbecco's modified Eagle's medium (DMEM) containing 10% of Foetal Bovine Serum (FBS) supplemented with 400 μg/ml hygromicin (hSERT) or geneticin at 500 μg/ml (hNET) or at 1000 μg/ml (hDAT). Cells are maintained at 37°C in a humidified environment containing 5% CO2 in air.

When cells are at 70-80% of confluence, the culture medium is removed and the cells harvested with phosphate buffered saline (PBS) containing 5 mM EDTA. The cell suspension is centrifuged at 90Og for 5 minutes at 4⁰C. The resultant pellets are re-suspended in 30-50 volumes of Assay Buffer (5OmM Tris pH 7.7 containing 12OmM NaCI, 5mM KCI, 10μM pargyline and 0.1% ascorbic acid) and homogenized using a glass-teflon Potter homogeniser and centrifuged at 4800Og for 20 minutes at 4⁰C. The resultant membrane pellets are re-suspended in the same volume of Assay Buffer, incubated for 20 minutes at 37⁰C and centrifuged as before at 4800Og. The final protein concentration for each preparation is adjusted to give approximately 480μg protein/ml for hSERT-LLCPK, hDAT-LLCPK and hNET-LLCPK, as determined by the Bio-Rad Protein Assay kit. Membranes are stored at -80⁰C as 1 ml aliquots until required.

- Assay Protocol (For general references to monoamine transporters filtration binding assays see: Michael J. Owens, et al, Neurotransmitter receptor and transporter binding profile of antidepressants and their metabolites, JPET, 283:1305-1322, 1997; PerAllard, Jan O. Marcusson, Svate B. Ross, [3H]WIN-35,428 binding in the human brain, Brain Res., 706 :347-350, 1996.)

The affinity of the compounds of the invention to bind the re-uptake site of SERT may be assessed using [3|H]citalopram filtration binding assay performed on hSERT- LLCPK cell membranes. The competition binding assay is conducted in deep-well 96 well plates (1 ml, NUNC, cod.260252) in a total volume of 400μl, with each concentration in duplicate. 4μl of test compound (100X solution in neat DMSO as 7 point curve ranging from 10^"6 to 10^"12M, final concentration) or DMSO (to define total binding) or a final concentration of 10μM fluoxetine in DMSO (to define non-specific binding, NSB) are added to wells; after this, 200μl of [N-Methyl-³H]citalopram (Amersham Biosciences, 80 Ci/mmol) at the final concentration of 0.25nM in Assay Buffer, is added to all wells and finally the reaction is started by adding 200μl/well of membranes diluted 1 :80 in Assay Buffer at concentration of about 2.5μg/well of protein. The reaction is carried out at room temperature for 2 hours and then stopped by rapid filtration through GF/B Unifilter 96-filterplate (Perkin-Elmer) pre-soaked in 0.5% polyethylenimmine (PEI) using a Perkin-Elmer FilterMat-196 harvester. Filterplate is washed 3 times with 1 ml/well ice-cold 0.9% NaCI solution. The plate is dried in an oven for 60 min at 50⁰C then opaque bottom-seal is placed on the underside of the plate and 50μl of Microscint 20 (Perkin-Elmer) added to each well. Plate is sealed with a TopSeal and the radioactivity in the samples is counted for 4 min using TopCount liquid scintillation counter (Packard-Perkin-Elmer) and recorded as counts per minute (CPM).

Competition binding assay for hNET may be conducted essentially as previously reported for hSERT in 96 well format and in a final assay volume of 400μl, except for the use of hNET-LLCPK cell membranes (1 :40 dilution i.e. 4.8μg of protein/well) and [³H]nisoxetine as radioligand (1.5nM [N-methyl-³H]nisoxetine, Amersham Biosciences, 84 Ci/mmol). 10μM desipramine is used for NSB.

