HK1116162A

HK1116162A - Morpholine derivatives for use as dopamine agonists in the treatment of i.a. sexual dysfunction

Info

Publication number: HK1116162A
Application number: HK08105010.7A
Authority: HK
Inventors: C．M．N．阿勒顿; A．D．巴克斯特; A．S．库克; D.赫普沃思; S．K-F．翁
Original assignee: 辉瑞大药厂
Priority date: 2002-12-10
Filing date: 2008-05-06
Publication date: 2008-12-19

Description

Morpholine derivatives as dopamine agonists in the treatment of sexual dysfunction

The present application is a divisional application of the chinese patent application having application number 200380105677.1(PCT/IB 2003/005683), application date 12/2/2003, entitled "morpholine derivatives as dopamine agonists for the treatment of sexual dysfunction".

The invention relates to a dopamine agonist, in particular to an agonist which is selective to D3 and better than D2. These compounds are useful for the treatment and/or prevention of sexual dysfunction, such as Female Sexual Dysfunction (FSD), in particular Female Sexual Arousal Disorder (FSAD), and male sexual dysfunction, in particular Male Erectile Dysfunction (MED). Male sexual dysfunction as referred to herein includes ejaculatory disorders such as premature ejaculation, loss of sexual arousal (inability to achieve orgasm), or sexual desire disorders such as hypoactive sexual desire disorder (HSDD; lack of interest in sex). These compounds are also useful in the treatment of neuropsychiatric and neurodegenerative disorders.

The present invention provides compounds of formula (I), (Ia) and (Ib)

Wherein:

a is selected from the group consisting of C-X and N,

b is selected from the group consisting of C-Y and N,

R¹is selected from H and (C)₁-C₆) An alkyl group, a carboxyl group,

R²is selected from H and (C)₁-C₆) An alkyl group, a carboxyl group,

x is selected from H, HO, C (O) NH₂、NH₂，

Y is selected from H, HO, NH₂Br, Cl and F, in the presence of a catalyst,

z is selected from H, HO, F, CONH₂And CN;

and pharmaceutically acceptable salts, solvates and prodrugs thereof;

the conditions are as follows:

for compounds of formula (I), (Ia) or (Ib), if A is C-X, B is C-Y, R¹Is H or (C)₁-C₆) Alkyl radical, R²Is H or (C)₁-C₆) Alkyl, at least one of X, Y and Z must be OH;

for compounds of formula (I), when A is C-X, B is C-Y, Y is H, Z is H, R¹Is H, R²Is H, then X cannot be OH.

Pharmaceutically acceptable salts of the compounds of formula (I) include acid addition and base salts thereof.

Pharmaceutically acceptable salts of the compounds of formula (I) can be readily prepared by suitably mixing together a solution of the compound of formula (I) and the desired acid or base. The salt may be precipitated from solution, collected by filtration, or may be recovered by evaporation of the solvent.

Suitable acid addition salts are formed from acids which form non-toxic salts, examples being hydrochloride, hydrobromide, hydroiodide, sulphate, bisulphate, nitrate, phosphate, hydrogen phosphate, acetate, maleate, fumarate, lactate, tartrate, citrate, gluconate, succinate, saccharate (saccharate), benzoate, methanesulphonate, ethanesulphonate, benzenesulphonate, p-toluenesulphonate and pamoate.

Suitable base salts may be derived from bases which form non-toxic salts, examples being the salts of sodium, potassium, aluminium, calcium, magnesium, zinc and diethanolamine.

For a review of suitable salts, see Berge et al, j.pharm.sci., 66, 1-19, 1977.

Pharmaceutically acceptable solvates of the compounds of formula (I) include hydrates thereof.

Also included within the scope of compounds of formula (I) are polymorphic forms thereof.

The compounds of formula (I) contain one or more asymmetric carbon atoms and thus exist in two or more stereoisomeric forms. Separation of the diastereomers may be effected by subjecting the mixture of stereoisomers of the compound of formula (I) or a suitable salt or derivative thereof to conventional techniques, for example fractional crystallisation, chromatography or h.p.l.c.. The individual enantiomers of the compounds of formula (I) may also be prepared from the corresponding optically pure intermediates, or by resolution of the corresponding racemates, for example h.p.l.c., using a suitable chiral support, or by reaction of the corresponding racemates with a suitable optically active acid or base, as the case may be, and fractional crystallisation of the diastereomeric salts formed.

Preferred compounds of the invention are compounds of the formulae (Ia) and (Ib).

Particularly preferred are compounds of formula (Ia).

Preferably, A is C-X or N and B is C-Y. More preferably, A is N and B is C-Y. More preferably, A is C-X and B is C-Y.

Preferably, R¹Is selected from H and (C)₁-C₄) An alkyl group. More preferably, R¹Are H, methyl and ethyl. Even more preferably, R¹Is H or methyl. Most preferably, R¹Is H.

Preferably, R²Is selected from H and (C)₁-C₄) An alkyl group. More preferably, R²Selected from H, methyl and ethyl. Most preferably, R²Selected from H and methyl.

In a particularly preferred embodiment, R²Is H. In a further particularly preferred embodiment, R²Is methyl.

Preferably, X is selected from H, OH and NH₂. Most preferably, X is selected from H and OH.

In a particularly preferred embodiment, X is H. In another particularly preferred embodiment, X is OH.

Preferably, Y is selected from H, NH₂Cl and F. Most preferably, Y is selected from H and NH₂。

In a particularly preferred embodiment, Y is H. In a further particularly preferred embodiment, Y is NH₂。

Preferably, Z is selected from H, HO and F. Most preferably, Z is selected from H or HO.

In a particularly preferred embodiment, Z is H. In a further particularly preferred embodiment, Z is HO.

Particularly preferred are the compounds of the invention (and salts thereof) as exemplified herein; more preferably:

r- (-) -3- (4-Propylmorpholin-2-yl) phenol (example 7A)

S- (+) -3- (4-propylmorpholin-2-yl) phenol (example 7B)

R- (-) -3- (4-Propylmorpholin-2-yl) phenol hydrochloride (example 8)

R-5- (4-Propylmorpholin-2-yl) benzene-1, 3-diol (example 15A)

S-5- (4-Propylmorpholin-2-yl) benzene-1, 3-diol (example 15B)

R- (+) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol (example 23A)

S- (-) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol (example 23B)

2-bromo-4- (4-propylmorpholin-2-yl) phenol (example 30)

2-hydroxy-5- (4-propylmorpholin-2-yl) benzamide (example 35)

2-Nitro-4- (4-propylmorpholin-2-yl) phenol (example 36)

2-amino-4- (4-propylmorpholin-2-yl) phenol (example 37)

5- (4-Propylmorpholin-2-yl) pyridin-2-ylamine (examples 44A and 44B)

2-chloro-5- (4-propylmorpholin-2-yl) phenol (example 54)

3- [ (5S) -5-methyl-4-propylmorpholin-2-yl ] phenol (example 60)

5- [ (2S, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine (example 66)

5- [ (2R, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine (example 67)

Most preferred are:

r- (-) -3- (4-Propylmorpholin-2-yl) phenol (example 7A)

S- (+) -3- (4-propylmorpholin-2-yl) phenol (example 7B)

R- (-) -3- (4-Propylmorpholin-2-yl) phenol hydrochloride (example 8)

R-5- (4-Propylmorpholin-2-yl) benzene-1, 3-diol (example 15A)

S-5- (4-Propylmorpholin-2-yl) benzene-1, 3-diol (example 15B)

R- (+) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol (example 23A)

S- (-) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol (example 23B)

5- (4-Propylmorpholin-2-yl) pyridin-2-ylamine (examples 44A and 44B)

2-chloro-5- (4-propylmorpholin-2-yl) phenol (example 54)

5- [ (2S, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine (example 66)

5- [ (2R, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine (example 67)

The compounds of the invention can be prepared in a variety of ways in a known manner. The following route illustrates a method of synthesizing the compound of formula (I); the skilled person will appreciate that the compounds of formula (Ia) and (Ib) may be isolated using appropriate resolution techniques.

Compounds of formula (I) wherein A is C-X, B is C-Y, R can be prepared according to scheme 1¹Is H or (C)₁-C₆) Alkyl radical, R²Is H, wherein X, Y and Z are as described herein.

Scheme 1

The compounds of formula (III) may be prepared by reacting an aldehyde of formula (II) with i) a cyanide source or nitromethane, followed by II) reduction with borane, lithium aluminum hydride or hydrogenation. Some compounds of formula (II) and (III) are also commercially available.

The compound of formula (IV) may be prepared by reacting a compound of formula (III) with III) an acid chloride in the presence of a suitable base such as triethylamine or 4-methylmorpholine. Typical reaction conditions comprise 1.0 equivalent of amine (III), 1.2 to 2.0 equivalents of base (preferably triethylamine), 1.1 to 1.3 equivalents of acid chloride in dichloromethane at 25 ℃.

The compound of formula (V) may be prepared by reducing the compound of formula (IV) with IV) a reducing agent such as borane or lithium aluminium hydride. Typical conditions comprise 1.0 equivalent of amide (IV), 1.2 to 3.0 equivalents of borane in THF at reflux. The compounds of formula (V) may also be prepared by reductive amination of a compound of formula (III) with a suitable aldehyde in the presence of sodium cyanoborohydride.

The compound of formula (VI) may be prepared by reacting a compound of formula (V) with V) chloroacetyl chloride or a 2-substituted chloroacetyl chloride, such as 2-chloropropionyl chloride or 2-chlorobutyryl chloride, in the presence of a base, such as triethylamine, sodium carbonate and potassium hydroxide. Typical conditions comprise 1.0 equivalent of amine (IV), 1.0 to 1.3 equivalents of acid chloride, 1.2 to 2.0 equivalents of triethylamine in dichloromethane at 25 deg.C, and the crude reaction mixture is then dissolved in IPA containing 1.2 to 3.0 equivalents of aqueous potassium hydroxide.

The compounds of formula (I) may be prepared by reacting a compound of formula (VI) with VI) a reducing agent such as borane or lithium aluminum hydride. Typical conditions comprise 1.0 equivalent of amide (VI), 1.2 to 3.0 equivalents of borane in THF at reflux.

The skilled artisan will appreciate that since either X, Y or Z is a hydroxyl group, it will be necessary to protect the hydroxyl group with a suitable protecting group during the conversion of scheme 1, and then remove the protecting group. The method of deprotection of the phenolic group depends on the protecting group. For examples of protection/deprotection methods, see "Protective groups in Organic synthesis", TW Greene and PGMWutz. For example, if the hydroxyl group is protected as a methyl ether, the deprotection conditions comprise refluxing in 48% aqueous HBr for 1 to 24 hours, or stirring with borane tribromide in dichloromethane for 1 to 24 hours. Alternatively, if the hydroxy group is protected as a benzyl ether, the deprotection conditions comprise hydrogenation with a palladium catalyst under a hydrogen atmosphere.

Compounds of formula (I) wherein one of A or B is N, R can be prepared according to scheme 2¹Is H or (C)₁-C₆) Alkyl radical, R²Is H, X, Y and Z are as described herein, with the proviso that either X, Y or Z is NH₂. This scheme illustrates where B is C-Y, Y is NH₂The case (1); the skilled person will appreciate that other compounds are equally achievable.

Scheme 2

The compound of formula (VII) may be prepared using the process described in JP 2001048864.

The compounds of formula (VIII) can be prepared by reacting epoxide (VII) with VII) propylamine. Typical reaction conditions involve stirring the epoxide with excess amine, either alone or in dimethylsulfoxide.

The compound of formula (IX) may be prepared by reacting a compound of formula (VIII) with v) chloroacetyl chloride or a 2-substituted chloroacetyl chloride (e.g. 2-chloropropionyl chloride or 2-chlorobutyryl chloride) in the presence of a base (e.g. triethylamine, sodium carbonate and potassium hydroxide). Typical conditions comprise 1.0 equivalent of amine (VIII), 1.2 to 2.0 equivalents of triethylamine in dichloromethane at 25 ℃ and the crude reaction mixture is then dissolved in IPA containing 1.2 to 3.0 equivalents of aqueous potassium hydroxide.

The compound of formula (X) may be prepared by reacting a compound of formula (IX) with a reducing agent such as lithium aluminium hydride. Typical conditions comprise 1.0 equivalent of amide (X), 1.2 equivalents of lithium aluminum hydride in THF at reflux.

The compounds of formula (I) can be prepared by ix) deprotection. Typical conditions comprise 1.0 equivalent of compound (X) and 5 equivalents of hydroxylamine hydrochloride in ethanol under reflux.

Compounds of formula (I) wherein A is C-X, B is C-Y, R can be prepared according to reaction scheme 3¹Is H, R²Is H or (C)₁-C₆) Alkyl, wherein X, Y and Z are as described herein.

Scheme 3

The compounds of formula (XII) can be prepared by reacting an amino acid ester of formula (XI) with an acid chloride of formula x) in the presence of a suitable base such as triethylamine and 4-methylmorpholine. Typical reaction conditions comprise 1 equivalent of amino acid ester (XI), 1 equivalent of acid chloride and 3 equivalents of base in dichloromethane at 25 ℃. Some compounds of formula (XI) are commercially available.

The compound of formula (XIII) may be prepared by reacting a compound of formula (XII) with xi) borane-THF complex followed by acid decomposition of the boron-nitrogen complex and tert-butoxycarbonyl protection of the resulting amine. Typical reaction conditions comprise 1 equivalent of amide (XII) with 3 equivalents of BH₃THF, in THF, under reflux, cooling, carefully adding 6M aqueous HCl, heating to reflux for another 6 hours. The solvent was then evaporated, dissolved in a methanol/water (8: 1) mixture and 5 equivalents addedA base, such as potassium hydroxide, and 1.5 equivalents of di-tert-butyl dicarbonate, the mixture is stirred for 72 hours.

The compound of formula (XIV) may be prepared by reacting a compound of formula (XIII) with xii) an organic solution of HCl. Typical reaction conditions comprise a solution of 1 equivalent of carbamate (XIII) and 1-10 equivalents of 4M HCl in dioxane at 25 ℃.

The compound of formula (XV) may be prepared by reacting a compound of formula (XIV) with xiii) 2-bromoacetophenone in the presence of a base such as triethylamine or 4-methylmorpholine. 2-bromoacetophenone is available from commercial sources or, alternatively, can be prepared from the parent acetophenone by standard bromination methods well known to those skilled in the art. Typical conditions comprise 1 equivalent of amino alcohol (XIV) with 1-3 equivalents of triethylamine and 1 equivalent of 2-bromoacetophenone at 65 ℃.

The compound of formula (I) may be prepared by reacting a compound of formula (XV) with xiv) triethylsilicon and trimethylsilyl triflate (trimethlilyltriflate). Typical conditions comprise the addition of 5-10 equivalents triethylsilane to 1 equivalent of morpholinol (XV) followed by the addition of 2 equivalents trimethylsilyl triflate in dichloromethane at-78 ℃.

The skilled artisan will appreciate that since either X, Y or Z is a hydroxyl group, it will be necessary to protect the hydroxyl group with a suitable protecting group during the conversion of scheme 3, and then remove the protecting group. The method of deprotection of the phenolic group depends on the protecting group. For examples of protection/deprotection methods, see "Protective groups in Organic synthesis", TW Greene and PGMWutz. For example, if the hydroxy group is protected as a methyl ether, the deprotection conditions comprise refluxing in 48% aqueous HBr for 1 to 24 hours, or stirring with borane tribromide in dichloromethane for 1 to 24 hours. Alternatively, if the hydroxy group is protected as the benzyl ether, the deprotection conditions comprise hydrogenation with a palladium catalyst under a hydrogen atmosphere.

The compounds of formula (I) wherein the stereogenic center located alpha to the morpholine nitrogen is absolutely defined can start from homochiral compounds of formula (XI) which are either commercially available or prepared by chemical literature methods readily available to the skilled person. The resulting compound of formula (I) will contain a mixture of diastereomers, which can be separated on an HPLC column. Typical conditions comprise passing through a Chiralcel OJ-H column, eluting with a mobile phase of 100% MeOH.

Compounds of formula (I) wherein one of A or B is N, R can be prepared according to reaction scheme 4¹Is H, R²Is H or (C)₁-C₆) Alkyl, X, Y and Z are as described herein, with the proviso that either X, Y or Z is NH₂. The scheme illustrates where B is C-Y, Y is NH₂The case (1); the skilled person will appreciate that other compounds may be suitable as well.

Scheme 4

The compound of formula (XVIII) can be prepared by reacting a compound of formula (XVI) with xv) an amino alcohol of formula (XIV) in the presence of a base such as triethylamine or 4-methylmorpholine. Typical conditions comprise 1 equivalent of amino alcohol (XIV) with 1-3 equivalents of triethylamine and 1 equivalent of the compound of formula (XVI) using toluene as solvent at room temperature or higher. The compounds of formula (XVI) are commercially available.

The compound of formula (IXX) may be prepared by reacting a compound of formula (XVIII) with xvi) an organometallic reagent generated from a bromide of formula (XVII). Suitable organometallic reagents include Grignard (organomagnesium) or organolithium reagents, which can be prepared from bromides via halogen metal displacement. Typical conditions include the addition of isopropyl magnesium chloride (for halogen metal displacement) to bromide (XVII) followed by the addition of morpholinone (XVIII) in an anhydrous ether solvent such as tetrahydrofuran at room temperature. The bromide (XVII) can be prepared using the process described in WO 99/32475.

Morpholinol (IXX) can be reduced to diol (XX) by reaction with xvii) a hydride reducing agent, such as sodium borohydride, in an alcoholic solvent, such as methanol.

The compounds of formula (XXI) can be prepared by deprotection via ix) from a diol (XX). Typical conditions comprise 1.0 equivalent of compound (XX) and 5 equivalents of hydroxylamine hydrochloride in ethanol at reflux.

The compound of formula (I) may be prepared by cyclisation of the compound of formula (XXI) by treatment with acid xviii). Typical conditions employ concentrated sulfuric acid and methylene chloride solvent at room temperature or higher.

All the above reactions and the preparation of the novel starting materials for use in the aforementioned processes are conventional and the skilled person is familiar with the reagents and reaction conditions suitable for carrying out the reactions or preparations and the procedures for isolating the desired products with reference to the cited documents and the examples and preparations below.

The compounds of the present invention may be used as selective D3 agonists in the treatment of disease states. There are a large number of compounds with agonist activity of both D2 and D3; however, the use of such compounds is associated with a number of side effects, including nausea, vomiting, fainting, hypotension and bradycardia, some of which can cause serious problems.

It was previously believed that the efficacy of the prior art compounds derives from their ability to agonize D2; however, the D2 agonism is responsible for the above side effects.

The present invention provides a class of selective D3 agonists. They were found to be effective by chance while reducing the side effects associated with non-selective prior art compounds.

The compounds of the invention are useful in the treatment of sexual dysfunction, female sexual dysfunction (including hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder and dyspareunia), male erectile dysfunction, hypertension, neurodegeneration, psychosis, depression (e.g., cancer patient depression, Parkinson's disease depression, post-myocardial infarction depression, sub-syndrome symptomatic depression, infertile female depression, pediatric depression (paediatric depression), major depression, single episode depression, recurrent depression, malaria-induced depression (child indedense depression), postpartum depression and geriatric dysphoric syndrome), generalized anxiety disorder, phobias (e.g., agoraphobia, social phobia and simple phobia), post-traumatic stress syndrome, avoidant personality disorder, premature ejaculation, eating disorders (e.g., anorexia nervosa and bulimia nervosa), obesity, sexual impotence, post-sexual arousal syndrome, post-sexual impotence, Chemical dependence (e.g., addiction to alcohol, cocaine, heroin, phenobarbital, nicotine and benzodiazepines *), cluster headache, migraine, pain, Alzheimer's disease, obsessive-compulsive disorders, panic disorders, memory disorders (e.g., dementia, amnesia and age-related cognitive decline (ARCD)), Parkinson's disease (e.g., Parkinson's dementia, neuroleptic-induced parkinsonism and tardive dyskinesia), endocrine disorders (e.g., hyperprolactinemia), vasospasm (particularly in the cerebral vasculature), cerebellar ataxia, gastrointestinal disorders (involving alterations in motility and secretion), negative symptoms of schizophrenia, premenstrual syndrome, fibromyalgia syndrome, stress urinary incontinence, Tourette's syndrome, trichotillomania, craving, male impotence, sexual impotence, psychoactive syndrome, psychoactive trichotillomania, crasia, craving for humans, and humans, Attention Deficit Hyperactivity Disorder (ADHD), chronic paroxysmal migraine, headache (associated with vascular disorders), emotional instability, pathological crying, sleep disorders (cataplexy), and shock.

