Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/17—Amides, e.g. hydroxamic acids having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/18—Sulfonamides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
  • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/445—Non condensed piperidines, e.g. piperocaine
  • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
  • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
  • A61P15/10—Drugs for genital or sexual disorders; Contraceptives for impotence
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• the present invention relates to methods for the treatment of sexual dysfunction and to the preparation of medicaments for the treatment of sexual dysfunction.
• Sexual dysfunctions are relatively common in the general population (see O'Donohue W, et al, Clin. Psychol Rev. 1997; 17: 537-566).
• the disorder may relate to seeking sexual behaviour (proceptivity) and/or to acceptance of sexual behaviour, accompanied by sexual arousal (receptivity).
• the prevalence of sexual problems is higher in populations receiving medicaments, in particular antidepressants and antihypertensives.
• a need for pharmacotherapy for sexual dysfunction is increasing, but there has been very little research effort directed at finding drugs to treat sexual dysfunction.
• Sexual dysfunctions include erectile dysfunctions of organic and psychogenic origin (Benet A.E and Melman A, 1995, Urol. Clin. N. Amer. 22: 699-709) as well as hypoactive sexual desire disorders, sexual arousal disorders, anorgasmy and sexual pain disorders (Berman et al., 1999, Urology 54: 385-391).
• impotence can be defined as an inability to achieve penile erection or ejaculation. Its prevalence is claimed to be between 2% and 7% of the human male population, increasing with age up to 50 years and between 18% and 80% between 55 and 80 years of age. In the USA alone, for example, it has been estimated that there are up to 10 million impotent males, with the majority suffering from problems of organic rather than of psychogenic origin. Although many different drags have been shown to induce penile erection, they were only effective after direct injection into the penis e.g. intraurethrally or intracavernosally (i.e.) and were not approved for erectile dysfunction.
• US-A-5576290 discloses peptides which are stated to induce erection, but they have to be given subcutaneously e.g. by injection, and if an excessive dose is given they produce an exaggerated erectile response and stomach discomfort. Impotence treatment was revolutionized by the unexpected discovery that cGMP PDE inhibitors, e.g. pyrazolo[4,3-d]pyrimidin-7-ones were useful in the treatment of erectile dysfunction and could be administered orally, therefore obviating the disadvantages associated with i.e. administration.
• cGMP PDE inhibitors e.g. pyrazolo[4,3-d]pyrimidin-7-ones were useful in the treatment of erectile dysfunction and could be administered orally, therefore obviating the disadvantages associated with i.e. administration.
• sildenafil Viagra
• This invention is based on the realisation that substances that act as bombesin receptor antagonists have utility in the treatment of sexual dysfunction, including the behavioural component thereof, in both male and female subjects.
• they can provide a treatment for erectile dysfunctions of organic and psychogenic origin as well as hypoactive sexual desire disorders, sexual arousal disorders, anorgasmy and sexual pain disorders.
• the invention therefore provides a method of treating sexual dysfunction which comprises administering to a subject suffering therefrom and in need of treatment an effective amount of a bombesin receptor antagonist.
• the invention further provides the use of a bombesin receptor antagonist in the manufacture of a medicament for preventing or treating male sexual dysfunction or female sexual dysfunction.
• many of the compounds of this invention have both the property of binding to bombesin receptors and the property that an effective dose can be administered orally.
• Figure 1 Effect of (S) 3-(lH-Indol-3-yl)-N-[l-(5-methoxy-pyridin-2-yl)-cyclohexyl- methyl]-2-methyl-2-[3-(4-nitro-phenyl)-ureido]-propionamide (Compound (1)) on female rat sexual proceptivity.
• Figure 2 Effect of Compound (1) on female rat sexual receptivity.
• Figure 3 Effect of repeated administration of Compound (1) on female rat proceptivity.
• Figure 4 Effect of intracerebro entricular administration of Compound (i) on female rat sexual proceptivity.
• Figure 5 Inhibitory effect of NMB on female rat sexual proceptivity and antagonism of this effect by Compound (1) .
• Figure 6 Results of an investigation to show whether the effect of Compound (1) on female sexual behaviour is mediated through progesterone.
• Figure 7 Results of an investigation to show whether the effect of Compound (1) on female sexual behaviour is mediated through oestradiol.
• Figure 8 Results of an investigation to show whether the effect of Compound (1) on female sexual behaviour is mediated through prolactin.
• Figure 9 Results of an investigation to show whether the effect of Compound (1) on female sexual behaviour is mediated through LH.
• Figure 10 Results of an investigation to show whether the effect of Compound (T) on female sexual behaviour is mediated through FSH.
• Figure 11 Effect of Compound (1) on the sexual behaviour of normal male rats (Mount Latency).
• Figure 12 Effect of Compound (1) on the sexual behaviour of normal male rats (Intromission Latency).
• Figure 13 Effect of Compound (1) on the sexual behaviour of normal male rats (Number of Mounts + Intromission).