Competition binding assay for hDAT may also be conducted essentially as previously reported for hSERT and hNET in 96 well format and in a final assay volume of 400μl, except for the use of hDAT-LLCPK cell membranes (1 :20 i.e. 9.6μg of protein/well) and [³H]WI N-35,428 as radioligand (1OnM [N-Methyl-³H]WIN-35,428, Perkin Elmer, 85.6 Ci/mmol). Furthermore, 10μM GBR-12909 is used for NSB and the incubation time of the binding reaction is 1 hour at room temperature.

b) Scintillation Proximity Assay (SPA) for human DAT, NET and SERT binding

- Generation of BacMam viruses for the expression of hSERT, hNET, and hDAT in mammalian cells

Membranes for the SPA-binding assays are produced by HEK-293F cell infection with BacMam viruses generated for each single human SERT, NET, and DAT transporter. hSERT and hDAT are cloned into pFBMRfA vector whereas hNET is cloned into pFASTBacMami vector. The generation and use of BacMam viruses is described in Condreay JP et al, Proc. Natl. Acad. Sci. USA, 1999, 96:127-132 and Hassan NJ et al, Protein Expression and Purification, 47(2): 591-598, 2006.

- Transduction of HEK-293F cells with hSERT/hDAT/hNET BacMam viruses

The HEK-293F suspension cell line (Invitrogen) is routinely grown in 293_Freestyle Expression media (Invitrogen) in shake flask suspension culture. The culture is transduced with the appropriate transporter BacMam at a MOI (multiplicity of infection) of 100 virus particles per cell and incubated for 48hrs at 37⁰C, 5% CO₂ in air, shaken at 90rpm in a humidified shaker incubator. The culture is then harvested by centrifugation at 1000g, 4⁰C, for 10 minutes and the cell pellet stored at -8O⁰C until required.

- Preparation of BacMam hSERT/hDAT/hNET-HEL293F cell membranes

Transduced cell pellets are re-suspended to 10x volume with buffer-A (5OmM HEPES, 1 mM EDTA, 1 mM leupeptin, 25ug/ml_ bacitracin, 1 mM phenylmethylsulfonylfluoride, PMSF, 2μM pepstatin A, pH 7.7) and homogenised with 2x 15 second bursts in a glass Waring blender. The homogenate is then centrifuged for 20 minutes at 50Og. Following this, the supernatant is pooled and centrifuged at 13,00Og for 30 minutes. Pellets are then re-suspended to 4x original pellet volume with buffer-B (5OmM TRIS pH 7.4, 13OmM NaCI) and forced through a 0.8mm needle to give a homogeneous suspension. Membrane aliquots are stored at -8O⁰C until required. The protein concentration is quantified by Bradford assay.

- Assay Protocol

The affinity of the compounds of the invention for hSERT, hNET or hDAT may also be assessed by using the [³H]citalopram, [³H]nisoxetine or [³H]WI N-35, 428 binding assays with the SPA technology on BacMam-recombinant human SERT, NET and DAT membranes produced as described before. With the SPA technology (GE

Healthcare, Amersham) only transporter-bound radioactivity can elicit bead excitation thus no separation of the bound/ unbound radioligand is required.

The protocol for hSERT binding SPA is based on Trilux beta-counter (Wallac, Perkin- Elmer). Briefly, 0.5μl_ of test compound in neat DMSO (or 1 μM fluoxetine as positive control) is added by 50μl_ of the SPA mixture, containing 2mg/ml_ SPA beads (Amersham RPNQ0001 ), 4μg/ml_ hSERT Bacmam membranes, 0.01% pluronic F- 127, 2.5nM [³H]citalopram in the assay buffer (2OmM HEPES, 145mM NaCI, 5mM KCI, pH 7.3). Incubation are performed at room temperature for at least 2 hours. Counts are stable and could be read up to 3 days.