The compounds of the present invention are particularly suitable for the treatment of female sexual dysfunction, male erectile dysfunction, neurodegeneration, depression and psychiatric disorders.

The compounds of the invention are useful for male sexual dysfunction, particularly male erectile dysfunction. Male Erectile Dysfunction (MED), also known as male erectile dysfunction, is defined as:

"penile erection failure to achieve and/or maintain satisfactory sexual performance" (NIHConsenssus Development Panel on Impentance, 1993)

According to estimates, the prevalence of Erectile Dysfunction (ED) in various degrees (mild, moderate and complete impotence) is 52% in men between 40 and 70 years of age, and higher in people over 70 years of age (Melman et al 1999, J.urology, 161, p 5-11). This condition has a significant negative impact on the quality of life of individuals and partners, often resulting in increased anxiety and stress, causing depression and reduced self-esteem. Whereas MED was primarily considered a psychological disorder two decades ago (Benet et al 1994 Comp. Ther., 20: 669-. As a result, great progress has been made in identifying the mechanism of normal penile erection and the pathophysiology of MED.

Penile erection is a hemodynamic event that depends on the contraction and relaxation balance of the cavernous smooth muscle of the penis and the vasculature of the penis (Lerner et al1993, J.Urology, 149, 1256-42 1255). The corpus cavernosum smooth muscle is also referred to herein as the corpus cavernosum smooth muscle or corpus cavernosum. Relaxation of the smooth muscle of the corpora cavernosa causes increased blood flow into the trabecular space of the corpora cavernosa, causing them to expand toward the surrounding capsule, compressing the drainage veins. This causes a large rise in blood pressure, resulting in an erection (Naylor, 1998, j. urology, 81, 424-.

The changes that occur during the erectile process are complex and require a high degree of coordinated control involving the peripheral and central nervous and endocrine systems (Naylor, 1998, j. urology, 81, 424-. Cavernosal smooth muscle contraction is regulated by sympathetic norepinephrine innervation via activation of postsynaptic alpha 1 adrenoreceptors. MED may be associated with an increase in the intrinsic smooth muscle tone of the corpus cavernosum. However,the process of cavernous smooth muscle relaxation is mediated in part by non-adrenergic non-cholinergic (NANC) neurotransmission. A number of other NANC neurotransmitters than NO are found in the penis, such as calcitonin gene-related peptide (CGRP) and Vasoactive Intestinal Peptide (VIP). The major relaxant responsible for mediating this relaxation is Nitric Oxide (NO), which is synthesized from L-arginine by Nitric Oxide Synthase (NOS) (Taub et al1993 Urology, 42, 698-. It is thought that reducing cavernosal smooth muscle tone may contribute to NO-induced cavernosal relaxation. During male sexual arousal, NO is released from neurons and endothelium, binds to and activates soluble guanylate cyclase (sGC) located in smooth muscle cells and endothelium, causing an increase in intracellular cyclic guanosine 3 ', 5' -monophosphate (cGMP) levels. This cGMP increase is due to intracellular calcium concentration ([ Ca ]²⁺]_i) The mechanism of this decrease, leading to cavernous relaxation, is unknown, but is thought to be involved in activation of protein kinase G (probably due to Ca)²⁺Activation of pumps and Ca²⁺Activated K⁺A channel).

Multiple sites of potential regulatory behavior have been identified within the central nervous system. The key neurotransmitters are believed to be serotonin, norepinephrine, oxytocin, nitric oxide and dopamine. By mimicking the action of one of these key neurotransmitters, sexual function can be modulated. Dopamine D3 receptors are expressed almost exclusively in the limbic regions, which are involved in reward, emotional, and cognitive processes.

Without being bound by any theory, it appears that "due to its role in motor activity control, the integrity of the nigrostriatal dopaminergic pathway is also essential for the manifestation of interpersonal behavior. Somehow, particularly with respect to sexual function, it is likely that dopamine can cause penile erection by acting on oxytocin neurons located in the subtenon of the hypothalamus, perhaps also on the erectogenic sacral parasympathetic nucleus within the spinal cord. Now, the site that appears to be significant is D3, rather than the previously thought D2.

In essence, D3 is the initiator of sexual activity.

Accordingly, the present invention provides the use of a compound of formula (I) for the manufacture of a medicament for the treatment or prevention of erectile dysfunction.

Mild to moderate MED patients should benefit from treatment with the compounds of the invention, and severe MED patients may also respond. However, early studies suggest that the response rates of mild, moderate and severe MED patients may be greater in the selective D3 agonist/PDE 5 inhibitor combination. Mild, moderate and severe MED will be terms known to those skilled in The art, but knowledge can be found in The Journal of Urology, vol.151, 54-61(Jan 1994).

Early studies suggest that the group of MED patients described below should benefit from treatment with a selective D3 agonist and a PDE5 inhibitor (or other combination described below). These patient groups are described in detail in Clinical android vol.23, No.4, p773-782 and the book "Erectile dye function-Current Investigation and management" of I.Eardley and K.Sethia, chapter 3 of published by Mosby-Wolfe, as follows: psychogenic, organic, vascular, endocrine, neurological, arterial, drug-induced sexual dysfunction (lactogenesis) and sexual dysfunction associated with cavernous factors, particularly venous causes.

Accordingly, the present invention provides the use of a compound of formula (I), (Ia) or (Ib) in combination with a PDE5 inhibitor for the manufacture of a medicament for the treatment of erectile dysfunction.

Suitable PDE5 inhibitors are as described herein.

The compounds of the invention are useful for treating or preventing Female Sexual Dysfunction (FSD), in particular FSAD.

According to the present invention, FSD can be defined as a female being difficult or unable to find satisfaction in sexual expression. FSD is a general term for a number of different Female sexual disorders (Leiblum, S.R. (1998) -Definition and classification of Female sextual disorders. Int.J.Impower Res., 10, S104-S106; Berman, J.R., Berman, L. & Goldstein, I. (1999) -Female sextual dynamic: index, pathology, evaluation and treatment options. Urology, 54, 385- > 391). Women may have hypoactive libido, arousal or orgasm difficulties, dyspareunia, or a combination of these problems. Several types of disease, medications, injuries or psychological problems can lead to FSD. The therapeutic approaches under development are aimed at treating specific FSD subtypes, mainly sexual desire and arousal disorders.

The classification of FSD is preferably compared to the stage of normal female sexual response: sexual desire, arousal and orgasm (leiplus, s.r. (1998) -Definition and classification of the functional sextual disorders. int. j. immunity res., 10, S104-S106). Sexual desire or sexual impulse is the driving force for sexual expression. Its performance often includes sexual thoughts when a companion of interest accompanies or when exposed to other sexual stimuli. Arousal is a vascular response to sexual stimulation, an important component of which is genital engorgement, including increased vaginal lubricity, vaginal elongation, and increased genital sensation/sensitivity. Orgasm is the release of sexual tension that has reached orgasm during arousal.

Thus, FSD occurs when a woman does not respond adequately or satisfactorily in any of these phases (usually sexual desire, arousal or orgasm). The FSD classification includes hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder and dyspareunia. While the compounds of the present invention will improve the genital response to sexual stimulation (as in female sexual arousal disorder), at the same time it may also improve the pain, tension and discomfort associated with sexual intercourse, and thus also treat other female sexual disorders.

Hyposexuality exists if a woman has no or little desire for sex, and no or little thought or fantasy for sex. This type of FSD may be due to low testosterone levels resulting from natural menopause or surgical menopause. Other causes include disease, medication, fatigue, depression, and anxiety.

Female Sexual Arousal Disorder (FSAD) is characterized by an inadequate response of the genitals to sexual stimuli. The genitals do not experience congestion typical of normal sexual arousal. The lubricity of the vaginal wall is so poor that sexual intercourse is painful. Orgasm may also be hindered. Arousal disorders may result from estrogen deprivation during menopause or post partum and lactation, and may be due to diseases with vascular components, such as diabetes and atherosclerosis. Other causes include treatment with diuretics, antihistamines, antidepressants (e.g., Selective Serotonin Reuptake Inhibitors (SSRIs)), or antihypertensive agents.

Sexual pain disorders (including dyspareunia and vaginismus) are characterized by pain from insertion, which may be caused by drugs that reduce lubricity, endometriosis, pelvic inflammatory disease, inflammatory bowel disease or urinary tract problems.

As discussed previously, D3 is considered to be the initiator of sexual activity. The clitoris are considered to be homologous to the penis (Levin, R.J (1991), exp. clin. endocrinol., 98, 61-69); the same mechanism provides an erectile response in men and an increase in genital blood flow in women, with a related effect on FSD. In addition, proceptivity and sensitivity also vary.

Thus, according to a preferred aspect of the present invention there is provided the use of a compound of formula (I), (Ia) or (Ib) in the manufacture of a medicament for the treatment or prevention of female sexual dysfunction, more specifically hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder and dyspareunia.

Preferably, the compounds of formula (I) are useful for the treatment or prevention of sexual arousal disorders, orgasmic disorders and hypoactive sexual desire disorders, most preferably for the treatment or prevention of sexual arousal disorders.

In a preferred embodiment, the compounds of formula (I), (Ia) and (Ib) are useful for treating a subject suffering from female sexual arousal disorder with hypoactive sexual desire disorder.

The Diagnostic and Statistical Manual (DSM) IV of The American psychiatric Association defines Female Sexual Arousal Disorder (FSAD) as:

"… … failure to reach or maintain adequate lubrication of sexual excitement continuously or repeatedly-the swelling reaction is to complete sexual activity. This disorder certainly leads to significant distress or dyspareunia … … ".

Evoked responses consist of vascular engorgement of the pelvis, vaginal lubrication and distension, and swelling of the external genitalia. The disorder results in significant pain and/or dyspareunia.

FSAD is a very common sexual disorder affecting women during premenopausal, menopausal, and postmenopausal (+ Hormone Replacement Therapy (HRT)). It is associated with concomitant disorders such as depression, cardiovascular disease, diabetes and genitourinary (UG) disorders.

Preliminary consequences of FSAD are a lack of congestion/swelling, lack of lubrication, and lack of genital pleasure. Secondary consequences of FSAD are decreased libido, pain during intercourse, and difficulty reaching orgasm.

Recent hypothesis suggests that at least a portion of patients with FSAD symptoms have vascular-based problems (Goldstein et al, int.j. impot.res., 10, S84-S90, 1998), and animal data support this view (Park et al, int.j. impot.res., 9, 27-37, 1997).

Levin teaches us that because "… … male and female genitalia develop embryologically from a common tissue primordium, the male and female genital structures are said to be homologous to each other. Thus, the clitoris is a homolog of the penis, and the labia are homologous to the scrotum … … "(Levin, R.J. (1991), exp.

Efficacy drug candidates for FSAD under investigation are primarily erectile dysfunction therapies that promote blood circulation in the male genitalia.

The compounds of the present invention provide a means for restoring normal sexual arousal response, i.e., increased genital blood flow, leading to vaginal, clitoral and labial engorgement, and are therefore advantageous. This will result in increased vaginal lubricity via mucus leakage, increased vaginal compliance and increased genital sensitivity. Thus, the present invention provides a means to restore or enhance normal sexual arousal responses.

Thus, according to a preferred aspect of the present invention there is provided the use of a compound of formula (I), (Ia) or (Ib) in a medicament for the treatment or prevention of sexual arousal disorder in a female.

In this context, female genitalia means: the "reproductive organs consist of an inner and an outer part. Internal organs are located within the pelvis, consisting of the ovaries, uterine tube, uterus and vagina. The external organs are located outside the urogenital diaphragm and below the pelvic arch. They comprise the mons pubis, labia majora and labia minora, the clitoris, the vestibule, the bulbar vestibule, and the large vestibular gland "(Gray's Anatomy, c.d. clement, 13th American Edition).

The compounds of the invention can be applied to the following subpopulations of FSD patients: young, elderly, pre-menopausal, post-menopausal women, received or not had hormone replacement therapy.

The compounds of the invention may be used in patients with FSD for the following reasons:

i) vascular etiologies such as cardiovascular or atherosclerotic disease, hypercholesterolemia, smoking, diabetes, hypertension, radiation, perineal trauma, traumatic injury to the sub-iliac, ventral, pudendal vasculature.

ii) neurological etiologies such as spinal cord injury or central nervous system disease including multiple sclerosis, diabetes, parkinsonism, cerebrovascular accident, peripheral neuropathy, trauma or radical pelvic surgery.

iii) hormonal/endocrine etiologies such as dysfunction of the hypothalamic/pituitary/genital axis, dysfunction of the ovary, dysfunction of the pancreas, surgical or pharmaceutical castration, androgen deficiency, high circulating levels of prolactin (e.g. hyperprolactinemia), natural menopause, premature ovarian failure, hyperthyroidism and hypothyroidism.

iv) psychiatric etiology such as depression, obsessive compulsive disorder, anxiety, postpartum depression/"infant depression", emotional and relational problems, behavioral anxiety, marital discordance, dysfunctional attitudes, sexual phobia, religious contraindications, or traumatic past experiences.

v) drug-induced sexual dysfunction, caused by Selective Serotonin Reuptake Inhibitors (SSRis) and other antidepressant therapies (tricyclic and primary sedative tranquilizers), antihypertensive therapy, anti-sympathetic agents, long-term oral contraceptive pill therapy.

The compounds of the invention are also useful in the treatment of depression.

Dopamine D3 receptors are expressed almost exclusively in the limbic regions, which are involved in reward, emotional, and cognitive processes. Chronic treatment with several classes of antidepressants is known to increase D3 expression in the limbic region, and the antidepressant effect of desipramine can be blocked by sulpiride (a D2/D3 antagonist) but injected into the nucleus accumbens (region enriched in D3) rather than the caudate putamen (region enriched in dopamine D2 receptors). In addition, an antidepressant effect was observed in patients treated with pramipexole, an agonist preferring D3, and in preclinical models of depression. Available information suggests that the D3 receptor mediates antidepressant activity and that selective D3 receptor agonists represent a new class of antidepressants. As antidepressants are known to be effective in other psychotic disorders, D3 agonists would have potential for the treatment of psychosis.

The present invention provides the use of a selective D3 agonist in the manufacture of a medicament for the treatment of depression and psychiatric disorders.

Preferably, the D3 agonist exhibits functional potency at the D3 receptor with an EC50 of less than 1000nM, more preferably less than 100nM, even more preferably less than 50nM, most preferably less than 10 nM.

Preferably, the D3 agonist is more selective for D3 than D2, wherein the dopamine D3 receptor agonist is at least about 15-fold, preferably at least about 27-fold, more preferably at least about 30-fold, and most preferably at least about 100-fold more functionally selective for the dopamine D3 receptor than the dopamine D2 receptor.

Suitable conditions include depression (e.g., depression in cancer patients, depression in Parkinson's disease patients, post-myocardial infarction depression, sub-syndromic depression, depression in infertile women, childhood depression, major depression, single-episode depression, recurrent depression, depression induced by children with malaria, postpartum depression, and geriatric temperament syndrome), generalized anxiety disorder, phobias (e.g., agoraphobia, social phobia, and simple phobias), post-traumatic stress syndrome, avoidant personality disorder, eating disorders (e.g., anorexia nervosa and bulimia nervosa), obesity, chemical dependence (e.g., addiction to alcohol, cocaine, heroin, phenobarbital, nicotine, and benzodiazepines *), Alzheimer's disease, obsessive-compulsive disorder, panic disorder, memory disorders (e.g., dementia, amnesia, and age-related cognitive decline (ARCD)), depression in infertility, and other patients, Parkinson's disease (e.g. parkinson's dementia, neuro-inhibitor induced parkinsonism and tardive dyskinesia), endocrine disorders (e.g. hyperprolactinemia), vasospasm (particularly in the cerebral vasculature), cerebellar ataxia, negative symptoms of schizophrenia, stress urinary incontinence, tourette's syndrome, trichotillomania, kleptomania, Attention Deficit Hyperactivity Disorder (ADHD), chronic paroxysmal migraine, mood swings, pathological crying, sleep disorders (cataplexy) and shock.

In a preferred embodiment, the present invention provides the use of a compound of formula (I), (Ia) and (Ib) for the manufacture of a medicament for the treatment of depression or a psychiatric disorder.

Suitable depressive disorders and psychiatric disorders are as described above.

The compounds of the invention also have utility in the treatment of neurodegeneration; sources of neurodegeneration include neurotoxin intoxication; vision loss due to neurodegeneration of the visual pathway, such as stroke in the visual pathway (e.g., retina, optic nerve, and/or occipital lobe); seizures; the glucose and/or oxygen supply to the brain is reduced.

Accordingly, the present invention provides the use of a selective D3 agonist in the manufacture of a medicament for the treatment of neurodegeneration.

Preferably, the D3 agonist exhibits a functional potency, expressed as EC50, at the D3 receptor of less than 1000nM, more preferably less than 100nM, even more preferably less than 50nM, and most preferably less than 10 nM.

In a preferred embodiment, the D3 agonist is a compound of formula (I), (Ia) or (Ib).

In addition to their role in the treatment of sexual dysfunction, depression, neurodegeneration and psychiatric disorders, it is likely that the compounds of the present invention will be effective for a number of other indications.

Accordingly, the present invention provides the use of a compound of formula (I), (Ia) or (Ib) in the manufacture of a medicament for the treatment of hypertension, premature ejaculation, obesity, cluster headache, migraine, pain, endocrine disorders (e.g. hyperprolactinemia), vasospasm (particularly in the cerebral vasculature), cerebellar ataxia, gastrointestinal disorders (involving alterations in motility and secretion), premenstrual syndrome, fibromyalgia syndrome, stress urinary incontinence, trichotillomania, chronic paroxysmal migraine, headache (associated with vascular disorders).

D3/D2 agonist binding assays

Gonazalez et al (Eup. J Pharmacology 272(1995) R1-R3) disclose assays for determining the binding capacity of compounds to the D3 and/or D2 dopamine receptors, and thus the binding selectivity of such compounds. Accordingly, such assays are referred to herein as binding assays.

D3/D2 agonist functional assay

Assays suitable for functionally determining the activity of compounds at D3 and/or D2 dopamine receptors are described in detail below.

cAMP levels in GH4Cl and CHO cell lines expressing the human D2 and D3 receptors, respectively, were observed and the compounds were evaluated as agonists or antagonists of the dopamine D2 and D3 receptors.

Experimental process

Inhibition of forskolin-stimulated adenylate cyclase activity material via dopamine D3 receptor

Cell culture medium:

hD₃CHO culture medium
hD₃CHO culture medium	DMEM, high glucose (Sigma D5671)
2mM L-Glutamine (Sigma G7513)	DMEM, high glucose (Sigma D5671)
2mM L-Glutamine (Sigma G7513)	10% dialyzed FBS (Sigma F0392)

hD expressing human dopamine D3 receptor₃CHO (chinese hamster ovary) cells are generated in the laboratory. These cells are deficient in the dihydrofolate reductase gene.

Fresh media was prepared weekly as follows and filtered through a 0.22 μ M filter before use. The medium was stored at 4 ℃ and warmed to 37 ℃ before addition of the cells.

Cell Dispersion Solution (CDS): (SigmaC-5914)

5mL of the solution is used from 225cm²Cells were harvested from flasks (37 ℃ C., for hD2LGH4Cl cells)5min for hD3CHO cells 10 min).

Phosphate Buffered Saline (PBS): (Gibco 14040-

Trypan blue: (Sigma T8154)

Forskolin (Calbiochem 344273)

Dissolved in distilled water to a concentration of 20mM (the stock solution was stored at +4 ℃). A500-fold dilution in PBS buffer was made to prepare 40. mu.M stock solutions for 4 × assay. 25 μ L of 40 μ M stock solution was added to a final assay volume of 100 μ L to give a final assay concentration of 10 μ M.

Test compounds

Dissolved in 100% DMSO to give a stock concentration of 10 mM.