• Figure 14 Effect of Compound (1) on the sexual behaviour of normal male rats (Ejaculation Latency).
• Figure 15 Effect of Compound (1) on the sexual behaviour of normal male rats (Refractory Period).
• Figure 16 Effect of Compound (1) on the sexual behaviour of sexually dysfunctional male rats (Mount Latency).
• Figure 17 Effect of Compound (1) on the sexual behaviour of sexually dysfunctional male rats (Ejaculation Latency).
• Figure 18 Effect of Compound (I) on the sexual behaviour of sexually dysfunctional male rats (% animals ejaculating).
• Figure 19 Effect of (S)-3-(lH-Indol-3-yl)-N-[l-(5-methoxy-pyridin-2-yl)- cyclohexyhnethyl] -2-methyl-2- [4-(4-nitro-phenyl)-oxazol-2-ylamino] -propionamide (Compound (2)) in PEG 200 on female rat sexual proceptivity.
• Figure 20 Effect of Compound (2) in methylcellulose on female rat sexual proceptivity.
• Figure 21 Effect of Compound (2) in PEG 200 on female rat sexual receptivity.
• Bombesin receptors are present in hypothalamic areas. We have found that they can exert a neuromodulatory effect on sexual behaviour.
• Female sexual dysfunction can be grouped into four classes (Scrip's Complete Guide to Women's Healthcare, p.194-205, 2000), which include hypoactive sexual desire disorders, sexual arousal disorders, orgasmic disorders or anorgasmy and sexual pain disorders.
• Hypoactive sexual desire disorders can be characterised as persistent or recurrent lack of sexual thoughts/fantasies and lack of receptivity to sexual activity, causing personal distress.
• Common problems include sexual aversion disorders.
• Sexual arousal disorders can be characterised as persistent or recurrent inabihty to achieve or maintain adequate sexual excitement, causing personal distress.
• Orgasmic disorders can be characterised as persistent or recurrent difficulty or delay in attaining orgasm after adequate sexual stimulation and arousal, causing personal distress.
• sexual pain disorders can be characterised by dyspareunia, (characterised by recurrent or persistent genital pain associated with sexual intercourse), vaginismus (characterised by recurrent or persistent involuntary spasm of the muscles of the outer third of the vagina which interferes with vaginal penetration, causing personal distress) and other pain disorders (characterised by recurrent or persistent genital pain induced by non coital sexual stimulation).
• the compounds of this invention are useful in the treatment of female sexual dysfunction, and this includes female sexual dysfunction associated with hypoactive sexual desire disorders, sexual arousal disorders, orgasmic disorders or anorgasmy, or sexual pain disorders.
• the psychogenic component of male sexual dysfunction has been classified by the nomenclature committee of the International Society for Impotence Research (and is illustrated in Sachs B. D., Neuroscience and Biobehavioral Review , 2000, 24 541-560) as generalised type, characterised by a general unresponsiveness or primary lack of sexual arousal, and ageing-related dechne in sexual arousability, characterised by generalised inhibition or chronic disorders of sexual intimacy.
• the inventors beheve that there are common mechanisms underlying the pathologies of male and female psychogenic sexual dysfunctions.
• the compounds of this invention are useful in the treatment of male sexual dysfunction, especially drug-induced male sexual dysfunction and psychogenic male sexual dysfunction associated with generalised unresponsiveness and ageing-related dechne in sexual arousability.
• proceptive behaviour is under hormonal control, progesterone being essential for induction of proceptive behaviour in combination with oestrogen (Johnson M and Everitt B., Essential Reproduction (3 rd edn), Blackwell, Oxford, 1988).
• the evidence for the hormonal control of proceptive behaviour in primates is conflicting, but on the whole oestrogens and/or androgens appear to enhance proceptive behaviour (Baum M.J., J. Biosci., 1983; 33:578-582).
• proceptive behaviour in the rat includes "hopping and darting" movement, with rapid vibration of the ears.
• Tests to assess the eagerness to seek sexual contact (sexual motivation) have been reported as the most appropriate way to measure proceptivity (Meyerson BJ, Lindstrom L.H., Acta Physiol. Scand., 1973; 389 (Suppl.): 1-80).
• Receptivity, in the rat is demonstrated when the female assumes a lordotic position. This occurs when, on mounting, the male exerts pressure with his forepaws on the flanks of the receptive female.
• NPN ventromedial nucleus
• MCG midbrain central grey area
• Bombesin is a 14-amino acid peptide originally isolated from the skin of the European frog Bombina bombina (Anastasi A. et al., Experientia, 1971; 27: 166). It belongs to a class of peptides which share structural homology in their C-terminal decapeptide region (DuttaA.S., Small Peptides; Chemistry, Biology, and Clinical Studies, Chapter 2, pp 66-82). At present, two mammalian bombesin-like peptides have been identified, the decapeptide neuromedin B (NMB) and a 23-residue amino acid, gastrin-releasing peptide (GRP).