Alternatively, hDAT hNET and hSERT SPA-binding assays are performed by using a Viewlux beta-counter (Wallac, Perkin-Elmer) with imaging PS-WGA beads (Amersham RPNQ0260) in a final assay volume of 30μl_ and in a 384-well plate format (Greiner 781075). Briefly, 0.3μl_ of test compound in neat DMSO and 0% and 100% effect controls (DMSO for total binding and 10 or 1 μM indatraline as positive control) are added to the wells by using a Hummingbird (Genomic Solutions), followed by the addition of 30μl_ of the SPA mixture, containing 1 mg/mL SPA beads (hSERT) or 2mg/ml SPA beads (hDAT and hNET), 40μg/ml or 20μg/ml or 6 μg/ml of hDAT or hNET or hSERT BacMam membranes, 0.02% pluronic F-127, 1OnM

[³H]WI N-35,428 or 1OnM [³H]nisoxetine or 3nM [³H]citalopram for hDAT or hNET or hSERT binding SPA in the assay buffer (2OmM HEPES, 145mM NaCI, 5mM KCI, pH 7.3-7.4). Incubation is performed at room temperature for at least 2 hours, best overnight in the dark. Bound radioactivity is recorded by using a 600s 6x binning and 613nm emission filter with the Viewlux instrument. Calculation

The affinity of the compounds of the invention for a particular transporter may be calculated from the IC₅₀ obtained in competition experiments as the concentration of a compound necessary to displace 50% of the radiolabeled ligand from the transporter, and is reported as a "K," value calculated by the following equation: κ _ IC₅₀

¹ 1 + L / K_D where L = radioligand and K₀ = affinity of radioligand for transporter (Cheng and Prusoff, Biochem. Pharmacol. 22:3099, 1973). Results are presented below as the pKi (i.e. the antilogarithm of Ki).

Compounds 1 to 10 were tested in assay b) using hSERT, hNET and hDAT. Each test was performed at least twice for each compound against each receptor. Using this assay, all the compounds gave an average pKi equal to or greater than 8.5 for hSERT, equal to or greater than 8.2 against hNET and equal to or greater than 8.0 against hDAT.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof

(I) wherein

R5 is C-|_4alkyl; or

2. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R^ is 3,4-dichlorophenyl.

3. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R^ and R^ are H.

4. A compound according to any preceding claim or a pharmaceutically acceptable salt thereof wherein R^ is C-_|_4alkyl or C2-4alkenyl.

5. A compound according to claim 1 selected from the list: (1 R,6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)1-(methyloxy)ethyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 1 );

(1 R,6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)propyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 2);

(1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[(R or S)-1-(methyloxy)butyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 5); (1 R.6S or 1 S,6R)-6-(3,4-dichlorophenyl)-1-[3-methyl-(R or S)-1-(methyloxy)butyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 6);

(1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(1 R or 1S)-methyloxy)-3-buten-1-yl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 8);

(1 S.6R or 1 R,6S)-6-(3,4-dichlorophenyl)-1-[(2R or 2S)-tetrahydro-2-furanyl]-3- azabicyclo[4.1.0]heptane (single enantiomer) (Compound 9);

6. A pharmaceutical composition comprising a) a compound defined in any preceding claim or a pharmaceutically acceptable salt thereof and b) one or more pharmaceutically-acceptable excipients.

7. A compound defined in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof for use in treating a disease or condition.

8. A compound defined in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof for use in treating a disease or condition for which inhibition of monoamine neurotransmitter re-uptake is beneficial.

9. A method of treating a disease or condition for which inhibition of monoamine neurotransmitter re-uptake is beneficial in a human comprising administering an effective amount of a compound defined in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof.

10. A compound according to claim 8 or a pharmaceutically acceptable salt thereof for use in treating depression or an anxiety disorder.

11. A compound according to claim 8 or a pharmaceutically acceptable salt thereof for use in treating an eating disorder.

12. A method according to claim 9 for treating depression or an anxiety disorder.

13. A method according to claim 9 for treating an eating disorder.