Pramipexole standard

Dissolved in 100% DMSO to give a stock concentration of 10 mM.

Cyclase activation Flashplate assay (NEN SMP004B)

Supplied by Perkin-Elmer Life Sciences, Inc.

[125I] Cyclic adenosine monophosphate (cAMP) (NEX 130)

Supplied by Perkin-Elmer Life Sciences, Inc.

Concrete equipment

Westbart micro-titration flat-plate shaking table/incubator

Packard Topcount NXT (ECADA compatibility program)

Tecan Genesis

Labsystems Multi-drop DW

By hD₃Protocol for testing compound activity in CHO cells

Compound diluent

Pramipexole was included as a reference standard. A 10 point semilogarithmic curve was generated from every 4 plates. Compound results were normalized to the minimum (0nM pramipexole) and maximum (100nM pramipexole) responses produced by cells. All test compounds can also be tested via a 10-point (semi-logarithmic) curve.

Test compounds were dissolved in 100% DMSO to give a stock concentration of 10 mM. They were further diluted to 1mM in 100% DMSO via 10-fold dilution (1000x the desired final assay concentration, e.g. 1mM would give a 1 μ M apical concentration).

Pramipexole was dissolved in 100% DMSO to give a concentration of 10 mM. Pramipexole was further diluted to 0.1mM in 100% DMSO via 100-fold dilution.

Further dilutions and additions were made in 0.4% DMSO/PBS using a suitable Tecan Genesis protocol, which enables serial dilutions (semilog units) 3.159-fold.

TECAN GENESIS Diluent

Add 10. mu.L of test compound to column 1 of the microplate. 240 μ L of 0.4% DMSO/PBS was added to give a 25-fold dilution (0.04 mM). mu.L of 0.04mM dilution was transferred to column 2 wells and 180. mu.L of 0.4% DMSO/PBS was added to give a further 10-fold dilution to reach 4 Xthe top assay concentration (0.004 mM).

Serial dilutions (3.159-fold) were performed to achieve a semilog dilution series: 4 μ M, 1.27 μ M, 400nM, 127nM, 40nM, 13nM, 4nM, 1.27nM, 0.4nM, 0.1nM

25 μ L (in duplicate) serial dilutions were transferred to Flashplate at columns 2-11 (see appendix). Since the final assay volume is 100 μ L, the final assay concentration will be: 1000. mu.M, 317nM, 100nM, 32nM, 10nM, 3.2nM, 1nM, 0.3nM, 0.1nM, 0.03nM

Minimum control (low control): to column 1 wells E-H and column 2 wells A-D was added 25. mu.L of 0.4% DMSO/PBS (vehicle). Later cells + forskolin were added.

Maximum control (high control): pramipexole at 10mM was diluted in PBS (10 μ L +2490 μ L PBS) via 250-fold dilution, yielding 40 μ M pramipexole. Further 40 μ M pramipexole was diluted in 0.4% DMSO/PBS (100 μ L +9900 μ L vehicle) via 100-fold dilution, yielding 400nM (4x measured concentration of standard pramipexole). 25 μ L of 400nM pramipexole was added to Flashplate column 1 wells A-D and column 12 wells E-H to give the final 100nM pramipexole. Later cells + forskolin were added.

Cyclase activation Flashplate assay (NEN SMP004B)

Forskolin was dissolved in distilled water to a stock solution concentration of 20mM as described in material section. Further diluted to 40 μ M with PBS (4 × assay concentration). 25 μ L of 40 μ M stock solution was added to all wells using a Multi-drop to give a final concentration of 10 μ M. The plates were then sealed, incubated in a 37 ℃ Westbart incubator, and the cells were harvested.

Cells were harvested from flasks with a confluency between 70% and 80%. It is necessary to warm all the components added to the wells to 37 ℃. 5mLCDS was added to each T225 flask, incubated at 37 ℃ for 5 minutes, and then neutralized with 5mL PBS. The cells were then centrifuged at 160g (1000rpm) for 5 minutes. The resulting supernatant was discarded and the cells were resuspended in stimulation buffer (warmed to 37 ℃) to 5X 10⁵cells/mL. Then 50. mu.L of the cell suspension was dispensed into all wells of the Flashplate.

Immediately incubate the plate on a shaking incubator at 37 ℃ for 15 minutes. The reaction was stopped in all wells with 100. mu.L of assay mix (100. mu.L per plate)¹²⁵I cAMP: 11mL of detection buffer).

Resealing the plate and incubating in the dark for 3 hours to allow for incubation of the anti-cAMP antibody (coating each well)¹²⁵I]Equilibrium is reached between cAMP tracer and cellular cAMP.

Plates were counted on a PackardTopcount NXT using the appropriate ECADA compatibility Protocol (Protocol 75).

Resuscitation of cryo ampoules

The ampoules were removed from the liquid nitrogen and allowed to equilibrate for 2 minutes as the trapped gas or liquid could cause the ampoules to rapidly expand and explode. They may also be left at-20 ℃ for 2 minutes prior to thawing.

The ampoule was rapidly and completely thawed in a 37 ℃ water bath.

Transfer the cell suspension to 75cm containing 10mL of growth medium²In a flask, 5% CO at 37 ℃₂And then incubated for 24 hours. After cell attachment (3-6 hours), the medium was removed and replaced with fresh medium (to remove DMSO). After 24 hours, if fusion is approached, the cells are transferred to 225cm²In a flask. If not, the cells are continued to be cultured until they reach 70% -80% confluence.

Cell harvesting and bottling

Cells were bottled on friday to provide cells for the assay on monday and tuesday. The cells needed for the rest of the week were in weekly flasks.

It is necessary not to make hD₃CHO cells grow more than 80% confluent, or at a split flask ratio > 1: 20, as this has a deleterious effect on their proliferative response and will subsequently affect the ability of the cells to perform assays.

The cells were grown at 225cm²In a flask (Jumbos). Each component added to the cells must be warmed to 37 ℃ prior to use.

Cell harvesting

Growth medium was removed from the flasks and the cells were removed by washing with warm PBS (Gibco.14040-091).

5mL of cell dispersion buffer was added to the cells, and the cells were placed in an incubator for about 5 minutes.

The flasks were tapped sharply to tap off all remaining cells from the tissue culture plastic.

Add 5mL PBS to the cells for washing the substrate and flask. The cells were centrifuged at 160g (1000rpm) for 5 minutes to pellet the cells.

Discard the supernatant and resuspend the cells in 5mL of stimulation buffer. Trypan blue exclusion assay was performed to determine the number of viable cells.

Cells were diluted in stimulation buffer to a concentration of 5X 10⁵cells/mL.

When passaging cells, the centrifugation step was omitted and the cell suspension was dispensed into new T225 flasks containing 50mL of medium.

Bottle dividing ratio

hD₃The bottle division ratio of CHO is 1: 5 to 1: 10. The culture cannot be continued for more than 30 passages because the cell line characteristics are lost as the number of passages increases.

Cryopreservation of cell lines

It is necessary to create a cell bank of your own cells to revive for further use.

Harvest cells as described in the previous section. After trypan blue exclusion assay, cells were diluted in medium containing 10% DMSO to 2 to 4 × 10⁶cells/mL.

Cells were divided into 1mL aliquots, immediately frozen in "Mr frost" (containing fresh IPA), gradually cooled to-80 ℃, and then transferred to a gas phase liquid nitrogen storage vessel (cells can be stored in "Mr frost" for up to 2 days).

It is also desirable to thaw an ampoule after freezing to test cell viability. Viability below 70% may cause recovery problems due to low cell numbers and debris.

Data analysis

Data were analyzed using ECADA.

Normalized% (relative to pramipexole) for all compounds was obtained via the following formula:

normalized% (X-B0)/(Max-B0) × 100

Wherein

X-the average net cell number for a given concentration of test compound,

b0 (average net cell number of min control (0nM pramipexole),

max is the average net cell number of the maximum control (100nM pramipexole).

Normalized% (y) was plotted against agonist concentration nm (x) to generate a curve. The slope was approximately 1 using a nonlinear regression fit to the data. The EC50 and% Emax of the test compounds were thus determined.

Assay plate layout (10 point EC 50):

	1	2	3	4	5	6	7	8	9	10	11	12
	1	2	3	4	5	6	7	8	9	10	11	12	A	MAX	C1	MIN
B	MAX	C1										MIN	A	MAX	C1	MIN
B	MAX	C1										MIN	C	MAX	C2	MIN
D	MAX	C2										MIN	C	MAX	C2	MIN
D	MAX	C2										MIN	E	MIN	C3	MAX
F	MIN	C3										MAX	E	MIN	C3	MAX
F	MIN	C3										MAX	G	MIN	C4	MAX
H	MIN	C4										MAX	G	MIN	C4	MAX

column 1: and the small hole A-D is MAX: high control (cell + forskolin +100nM pramipexole)

Small hole E-H ═ MIN: low control (cell + forskolin + vehicle)

Column 12: and (4) small hole A-D is MIN: low control (cell + forskolin + vehicle)

Small hole E-H MAX: high control (cell + forskolin +100nM pramipexole)

Columns 2-11: serial 10-point dilutions (in duplicate) of test compounds. Concentrations decreased from column 2 to column 11 (1000nM to 0.03 nM). In the first plate C1 was replaced with pramipexole.

Inhibition of forskolin-stimulated adenylate cyclase activity via dopamine D2 receptor

Material

Cell culture medium:

HD2 GH4C1/hD_2Lculture medium
HD2 GH4C1/hD_2Lculture medium	Hams F-12(Sigma N6013)
2mM L-Glutamine (Sigma G7513)	Hams F-12(Sigma N6013)
2mM L-Glutamine (Sigma G7513)	10％FBS(Gibco 10106-169)
700. mu.g/mL Geneticin (Gibco 10131-	10％FBS(Gibco 10106-169)

GH4Cl/hD_2LIs expressing human dopamine D2_{Long and long}Rat pituitary cells of the recipient.

Cell Dispersion Solution (CDS): (Sigma C-5914)

5mL of the solution is used from 225cm²The cells were harvested from the flask.

Phosphate Buffered Saline (PBS): (Gibco 14040-

Trypan blue: (Sigma T8154)

Forskolin (Calbiochem 344273)

Dissolved in distilled water to a concentration of 20mM (the stock solution was stored at +4 ℃). A1000-fold dilution in PBS buffer was made to prepare 20. mu.M stock solutions for 4 × assay. 25 μ L of 20 μ M stock solution was added to a final assay volume of 100 μ L to give a final assay concentration of 5 μ M.

Test compounds

Dissolved in 100% DMSO to give a stock concentration of 10 mM.

Pramipexole standard

Dissolved in 100% DMSO to give a stock concentration of 10 mM.

Cyclase activation Flashplate assay (NEN SMP004B)

Supplied by Perkin-Elmer Life Sciences, Inc.

[¹²⁵I]Cyclic adenosine monophosphate (cAMP) (NEX 130)

Supplied by Perkin-Elmer Life Sciences, Inc.

Concrete equipment

Westbart micro-titration flat plate shaker/incubator

Packard Topcount NXT (ECADA compatibility program)

Tecan Genesis

Labsystems Multi-drop DW

Scheme(s)

Compound diluent

Pramipexole was included as a reference standard. A 10 point semilogarithmic curve was generated for each 4 plates. Compound results were normalized to the minimum (0nM pramipexole) and maximum (1000nM pramipexole) responses produced by the cells. All test compounds can also be tested via a 10-point (semi-logarithmic) curve.

Test compounds were dissolved in 100% DMSO to give a stock concentration of 10mM (1000x the desired final assay concentration, e.g. 10mM would give a top concentration of 10000 nM).

Pramipexole was dissolved in 100% DMSO to give a concentration of 10 mM. Pramipexole was further diluted to 1mM in 100% DMSO via a 10-fold dilution.

TECAN GENESIS Diluent

Add 10. mu.L of test compound to column 1 of the microplate. To this was added 240. mu.L of 90 in 0.4% DMSO/PBS to give a 25-fold dilution (0.4 mM). mu.L of 0.4mM dilution was transferred to column 2 wells and 180. mu.L of 0.4% DMSO/PBS was added to give a further 10-fold dilution to reach 4 Xthe top assay concentration (0.04 mM).

Serial dilutions (3.159-fold) were performed to achieve a semilog dilution series: 40 μ M, 12.7 μ M, 4 μ M, 1.27 μ M, 400nM, 130nM, 40nM, 13nM, 4nM, 1.3nM

25 μ L (in duplicate) serial dilutions were transferred to Flashplate at columns 2-11 (see appendix). Since the final assay volume is 100 μ L, the final assay concentration will be: 10,000. mu.M, 3170nM, 1000nM, 320nM, 100nM, 32nM, 10nM, 3nM, 1nM, 0.3nM

Maximum control (high control): pramipexole at 10mM was diluted in PBS (10 μ L +2490 μ L PBS) via 250-fold dilution, yielding 40 μ M pramipexole. Further dilution of 40 μ M pramipexole in 0.4% DMSO/PBS (100 μ L +990 μ L vehicle) via 10-fold dilution yielded 4000nM (4x measured concentration of standard pramipexole). 25 μ L of 4000nM pramipexole was added to Flashplate column 1 orifice A-D and column 12 orifice E-H to obtain the final 1000nM pramipexole. Then cell + forskolin was added.

Cyclase activation Flashplate assay (NEN SMP004B)

Forskolin was dissolved in distilled water to a stock solution concentration of 20mM as described in material section. Further diluted to 20 μ M with PBS (4 × assay concentration). 25 μ L was added to all wells using a Multi-drop to give a final concentration of 5 μ M. The plates were then sealed and incubated in a Westbart incubator at 37 ℃ while the cells were harvested.

Harvest cells from flasks with fusion rate between 70% and 80%. It is necessary to warm all the components added to the cells to 37 ℃. To each 225cm²The flask was charged with 5mL CDS, incubated at 37 ℃ for 5 minutes, and then neutralized with 5mL PBS. The cells were then centrifuged at 160g (1000rpm) for 5 minutes. The resulting supernatant was discarded and the cells were resuspended in stimulation buffer (warmed to 37 ℃) to 1X 10⁵cells/mL. Then 50. mu.L of the cell suspension was dispensed into all wells of the Flashplate.

Resealing the plate and incubating in the dark for 3 hours to allow for incubation in the anti-cAMP antibody (coated wells)¹²⁵I]Equilibrium is reached between cAMP tracer and cellular cAMP.

Resuscitation of cryo ampoules

The ampoule was rapidly and completely thawed in a 37 ℃ water bath.

Transfer the cell suspension to 75cm containing 10mL of growth medium²In a flask, 5% CO at 37 ℃₂And then incubated for 24 hours. After cell attachment (3-6 hours), the medium was removed and replaced with fresh medium (to remove DMSO). After 24 hours, if fusion is approached, the cells are transferred to 225cm²In a flask. If not, the cells are cultured until their fusion rate is 60%.

Cell harvesting and bottling

Cells were bottled on friday to provide cells assayed on monday and tuesday. The cells needed for the rest of the week were in weekly flasks.

It is necessary to avoid growing cells beyond 60% confluency, as this has a deleterious effect on their proliferative response and will subsequently affect the ability of the cells to perform assays.

Cell harvesting

Add 5mL of cell dispersion buffer to the cells and place in the incubator for approximately 5 minutes.

The flask was swiped sharply to tap off all remaining cells from the tissue culture plastic bottle.

Cells were diluted in stimulation buffer to a concentration of 1X 10⁵cells/mL.

For passaging cells, the centrifugation step is omitted and the cell suspension is dispensed into new T225 flasks containing 50mL of medium.

Bottle dividing ratio

The flask division ratio of GH4Cl/D2 is between 1: 3 and 1: 5.

Cryopreservation of cell lines

Data analysis

Data were analyzed using ECADA.

All compounds were normalized (relative to pramipexole) via the following formula:

normalized% (X-B0)/(Max-B0) × 100

Wherein

X-the average net cell number for a given concentration of test compound,

b0 (average net cell number of min control (0nM pramipexole),

max is the average net cell number of the maximum control (100nM pramipexole).

Normalized% (y) was plotted against agonist concentration nm (x) to generate a curve. The slope was approximately 1 using a nonlinear regression fit to the data. Thus, the test compounds were tested for EC50 and% Emax.

Assay plate layout (10 point EC 50):

Small hole E-H ═ MIN: low control (cell + forskolin + vehicle)

Small hole E-H MAX: high control (cell + forskolin +100nM pramipexole)

Using the above assay, the compounds of the invention all had functional potency at the D3 receptor (expressed as EC50, below 1000nM) and were more selective for D3 than D210-fold.

The compound described in example 8 has a functional potency at the D3 receptor expressed as EC50 of 7.6nM, with a selectivity for D3 that is greater than that of D21315.8-fold. Selectivity was calculated by dividing the D2 EC50 value by the D3 EC50 value. If D2 EC50 is greater than 10000, 10000 is taken for calculation.

It is understood that all references herein to treatment include curative, palliative and prophylactic treatment.

Auxiliary active ingredients suitable for use in the combinations of the invention include:

1) naturally occurring or synthetic prostaglandins or esters thereof. Prostaglandins suitable for use herein include compounds such as alprostadil, prostaglandin E₁Prostaglandin E₀13, 14-dihydroprostaglandin E₁Prostaglandin E₂Natural, synthetic and semi-synthetic prostaglandins and derivatives thereof, including those described in WO 00/33825 and/or US 6,037,346 issued 3/14 of 2000 (all of which are incorporated herein by reference), PGE₀、PGE₁、PGA₁、PGB₁、PGF₁Alpha, 19-hydroxy-PGA₁19-hydroxy-PGB₁、PGE₂、PGB₂19-hydroxy-PGA₂19-hydroxy-PGB₂、PGE₃Alpha, carboprost tromethamine, dinoprost tromethamine, dinoprostone, lipoprost, gemeprost, meprost (metaloprost), sulprostone (sulprostune), tiaprost and moxider;

2) alpha-adrenergic receptor antagonist compounds, also known as alpha-adrenergic receptors or alpha-blockers. Suitable compounds for use herein include: alpha-renal as described in PCT application WO 99/30697 published on 6/14 of 1998Adrenergic receptor blockers (the disclosure of which is related to alpha-adrenergic receptors is incorporated herein by reference), including selective alpha₁-adrenoceptor or alpha₂-adrenoceptor blockers, and non-selective adrenoceptor blockers, suitably alpha₁-adrenoceptor blockers comprise: phentermine, phentermine mesylate, trazodone, alfuzosin, indoramine, naftopidil, tamsulosin, dapiprazole, phenoxybenzamine, idazoxan, efaroxan, yohimbine, rauvolfia alkaloids, Recordati 15/2739, SNAP 1069, SNAP 5089, RS17053, SL 89.0591, doxazosin, terazosin, abanoquine, and prazosin; alpha from US 6,037,346 (3.2000, 14 days)₂-the blockers diphenylamin, tolazoline, tramazosin and diphenylamin; the α -adrenergic receptors are described in the following U.S. patents: 4,188,390, 4,026,894, 3,511,836, 4,315,007, 3,527,761, 3,997,666, 2,503,059, 4,703,063, 3,381,009, 4,252,721, and 2,599,000, each of which is incorporated herein by reference; alpha is alpha₂-adrenoceptor blockers comprise: clonidine, papaverine hydrochloride, optionally in the presence of a cardiotonic agent such as pirxamine;

3) NO-donor (NO-agonist) compounds. NO-donor compounds suitable for use herein include organic nitrates, such as mono-, di-or tri-nitrates, or organic nitrates, including glyceryl trinitrate (also known as nitroglycerin), isosorbide 5-mononitrate, isosorbide dinitrate, pentaerythritol tetranitrate, erythritol tetranitrate, Sodium Nitroprusside (SNP), 3-morpholino sydnomine, molsidomine, S-nitroso-N-acetylpenicillamine (SNAP), S-nitroso-N-glutathione (SNO-GLU), N-hydroxy-L-arginine, amyl nitrate, linsidomine chloride hydrate (SIN-1), S-nitroso-N-cysteine, diazoniadiolates (NONO acid esters), 1, 5-pentane dinitrate, L-arginine, ginseng, jujube, molsidomine, Re-2047, nitrosylated derivatives of moxideril, such as NMI-678-11 and NMI-937, as described in published PCT application WO 00/12075;