• NMB decapeptide neuromedin B
• GFP 23-residue amino acid, gastrin-releasing peptide
• BBi receptor binds neuromedin B (NMB) with higher affinity than gastrin-related peptide (GRP) and neuromedin C (NMC) and BB 2 receptors bind GRP and NMC with greater affinity than NMB. More recently evidence has emerged of two more receptor subtypes denoted BB 3 and BB 4 but due to limited pharmacology, little is known of their function at present. BBi and BB 2 receptors have a heterogeneous distribution within the central nervous system indicating that the endogenous ligands for these receptors may differentially modulate neurotransmission. Among other areas, BB t receptors are present in the ventromedial hypothalamus (Ladenheim E.E et al, Brain Res., 1990; 537: 233-240).
• Bombesin receptor antagonists to which this invention is applicable include both non-peptide compounds and peptide compounds.
• Compounds that can be formulated into compositions for oral administration, especially human oral administration, without substantial loss of activity are preferred.
• Many non-peptide compounds having the desired properties fall into this category.
• One preferred genus of compounds for use in the invention comprises bombesin receptor antagonists of the formula (I)
• n 0, 1 or 2;
• Ar is phenyl, pyridyl or pyrimidyl, each unsubstituted or substituted by from 1 to 3 substituents selected from alkyl, halogen, alkoxy, acetyl, nitro, amino, - CH 2 NR 10 R n , cyano, -CF 3 , -NHCONH , and -CO 2 R 12 ;
• R* is hydrogen or straight, branched, or cyclic alkyl of from 1 to 7 carbon atoms
• R is hydrogen or forms a ring with Rl of from 3 to 7 carbon atoms
• R 2 is hydrogen or straight, branched, or cyclic alkyl of from 1 to 8 carbon atoms which can also contain 1 to 2 oxygen or nitrogen atoms;
• R9 is hydrogen or forms with R 2 a ring of from 3 to 7 carbon atoms which can contain an oxygen or nitrogen atom; or R 2 and R ⁇ can together be a carbonyl;
• Arl can be independently selected from Ar and can also include pyridyl-N- oxide, indolyl, imidazolyl, and pyridyl;
• R4, R5, R6 S and are each independently selected from hydrogen and lower alkyl;
• R 4 can also form with R ⁇ a covalent link of 2 to 3 atoms which may include an oxygen or a nitrogen atom;
• R3 can be independently selected from Ar or is hydrogen, hydroxy, -NM ⁇ 2, N-methyl-pyrrolyl, imidazolyl, N-methyl-imidazolyl, tetrazolyl, N-methyl- tetrazolyl, thiazolyl, -CONR 13 R 14 , alkoxy,
• Rl , RU, R!2, Rl3 and R*4 are each independently selected from hydrogen or straight, branched, or cyclic alkyl of from 1 to 7 carbon atoms.
• Preferred compounds are those of Formula (la)
• Ar is phenyl unsubstituted or substituted with 1 or 2 substituents selected from isopropyl, halo, nitro, and cyano; 4, R5, and are hydrogen;
• R is methyl or hydrogen
• R3 is 2-pyridyl or hydroxy
• Arl is indolyl, pyridyl, pyridyl-N-oxide, or imidazolyl.
• Ar is unsubstituted phenyl
• R! is cyclopentyl or tert-butyl
• R4 and R are hydrogen
• R is methyl
• R6 is hydrogen
• R3 is phenyl with two isopropyl substituents, unsubstituted phenyl, or
• Arl is indolyl.
• Ar is 2,6-diiso ⁇ ropyl-phenyl, 4-nitro-phenyl, and 4-cyano-phenyl;
• R4, R5, and R6 are hydrogen; • R is methyl;
• R 2 is hydrogen or cyclohexyl
• R3 is hydroxyl, pyridyl,
• Ar is phenyl, pyridyl, pyrimidyl, thienyl, furyl, imidazolyl, pyrrolyl or thiazolyl each unsubstituted or substituted by from 1 to 3 substituents selected from acetyl, alkoxy, alkyl, amino, cyano, halo, hydroxy, nitro, sulfonamido, sulfonyl, -CF 3 , -OCF 3 , -CO 2 H, -CH 2 CN, -SO 2 CF 3 , -CH 2 CO 2 H and -
• R 7 and R 8 are each independently selected from H, straight or branched alkyl of up to 6 carbon atoms, or R and R 8 together with the nitrogen atom to which they are linked can form a 5- to 7- membered aliphatic ring which may contain 1 or 2 oxygen atoms;
• R is hydrogen, straight or branched alkyl of up to 6 carbon atoms or cycloalkyl of between 5 and 7 carbon atoms which may contain 1 or 2 nitrogen or oxygen atoms;
• R is hydrogen, methyl, or forms with R an aliphatic ring of from 3 to 7 atoms which can contain an oxygen or nitrogen atom, or together with R 1 is a carbonyl group;
• Ar is independently selected from Ar or is indolyl or pyridyl-N-oxide; 3 4 5
• R , R , and R are each independently selected from hydrogen and lower alkyl
• R is independently selected from Ar or is hydrogen, hydroxy, alkoxy,
• R and R are each mdependently selected from hydrogen, straight or branched alkyl of up to 6 carbon atoms, or R 9 and R 10 together with the nitrogen atom to which they are linked can form a 5-to 7- membered aliphatic ring which may contain 1 or 2 oxygen or nitrogen atoms, or R 2 is
• p is 0, 1 or 2 and Ar 2 is phenyl or pyridyl;
• X is a divalent radical derived from any of the following
• R 11 and R 12 are independently selected from H, halogen, hydroxy, alkoxy, acetyl, nitro, cyano, amino, CF 3 and -(CH 2 ) t NR 13 R 14 where t can be 0 or 1, R 13 and R 14 are each independently selected from hydrogen, straight or branched alkyl of up to 6 carbon atoms or cycloalkyl of 5 to 7 carbon atoms, containing up to 2 oxygen or nitrogen atoms.
• a preferred species of compounds within the genus defined by formula (II) is represented by the formula (Ha), and includes pharmaceutically acceptable salt thereof: Ar -
• n O or l
• Ar is phenyl or pyridyl which may be unsubstituted or substituted with from 1 to 3 substituents selected from halogen, alkoxy, nitro and cyano;
• Ar 1 is independently selected from Ar or is pyridyl-N-oxide or indolyl;
• R 6 forms with R 1 an aliphatic ring of from 3 to 7 atoms which can contain an oxygen or nitrogen atom, or together with R 1 is a carbonyl group;
• R 2 is independently selected from Ar or is hydrogen, hydroxy, alkoxy,
• R and R are each independently selected from hydrogen or methyl or R 2 is any of
• R 3 , R 4 and R 5 are each independently selected from hydrogen and methyl
• R 11 and R 12 being independently selected from H, halogen, hydroxy, alkoxy, acetyl, nitro, cyano, amino, CF 3 and (CH 2 ) t NR 13 R 14 wherein t is 0 or 1 and R 13 and R 14 are independently selected from hydrogen and methyl.
• a sub-species of preferred compounds within the general formula (II) has the formula (lib) or (He):
• Ar and R 2 independently represent phenyl or pyridyl which may be unsubstituted or substituted with from 1 to 3 substituents selected from halogen, alkoxy, nitro and cyano, and pharmaceutically acceptable salts thereof.
• a particularly preferred compound falling within formula (II) is (S)-3-(lH- indol-3 -yl)-N-[ 1 -(5 -methoxy-pyridin-2-yl)-cyclohexylmethyl] -2-methyl-2- [4-(4-nitro- phenyl)-oxazol-2-ylamino]-propionamide (also referred to as Compound 2) and its pharmaceutically acceptable salts.
• a third genus of bombesin receptor antagonists according to the invention has the formula (III) and include pharmaceutically acceptable salts thereof:
• Ar is benzimidazolyl, benzofuryl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzopyrazinyl, benzotriazolyl, benzoxadiazolyl, furyl, imidazolyl, indanyl, indolyl, isoquinolyl, isoxazolyl, naphthyl, oxazolyl, phenyl, pyrazinyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidyl, pyrrolyl, quinolinyl, tetralinyl, tetrazolyl, thiazolyl, thienyl or triazolyl each unsubstituted or substituted with from 1 to 3 substituents selected from amino, acetyl, alkyl (straight chain or branched with from 1 to 6 carbon atoms), alkoxy, cyano, halogen,
• Ar 1 is independently selected from Ar and can also be pyridyl-N-oxide
• R 1 is hydrogen or straight or branched alkyl of up to 6 carbon atoms or cyclic alkyl of between 5 and 7 atoms which may contain 1 or 2 oxygen or nitrogen atoms;
• R 2 is independently selected from Ar or is hydrogen, hydroxy, alkoxy, - ⁇ Me 2 ,
• Ar 2 is phenyl or pyridyl; and, R* 2 and Rl3 are each independently selected from hydrogen, straight or branched alkyl of up to 6 carbon atoms or cyclic alkyl of between 5 and 7 carbon atoms;
• R 3 , R 4 and R 5 are each independently selected from hydrogen and lower alkyl
• R 6 is hydrogen, methyl or forms with R 1 a ring of from 3 to 7 carbon atoms which can contain an oxygen or nitrogen atom, or R 1 and R 6 can together be carbonyl.