4) potassium channelsOpeners or modulators. Potassium channel openers/modulators suitable for use herein include nicorandil, chromancalin, levochromancalin, ramacarin, pinacidil, cliazoxide, minoxidil, charybdotoxin, glyburide, 4-aminopyridine, BaCl₂；

5) A vasodilator. Vasodilators suitable for use herein include nimodipine, pinadil, cyclandelate, isoxsuprine, chloroprumazine, haloperidol, Rec 15/2739, trazodone;

6) thromboxane a2 agonist;

7) a CNS active agent;

8) an ergot alkaloid. Suitable ergot alkaloids are described in us patent 6,037,346 issued 3.14.2000, including aceergamine, bromoergoline, bromoergourea, cyanoergoline, diorgoline (delorgotrile), disulerine, ergonovine maleate, ergotamine tartrate, ethergine, ergonitrile, ergodiethylamine, mesuler, ergoline, metergoline, nicergoline, pergolide, prolide, proterguride, and terguride;

9) compounds which modulate the action of natriuretic factors, in particular atrial natriuretic peptides (also known as atrial natriuretic peptides), natriuretic factors of type B and type C, such as neutral endopeptidases or inhibitors;

10) angiotensin converting enzyme inhibiting compounds, such as enalapril, and angiotensin converting enzyme in combination with neutral endopeptidase inhibitors, such as omatrazole;

11) angiotensin receptor antagonists such as losartan;

12) NO synthase substrates, such as L-arginine;

13) calcium channel blockers such as amlodipine;

14) endothelin receptor antagonists and endothelin converting enzyme inhibitors;

15) cholesterol lowering agents, such as statins (e.g. atorvastatin/Lipitor-trademark) and fibrates;

16) antiplatelet and antithrombotic agents, such as tPA, uPA, warfarin, hirudin and other thrombin inhibitors, heparin, tissue thromboplastin activator inhibitors;

17) insulin sensitizers, such as troglitazone tablets, and hypoglycemic agents, such as glipizide;

18) L-DOPA or carbidopa;

19) acetylcholinesterase inhibitors, such as donezipil;

20) steroidal or non-steroidal anti-inflammatory agents;

21) estrogen receptor modulators and/or estrogen agonists and/or estrogen antagonists, preferably raloxifene or lasofoxifene, (-) -cis-6-phenyl-5- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl ] -5, 6, 7, 8-tetrahydronaphthalene-2-ol and pharmaceutically acceptable salts thereof, the preparation of which is described in WO 96/21656;

22) PDE inhibitors, more precisely PDE2, 3, 4, 5, 7 or 8 inhibitors, preferably PDE2 or PDE5 inhibitors, most preferably PDE5 inhibitors (see below), which preferably have an IC50 of less than 100nM for the corresponding enzyme (with the proviso that the PDE3 and 4 inhibitors are administered only locally or by penile injection);

23) vasoactive Intestinal Peptide (VIP), VIP mimetics, VIP analogues, are more specifically mediated by: one or more VIP receptor subtypes VPAC1, VPAC or PACAP (pituitary adenylate cyclase activating peptide), one or more VIP receptor agonists or VIP analogs (e.g., Ro-125-1553) or VIP fragments, one or more alpha-adrenoceptor antagonists in combination with VIP (e.g., invicor, Aviptadil);

24) melanocortin (melanocortin) receptor agonists or modulators or melanocortin enhancers, e.g., melanotan II, PT-14, PT-141 or as claimed in WO 99/64002, WO00/74679, WO 99/55679, WO 01/05401, WO 00/58361, WO 01/14879, WO 01/13112, WO 99/54358;

25) serotonin receptor agonists, antagonists or modulators, more specifically agonists, antagonists or modulators of the 5HT1A (including VML 670), 5HT2A, 5HT2C, 5HT3 and/or 5HT6 receptors, including those described in WO 99/02159, WO 00/02550 and/or WO 00/28993;

26) testosterone substitutes (including dehydroandrosterone), testosterone (Tostrelle), dihydrotestosterone, or testosterone implants;

27) estrogens, estrogens with medroxyprogesterone or medroxyprogesterone acetate (MPA) (i.e., as a combination) or estrogens with methyltestosterone hormone replacement therapies (e.g., HRT, especially Premarin, Cenestin, oestrofemal, Equin, Estrace, Estrofem, eleste Solo, Estring, eastaderm TTS, eastaderm Matrix, Dermestril, primrose, Preempro, primak, primique, estempnext, estest HS, Tibolone);

28) modulators of norepinephrine, dopamine, and/or serotonin transporters, such as bupropion, GW-320659;

29) purinergic receptor agonists and/or modulators;

30) neurokinin (NK) receptor antagonists including those described in WO 99/64008;

31) an opioid receptor agonist, antagonist or modulator, preferably an agonist of the ORL-1 receptor;

32) an oxytocin/vasopressin receptor agonist or modulator, preferably a selective oxytocin agonist or modulator;

33) cannabinoid receptor modulators;

34) SEP inhibitors (SEPi), for example SEPi with an IC50 of less than 100nM, more preferably less than 50 nM. Preferably, the SEP inhibitors according to the invention have a selectivity for SEP of more than 30-fold, more preferably more than 50-fold over neutral endopeptidase NEP EC 3.4.24.11 and Angiotensin Converting Enzyme (ACE) selection. Preferably, the SEPi also has a selectivity greater than 100 times better than Endothelin Converting Enzyme (ECE).

By cross-reference herein to compounds contained in patents and patent applications that may be employed in accordance with the present invention, we define the therapeutically active compounds in the claims (especially claim 1) and the specific examples (all incorporated herein by reference).

If a combination of active ingredients is administered, they may be administered simultaneously, separately or sequentially.

Auxiliary ingredient PDE5 inhibitor

The suitability of any particular cGMP PDE5 inhibitor can be readily determined by evaluating its potency and selectivity using literature procedures, followed by evaluating its toxicity, absorption, metabolism, pharmacokinetics, and the like in accordance with standard pharmaceutical practice.

The IC50 values of cGMP PDE5 inhibitors can be determined using the PDE5 assay (see below).

Preferably, the cGMP PDE5 inhibitor used in the pharmaceutical combination according to the invention is selective for the PDE5 enzyme. Preferably (when used orally) they have a selectivity over PDE3, more preferably over PDE3 and PDE 4. Preferably (when administered orally) the cGMPPDE5 inhibitor of the invention has a selectivity ratio over PDE3, more preferably over PDE3 and PDE4 of more than 100, more preferably more than 300.

The skilled person can easily determine the selectivity ratio. IC50 values for the PDE3 and PDE4 enzymes can be determined using established literature methods, see S A Ballard et al, Journal of urology, 1998, vol.159, pages 2164-2171 and described below.

cGMP PDE5 inhibitors suitable for use in the present invention include:

pyrazolo [4, 3-d ] pyrimidin-7-ones as disclosed in EP-A-0463756; pyrazolo [4, 3-d ] pyrimidin-7-ones as disclosed in EP-A-0526004; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international patent application WO 93/06104; the isomeric pyrazolo [3, 4-d ] pyrimidin-4-ones as disclosed in published international patent application WO 93/07149; quinazolin-4-one as disclosed in published international patent application WO 93/12095; pyrido [3, 2-d ] pyrimidin-4-one as disclosed in published international patent application WO 94/05661; purin-6-ones disclosed in published international patent application WO 94/00453; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international patent application WO 98/49166; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international patent application WO 99/54333; pyrazolo [4, 3-d ] pyrimidin-4-ones as disclosed in EP-A-0995751; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international patent application WO 00/24745; pyrazolo [4, 3-d ] pyrimidin-4-ones as disclosed in EP-A-0995750; the compounds disclosed in published international application WO 95/19978; the compounds disclosed in published international application WO 99/24433; the compounds disclosed in published international application WO 93/07124; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international application WO 01/27112; pyrazolo [4, 3-d ] pyrimidin-7-one as disclosed in published international application WO 01/27113; the compounds disclosed in EP-A-1092718; the compounds disclosed in EP-A-1092719.

PDE5 inhibitors further suitable for use in the present invention include:

5- [ 2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (sildenafil), also known as 1- [ [3- (6, 7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4, 3-d ] pyrimidin-5-yl) -4-ethoxyphenyl ] sulfonyl ] -4-methylpiperazine (see EP-A-0463756); 5- (2-ethoxy-5-morpholinoacetylphenyl) -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see EP-A-0526004); 3-ethyl-5- [5- (4-ethylpiperazin-1-ylsulfonyl) -2-n-propoxyphenyl ] -2- (pyridin-2-yl) methyl-2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 98/49166); 3-ethyl-5- [5- (4-ethylpiperazin-1-ylsulfonyl) -2- (2-methoxyethoxy) pyridin-3-yl ] -2- (pyridin-2-yl) methyl-2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 99/54333); (+) -3-ethyl-5- [5- (4-ethylpiperazin-1-ylsulfonyl) -2- (2-methoxy-1 (R) -methylethoxy) pyridin-3-yl ] -2-methyl-2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, also known as 3-ethyl-5- {5- [ 4-ethylpiperazin-1-ylsulfonyl ] -2- ([ (1R) -2-methoxy-1-methylethyl ] oxy) pyridin-3-yl } -2-methyl-2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 99/54333); 5- [ 2-ethoxy-5- (4-ethylpiperazin-1-ylsulfonyl) pyridin-3-yl ] -3-ethyl-2- [ 2-methoxyethyl ] -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one, also known as 1- { 6-ethoxy-5- [ 3-ethyl-6, 7-dihydro-2- (2-methoxyethyl) -7-oxo-2H-pyrazolo [4, 3-d ] pyrimidin-5-yl ] -3-pyridylsulfonyl } -4-ethylpiperazine (see WO 01/27113, example 8); 5- [ 2-isobutoxy-5- (4-ethylpiperazin-1-ylsulfonyl) pyridin-3-yl ] -3-ethyl-2- (1-methylpiperidin-4-yl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 01/27113, example 15); 5- [ 2-ethoxy-5- (4-ethylpiperazin-1-ylsulfonyl) pyridin-3-yl ] -3-ethyl-2-phenyl-2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 01/27113, example 66); 5- (5-acetyl-2-propoxy-3-pyridyl) -3-ethyl-2- (1-isopropyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d1 pyrimidin-7-one (see WO 01/27112, example 124); 5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (see WO 01/27112, example 132); (6R, 12aR) -2, 3, 6, 7, 12, 12 a-hexahydro-2-methyl-6- (3, 4-methylenedioxyphenyl) -pyrazino [2 ', 1': 6, 1] pyrido [3, 4-b ] indole-1, 4-dione (IC-351), also known as the compound of examples 78 and 95 in published International application WO 95/19978, and as the compound of examples 1, 3, 7 and 8; 2- [ 2-ethoxy-5- (4-ethyl-piperazin-1-yl-1-sulfonyl) -phenyl ] -5-methyl-7-propyl-3H-imidazo [5, 1-f ] [1, 2, 4] triazin-4-one (vardenafil), also known as 1- [ [3- (3, 4-dihydro-5-methyl-4-oxo-7-propylimidazo [5, 1-f ] -as-triazin-2-yl) -4-ethoxyphenyl ] sulfonyl ] -4-ethylpiperazine, i.e. the compounds of examples 20, 19, 337 and 336 in published international application WO 99/24433; the compound of example 11 (EISAI) in published international application WO 93/07124; and compounds 3 and 14 from Rotella D P, j.med.chem., 2000, 43, 1257.

Still other suitable PDE5 inhibitors include:

4-bromo-5- (pyridylmethylamino) -6- [3- (4-chlorophenyl) -propoxy ] -3(2H) pyridazinone; 1- [4- [ (1, 3-benzodioxol-5-ylmethyl) amino ] -6-chloro-2-quinazolinyl ] -4-piperidine-carboxylic acid, monosodium salt; (+) -cis-5, 6a, 7, 9, 9, 9 a-hexahydro-2- [4- (trifluoromethyl) -phenylmethyl-5-methyl-cyclopent [4, 5] imidazo [2, 1-b ] purin-4 (3H) one; furazlocillin; cis-2-hexyl-5-methyl-3, 4, 5, 6a, 7, 8, 9, 9 a-octahydrocyclopent [4, 5] -imidazo [2, 1-b ] purin-4-one; 3-acetyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylate; 3-acetyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylate; 4-bromo-5- (3-pyridylmethylamino) -6- (3- (4-chlorophenyl) propoxy) -3- (2H) pyridazinone; 1-methyl-5- (5-morpholinoacetyl-2-n-propoxyphenyl) -3-n-propyl-1, 6-dihydro-7H-pyrazolo (4, 3-d) pyrimidin-7-one; 1- [4- [ (1, 3-benzodioxol-5-ylmethyl) amino ] -6-chloro-2-quinazolinyl ] -4-piperidinecarboxylic acid, monosodium salt; pharmaprojects No.4516 (GlaxoWellcome); pharmaprojects No.5051 (Bayer); pharmaprojects No.5064(Kyowa Hakko; see WO 96/26940); pharmaprojects No.5069(Schering Plough); GF-196960 (GlaxoWellcome); e-8010 and E-4010 (Eisai); bay-38-3045 & 38-9456(Bayer) and Sch-51866.

The compounds of formula (I) may be administered alone but will generally be administered in admixture with a suitable pharmaceutical excipient, diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice.

Accordingly, the present invention provides a composition comprising a compound of formula (I), (Ia) or (Ib) and a pharmaceutically acceptable diluent or carrier.

For example, the compounds of formula (I), (Ia) or (Ib) may be administered orally, buccally or sublingually in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavouring or colouring agents, for immediate, delayed, modified, sustained, pulsed or controlled release applications.

Such tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, Hydroxypropylmethylcellulose (HPMC), Hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Additionally, lubricants such as magnesium stearate, stearic acid, glycerol behenate and talc may be included.

Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, cellulose, milk sugar or high molecular weight polyethylene glycols. For aqueous suspensions and/or elixirs, the compounds of formula (I), (Ia) or (Ib) may be combined with various sweetening or flavoring agents, coloring matter or dyes, with emulsifying and/or suspending agents, with diluents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.

The compounds of formula (I), (Ia) or (Ib) may also be administered parenterally, for example intravenously, intraarterially, intraperitoneally, intrathecally, intraventricularly, intraurethrally, intrasternally, intracranially, intramuscularly or subcutaneously, or they may be administered by means of infusion techniques. For such parenteral administration, they are best used in the form of sterile aqueous solutions which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solution should be suitably buffered (preferably to a pH of 3-9) if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

The compounds of formula (I), (Ia) or (Ib) may also be administered intranasally or by inhalation, typically in the form of a dry powder (either by itself or as a dry mixture with lactose or as a mixture of component particles, for example mixed with phospholipids) delivered as a dry powder inhaler or aerosol spray from a pressurised container, pump, nebuliser or nebuliser (preferably a nebuliser in which a fine mist is generated using electro-hydraulics), with or without the use of a suitable propellant, for example dichlorofluoromethane.

The pressurised container, pump, spray or nebuliser contains a solution or suspension of the active compound, e.g. comprising ethanol (optionally aqueous ethanol) or a suitable substitute for the dispersed, dissolved or time-delayed release of the active ingredient, a propellant as solvent and optionally a surfactant, e.g. sorbitan trioleate or oligolactic acid.

Prior to use in dry powder or suspension formulations, the drug product is comminuted to a size suitable for delivery by inhalation (typically less than 5 microns). This can be achieved by any suitable comminution method, such as spiral jet milling, fluidized bed jet milling, supercritical fluid processing into nanoparticles, high pressure homogenization, or spray drying.

Solution formulations suitable for use in atomizers that generate fine mist using electro-hydraulics may contain 1 μ g to 10mg of the compounds of the present invention per press, and may vary from 1 μ L to 100 μ L per press volume. Typical formulations may comprise a compound of formula (I), (Ia) or (Ib), propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents may be used in place of propylene glycol, including glycerol and polyethylene glycol.

Capsules, blisters and cartridges (made, for example, from gelatin or HPMC) for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention, a suitable powder base (for example, lactose or starch) and a modifying agent (for example, 1-leucine, mannitol or magnesium stearate).

Formulations for inhalation/intranasal administration may be formulated for immediate and/or modified release.

Alternatively, the compounds of formula (I), (Ia) or (Ib) may be administered in the form of suppositories or pessaries, or they may be administered topically in the form of gels, hydrogels, lotions, solutions, creams, ointments or dusting powders. The compounds of formula (I), (Ia) or (Ib) may also be administered transdermally or transdermally, for example using a skin patch. They may also be administered by the pulmonary or rectal route.

They may also be administered by the ocular route. For ophthalmic purposes, the compounds may be formulated as micronized suspensions in isotonic, pH adjusted, sterile saline, or preferably as solutions in isotonic, pH adjusted, sterile saline, optionally in combination with preservatives, such as benzalkonium chloride. Alternatively, they may be formulated in ointments, such as vaseline.

For topical application to the skin, the compounds of formula (I), (Ia) or (Ib) may be formulated in a suitable ointment containing the active compound suspended or dissolved in a mixture of one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, they may be formulated in a suitable lotion or cream, suspended or dissolved in a mixture of one or more of the following: mineral oil, sorbitan monostearate, polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, cetyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The compounds of formula (I), (Ia) or (Ib) may also be used in combination with cyclodextrins. Cyclodextrins are known to form entrapped and non-entrapped complexes with drug molecules. The formation of drug-cyclodextrin complexes can alter the solubility, dissolution rate, bioavailability, and/or stability of the drug molecule. Drug-cyclodextrin complexes are generally useful in most dosage forms and routes of administration. As an alternative to direct complexation with the drug, the cyclodextrin may be used as an auxiliary additive, such as a carrier, diluent or solubiliser. alphA-, betA-and gammA-cyclodextrins are the most commonly used, suitable examples being described in WO-A-91/11172, WO-A-94/02518 and WO-A-98/55148.

The following non-limiting examples further illustrate the invention.

The invention is illustrated by the following non-limiting examples in which the following abbreviations and definitions are used:

optical rotation at alpha D587 nm

Arbacel ® filtering agent

b broad peak

Boc tert-butyloxycarbonyl group

CDCl₃Chloroform-d 1

CD₃OD methanol-d 4

Chemical shift of delta

d double peak

dd double doublet

DCM chloromethane

DMF N, N-dimethylformamide

DMSO dimethyl sulfoxide

h hours

HCl hydrogen chloride

LRMS low resolution mass spectrometry

m multiplet

Peak of mass m/z

min is divided into different species

Mpt melting Point

NaOH sodium hydroxide

NMR nuclear magnetic resonance

q quartet peak

s single peak

t triplet peak

Tf trifluoromethanesulfonyl

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

Melting points were determined using a Perkin Elmer DSC7 with a heating rate of 20 deg.C/min.

X-ray diffraction data were recorded at room temperature using a Bruker Axs Smart-Apex CCD area detection diffractometer (MO K α radiation). The intensity is integrated from several series of exposures. Each exposure covers 0.3 IN ω, the exposure time is 60 seconds, and the total exposure data set has more than one sphere.

Example 1

2-amino-1- (3-methoxyphenyl) ethanol

A stirred solution of 3N HCl (aq) (150ml, 0.3mol) and sodium sulfite (37.8g, 0.3mol) was added to a solution of 3-methoxybenzaldehyde (27.2g, 0.2mol) in THF (150ml) at room temperature. After 10 minutes, potassium cyanide (19.53g, 0.3mol) was added in portions and the reaction mixture was stirred for 30 minutes. Diethyl ether (800ml) and water (300ml) were added, followed by layer separation. The aqueous layer was back-extracted with diethyl ether (500ml), and the organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and then concentrated in vacuo to giveTo the cyanohydrin intermediate as a colourless oil (35.57g, 0.22mol, > 100%). The borane-tetrahydrofuran complex (1M THF solution) (400ml, 0.4mol) was then carefully added to a THF solution of cyanohydrin (100 ml). Once effervescence ceased, stirring under nitrogen atmosphere at reflux was continued for 1.5 hours. The reaction mixture was cooled and then quenched with methanol (40ml) and concentrated in vacuo to give a colorless oil. 6M HCl (aq) (200ml) was added and the reaction stirred at reflux for 2h then concentrated in vacuo to give a white solid. This was pre-adsorbed onto silica and then purified by column chromatography eluting with dichloromethane: methanol: ammonia (90: 10: 1) to give the title compound as a colourless oil (31.3g, 0.19mol, 94%).¹H NMR(CDCl₃，400MHz)δ：1.60(bs，2H)，2.80(dd，1H)，3.02(dd，1H)，3.46(s，1H)，3.81(s，3H)，4.60(dd，1H)，6.81(d，1H)，6.91(d，1H)，6.93(s，1H)，7.22(t，1H).LRMS：m/z 168(M-H⁺) Analyze found C, 56.66; h, 8.28; n, 6.91%. C₉H₁₃NO₂·1.33H₂Theoretical value of O, C, 56.33; h, 8.27; and N.7.30 percent.