• Ar is benzofiiryl, furyl, indolyl, isoquinolyl, naphthyl, phenyl, pyridyl, quinolyl or thienyl each unsubstituted or substituted with 1 or 2 substituents selected from alkoxy, cyano, halogen, nitro, phenyl, phenoxy, -CF 3 ,
• R 7 R 7 R -(CH 2 ) q NR R wherein R and R can form a ring of between 5 to 7 atoms
• R 7 R which may contain 1 or 2 oxygen or nitrogen atoms, or R and R can be independently selected from hydrogen, straight or branched alkyl of up to 4 carbon atoms or cyclic alkyl of 5 carbon atoms;
• Ar 1 is independently selected from Ar, preferably indolyl, and can also be pyridyl-N-oxide;
• R 1 and R 6 can form a cyclic alkyl of from 5 to 7 carbon atoms or R 1 and R 6 together are carbonyl;
• R 2 is independently selected from unsubstituted or substituted pyridyl or is hydrogen, hydroxy, alkoxy, -NMe 2 , -CONR 12 R 13 wherein R 12 andR 13 are each independently selected from H and CH 3 ;
• R 3 , R 4 and R 5 are each independently selected from hydrogen and methyl.
• R 2 is 2-pyridyl
• R 6 forms a cyclohexyl with R 1 .
• a particularly preferred group of compounds is of formula (Ilia): wherein Ar, k and X have the meanings given above in first, and the pyridine ring is optionally substituted by with 1 or 2 substituents, R and R', independently selected from alkoxy, cyano, halogen, nitro, phenyl, phenoxy, - CF 3 , -(CH 2 ) q NR 7 R 8 , wherein R 7 and R 8 together with the nitrogen atom to which they are linked can form a 5- to 7-membered aliphatic ring which may contain 1 or 2 oxygen or nitrogen atoms, or R 7 and R 8 can be independently selected from hydrogen or cyclic alkyl of between 5 to 7 carbon atoms, and their pharmaceutically acceptable salts thereof.
• Ar is benzofuryl, furyl, indolyl, isoquinolyl, naphthyl, phenyl, pyridyl, quinolyl or thienyl each unsubstituted or substituted with 1 or 2 substituents selected from alkoxy, cyano, halogen, nitro, phenyl, phenoxy, -CF 3 , -(CH 2 ) q NR 7 R 8 , wherein R 7 and R 8 can form a ring of between 5 to 7 atoms which may contain 1 or 2 oxygen or nitrogen atoms, or R 7 or R 8 can be a independently selected from hydrogen or cyclic alkyl of 5 carbon atoms, and X is -C(O)-, -OC(O)- or -SO 2 .
• N-terminal amide derivatives i.e. compounds of formula (UI) wherein X is -C(O)-
• the following compounds are most preferred: N- ⁇ (S)-2-(lH-indol-3-yl)-l-methyl-l-[(l-pyridin-2-yl-cyclohexylmethyl)- carbamoyl]-ethyl ⁇ -4-mfro-benzamide;
• N-terminal amide derivatives of formula (DI) include the following: N- ⁇ (S)-2-(lH-indol-3-yl)-l-methyl-l-[(l- ⁇ yridin-2-yl-cyclohexyhnethyl)- carbamoyl]-ethyl ⁇ -benzamide;
• the following compounds are particularly preferred:
• N-te ⁇ riinal urethane derivatives of formula (DT) include the following: ⁇ (S)-2-(lH-indol-3-yl)- 1 -methyl- 1-[(1 - ⁇ yridin-2-yl-cyclohexyhnethyl)- carbamoyl]-ethyl ⁇ -carbamic acid 3,4-dimethoxy-benzyl ester;
• N-terminal sulfonamide derivatives of formula (ID) (compounds of formula (ID) wherein X is -SO 2 -) the following compounds are particularly preferred: (S)-3-(lH-indol-3-yl)-2-methyl-2-phenyhnethanesulfonylamino-N-(l-pyridin- 2-yl-cyclohexylmethyl)- ⁇ ropionamide; (S)-2-(2-clhoro-benzenesulfonylamino)-3-(lH-indol-3-yl)-2-methyl-N-(l- pyridin-2-yl-cyclohexylmethyl)-propionamide;
• N-terminal sulfonamide derivatives of formula (ID) include the following:
• the compounds of the general formulae above are optically active.
• the scope of the invention therefore also includes: • All stereoisomers of the compounds of the above general formulae.
• alkyl groups contemplated by the invention include straight, branched, or cyclic carbon chains of from 1 to 8 carbon atoms except where specifically stated otherwise. Representative groups are methyl ethyl, propyl, isopropyl, n-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, 2-methylhexyl, n-pentyl, 1-methylbutyl,
• the lower alkyl groups include carbon chains of up to 6 carbon atoms.
• the cycloalkyl groups contemplated by the invention comprise those having 3 to 7 carbon atoms including cyclopentyl and cyclohexyl. They may be substituted with from 1 to 3 groups selected from halogens, nitro, alkyl, and alkoxy.
• alkoxy groups contemplated by the invention comprise both straight and branched carbon chains of from 1 to 6 carbon atoms unless otherwise stated. Representative groups are methoxy, ethoxy, propoxy, i-propoxy, t-butoxy, and hexoxy.
• halogen is intended to include fluorine, chlorine, bromine, and iodine.
• amine is intended to include free amino, alkylated amines, and acylated amines.
• subject includes animals, particularly mammals and more particularly humans.
• the compounds of the above general formulae all have at least one chiral centre and some have multiple chiral centres depending on their structure.
• the compounds of the present invention may exist as diastereoisomers, mixtures of diastereoisomers, or as the mixed or the individual optical enantiomers.
• the present invention contemplates all such forms of the compounds.
• the mixtures of diastereoisomers are typically obtained as a result of the reactions described more fully below.
• Individual diastereoisomers maybe separated from mixtures of the diastereoisomers by conventional techniques such as column chromatography or repetitive recrystallization.
• Individual enantiomers may be separated by conventional methods well known in the art such as conversion to a salt with an optically active compound, followed by separation by chromatography or recrystallization and reconversion to the non-salt form.
• the pharmaceutically acceptable salts include acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium acetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycoloylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate/diphosphate, polygalacturonate, salicylate
• Preferred salts are made from strong acids. Such salts include hydrochloride, mesylate, and sulfate.
• non-peptide bombesin antagonists which are believed to be suitable for use in the present invention are described and claimed in the following documents, the contents of which are inco ⁇ orated herein by reference: WO 00/09115, WO 00/09116, WO 92/07830, JP 07258081 and WO 98/07718.
• a primary urea 2a is cyclised with an appropriate bromomethyl ketone containing the group Z3 to form an oxazole ring (Intermediate 5). Hydrolysis of the methyl ester protecting group of the resulting Intermediate 5a, 5b or 5c gives the Intermediates 6 a-c.
• Example 23 In the above scheme: i) a) 10% Cul, K 2 CO 3 , DMF, 130°C b) HBTU, DIPEA, DMF, and [l-(5-methoxy-2-pyridyl)cyclohexyl]methan- amine (described in WO 98/07718) or [l-(2-pyridyl)cyclohexyl]methylamine (described in WO 98/07718) ii) a) 5-10% Cul, K 2 CO 3 , TEBA, Pd(P( ⁇ -tolyl) 3 )Cl 2 , DMF, 130°C b) HBTU, DIPEA, DMF, and [l-(5-methoxy-2- ⁇ yridyl)cyclohexyl]methan- amine (described in WO 98/07718) or [l-(2-pyridyl)cyclohexyl]methylamine (described in WO 98/0771
• the resulting nitrile is reduced using Raney nickel under an atmosphere of hydrogen.
• the condensation may be carried out in dimethylformamide using O-benzotriazol-l-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) and NN-diisopropyl-ethylamine (DIPEA) as catalyst.
• HBTU O-benzotriazol-l-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate
• DIPEA NN-diisopropyl-ethylamine
• the compound of formula (DI-3) may be reacted with 4- nitrophenyl chloroformate in dichloromethane using pyridine as catalyst, and the resulting carbonate may be reacted with the amine of formula (HI-2) in dimethyl formamide using N, N-dimethyl-4-amino pyridine as catalyst.
• the condensation may be carried out in dimethylformamide in the presence of N,N-diisopropylethylamine and NN-dimethyl- 4-aminopyridine.
• the amine of formula (ITI-2) is preferably a chiral amine of formula (ID-5) wherein the pyridine ring is optionally substituted by with 1 or 2 substituents R and R' selected from alkoxy, cyano, halogen, nitro, phenyl, phenoxy, - CF 3 , - (CH 2 ) q NR 7 R 8 , wherein R 7 and R 8 can form a ring of between 5 to 7 atoms, which may contain 1 or 2 oxygen or nitrogen atoms, or R 7 and R 8 can be independently selected from hydrogen or cyclic alkyl of from 1 to 5 carbon atoms, methoxy being a particularly preferred substituent, as in the chiral amine (DI-6):
• Bombesin antagonists which are peptides and which are believed to be suitable for use in the present invention are described in the following documents, the contents of which are incorporated herein by reference:
• inert, pharmaceutically acceptable carriers can be either solid or liquid.
• Solid form preparations include powders, tablets, dispersible granules, capsules, sachets, and suppositories.
• a solid carrier can be one or more substances which may also act as diluents, flavouring agents, solubihzers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.
• the carrier is a finely divided solid which is in a mixture with the finely divided active component.
• the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
• the powders and tablets preferably contain 5% to about 70% of the active component.
• Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.
• Liquid form preparations include solutions, suspensions, and emulsions.
• Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavouring agents, stabilizers, and thickening agents as desired.
• Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.
• the pharmaceutical preparation is in unit dosage form.
• the preparation is divided into unit doses containing appropriate quantities of the active component.
• the unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparation, for example, packeted tablets, capsules, and powders in vials or ampoules.
• the unit dosage form can also be a capsule, sachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.
• a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized moulds and allowed to cool and solidify.
• compositions that are adapted for oral administration to humans are preferred, especially such compositions in unit dosage form.
• bombesin receptor antagonists could be used as part of a medicament in combination with one or more vasodilator, hormone therapy or neurotransmitter modulator. Such products are used or tested in the treatment of sexual dysfunction.
• Vasodilators for the treatment of sexual dysfunctions of organic (rather than psychogenic) origin act at the penis, clitoris or vagina level on local blood flow or lubricant secretions.