Example 2

N- [ 2-hydroxy-2- (3-methoxyphenyl) ethyl ] propanamide

Triethylamine (52ml, 0.37mol) was added to a solution of the amine of example 1 (31.3g, 0.19mol) in dichloromethane (400ml) and the reaction mixture was stirred at 0 ℃ under nitrogen for 10 minutes. Propionyl chloride (16.3ml, 0.19mol) was added, followed by stirring for 30 minutes, and the reaction temperature was allowed to rise to room temperature for another 5 hours. The reaction mixture was quenched with 1N HCl (aq) (100ml) and then extracted with dichloromethane (2X 50 ml). The organic fractions were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a colorless oil that crystallized upon standing as white crystals (28g, 0.13mol, 67%).

¹H NMR(CDCl₃400MHz) δ: 1.18(t, 3H), 2.22(q, 2H), 2.51(bs, 1H), 3.31(m, 1H), 3.71(dd, 1H), 3.80(s, 3H), 4.81(m, 1H), 5.95(bs, 1H), 6.80(d, 1H), 6.90(d, 1H), 6.91(s, 1H), 7.22(t, 1H). LRMS: m/z 224. Mpt: 77-78 deg.C, analysis measured value

C，63.86；H，7.82；N，6.28％.C₁₂H₁₇NO₃·0.1H₂Theoretical value of O, C, 64.04; h, 7.70; n, 6.22 percent.

Example 3

1- (3-methoxyphenyl) -2-propylaminoethanol

Borane-tetrahydrofuran complex (1M THF solution) (376ml, 0.4mol) was added to a solution of the amide of example 2 (28g, 0.13mol) in anhydrous THF (100ml) and the reaction mixture stirred under nitrogen was refluxed for 2.5 hours. The reaction mixture was cooled and then quenched with methanol (40ml) and concentrated in vacuo to give an opaque white oil. 6N HCl (aq) (200ml) was added and the reaction mixture was stirred at reflux for 2 h. The reaction mixture was cooled, then dichloromethane (200ml) was added and the layers were separated. Potassium carbonate was added to the aqueous layer to make it basic, followed by back extraction with dichloromethane (2X 200 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a colorless oil that crystallized upon standing to give colorless crystals (15.3g, 0.07mol, 59%).

¹HNMR(CDCl₃，400MHz)δ：0.93(t，3H)，1.62(q，2H)，2.71(q，2H)，2.81(t，2H)，3.00(d，1H)，3.80(s，3H)，4.30(bs，1H)，4.89(d，1H)，6.81(d，1H)，6.91(d，1H)，6.93(s，1H)，7.22(t，1H).LRMS：m/z 210.Mpt: analysis found value C, 67.47, at 50-51 ℃; h, 9.02; n, 6.45%. C₁₂H₁₉NO₂·0.2H₂Theoretical value of O, C, 67.70; h, 9.19; n, 6.58 percent.

Example 4

2-chloro-N- [ 2-hydroxy-2- (3-methoxyphenyl) ethyl ] -N-propylacetamide

A solution of sodium hydroxide (15.1g, 0.38mol) in water (180ml) was added to a solution of the amine of example 3 (15.8g, 0.08mol) in dichloromethane (500ml) and the solution was stirred vigorously at room temperature. Chloroacetyl chloride (7.22ml, 0.09mol) was then added and the reaction mixture was stirred for an additional 30 minutes. The layers were separated and the aqueous layer was back-extracted with dichloromethane (200 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a colorless oil (17.8g, 0.06mol, 83%).¹H NMR(CDCl₃400MHz) δ: 0.96(t, 3H), 1.62(q, 2H), 3.21(q, 2H), 3.57-3.71(m, 2H), 3.82(s, 3H), 4.01-4.21(bq, 1H), 4.16(s, 2H), 5.00(m, 1H), 6.82(m, 1H), 6.91-6.99(m, 2H), 7.22(m, 1H). LRMS: m/z286. assay found C, 57.38; h, 6.95; n, 4.67%. C₁₄H₂₀NO₃Cl·0.33H₂Theoretical value of O, C, 57.64; h, 7.14; n, 4.80 percent.

Example 5

6- (3-methoxyphenyl) -4-propylmorpholin-3-one

Potassium hydroxide (4.2g, 0.07mol), isopropanol (500ml) and the amide of example 4 (17) were added.8g, 0.06mol) was stirred with water (15ml) to an opaque solution for 2 hours. The reaction mixture was concentrated in vacuo and the yellow residue was dissolved in ethyl acetate (200 ml). It was dispensed with water (200ml) followed by brine (200 ml). The organic portion was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a yellow oil (15.8g, 0.06mol, 100%).¹H NMR(CDCl₃400MHz) δ: 0.96(t, 3H), 1.62(m, 2H), 3.36(m, 2H), 3.51(q, 2H), 3.81(s, 3H), 4.30-4.62(bq, 2H), 4.79(d, 1H), 6.85(d, 1H), 6.91(d, 1H), 6.95(s, 1H), 7.29(t, 1H). LRMS: m/z272. assay found C, 66.80; h, 7.78; n, 5.52%. C₁₄H₁₉NO₃·0.1H₂Theoretical value of O, C, 66.96; h, 7.71; n, 5.58 percent.

Example 6

2- (3-methoxyphenyl) -4-propylmorpholine

Borane-tetrahydrofuran complex (1M in THF) (200ml, 0.19mol) was added dropwise over 30 minutes to a solution of morpholin-3-one described in example 5 (15.8g, 0.06mol) in anhydrous THF (100ml) under a nitrogen atmosphere. The reaction mixture was refluxed for 3 hours, then cooled and quenched by addition of methanol (30 ml). The reaction mixture was then concentrated in vacuo and the colorless residue was carefully suspended in 4N HCl (aq) (400ml) and then refluxed for 2.5 hours. The reaction mixture was cooled and dichloromethane (200ml) was added. The layers were separated and the aqueous layer was made alkaline by addition of potassium carbonate and back-extracted with dichloromethane (3X 100 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a colorless oil (12.51g, 0.05mol, 84%).

¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.59(q，2H)，2.05(t，1H)2.23(t, 1H), 2.40(t, 2H), 2.81(d, 1H), 2.98(d, 1H), 3.80(s, 3H), 3.85(t, 1H), 4.05(d, 1H), 4.60(d, 1H), 6.81(d, 1H), 6.91(d, 1H), 7.21(t, 1H), 7.23(s, 1H). LRMS: m/z236. assay found C, 68.94; h, 8.80; n, 5.79%. C₁₄H₂₁NO₂·0.5H₂Theoretical value of O, C, 68.82; h, 9.08; n.5.73%.

Example 7A

R- (-) -3- (4-propylmorpholin-2-yl) phenol

Example 7B

S- (+) -3- (4-propylmorpholin-2-yl) phenol

Hydrobromic acid (250ml) and anisole from example 6 (8.62g, 0.03mol) were heated together to reflux for 1 hour. After cooling, the reaction mixture was diluted with water (100ml) and NH was added₄OH (20ml) was neutralized. The yellow opaque solution was then extracted with dichloromethane (2X 100 ml). The organic extracts were combined, then dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give a racemic mixture of the title compound as a yellow oil (7.78g, 0.03mol, 96%). Enantiomers were separated by chiral chromatography (Chiralpak AD 250 x 20mm column) eluting with hexane, isopropanol, diethylamine (70: 30: 0.05) to give enantiomer 1(ee > 99.5%) and enantiomer 2(ee > 99%). Each enantiomer was purified by column chromatography on silica eluting with methylene chloride: methanol (95: 5) to give enantiomer 1(7a) (3.02g, 0.014mol, 39%) and enantiomer 2(7b) (3.15g, 0.014mol, 40%) as a colorless oil. Enantiomer 1(7 a):¹H NMR(CDCl₃，400MHz)δ：0.96(t，3H)，1.60(q，2H)，2.13(t，1H)，2.31(t，1H)，2.41(t，2H)，2.85(d，1H)，3.02(d，1H)，3.90(t，1H)，4.02(dd，1H)，4.60(d，1H)，6.78(d，1H)，6.80(s，1H)，6.91(d，1H)，7.20(t，1H).LRMS：m/z 222(M-H⁺) Enantiomer 2(7 b):¹H NMR(CDCl₃，400MHz)δ：0.96(t，3H)，1.60(q，2H)，2.13(t，1H)，2.31(t，1H)，2.41(t，2H)，2.85(d，1H)，3.02(d，1H)，3.90(t，1H)，4.02(dd，1H)，4.60(d，1H)，6.78(d，1H)，6.80(s，1H)，6.91(d，1H)，7.20(t，1H).LRMS：m/z 222(M-H⁺).

example 8

R- (-) -3- (4-propylmorpholin-2-yl) phenolate

Enantiomer 1(7a) of example 7 (3.00g, 0.014mol) was dissolved in diethyl ether (180ml), and hydrogen chloride (2.0M diethyl ether solution) (10ml) was added. The reaction mixture was stirred at room temperature for 30 minutes, then the solvent was decanted and dried in vacuo to give the title compound as a white solid (3.115g, 0.012mol, 90%).

¹H NMR(CD₃OD，400MHz)δ：1.06(t，3H)，1.81(m，2H)，3.02(t，1H)，3.16(t，2H)，3.20(t，1H)，3.60(t，2H)，4.01(t，1H)，4.26(d，1H)，4.71(d，1H)，6.78(d，1H)，6.82(s，1H)，6.83(d，1H)，7.21(t，1H).LRMS：m/z 222(M-H⁺) Analyze found C, 59.74; h, 7.98; n, 5.25%. C₁₃H₁₉NO₂·0.18H₂Theoretical value of O, C, 59.82; h, 7.86; n, 5.37%. alpha._D＝-5.66°(Methanol 10.6mg/10ml).

A sample of the title compound was recrystallized from a methanol: diethyl ether mixture by vapor diffusion to give an X-ray crystal structure. By means of Flack¹The absolute stereochemistry of the title compound was determined from the diffraction data and was shown to have the R configuration.

Reference 1: H.D. FLACK, ACTA CRYST.1983, 439, 876-one 881

Example 9

2-amino-1- (3, 5-dimethoxyphenyl) ethanol

The preparation was carried out in the same manner as in example 1, starting from 3, 5-dimethoxybenzaldehyde (5.00g, 0.03 mol). After refluxing in 6M HCl (aq), the reaction mixture was cooled and extracted with diethyl ether (2X 80 ml). The organic layer was discarded, and potassium carbonate was added to the aqueous layer to alkalify. The aqueous residue was then extracted with ethyl acetate (3X 70 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a pale yellow oil (3.47g, 0.018mol, 59%).¹H NMR(CD₃OD，400MHz)δ：2.77-2.86(m，2H)，3.78(s，6H)，4.60(m，1H)，6.38(s，1H)，6.52(s，2H).LRMS：m/z 198(M-H⁺).

Example 10

N- [2- (3, 5-dimethoxyphenyl) -2-hydroxyethyl ] propionamide

Following the same procedure as in example 2, starting from the amine described in example 9 (3.41g, 0.017 mol). The crude reaction mixture was purified by column chromatography on silica eluting with dichloromethane: methanol (95: 5) to give the title compound as a pale yellow oil (3.08g, 0.012mol, 70%).

¹HNMR(CDCl₃，400MHz)δ：1.18(m，3H)，2.24(m，2H)，3.34(m，1H)，3.68(m，1H)，3.81(s，6H)，4.80(dd，1H)，5.95(bs，1H)，6.39(s，1H)，6.51(s，2H).LRMS：m/z 252(M-H^-).

Example 11

1- (3, 5-Dimethoxyphenyl) -2-propylaminoethanol

Following the procedure of example 3, starting from the amide described in example 10 (3.06g, 0.012mol), the title compound was obtained as an orange oil (2.72g, 0.011mol, 94%).

¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.56(m，2H)，2.61(m，2H)，2.77(d，2H)，3.78(s，6H)，4.70(t，1H)，6.38(s，1H)，6.51(s，2H).LRMS：m/z 240(M-H⁺).

Example 12

2-chloro-N- [2- (3, 5-dimethoxyphenyl) -2-hydroxyethyl ] -N-propylacetamide

Following the same procedure as in example 4, starting from the amine described in example 11 (2.70g, 0.011mol), the title compound was obtained as a yellow oil (3.56g, 0.011mol, 100%).¹H NMR(CDCl₃，400MHz)δ：0.92(t，3H)，1.61(m，2H)，3.20(m，2H)，3.5)-3.64(m，2H)，3.80(d，6H)，4.13(s，2H)，4.95(m，1H)，6.40(m，1H)，6.55(s，2H).LRMS：m/z 316(M-H⁺).

Example 13

6- (3, 5-Dimethoxyphenyl) -4-propylmorpholin-3-one

Following the same procedure as in example 5, starting from the amide described in example 12 (3.54g, 0.011mol), the title compound was obtained as a yellow oil (2.44g, 0.009mol, 78%).¹H NMR(CDCl₃，400MHz)δ：0.94(t，3H)，1.61(m，2H)，3.30(m，2H)，3.49(m，2H)，3.80(s，6H)，4.30(d，1H)，4.42(d，1H)，4.73(dd，1H)，6.42(s，1H)，6.53(s，2H).LRMS：m/z 280(M-H⁺).

Example 14

2- (3, 5-dimethoxyphenyl) -4-propylmorpholine

The preparation was carried out following the procedure of example 6, starting from the amide described in example 13 (2.42g, 0.009 mol). After refluxing in 6M HCl (aq), the cooled reaction mixture was extracted with diethyl ether (2X 80 ml). The organic layer was discarded, and potassium carbonate was added to the aqueous layer to alkalify. The aqueous residue was then extracted with ethyl acetate (3 × 80ml), the organic extracts combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a pale orange oil (2.14g, 0.008mol, 93%).¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.58(m，2H)，2.01(m，1H)，2.22(dt，1H)，2.38(t，2H)，2.83(d，1H)，2.93(d，1H)，3.78(m，7H)，4.01(dd，1H)，4.45(dd，1H)，6.39(s，1H)，6.49(s，2H).LRMS：m/z 266(M-H⁺).

Example 15A

R-5- (4-propylmorpholin-2-yl) benzene-1, 3-diol

Example 15B

S-5- (4-propylmorpholin-2-yl) benzene-1, 3-diol

Following the same route as example 7, starting from the 3, 5-dimethoxyphenyl compound described in example 14 (1.00g, 0.004mol), the title compound was obtained as a brown oil (145mg, 0.61mmol, 16%). The enantiomers were separated by chiral chromatography (Chiralpak AD 250 x 20mm column) eluting with hexane isopropanol (80: 20) to give enantiomer 1(15a) (5.2mg) (ee > 98.94%) and enantiomer 2(15b) (5.1mg) (ee > 96.46%) as brown oil. Enantiomer 1(15 a):

¹H NMR(CD₃OD，400MHz)δ：0.96(t，3H)，1.58(m，2H)，2.01(t，1H)，2.20(dt，1H)，2.37(t，2H)，2.81-2.92(m，2H)，3.89(dt，1H)，3.99(dd，1H)，4.38(dd，1H)，6.18(t，1H)，6.26(s，2H).LRMS：m/z 238(M-H⁺) Enantiomer 2(15 b):¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.58(m，2H)，2.01(t，1H)，2.20(dt，1H)，2.38(1，2H)，2.80-2.92(q，2H)，3.78(dt，1H)，3.98(dd，1H)，4.38(dd，1H)，6.18(s，1H)，6.25(s，2H).LRMS：m/z 238(M-H⁺).

example 16

4-fluoro-3-methoxybenzaldehyde

(4-fluoro-3-methoxyphenyl) methanol (5.00g, 0.03mol) and manganese dioxide (33.4g, 0.38mol) were stirred in dichloromethane (100ml) under nitrogen at gentle reflux for 16 h. The cooled reaction mixture was then filtered through arbacell and concentrated in vacuo to give the title compound as a white solid (4.18g, 0.027mol, 85%).

¹H NMR(CDCl₃400MHz) δ: 3.96(s, 3H), 7.23(d, 1H), 7.43(m, 1H), 7.50(d, 1H)9.91(s, 1H). Mpt: analysis found value C, 62.18, at 61-63 ℃; h, 4.54%. C₈H₇FO₂Theoretical value C, 62.34; h, 4.58 percent.

Example 17

2-amino-1- (4-fluoro-3-methoxyphenyl) ethanol

Prepared by following the same procedure as in example 1, starting from 4-fluoro-3-methoxybenzaldehyde (4.17g, 0.03 mol). After refluxing in 6M HCl (aq), the reaction mixture was cooled and extracted with diethyl ether (2X 60 ml). The organic layer was discarded, and potassium carbonate was added to the aqueous layer to alkalify. The aqueous residue was then extracted with ethyl acetate (3X 80 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as an orange oil (2.36g, 0.013mol, 47%).

¹H NMR(CD₃OD，400MHz)δ：2.80-2.91(m，2H)，3.86(s，3H)，4.64(m，1H)，6.89(m，1H)，7.03(t，1H)，7.11(dd，1H).LRMS：m/z186(M-H⁺).

Example 18

N- [2- (4-fluoro-3-methoxyphenyl) -2-hydroxyethyl ] propionamide

The preparation was carried out following the same procedure as in example 2, starting from the amine described in example 17 (1.32g, 0.007 mol). The crude reaction mixture was purified by column chromatography on silica eluting with ethyl acetate to pentane (2: 1) to give the title compound as a yellow oil which crystallized upon standing (0.59g, 0.002mol, 35%).

¹H NMR(CDCl₃，400MHz)δ：1.18(t，3H)，2.24(q，2H)，2.58(bs，1H)，3.34(m，1H)，3.63(m，1H)，3.88(s，3H)，4.82(dd，1H)，5.98(bs，1H)，6.82(m，1H)，7.01(m，2H).LRMS：m/z 242(M-H⁺).

Example 19

1- (4-fluoro-3-methoxyphenyl) -2-propylaminoethanol

The preparation was carried out following the same procedure as in example 3, starting from the amide described in example 18 (585mg, 2.42 mmol). After refluxing in 6M HCl (aq), the reaction mixture was cooled and extracted with diethyl ether (2X 50 ml). The organic layer was discarded, and potassium carbonate was added to the aqueous layer to alkalify. The aqueous residue was then extracted with ethyl acetate (3X 50 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a pale yellow oil (448mg, 1.97mmol, 81%).¹H NMR(CD₃OD，400MHz)δ：0.96(t，3H)，1.58(m，2H)，2.63(m，2H)，2.79(d，2H)，3.96(s，3H)，4.77(t，1H)，6.90(m，1H)，7.03(t，1H)，7.11(d，1H).LRMS：m/z 228(M-H⁺).

Example 20

2-chloro-N- [2- (4-fluoro-3-methoxyphenyl) -2-hydroxyethyl ] -N-propylacetamide

Following the same procedure as in example 4, starting from the amine described in example 19 (0.84g, 4.00mmol) the title compound was obtained as a yellow oil (0.97g, 3.00mmol, 87%). LRMS: m/z 304 (M-H)⁺). The crude product can be used.

Example 21

6- (4-fluoro-3-methoxyphenyl) -4-propylmorpholin-3-one

Following the same procedure as in example 5, starting from the amide described in example 20 (0.96g, 3.00mmol) the title compound was obtained as a yellow oil (0.64g, 2.40mmol, 75%).¹H NMR(CDCl₃，400MHz)δ：0.94(t，3H)，1.62(m，2H)，3.33(m，2H)，3.48(m，2H)，3.91(s，3H)，4.34(d，1H)，4.43(d，1H)，4.76(dd，1H)，6.85(m，1H)，7.01-7.08(m，2H).LRMS：m/z 268(M-H⁺).