• Vasodilators useful for the treatment of sexual dysfunction include alprostadil or phentolamine, NO (nitric oxide) enhancers such as L-arginine, and PDE5 inhibitors such as sildenafil or a pharmaceutically acceptable salt thereof (Scrip 's Complete Guide to Women's Healthcare, p.194-205, 2000)(Sachs B.D., Neuroscience and Biobehavioral Review , 2000, 24 541-560, Benet and Mehnan, 1995, Urol. Clin.N. Amer.
• NO nitric oxide
• PDE5 inhibitors such as sildenafil or a pharmaceutically acceptable salt thereof
• NIP Neuronal Peptide
• angiotensin-2 receptor antagonists such as losartan (American Heart Association meeting, New Jersey, 2000).
• Hormone therapies useful in the treatment of sexual dysfunction of organic and psychogenic nature include modulators of steroid hormones, steroid hormones or hormone product (including synthetic hormones) including oestrogen (Scrip's Complete Guide to Women's Healthcare, p.194-205, 2000), or androgens such as testosterone (Scrip's Complete Guide to Women's Healthcare, p.194-205, 2000, Sachs B.D., Neuroscience and Biobehavioral Review, 2000, 24 541-560), which act in areas of the CNS associated with sexual desire and sexual arousal (Wilson CA. Pharmacological targets for the control of male and female sexual behaviour. In: Sexual Pharmacology, Riley AJ, Peet M, Wilson CA (Eds), Clarendon Press, Oxford. 1993:1-58).
• Neurotransmitter modulators useful in the treatment of both psychogenic and organic sexual dysfunction include neurotransmitter agonists and antagonists such as catecholamine agonists such as the D2 agonist quinelorane, 5HT2 antagonists such as ritanserin, monoamine synthesis modifiers such as treatments that reduce endogenous 5HT activity, including inhibition of 5HT synthesis using para- chlorophenylalanine, monoamine metabolism or uptake modifiers that inhibit catecholamine metabolism or reuptake, such as tricyclic antidepressants, e.g. imipramine (Wilson CA. Pharmacological targets for the control of male and female sexual behaviour. In: sexual Pharmacology, Riley AJ, Peet M, Wilson CA (Eds), Clarendon Press, Oxford. 1993:1-58).
• neurotransmitter agonists and antagonists such as catecholamine agonists such as the D2 agonist quinelorane, 5HT2 antagonists such as ritanserin, monoamine synthesis modifiers such as treatments that reduce endogenous 5
• this combination therapy includes the preparation of therapies that would allow administration of both components of the medicament, i.e. bombesin receptor antagonists and a vasodilator, hormone therapy medicament or neurotransmitter modulator medicament in a single dose.
• a preferred formulation would allow oral administration.
• administration by suppository, cream, transdermal patch or injection is also part of this invention.
• the inventors envisage formulations which allow administration of the bombesin receptor antagonist via a separate route to that of the vasodilator, hormone therapy medicament or neurotransmitter modulator medicament.
• Such routes could include for example oral administration of the bombesin receptor antagonist and transdermal patch application of the vasodilator.
• kits in which unit doses of bombesin receptor antagonist occur in association with unit doses of the vasodilator, hormone therapy medicament or neurotransmitter modulator medicament.
• the two dosage forms could be provided in the form of a two-row tear-off strip in which compartments containing the tablets, etc. occur above compartments containing the transdermal patches.
• Other forms of packaging in which the two dosage forms are spatially associated so as to make it easy for patients to take them together and to be reminded when they have done so will readily occur to those skilled in the art.
• the kit will also contain instructions as to when and how the individual components of the kit should be administered.
• Ovariectomised adult female Sprague Dawley rats (180-200g, from Charles River) were housed in groups of 6 in a reversed lighting system of 12h light: dark (lights off 7.00-19.00h). Two weeks after ovariectomy they were used for sexual activity tests. The experiments started at least 5h into the dark period.
• Tests were carried out in a circular arena of 90cm diameter, surrounded by a 30cm high wall.
• Two small cages with wire-mesh front (15x15cm) are fixed into the wall such that the front of the cage is «flush» with the wall and the 2 cages are opposite each other.
• sexually naive test and control animals were used.
• test and control animals were primed with 5 ⁇ g oestradiol benzoate.
• progesterone 0.5mg/0.1ml
• s.c subcutaneously
• Test and control animals were introduced one at a time for 10 minute periods into the arena. During the lOmin test, the time that the test or positive control animal spent investigating each stimulus animal was noted. The arena was thoroughly cleaned between animals. The position of the male/female stimuli boxes was randomised between animals, in order to avoid place preference. The difference in the percentage of time spent investigating the male minus the female stimuli was calculated, out of the total time spent investigating stimuli animals.