Example 22

2- (4-fluoro-3-methoxyphenyl) -4-propylmorpholine

Following the same procedure as in example 6, starting from morpholin-3-one (633mg, 2.37mmol) as described in example 21. After refluxing in 6M HCl (aq), the reaction mixture was cooled and extracted with diethyl ether (2X 20 ml). The organic layer was discarded, and potassium carbonate was added to the aqueous layer to alkalify. The aqueous residue was then extracted with ethyl acetate (3X 20 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a yellow oil (552 mg)，2.18mmol，92％)。 ¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.58(m，2H)，2.02(t，1H)，2.22(dt，1H)，2.38(t，2H)，2.85(d，1H)，2.93(d，1H)，3.80(m，1H)，3.84(s，3H)，4.01(dd，1H)，4.50(dd，1H)，6.88(m，1H)，7.02(t，1H)，7.09(d，1H).LRMS：m/z 254(M-H⁺).

Example 23A

R- (+) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol

Example 23B

S- (-) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol

Prepared following the same procedure as in example 7, starting from anisole (200mg, 0.789mmol) as described in example 22. The crude reaction mixture was purified by column chromatography on silica eluting with methylene chloride: methanol (90: 10) to give the title racemic compound as a dark yellow viscous oil (149mg, 0.62mmol, 79%). The enantiomers were separated by chiral chromatography (ChiralpakAD 250 x 20mm column) eluting with hexane isopropanol (90: 10) to give enantiomer 1(23a) as an opaque oil (15mg) (ee > 99.5%) and enantiomer 2(23b) as a crystalline solid (16mg) (ee > 99%). Enantiomer 1(23 a):

¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.58(m，2H)，2.01(t，1H)，2.21(dt，1H)，2.37(t，2H)，2.82-2.97(bq，2H)，3.78(dt，1H)，3.99(dd，1H)，4.43(d，1H)，6.78(m，1H)，6.89-7.01(m，2H).LRMS：m/z 240(M-H⁺).α_Denantiomer 2(23 b):¹H NMR(CD₃OD，400MHz)δ：0.96(t，3H)，1.58(m，2H)，2.01(t，1H)，2.22(dt，1H)，2.38(t，2H)，2.78(dd，2H)，3.78(dt，1H)，4.00(dd，1H)，4.43(dd，1H)，6.78(m，1H)，6.91(d，1H)，6.98(t，1H).LRMS：m/z 240(M-H⁺).α_D＝-0.40(Ethanol 1.00mg/ml).

example 24

2-amino-1- (4-benzyloxyphenyl) ethanol

Potassium cyanide (20.15g, 0.31mol) and ammonium chloride (16.4g, 0.31mol) were dissolved in water (60ml), to which was added 4-benzyloxybenzaldehyde (32.9g, 0.155mol), followed by diethyl ether (100 ml). The reaction mixture was stirred vigorously at room temperature for 48 hours, then extracted with ethyl acetate (2X 200 ml). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the cyanohydrin intermediate as a yellow solid (34.2g, 0.14mol, 90%). The cyanohydrin was then dissolved in anhydrous THF (300ml) and borane-methyl sulfide complex (26.6ml, 0.28mol) was added. The reaction mixture was refluxed for 2 hours and then quenched with methanol (50 ml). Water (50ml) was added followed by concentrated HCl (40ml) and the reaction mixture was stirred for 2 hours until the exotherm ceased. The reaction mixture was then concentrated in vacuo and the residue was diluted with water (100 ml). NH is then added to the aqueous solution₄OH (30ml) was basified and extracted with ethyl acetate (3X 150 ml). The organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a white solid (24.8g, 0.10mol, 73%).

¹H NMR(CDCl₃，400MHz)δ：1.62(bs，3H)，2.81(dd，1H)，2.99(d，1H)，4.61(q，1H)，5.07(s，2H)，6.95(d，2H)，7.22-7.45(m，7H)LRMS：m/z 244(M-H⁺).

Example 25

N- [2- (4-benzyloxyphenyl) -2-hydroxyethyl ] propionamide

The amine described in example 24 (24.8g, 0.10mol) was dissolved in dichloromethane (700ml), to which triethylamine (20.86ml, 0.15mol) was added. The reaction mixture was stirred, cooled to 0 ℃ and propionyl chloride (7.12ml, 0.082mo1) was added dropwise. The reaction mixture was allowed to warm to room temperature over 16 hours and then quenched with 3M HCl (aq) (20ml) and water (100 ml). The reaction mixture was extracted with dichloromethane (3 × 200ml), the organic layers combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a clear viscous gum (27.5g, 0.092mol, 90%).¹H NMR(CDCl₃，400MHz)δ：1.10(t，3H)，2.19(q，2H)，3.32-3.43(m，4H)，4.81(s，2H)，5.11(m，1H)，6.99(d，2H)，7.25-7.42(m，7H).LRMS：m/z 298(M-H^-).

Example 26

1- (4-benzyloxyphenyl) -2-propylaminoethanol

To a solution of the amide described in example 25 (27.5g, 0.092mol) in anhydrous THF (100ml) was added borane-methyl sulfide complex (17.5ml, 0.18mol) and the reaction mixture was stirred at reflux for 2 hours. The reaction mixture was cooled and then quenched with methanol (30 ml). Water (50ml) and concentrated HCl (35ml) were added and the reaction mixture was stirred until no more bubbling occurred and then concentrated in vacuo. Water (250ml) was added to the residue followed by NH₄OH (30ml) was basified. The aqueous layer was extracted with ethyl acetate (3X 200ml) and the organic extracts were combined, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the title compound as a white solid (26.1g, 0.09mol, 99%).¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.58(q，2H)，2.62(m，2H)，2.81(m，2H)，4.72(dd，1H)，5.05(s，2H)，6.95(d，2H)，7.24(m，3H)，7.35(t，2H)，7.41(d，2H).LRMS：m/z 286(M-H⁺).

Example 27

6- (4-benzyloxyphenyl) -4-propylmorpholin-3-one

A solution of sodium hydroxide (22.5g, 0.56mol) in water (100ml) was added to a solution of the amine described in example 26 (26.0g, 0.09mol) in dichloromethane (400ml) and the solution was stirred vigorously at room temperature. Chloroacetyl chloride (8.6ml, 0.11mol) was then added and the reaction mixture was stirred for an additional 60 minutes. The layers were separated and the aqueous layer re-extracted with dichloromethane (200 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give a colorless oil. Potassium hydroxide (15.0g, 0.27mol), isopropanol (400ml) and colourless oil residue were stirred together with water (30ml) into an opaque solution for 2 hours. The reaction mixture was concentrated in vacuo and the yellow residue was dissolved in ethyl acetate (200 ml). It was dispensed with water (200ml) followed by brine (200 ml). The organic portion was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a white solid (19.9g, 0.06mol, 67%).

¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.62(m，2H)，3.34(m，2H)，3.51(m，2H)，4.32(d，1H)，4.41(d，1H)，4.72(dd，1H)，5.04(s，2H)，6.98(d，2H)，7.31-7.43(m，7H). LRMS：m/z 326(M-H⁺).

Example 28

2- (4-benzyloxyphenyl) -4-propylmorpholine

Following the same procedure as in example 26, starting from morpholin-3-one (19.9g, 0.061mol) as described in example 27, the title compound is obtained as a colourless oil (17g, 0.055mol, 90%).

¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.55(q，2H)，2.06(t，1H)，2.21(dt，1H)，2.35(dd，2H)，2.80(d，1H)，2.91(d，1H)，3.82(dt，1H)，4.02(dd，1H)，4.52(dd，1H)，5.05(s，2H)，6.98(t，2H)，7.24-7.42(m，7H).LRMS：m/z 312(M-H⁺).

Example 29

4- (4-propylmorpholin-2-yl) phenol

The benzyl ether described in example 28 (3.0g, 9.64mmol) was dissolved in methanol (150ml) and 10% palladium on carbon (800mg) was added. The reaction mixture was stirred for a few minutes, then ammonium formate (6.17g, 96.4mmol) was added in portions. The reaction mixture was heated carefully to 80 ℃ until no more gas was evolved. After cooling, the reaction mixture was filtered through arbacel, washed with methanol (50ml) and concentrated in vacuo to give the title compound as a white crystalline solid (1.51g, 6.83mmol, 71%).

¹H NMR(CDCl₃，400MHz)δ：0.91(t，3H)，1.58(q，2H)，2.10(t，1H)，2.22(t，1H)，2.40(dd，2H)，2.81(d，1H)，2.93(d，1H)，3.85(t，1H)，4.02(dd，1H)，4.57(d，1H)，6.79(d，2H)，7.21(d，2H).LRMS：m/z 222(M-H⁺).

Example 30

2-bromo-4- (4-propylmorpholin-2-yl) phenol

To a solution of the phenol described in example 29 (200mg, 0.9mmol) in dichloromethane (5ml) was added N-bromosuccinimide (161mg, 0.9 mmol). The reaction mixture was stirred at room temperature for 55 minutes, then concentrated in vacuo. The crude product was purified by column chromatography on silica eluting with dichloromethane: methanol (95: 5) to give the title compound as a white foam (117.5mg, 0.39mmol, 44%).¹H NMR(CDCl₃，400MHz)δ：0.96(t，3H)，1.59(q，2H)，2.03(t，1H)，2.23(t，1H)，2.40(t，2H)，2.81(d，1H)，2.98(d，1H)，3.82(t，1H)，4.01(d，1H)，4.56(d，1H)，6.96(d，1H)，7.20(d，1H)，7.49(s，1H).LRMS：m/z 302(M-H⁺，Brisotope).

Example 31

2- (4-benzyloxy-3-bromophenyl) -4-propylmorpholine

To a solution of the phenol described in example 30 (117.5mg, 0.39mmol) in anhydrous DMF (10ml) under nitrogen was added potassium carbonate (75mg, 0.54mmol) and benzyl bromide (0.07ml, 0.54 mmol). The reaction mixture was heated to 150 ℃ for 48 hours. After cooling, the reaction mixture was concentrated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (50 ml). The aqueous layer was then re-extracted with ethyl acetate (2X 20 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the crude product as a brown oil. Purification by silica column chromatography eluting with dichloromethane: methanol (98: 2) gave the title compound as a colourless oil (153mg, 0.39mmol, 100%).

¹H NMR(CDCl₃，400MHz)δ：0.93(t，3H)，1.56(q，2H)，2.05(t，1H)，2.25(t，1H)，2.37(t，2H)，2.82(d，1H)，2.92(d，1H)，3.85(t，1H)，4.02(d，1H)，4.52(d，1H)，5.15(s，2H)，6.87(d，1H)，7.20(d，1H)，7.30(d，1H)，7.37(t，2H)，7.45(d，2H)，7.58(s，1H).LRMS：m/z 392(M-H⁺).

Example 32

2-benzyloxy-5- (4-propylmorpholin-2-yl) benzoic acid methyl ester

To a solution of the bromide described in example 31 (153mg, 0.39mmol) in anhydrous DMF (4ml) were added triethylamine (2.1ml, 0.78mmol) and methanol (2ml) and the reaction mixture was stirred for 5 minutes. The [1, 1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) complex with dichloromethane (1: 1) (16mg, 0.02mmol) was added and then carbon monoxide (g) was passed over the reaction mixture (3 air-filled balloons). The reaction mixture was heated to 100 ℃ for 16 hours under a carbon monoxide atmosphere. After cooling, the reaction mixture was concentrated in vacuo and the residue partitioned between ethyl acetate (25ml) and water (20 ml). The organic layer was separated, washed with brine (20ml), dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give a black solid. Purification by silica column chromatography eluting with dichloromethane, methanol, ammonia (90: 10: 1) gave the title compound as a colourless oil (105mg, 0.28mmol, 73%).

¹H NMR(CDCl₃，400MHz)δ：0.94(t，3H)，1.60(m，2H)，2.18(s，4H)，2.43(m，2H)，3.00(m，2H)，3.90(s，3H)，4.04d，1H)，5.18(s，2H)，5.97(d，1H)，7.26-7.47(m，6H)，7.82(s，1H).LRMS：m/z 370(M-H⁺).

Example 33

2-benzyloxy-5- (4-propylmorpholin-2-yl) benzoic acid

To a solution of the methyl ester described in example 32 (105mg, 0.28mmol) in methanol (5ml) was added 10% sodium hydroxide (aq) (15ml) and the milky white suspension was refluxed for 2 hours. The now colorless reaction mixture was cooled and then neutralized by the addition of 2M HCl (aq) (a few drops). The reaction mixture was then concentrated in vacuo to give the title compound as an off-white solid (99mg, 0.28mmol, 100%). LRMS: m/z 355 (M-H)⁺). This material was used directly in crude form in example 34.

Example 34

2-benzyloxy-5- (4-propylmorpholin-2-yl) benzamide

To the crude benzoic acid described in example 33 (99mg, 0.28mmol) was added thionyl chloride (5ml) and the reaction mixture was heated to 50 ℃ for 2 h. The reaction mixture was cooled and excess thionyl chloride was removed in vacuo. The residue was then dissolved in dichloromethane (10ml) and ammonia (g) was bubbled through the reaction mixture for 10 minutes. The resulting suspension was stirred at room temperature for 1 hour, then concentrated in vacuo. The crude product was purified by column chromatography on silica eluting with dichloromethane, methanol, ammonia (95: 5: 0.5) to give the title compound as an off white solid (88mg, 0.25mmol, 90%).¹H NMR(CDCl₃，400MHz)δ：0.94(t，3H)，1.59(m，2H)，2.15-2.42(m，4H)，2.87(m，1H)，3.03(m，1H)，3.96(m，1H)，4.02(d，1H)，4.67(m，1H)，5.19(s，2H)，5.72(m，1H)，7.04(d，1H)，7.41(m，5H)，7.50(d，1H)，7.70(m，1H)，8.21(s，1H).LRMS：m/z 355(M-H⁺).

Example 35

2-hydroxy-5- (4-propylmorpholin-2-yl) benzamide

Prepared by the same method as example 29, starting from the benzyl ester described in example 34 (80mg, 0.22mmol) to give the title compound as an off-white solid (56mg, 0.21mmol, 96%).¹H NMR(CD₃OD，400MHz)δ：0.95(t，3H)，1.55(m，2H)，2.13(t，1H)，2.29(t，1H)，2.42(m，2H)，2.88(d，1H)，2.97(d，1H)，3.81(t，1H)，4.00(d，1H)，4.49(d，1H)，6.87(d，1H)，7.42(d，1H)，7.78(s，1H).LRMS：m/z 265(M-H⁺).

Example 36

2-nitro-4- (4-propylmorpholin-2-yl) phenol

The phenol (100mg, 0.45mmol) described in example 29 was dissolved in nitric acid: water (1: 3) (2ml) and stirred at room temperature for 10 minutes. The reaction mixture was then diluted with water (5ml) and NH₄OH (1ml) was basified and then extracted with ethyl acetate (3X 10 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a yellow solid (95mg, 0.35mmol, 79%).

¹H NMR(CDCl₃，400MHz)δ：0.97(t，3H)，1.33(t，2H)，1.43-1.79(bm，4H)，2.02(d，3H)，4.06(m，2H)，7.17(d，1H)，7.60(d，1H)，8.16(s，1H)，10.55(bs，1H).LRMS：m/z 267(M-H⁺).

Example 37

2-amino-4- (4-propylmorpholin-2-yl) phenol

To a solution of the nitro compound described in example 36 (95mg, 0.35mmol) in ethanol (10ml) were added 10% palladium on carbon (50mg) and ammonium formate (100mg, XS). The reaction mixture was heated slightly to 70 ℃, held at this temperature for 1 hour, and then cooled to room temperature. The reaction mixture was filtered through arbacel, washed with ethanol (20ml) then dichloromethane (20 ml). The combined organic washes were concentrated in vacuo to give the title compound as a yellow solid (65mg, 0.28mmol, 78%).

¹H NMR(CDCl₃，400MHz)δ：0.91(t，3H)，1.55(m，2H)，2.12(t，1H)，2.25(dt，1H)，2.40(t，2H)，2.81-2.92(dd，2H)，3.82(t，1H)，4.00(d，1H)，4.42(d，1H)，6.60(m，2H)，6.71(s，1H).LRMS：m/z 237(M-H⁺).

Example 38

5-bromo-2- (2, 5-dimethylpyrrol-1-yl) pyridine

5-Bromopyridin-2-yl-amine (13.8g, 0.08mol), acetonylacetone (14.1ml, 0.12mol) and p-toluenesulfonic acid (100mg) were dissolved in toluene (180ml) and refluxed under Dean Stark conditions for 14 hours. After cooling, the brown solution was poured into water (200ml) and extracted with toluene (2X 200 ml). The organic extracts were combined, washed with brine (50ml), then dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the crude product. Purification by silica column chromatography eluting with ethyl acetate: pentane (1: 3) gave the title compound as a brown oil (18.4g, 0.073mol, 92%).

¹H NMR(CDCl₃，400MHz)δ：2.18(s，6H)，5.90(s，2H)，7.11(d，1H)，7.92(d，1H)，8.62(s，1H).LRMS：m/z 253(M-H⁺，Br isotope).

Example 39

2-chloro-1- [6- (2, 5-dimethylpyrrol-1-yl) pyridin-3-yl ] ethanone

To a solution of the bromopyridine described in example 38 (2g, 8.0mmol) in dry THF (30ml) at-78 deg.C was added butyllithium (2.5M in hexane) (3.5ml, 8.8mmol) dropwise over 20 min. The reaction mixture was stirred for 30 minutes, then a solution of 2-chloro-N-methoxy-N-methylacetamide (1.2g, 8.8mmol) in dry THF (20ml) was added dropwise, maintaining the temperature at-78 ℃. Stirring at this temperature was continued for 30 min, then 1M HCl (aq) (50ml) was added and the reaction mixture was allowed to warm to room temperature. The organic layer was separated and the aqueous layer was washed with ethyl acetate (56 ml). The organic layers were combined and washed with 3M NaOH (aq) (10ml) and brine (10ml), dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the crude title compound as a brown oil (1.34g, 5.4mmol, 67%).

¹H NMR(CDCl₃，400MHz)δ：2.20(s，6H)，4.68(s，2H)，5.92(s，2H)，7.32(d，1H)，8.38(d，1H)，9.16(s，1H).LRMS：m/z249(M-H⁺).

Example 40

2- (2, 5-dimethylpyrrol-1-yl) -5-oxiranylpyridines

To a solution of the ketone described in example 39 (1.34g, 5.4mmol) in dry THF (20ml) cooled to 0 deg.C was added sodium borohydride (308mg, 8.1mmol) portionwise. The reaction mixture was stirred for 2 hours, then 3M NaOH (aq) (10ml) was added and stirring was continued for a further 16 hours. The reaction mixture was extracted with ethyl acetate (2X 20ml) and the organic extracts were combined, washed with brine (5ml), dried over anhydrous magnesium sulphate, filtered and concentrated in vacuo. The residue was purified by silica column chromatography eluting with ethyl acetate: pentane (1: 5) to give the title compound as a colourless oil (900mg, 4.2mmol, 78%).¹H NMR(CDCl₃，400MHz)δ：2.13(s，6H)，2.91(dd，1H)，3.25(t，1H)，3.98(t，1H)，5.90(s，2H)，7.20(d，1H)，7.62(dd，1H)，8.58(s，1H).LRMS：m/z 215(M-H⁺).

EXAMPLE 41

1- [6- (2, 5-dimethylpyrrol-1-yl) pyridin-3-yl ] -2-propylaminoethanol

Propylamine (4ml, 4.8mmol) was added to a solution of the epoxide described in example 40 (900mg, 4.2mmol) in DMSO (5ml) and the reaction mixture was heated to 40 ℃ for 4 days. The reaction mixture was then cooled, 3M HCl (aq) (10ml) and water (10ml) were added, and washed with diethyl ether (2X 10 ml). The organic layer was discarded. NH for aqueous layer₄OH (5ml) was basified and extracted with ethyl acetate (3X 10 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as an oil (1.15g, 4.2mmol, 100%).

¹H NMR(CDCl₃，400MHz)δ：0.93(t，3H)，1.62(m，2H)，2.11(s，6H)，2.69-2.82(m，3H)，3.06(dd，1H)，3.60(bs，2H)，4.92(dd，1H)，5.84(s，2H)，7.20(d，1H)，7.88(d，1H)，8.61(s，1H).LRMS：m/z 274(M-H⁺).

Example 42

6- [6- (2, 5-dimethylpyrrol-1-yl) pyridin-3-yl ] -4-propylmorpholin-3-one

Preparation was carried out following the same procedure as in example 27, starting from the amine described in example 41 (1.15g, 4.2 mmol). Purification by silica column chromatography eluting with dichloromethane: methanol (98: 2) gave the title compound as a brown membrane (191mg, 0.61mmol, 14%).

¹H NMR(CDCl₃，400MHz)δ：0.97(t，3H)，1.65(m，2H)，2.13(s，6H)，3.38(m，1H)，3.42-3.56(m，2H)，6.61(t，1H)，4.35(d，1H)，4.45(d，1H)，4.91(dd，1H)，6.91(s，2H)，7.22(d，1H)，7.89(d，1H)，8.61(s，1H).LRMS：m/z 314(M-H⁺).

Example 43

6- [6- (2, 5-dimethylpyrrol-1-yl) pyridin-3-yl ] -4-propylmorpholine

To a solution of morpholin-3-one as described in example 42 (191mg, 0.61mmol) in dry THF (5ml) was added lithium aluminium hydride (1M in diethyl ether) (1.25ml, 0.61mmol) and the reaction mixture was warmed to reflux for 2.5 h. The reaction mixture was cooled to room temperature, and 1M NaOH (1.25ml) was then added to give a white precipitate. The reaction mixture was filtered and concentrated in vacuo. The white solid was discarded. Concentrating the filtrate over silicaPurification by column chromatography eluting with dichloromethane: methanol (95: 5) gave the title compound as a white film (108mg, 0.36mmol, 59%).¹H NMR(CDCl₃，400MHz)δ：0.92(t，3H)，1.61(q，2H)，2.10(s，6H)，2.15(m，1H)，2.29(dt，1H)，2.40(t，2H)，2.82(d，1H)，3.02(d，1H)，3.90(t，1H)，4.08(d，1H)，4.71(d，1H)，5.89(s，2H)，7.20(d，1H)，7.81(d，1H)，8.60(s，1H).LRMS：m/z 300(M-H⁺).

Examples 44A and 44B

5- (4-propylmorpholin-2-yl) pyridin-2-ylamine

To a solution of 2, 5-dimethylpyrrole (45mg, 0.15mmol) described in example 43 in ethanol (3ml) was added hydroxylamine hydrochloride (52mg, 0.75mmol) and the reaction mixture was heated to 80 ℃ for 20 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by column chromatography on silica eluting with dichloromethane, methanol, ammonia (90: 10: 1) to give the racemic compound as a colorless film (31mg, 0.14mmol, 94%).

¹H NMR(CDCl₃，400MHz)δ：0.92(t，3H)，1.60(m，2H)，2.11(t，1H)，2.25(dt，1H)，2.41(t，2H)，2.82-2.91(dd，2H)，3.89(dt，1H)，4.01(dd，1H)，4.57(bd，3H)，6.49(d，1H)，7.42(d，1H)，8.02(s，1H).LRMS：m/z 222(M-H⁺).

A sample of the racemic product (580mg) was separated into its enantiomers by chiral HPLC. The conditions used were: chiralpak AD column (250X 21.2mm), eluent methanol: ethanol (1: 1), flow rate 15 mL/min.

The faster eluting enantiomer of example 44A (retention time 8.3min) > 99% ee was obtained.¹H NMR(CDCl₃400MHz) and outerRacemic identity LRMS: m/z 222. analysis found C, 63.54; h, 8.60; n, 18.38%. C₁₂H₁₉N₃O.3H₂Theoretical value of O, C, 63.58; h, 8.71; n, 18.53% [ alpha ]]²⁵ ₅₄₆-2.1(c＝0.12，MeOH)；[α]²⁵ ₄₃₆-8.9(c＝0.12，MeOH).

The slower eluting enantiomer of example 44B (retention time 9.4min) > 98.9% ee was obtained.¹H NMR(CDCl₃400MHz) is identical to the racemate LRMS: m/z 222. analysis found C, 63.53; h, 8.57; n, 18.36%. C₁₂H₁₉N₃O.3H₂Theoretical value of O, C, 63.58; h, 8.71; n, 18.53% [ alpha ]]²⁵ ₅₄₆+2.4(c＝0.12，MeOH)；[α]²⁵ ₄₃₆+7.2(c＝0.12，MeOH).

Example 45

2-Ethyl-6- (3-methoxy-phenyl) -4-propyl-morpholin-3-one

A solution of sodium hydroxide (0.48g, 12.0mmol) in water (2ml) was added to a solution of the product described in example 3 (0.50g, 2.4mmol) in dichloromethane (5ml) and the mixture was stirred at room temperature. 2-chlorobutyryl chloride (0.28ml, 2.87mmol) was then added dropwise and the reaction mixture was stirred for 60 hours. The reaction mixture was diluted with dichloromethane (10ml), and the aqueous layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the crude product as a clear oil (mixture containing cyclized and uncyclized products) (0.57 g). LRMS: m/z 314 (M-H of the uncyclized product)⁺) 296 (M-H with small amount of Water)⁺) 278 (cyclization product M-H)⁺). Potassium hydroxide (0.13g, 2.20mmol) was dissolved in water (1ml) and added to a solution of the crude product (0.57g, 1.83mmol) in isopropanol (5 ml). The reaction mixture was stirred at room temperature overnight, then the organic solvent was evaporated in vacuo. The residue was dissolved in ethyl acetateEster (10ml) and the aqueous layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the crude product as an oil. The residue was purified by silica column chromatography eluting with ethyl acetate: pentane (1: 5 to 1: 1) to give the title compound as a clear oil (326mg, 1.17mmol, 49%) as a mixture of diastereomers.

¹H NMR(CDCl₃，400MHz)δ：0.90(t，3H)，1.00(t，3H)，1.60(m，2H)，2.00(bm，2H)，3.10-3.60(m，4H)，3.80(s，3H)，4.20(d，0.5H)，4.25(d，0.5H)，4.75(d，0.5H)，4.90(d，0.5H)，6.80(d，1H)，6.90(m，2H)，7.25(m，1H).LRMS(APCI)：m/z 278(MH⁺)，276(MH^-).

Examples 46A and 46B

2-ethyl-6- (3-methoxy-phenyl) -4-propyl-morpholine

Borane-tetrahydrofuran complex (1M in THF) (3ml, 3mmol) was added dropwise to a solution of the product described in example 45 (0.33g, 1.18mmol) in dry THF (4ml) under a nitrogen atmosphere. The reaction mixture was heated at 85 ℃ for 3 hours, then cooled and quenched by addition of methanol (1 ml). The reaction mixture was then concentrated in vacuo and the residue suspended in 6N HCl (aq) (10ml) and heated to 60 ℃ for 1.5 h. The reaction mixture was cooled and extracted with diethyl ether (2X 10 ml). Solid potassium carbonate was added to the aqueous layer to make it basic (pH9-10) and re-extracted with dichloromethane (2X 15 ml). The dichloromethane extract was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the crude product as a clear oil. Purification by silica column chromatography eluting with ethyl acetate: pentane (1: 10) gave the two title compounds as single diastereomers.

Example 46A: clear oil (0.10g, 0.38mmol, 32%):

¹H NMR(CDCl₃，400MHz)δ：1.00(m6H)，1.60(bm，3H)，1.85(m，1H)，2.25(bt，2H)，2.35(s，1H)，2.45(m，1H)，2.60(m，1H)，2.65(m，1H)，3.70(s，1H)，3.80(s，3H)，4.80(s，1H)，6.80(d，1H)，7.00(m，2H)，7.25(m，1H).LRMS(APCI)：m/z 264(M-H⁺).

example 46B: clear oil (0.10g, 0.38mmol, 32%):

¹H NMR(CDCl₃，400MHz)δ：0.90(t，3H)，1.00(t，3H)，1.60(bm，4H)，1.80(bs，1H)，2.00(bs，1H)，2.35(bs，2H)，2.85(bd，1H)，2.95(bd，1H)，3.60(s，1H)，3.80(s，3H)，4.60(s，1H)，6.80(d，1H)，6.95(s，2H)，7.25(t，1H).LRMS(APCI)：m/z 264(MH⁺).

example 47A

3- (6-Ethyl-4-propyl-morpholin-2-yl) -phenol

Hydrobromic acid (48% aq., 5ml) and the product described in example 46A (0.10g, 0.38mmol) were heated at 80 ℃ for 16 h. After cooling, the reaction mixture was concentrated in vacuo. The residue was partitioned between aqueous ammonia (0.880, 15ml) and dichloromethane (15ml), the layers were separated and the aqueous layer re-extracted with dichloromethane (2X 15 ml). The combined organic extracts were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica eluting with methylene chloride and then methylene chloride: methanol (99: 1 to 95: 5) to give the title compound as a clear oil (65mg, 0.26mmol, 69%) as the single diastereomer.

¹H NMR(CDCl₃，400MHz)δ：0.95(m 6H)，1.60(m，3H)，1.85(m，1H)，2.25(m，2H)，2.45(m，2H)，2.55(q，1H)，2.75(d，1H)，3.75(s，1H)，4.80(m，1H)，6.70(d，1H)，6.90(s，1H)，7.00(1H，d)，7.25(t，1H).LRMS(APCI)：m/z 250(MH⁺) Analyzed found C, 70.94%; h, 9.16%; n, 5.53%. C₁₅H₂₃NO₂.0.3H₂Theoretical value of O, C, 70.72%; h, 9.34%; and N, 5.50%.

Example 47B

3- (6-Ethyl-4-propyl-morpholin-2-yl) -phenol

The preparation was carried out in the same manner as in example 47A, starting from the product described in example 46B (0.10 g; 0.38 mmol). Without purification by silica column chromatography. The title compound was obtained as a yellow oil (57mg, 0.23mmol, 60%) as a single diastereomer.

¹H NMR(CDCl₃，400MHz)δ：0.90(t，3H)，1.00(t，3H)，1.60(m，4H)，1.85(t，1H)，2.00(t，1H)，2.35(m，2H)，2.90(d，1H)，3.00(d，1H)，3.65(m，1H)，4.60(m，1H)，6.75(d，1H)，6.80(s，1H)，6.90(1H，d)，7.20(t，1H).LRMS(ESl)：m/z 250(MH⁺)，248(M-H^-) Found C, 71.63%; h, 9.19%; n, 5.55%. C₁₅H₂₃NO₂.0.1H₂Theoretical value of O, C, 71.73%; h, 9.31%; n, 5.58 percent.

Example 48

2-methyl-6- (3-methoxy-phenyl) -4-propyl-morpholin-3-one

Preparation was carried out following the same procedure as in example 45, starting from the product described in example 3 (0.44g, 2.10mmol) and 2-chloropropionyl chloride (0.25ml, 2.50 mmol). The title compound was not purified by silica column chromatography. The title compound was obtained as a clear oil (0.42g, 1.60mmol, 76%) as a mixture of diastereomers.¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.60(m，5H)，3.30(bm，2H)，3.50(bm，2H).3.80(s，3H)，4.40(q，0.5H)，4.55(q，0.5H)，4.80(dd，0.5H)，4.95(dd，0.5H)，6.85(d，1H)，6.95(s，2H)，7.25(m，1H).LRMS(APCI)：m/z 264(MH⁺)，262(MH^-).

Examples 49A and 49B

2-methyl-6- (3-methoxy-phenyl) -4-propyl-morpholine

The preparation was carried out in the same manner as in example 46, starting from the product described in example 48 (0.42 g; 1.6 mmol). Purification by silica column chromatography eluting with ethyl acetate to pentane (1: 6) gave the two title compounds as single diastereomers.

Example 49A: clear oil (0.10g, 0.40mmol, 25%):

¹H NMR(CDCl₃，400MHz)δ：0.95(t 3H)，1.30(d，3H)，1.60(m，2H)，2.20-2.35(m，3H)，2.50(d，1H)，2.60(m，1H)，2.65(d，1H)，3.80(s，3H)，4.00(s，1H)，4.85(s，1H)，6.80(d，1H)，7.05(m，2H)，7.25(m，1H).LRMS(APCI)：m/z 250(MH⁺).

example 49B: clear oil (0.10g, 0.40mmol, 25%):

¹H NMR(CDCl₃，400MHz)δ：0.90(t3H)，1.25(m，3H)，1.60(m，2H)，1.80(m，1H)，2.00(bm，1H)，2.35(s，2H)，2.80(d，1H)，2.90(d，1H)，3.80(s，3H)，3.85(s，1H)，4.60(s，1H)，6.80(d，1H)，7.00(m，2H)，7.25(m，1H).LRMS(APCI)：m/z 250(MH⁺).

example 50A

3- (6-methyl-4-propyl-morpholin-2-yl) -phenol

Preparation was carried out in the same manner as in example 47A, starting from the product of example 49A (0.10 g; 0.4 mmol). Purification by column chromatography on silica eluting with dichloromethane then dichloromethane: methanol (99: 1) gave the title compound as a clear oil (70mg, 0.30mmol, 74%) as the single diastereomer.

¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.35(d，3H)，1.55(m，2H)，2.25(m，2H)，2.35(m，1H)，2.50(m，1H)，2.55(m，1H)，2.75(d，1H)，4.05(s，1H)，4.85(m，1H)，6.70(d，1H)，6.90(s，1H)，7.00(1H，d)，7.20(t，1H).LRMS(APCI)：m/z 236(MH⁺) Analyzed found C, 70.62%; h, 8.89%; n, 5.95%. C₁₄H₂₁NO₂.0.1H₂Theoretical value of O, C, 70.91%; h, 9.01%; n, 5.91 percent.

Example 50B

3- (6-methyl-4-propyl-morpholin-2-yl) -phenol

Preparation was carried out following the same procedure as in example 47A, starting from the product described in example 49B (0.10g, 0.4 mmol). Without purification by silica column chromatography. The title compound was obtained as a yellow oil (100mg, 0.42mmol, 103% -3% starting material) as a single diastereomer.

¹H NMR(CDCl₃，400MHz)δ：0.90(t，3H)，1.25(d，3H)，1.60(m，2H)，1.85(m，1H)，2.00(m，1H)，2.35(m，2H)，2.85(d，1H)，3.00(d，1H)，3.85(s，1H)，4.60(d，1H)，6.75(d，1H)，6.80(s，1H)，6.90(1H，d)，7.20(m，1H).LRMS(APCI)：m/z 236(MH⁺) Analysis found C, 69.38%; h, 8.86%; n, 5.73%. C₁₄H₂₁NO₂.0.45H₂Theoretical value of O, C, 69.33%; h, 9.06%; n, 5.78%.

Example 51

1- (4-chloro-3-methoxy-phenyl) -2-propylamino-ethanol

Sodium triacetoxyborohydride (1.25g, 5.89mmol) was added carefully to a solution of 2-amino-1- (4-chloro-3-methoxy-phenyl) -ethanol (j.med. chem., 30(10), 1887, (1987)) (600mg, 2.98mmol) and propionaldehyde (0.22ml, 2.96mmol) in dichloromethane (10ml) and the reaction mixture was stirred at room temperature for 1 hour. Sodium bicarbonate solution (saturated aqueous solution, 10ml) was added dropwise, and then the reaction mixture was further diluted with water (20ml) and dichloromethane (20 ml). The aqueous layer was separated and re-extracted with dichloromethane (2X 20 ml). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica eluting with dichloromethane: methanol: 0.880 ammonia (95: 5: 0.5 to 92: 8: 0.8) to give the title compound as a solid (320mg, 1.31mmol, 44%).

¹H NMR(CDCl₃，400MHz)δ：0.90(t，3H)，1.50(q，2H)，2.50-2.70(m，5H)，2.90(dd，1H)，3.80(s，3H)，4.65(dd，1H)，6.85(d，1H)，7.00(1H，d)，7.30(bd，1H).LRMS(APCI)：m/z 244(MH⁺)，226(MH⁺ less H₂O).

Example 52

6- (4-chloro-3-methoxy-phenyl) -4-propyl-morpholin-3-one

Chloroacetyl chloride (0.11ml, 1.33mmol) was added to a solution of the product described in example 51 (0.31g, 1.27mmol) and triethylamine (0.19ml, 1.36mmol) in dichloromethane (10ml) and stirred at room temperature for 60 h. The reaction mixture was diluted with dichloromethane (20ml) and washed with hydrochloric acid (aq.1N, 10ml), water (10ml) and sodium bicarbonate solution (saturated aqueous solution, 10 ml). The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the acyclic product as an oil (0.40 g). Lrms (apci): m/z 320 (MH of the uncyclized product)⁺) 302 (MH with small amount of water)⁺) 284 (MH of the cyclization product)⁺). Potassium hydroxide (0.75g, 1.33mmol) was added to a solution of the uncyclized product (0.40g, 1.23mmol) in isopropanol (10ml) and water (0.4ml) and stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo and partitioned between dichloromethane (30ml) and water (30 ml). The layers were separated and the aqueous layer re-extracted with dichloromethane (2X 20 ml). The organic layers were combined, washed with water (30ml), dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as an oil (0.34g, 1.19mmol, 94%).¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.60-1.70(m，2H)，3.30-3.40(m，2H)，3.40-3.55(m，2H)，3.95(s，3H)，4.35(bd，1H)，4.42(bd，1H)，4.78(dd，1H)，6.85(dd，1H)，7.00(s，1H)，7.38(dd，1H).LRMS(APCI)：m/z 284(MH⁺).

Example 53

6- (4-chloro-3-methoxy-phenyl) -4-propyl-morpholine

Borane-tetrahydrofuran complex (1M in THF) (3.5ml, 3.5mmol) was added dropwise to a solution of the product described in example 52 (0.33g, 1.16mmol) in dry THF (3ml) under a nitrogen atmosphere. The reaction mixture was refluxed for 2.5 hours, then cooled and quenched by addition of methanol (1 ml). The reaction mixture was concentrated in vacuo and the residue suspended in 4N HCl (aq., 8ml) and refluxed for 2 hours. The reaction mixture was cooled and extracted with dichloromethane (2X 10 ml). The organic layer was discarded. Solid potassium carbonate was added to the aqueous layer to make it basic (pH9-10) and re-extracted with dichloromethane (2X 15 ml). The dichloromethane extract was washed with water (10ml), dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as an oil (0.31g, 1.15mmol, 99%).¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.45-1.60(m，2H)，2.00(t，1H)，2.20(t，1H)，2.35(t，2H)，2.80(d，1H)，2.90(d，1H)，3.80(t，1H)，3.90(s，3H)，4.03(dd，1H)，4.55(d，1H)，6.85(dd，1H)，7.00(s，1H)，7.30(dd，1H).LRMS(APCI)：m/z 270(MH⁺).

Example 54

2-chloro-5- (4-propyl-morpholin-2-yl) -phenol

The preparation was carried out following the same procedure as example 7b, except that reflux was continued for 2.5 h instead of 1h, starting from the product described in example 53 (0.28g, 1.02 mmol). Without purification by silica column chromatography. The title compound was obtained as light brown gum (0.21g, 0.82mmol, 81%).¹H NMR(CDCl₃，400MHz)δ：0.93(t，3H)，1.55(q，2H)，2.0(t，1H)，2.20(dt，1H)，2.30-2.40(m，2H)，2.80(bd，1H)，2.90(bd，1H)，3.80(dt，1H)，4.0(dd，1H)，4.30(d，1H)，6.87(dt，1H)，7.02(fd，1H)，7.25(s，1H).LRMS(APCI)：m/z 256(MH⁺) Analyzed found C, 60.71%; h, 7.10%; n, 5.45%. C₁₃H₁₈NO₂Theoretical value of Cl, 61.05%; h, 7.09%; and N, 5.48 percent.

Example 55

(2S) -2- (propionylamino) propionic acid methyl ester

L-alanine methyl ester hydrochloride (14g, 0.1mol) was dissolved in dichloromethane (150ml) and treated with triethylamine (30.45g, 0.3 mmol). The solution was stirred and propionyl chloride was added dropwise. After stirring overnight, the mixture was quenched by addition of 1M hydrochloric acid (200ml) and the organic layer was separated. The aqueous layer was re-extracted with dichloromethane (3X 200ml) and the organic layers were combined, dried over magnesium sulphate, filtered and evaporated to give a clear oil (16.0g, quant.).

¹H NMR(DMSO-d6，400MHz)δ：0.95(t，3H)，1.25(d，3H)，2.1(q，2H)，3.6(s，3H)，4.2(quin，1H)，8.2(bd，1H).LRMS(ESI+)m/z 160(MH⁺)

Example 56

(1S) -2-hydroxy-1-methylethyl (propyl) carbamic acid tert-butyl ester

The product described in example 55 was dissolved in tetrahydrofuran (200ml) and borane-tetrahydrofuran complex (300ml, 0.3mol) was added to the stirred solution at room temperature. The mixture is then refluxedHeat overnight. After cooling to room temperature, 6M hydrochloric acid (100ml) was carefully added to the reaction to quench the reaction, then heated to reflux for 6 hours. The reaction mixture was allowed to cool to room temperature overnight and then evaporated to dryness (11.77 g). The m/z of the crude mixture was 118, consistent with the desired amino alcohol intermediate. The crude mixture was then dissolved in methanol (50ml) and water (400ml) and potassium hydroxide (28.22g, 0.5mol) was added. Di-tert-butyl dicarbonate (32.87g, 0.15mol) was added to the mixture, and stirring was continued for 3 days. The reaction mixture was partitioned between DCM (500ml) and water (100ml), the organic layer was separated and the aqueous layer was extracted twice more with DCM. The organic fractions were combined, dried over magnesium sulfate, filtered and evaporated to give the crude product. Through SiO₂Flash chromatography with dichloromethane: methanol: 880NH₃(97: 3: 0.3) to give the desired product as a clear oil (4.5g, 21%), and an additional 10g of partially purified product.

¹H NMR(DMSO-d6，400MHz)δ：0.8(t，3H)，1.05(bs，3H)，1.4(m，11H)，2.95(bs，2H)，3.35(bm，3H)，4.6(bs，1H)LRMS(ESI+)m/z 240(MNa⁺)

Example 57

(2S) -2- (propylamino) propan-1-ol hydrochloride

The pure product described in example 56 (4.2g, 0.021mol) was dissolved in dioxane (10ml) and treated with 4M HCl in dioxane (30 ml). The mixture was stirred at room temperature for 16 h, then evaporated to give a white solid (2.74g, 92%).

¹H NMR(DMSO-d6，400MHz)δ：0.9(t，3H)，1.15(d，3H)，1.6(m，2H)，2.8(m，2H)，3.15(m，1H)，3.5(bm，1H)，3.6(m，1H)，5.4(bs，1H)，8.8(bd，2H).LRMS(APCI+)118(MH⁺)

Example 58

(5S) -2- (3-methoxyphenyl) -5-methyl-4-propylmorpholin-2-ol

The product described in example 57 (1.0g, 6.6mmol) was dissolved in toluene (10ml), treated with triethylamine (1.38g, 14mmol) and then 2-bromo-3' -methoxyacetophenone (1.5g, 6.6mmol) was added. The mixture was heated to 65 ℃ and stirred for 3 days. After cooling to room temperature, the mixture was partitioned between brine and ethyl acetate, the organic layer was separated, dried over magnesium sulfate, filtered and evaporated. The residue is passed through SiO₂Purification by flash chromatography eluting with ethyl acetate afforded the desired morpholinol (morpholinol) compound as a mixture of stereoisomers as a pale yellow oil (1.0g, 58%).

¹H NMR(DMSO-d6，400MHz)δ：0.8(m，3H)，0.95(d，3H)，1.35(m，2H)，2.1(m，2H)，2.4(bm，1H)，2.6(m，1H)，2.75(m，1H)，3.5(d，1H)，3.75(m，4H)，6.0(s，0.75H)，6.1(s，0.25H)，6.85(d，1H)，7.05(m，2H)，7.25(t，1H).LRMS(ESI+)m/z 248(M-H2O)，266(MH⁺)，288(MNa+)

Example 59

(5S) -2- (3-methoxyphenyl) -5-methyl-4-propylmorpholine

The product from example 58 (770mg, 2.9mmol) was dissolved in dichloromethane (20ml) and cooled to-78 ℃ under nitrogen. To the stirred mixture was added triethylsilane (3.7ml, 23mmol) followed by trimethylsilyl triflate (1.1ml, 5.8 mmol). Stirring was continued overnight to bring the reaction mixture to room temperature. To the direction ofThe reaction was quenched by addition of saturated aqueous sodium bicarbonate and extracted with dichloromethane (three times). The combined organic layers were dried over magnesium sulfate, filtered and evaporated. The crude product is passed through SiO₂Purification by flash chromatography eluting with dichloromethane: methanol: 880 ammonia (97: 3: 0.3) gave the desired morpholino compound (600mg, 83%).

¹H NMR(CDCl₃，400MHz)δ：0.95(m，3H)，1.1(b，d，3H)，1.6(bm，2H)，2.2-3.1(5H)，3.5(bm，1H)，4.85(m，4H)，4.6(b，1H)，6.8(d，1H)，6.95(m，2H)，7.25(m，1H+CHCl₃)

LRMS(APCI+)m/z 250(MH⁺)

Found analyzed C, 71.53%; h, 9.21%; n, 5.55%. C₁₅H₂₃NO₂.0.15H₂Theoretical value of O, C, 71.48%; h, 9.32%; n, 5.56 percent.

Example 60

3- [ (5S) -5-methyl-4-propylmorpholin-2-yl ] phenol

The product described in example 59 (400mg, 1.6mmol) was dissolved in 48% aqueous hydrobromic acid (8ml) and the mixture was heated to 80 ℃ overnight. After cooling to room temperature, the reaction was quenched by adding saturated aqueous sodium bicarbonate to the mixture, and the mixture was extracted with dichloromethane (three times). The organic layers were combined, dried over magnesium sulfate, filtered and evaporated to give the product as a white solid (285mg, 76%).

¹H NMR(CDCl₃，400MHz)δ：0.9(m，3H)，1.1+1.2(2×d，3H)，1.5(m，2H)，2.3(m，2H)，2.5(bm，1H)，2.8(bm，1H)，3.1(d，1H)，3.5(bm，1H)，3.85(bm，1H)，4.6(d，1H)，6.8(m，2H)，6.95(m，1H)，7.2(t，1H)

LRMS(APCI+)，236(MH⁺)

Found analyzed C, 70.61%; h, 9.00%; n, 5.86%. C₁₄H₂₁NO₂.0.1H₂Theoretical value of O, C, 70.91%; h, 9.01%; n, 5.91 percent.

The diastereomer mixture was separated on a Chiralcel OJ-H (250 × 21.2mm) HPLC column. The mobile phase 100% MeOH, flow 15 ml/min. Samples were prepared by dissolving 200mg in 4ml MeOH and injecting 250. mu.L. Two major peaks were obtained with retention times of 5.822min (example 60A, 57mg 28%) and 7.939min (example 60B, 12mg, 6%).

Example 60A:¹H NMR(CDCl₃，400MHz)δ：0.85(t，3H)，1.05(d，3H)，1.5(m，2H+H₂O)，2.2(m，2H)，2.4(m，1H)，2.8(m，1H)，3.0(d，1H)，3.4(t，1H)，3.9(dd，1H)，4.55(d，1H)，5.6(bs，1H)，6.75(d，1H)，6.85(s，1H)，6.95(d，1H)，7.2(t，1H)

HRMS m/z 236.1643(MH⁺)

example 60B:¹H NMR(CDCl₃，400MHz)δ：0.95(t，3H)，1.15(d，3H)，1.55(m，2H)，2.4(m，2H)，2.55(t，1H)，2.65(dd，1H)，2.95(bm，1H)，3.8(d，1H)，3.95(d，1H)，4.55(dd，1H)，6.75(d，1H)，6.85(s，1H)，6.95(d，1H)，7.2(t，1H)HRMS m/z 236.1643(MH⁺)

example 61

(S) -2-propylamino-propan-1-ol hydrochloride

To a solution of (S) - (+) -2-amino-1-propanol (19.6g, 0.26mol) in dichloromethane (500ml) was added propionaldehyde (20.9ml, 0.28mol) followed by pre-dried, crushed 4A molecular sieves (40g),the mixture was stirred at room temperature overnight. The mixture was filtered through a pad of celite, the pad was washed with dichloromethane and the solvent was evaporated to give a clear oil. The oil was dissolved in methanol (200ml) and NaBH added in portions over 15 minutes₄. The mixture was stirred at room temperature overnight, then quenched by careful addition of 2M HCl (aq) (200ml), basified by addition of 2M NaOH (200ml) and methanol evaporated. Di-tert-butyl dicarbonate (115g, 0.52mol) was added followed by 1, 4-dioxane (200ml), and the mixture was stirred at room temperature overnight. The 1, 4-dioxane was removed by evaporation to give a clear oil. To the oil was added 4M HCl in 1, 4-dioxane (200ml) and the mixture was stirred at room temperature overnight. The solvent was removed by evaporation to give a white solid (24 g).

¹H NMR(DMSO，400MHz)δ：0.95(t，3H)，1.2(d，3H)，1.6(m，2H)，2.8(m，2H)，3.15(m，1H)，3.5(bm，1H)，3.6(m，1H)，5.4(b，1H)，8.6-8.9(bd，2H)LRMS(APCI+)，118(MH⁺)

Example 62

(5S) -4-propyl-5-methylmorpholin-2-one

The product from example 61 (4g, 26mmol) was dissolved in benzene followed by the addition of N-ethyldiisopropylamine (9.07ml, 52mmol) and methyl bromoacetate (2.4ml, 26 mmol). The mixture was heated to reflux overnight with azeotropic removal of water. The solvent is removed by evaporation, the crude product is dissolved in methanol and pre-adsorbed onto SiO₂Through SiO₂Flash chromatography eluting with 40% EtOAc/pentane afforded the title morpholinone as a clear oil (1.78 g).

¹H NMR(CDCl₃，400MHz)δ：0.9(t，3H)，1.1(d，3H)，1.5(m，2H)，2.25(m，1H)，2.6(m，1H)，2.8(m，1H)，3.2(d，1H)，3.6(d，1H)，4.05(dd，1H)，4.3(dd，1H)t.l.c.Rf＝0.18(50％EtOAc/Pentane，UV visualisation)

Example 63

(5S) -2- [6- (2, 5-dimethyl-1H-pyrrol-1-yl) pyridin-3-yl ] -4-propyl-5-methylmorpholin-2-ol

5-bromo-2- (2, 5-dimethyl-pyrrol-1-yl) -pyridine (1.5g 5.9mmol) was azeotroped with toluene and dissolved in THF (20 ml). The mixture was cooled to-78 ℃ and tert-butyllithium (1.7M pentane solution, 7ml, 11.9mmol) was added while maintaining the temperature below-70 ℃. The product of example 62 was dissolved in THF (20ml) and added to the mixture immediately after the addition of tert-butyllithium. The mixture was stirred at-78 ℃ for 30 minutes, at which time NH was added₄Cl (10% aq, 150ml), the mixture was extracted with EtOAc (200ml), dried over magnesium sulfate, filtered and evaporated. Through SiO₂Flash chromatography with a step gradient of 25% EtOAc/pentane to 50% EtOAc/pentane afforded the title compound as a mixture of diastereomers in a ratio of about 3.5: 1 as a yellow oil (480 mg).

¹H NMR(CDCl₃400MHz) (diastereomer) δ: 0.95(m, 3H), 1.1, 1.2(2 xd, 3H)1.5(m, 2H), 2.15(s, 6H), 2.4(m, 1H), 2.5(d, 1H), 2.6(m, 1H), 2.75(m, 1H)3.85-3.95(m, 1H), 3.6, 3.75, 4.4(3 xm, 2H), 5.15(bs, 1H), 5.9(s, 2H), 7.2(d, 1H), 8.05(dd, 1H), 8.8(s, 1H)

LRMS(ES+)，330(MH⁺)，352(MNa+)

LRMS(ES-)，328(M-H)

Example 64

(2S) -2- [ { (2RS) -2- [6- (2, 5-dimethyl-1H-pyrrol-1-yl) pyridin-3-yl ] -2-hydroxyethyl } propyl ] amino ] propan-1-ol

Reacting (5S) -2- [6- (2, 5-dimethyl-1H-pyrrol-1-yl) pyridin-3-yl]-4-propyl-5-methylmorpholin-2-ol (480mg, 1.45mmol) is dissolved in ethanol (5ml) and water (2ml) and treated with sodium borohydride (220mg, 5.8 mmol). The reaction mixture was stirred at room temperature overnight, then quenched by addition of saturated aqueous NH4Cl solution (50ml) and extracted with ethyl acetate (2X 100 ml). The combined organic extracts were washed with MgSO₄Drying and evaporation gave 400mg fluffy white solid which was used without further purification.

¹H NMR(CDCl₃400MHz) diastereomer δ: 0.8-1.1(m, 6H), 1.15, 1.35(2 xd, 3H), 1.6-2.0(m, 2H)2.1(s, 6H), 2.5-4.05(m, 7H), 4.8-5.2(m, 1H), 5.9(s, 2H), 7.2(m, 1H), 7.8-8.1(m, 1H), 8.55(m, 1H).

LRMS(ES+)，332(MH⁺)

Example 65

(2S) -2- [ [ (2RS) -2- (6-aminopyridin-3-yl) -2-hydroxyethyl ] (propyl) amino ] propan-1-ol

Reacting (2S) -2- [ [ (2RS) -2- (6-aminopyridin-3-yl) -2-hydroxyethyl](propyl) amino group]Propan-1-ol (400mg, 1.2mmol) was dissolved in EtOH (5ml), hydroxylamine hydrochloride (419mg, 6mmol) was added and the mixture was heated to 80 ℃ overnight. The solvent was removed under vacuum and the residue was passed over SiO₂Purification by flash chromatography eluting with dichloromethane/methanol/880 ammonia (95: 5: 0.5, increasing polarity to 93: 7: 1) gave the title compound as a mixture of diastereomers (300mg, 98%).

¹H NMR(CDCl₃400MHz) (2 (diastereomers) δ: 0.82-0.97(6H, m), 2.40-2.77(2H, m), 3.27-3.51(2H, m), 4.51(1H, m), 6.58(1H, m), 7.49(1H, m), 7.86(1H, m)

LRMS(APCI+)，254(MH⁺)

Examples 66 and 67

5- [ (2S, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine

And

5- [ (2R, 5S) -5-methyl-4-propylmorpholin-2-yl ] pyridin-2-amine

The "diol" (300mg, 1.2mmol) of example 65 was dissolved in dichloromethane (3ml) and concentrated sulfuric acid (3ml) was added. The mixture was stirred at room temperature for 3 hours. The reaction was cooled to 0 ℃ and quenched by careful addition of 6M sodium hydroxide solution and then extracted with dichloromethane (4X 50 ml). The combined extracts were dried (MgSO)₄) Evaporation gave a brown gummy solid. Through SiO₂Purification by flash chromatography eluting with 10% methanol in ethyl acetate afforded 5mg of the product enriched in the less polar diastereomer (about 80% d.e.), 12mg of the product enriched in the less polar diastereomer (about 80% d.e.), and 150mg of an about 1: 1 mixture of diastereomers (total yield 167mg, 59%). The latter 1: 1 mixture was subjected to HPLC purification on a Chiralpak OD-H column (250X 21.2mm) eluting with methanol/ethanol (1: 1).

Faster eluting diastereomer was obtained (> 99% d.e. (60mg, 21%).

¹H NMR(CDCl₃，400MHz)0.88(3H，t)，1.01(3H，d)，1.26(3H，t)，1.37-1.58(2H，m)，2.18-2.28(2H，m)，2.36-2.47(1H，m)，2.69-2.77(1H，m)，2.90(1H，m)，3.38(1H，m)，3.72(2H，d)，3.82(1H，m)，4.40(2H，brs)，4.45(1H，dd)，6.48(1H，d)，7.45(1H，dd)，8.04(1H，d)

LRMS(ES⁺)：m/z 236(MH⁺)

[α]_D ²⁵ 46.28(c 0.13，MeOH)

The slower eluting diastereomer was obtained (> 99% d.e. (62mg, 22%).¹H NMR(CDCl₃，400MHz)0.93(3H，t)，1.11(3H，d)，1.49(2H，m)，2.38(2H，m)，2.50-2.56(2H，m)，2.89(1H，m)，3.75(1H，m)，3.89(1H，m)，4.40(2H，brs)，4.46(1H，m)，6.50(1H，d)，7.50(1H，dd)，8.07(1H，d)

LRMS(ES⁺)：m/z 236(MH⁺)

[α]_D ²⁵ 22.58(c 0.13，MeOH)

Claims

1. Compounds of formula (Ia) or (Ib)

Wherein:

a is selected from C-X,

b is selected from C-Y, and B is selected from C-Y,

R¹is selected from H and (C)₁-C₆) An alkyl group, a carboxyl group,

R²is selected from H and (C)₁-C₆) An alkyl group, a carboxyl group,

x is selected from H, HO, C (O) NH₂And NH₂，

Y is selected from H, HO, NH₂Br, Cl and F, in the presence of a catalyst,

z is selected from H, HO, F, CONH₂And CN;

or a pharmaceutically acceptable salt or solvate thereof;

the conditions are as follows:

x, Y and Z must be at least one OH.

2. A compound of formula (Ia) or (Ib) as claimed in claim 1, wherein R is¹Selected from H, methyl and ethyl.

3. A compound of formula (Ia) or (Ib) as claimed in claim 1 or 2 wherein R is²Selected from H and methyl.

4. A compound of formula (Ia) or (Ib) as claimed in claim 1 or 2 wherein X is selected from H, OH and NH₂。

5. A compound of formula (Ia) or (Ib) as claimed in claim 1 or 2 wherein Y is selected from H, NH₂Cl and F.

6. A compound of formula (Ia) or (Ib) as claimed in claim 1 or 2 wherein Z is selected from H, OH and F.

7. A compound according to claim 1 which is

R- (-) -3- (4-propylmorpholin-2-yl) phenol;

s- (+) -3- (4-propylmorpholin-2-yl) phenol;

r- (-) -3- (4-propylmorpholin-2-yl) phenol hydrochloride;

r-5- (4-propylmorpholin-2-yl) benzene-1, 3-diol;

s-5- (4-propylmorpholin-2-yl) benzene-1, 3-diol;

r- (+) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol;

s- (-) -2-fluoro-5- (4-propylmorpholin-2-yl) phenol;

2-chloro-5- (4-propylmorpholin-2-yl) phenol; or

A pharmaceutically acceptable salt or solvate thereof.

8. The compound according to claim 7, which is R- (-) -3- (4-propylmorpholin-2-yl) phenol.

9. A compound according to claim 8 in the form of a hydrochloride salt.

10. Use of a compound of formula (Ia) or (Ib) as claimed in any one of claims 1-8 in the manufacture of a medicament for the treatment of a condition selected from: sexual dysfunction, hypertension, neurodegeneration, depression, generalized anxiety disorder, phobias, post-traumatic stress syndrome, avoidant personality disorder, eating disorder, obesity, chemical dependence, pain, Alzheimer's disease, obsessive-compulsive disorder, panic disorder, memory disorders, Parkinson's disease, endocrine disorders, vasospasm, cerebellar ataxia, gastrointestinal disorders, negative symptoms of schizophrenia, premenstrual syndrome, fibromyalgia syndrome, stress urinary incontinence, Tourette's syndrome, trichotillomania, kleptomania, attention deficit hyperactivity disorder, mood instability, pathological crying, sleep disorders, cataplexy and shock.

11. The use as claimed in claim 10, wherein the condition is female sexual dysfunction, male erectile dysfunction, neurodegeneration and depression.

12. The use as claimed in claim 11, wherein the condition is male erectile dysfunction.

13. The use as claimed in claim 11, wherein the condition is hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder and dyspareunia in women.

14. The use as claimed in claim 13, wherein the condition is female sexual arousal disorder and concomitant hypoactive sexual desire disorder.

15. The use as claimed in claim 10, wherein the condition is pain.

16. The use as claimed in claim 10, wherein the condition is chronic paroxysmal pain.

17. A composition which comprises a compound of formula (Ia) or (Ib) as claimed in any one of claims 1 to 8 and a pharmaceutically acceptable diluent or carrier.