• Compound (I) was dissolved in 100% ⁇ -cyclodextrin and then diluted with saline to a final solution of 50% 2-hydroxypropyl- ⁇ -cyclodextrin. It was administered intraperitoneally (i.p.) at doses of 3 and lOmg/kg, in a dosing volume of lml/kg, lh before tests. Progesterone (0.5mg/0.1ml) was dissolved in com oil and administered subcutaneously (s.c), 4h before test, as a positive control.
• ovariectomised female rats (as described above), were primed with 5 ⁇ g oestradiol benzoate dissolved in corn oil and injected subcutaneously. This is a low dose of oestrogen that does not re-establish sexual behaviour in an ovariectomised female but provides a minimum hormonal background for pharmacological agents to stimulate sexual behaviour.
• the females were placed with a series of vigorous male rats and subjected to 10 mounts.
• the lordotic response of the animal was recorded and expressed as a percentage of the mounts (i.e. lordosis quotient, LQ), as previously described. Animals showing LQ ⁇ 20 were considered non-receptive and were included in the study. Each rat was tested prior to administration of the compound and then tested similarly post-injection. The pre-treatment times were lh for Compound (1) and vehicle (50% ⁇ -cyclodextrin, i.p.) or 90min for quinelorane.
• Ovariectomised adult female Sprague Dawley rats (180-200g) were housed in groups of 5 in a reversed lighting system of 12h lightdark (lights off 5.00-17.00h). They were used for the experiments at least two weeks after ovariectomy. Forty eight hours before tests, the animals were primed with oestradiol benzoate (5 ⁇ g/0.1ml in com oil, s.c). On day 1, progesterone (0.5mg/0.1ml, in com oil, s.c.) was administered to one of the groups 4h before tests, as a positive control.
• Compound (T) (15mg/kg, i.p.) was administered in 50% 2-hydroxypropyl- ⁇ -cyclodextrin, lh before tests. The test lasted 10 minutes and was carried out as described before. The difference in the percentage of time spent investigating the male minus the female stimuli was calculated, out of the total time spent investigating stimuli animals. Animals were submitted to a test on day 1 and on day 15. From day 2 to 14 the Compound (X) group received a daily injection of the compound (15mg/kg, i.p.), while both the vehicle and the progesterone groups received an injection of vehicle. On day 15 the test took place again, as described for day 1.
• Compound (1) intracerebroventricularly (i.c.v.).
• the animals were primed with 5 ⁇ g oestradiol benzoate (s.c, in com oil) and adapted to the apparatus (in the absence of stimuli animals) for lOmin on 2 consecutive days prior testing.
• the lOmin test was carried out as previously described. The difference in the percentage of time spent investigating the male minus the female stimuli was calculated, out of the total time spent investigating stimuli.
• Compound (1) was dissolved in 50% 2-hydroxypropyl- ⁇ -cyclodextrin in saline. It was administered i.c.v. over a 30sec period, with the aid of a pump set to deliver a flow of lO ⁇ l/min. The dosing volume was 5j-.l/rat. The compounds were administered lOmin before tests. Progesterone (0.5mg/0.1ml) was dissolved in com oil and administered subcutaneously (s.c), 4h before test, as a positive control. As shown in Figure 4, Compound (1) dose-dependently (3-30 ⁇ g/rat) increased the percentage of time spent investigating the male stimulus, with a MED of lO ⁇ g. The effect of this dose was similar to the effect of progesterone.
• Ovariectomised female rats (Sprague Dawley, obtained from Charles River, UK) were stereotaxically implanted (coordinates 0.89mm behind Bregma, 1.3mm lateral and 2.5mm vertical) with stainless steel cannulae (6 mm long, O.D. 0.75mm), held in place with dental cement. Animals were housed in groups of three and returned to a reversed lighting system of 12h lightdark (lights off 5.00-17. OOh). Correct placement of the cannulae was assessed post-mortem. Rats were used for tests two weeks after ovariectomy (one week after cannulation). The experiments started at least 5h into the dark period.
• OB oestradiol benzoate
• Progesterone Prog, 0.5mg/0.1ml was dissolved in com oil and administered subcutaneously (s.c), 4h before test, to induce proceptive behaviour.
• Compound (1) (15mg kg, i.p.) was dissolved in 50% 2-hydroxypropyl- ⁇ -cyclodextrin in saline and administered lh before the i.c.v. administration.
• Neuromedin B was obtained from Bachem, UK. It was dissolved in isotonic saline and administered i.c.v. over a 30sec period, with the aid of a pump set to deliver a flow of lO l/min, lOmin before tests.
• the dosing volume was 5j-.l/rat. Each rat received a total amount of lOOng.
• progesterone increased the percentage of time spent investigating the male stimulus, compared to the vehicle group, thus showing stimulation of proceptive behaviour.
• NMB (lOOng, i.c.v.) significantly reduced proceptivity in progesterone-treated rats.
• pre-treatment with Compound (V) which acts as an antagonist (15mg/kg, i.p.) prevented the inhibitory effect of NMB.
• the blockade obtained with the dose of Compound (1) used was not total. From the present study we can conclude that stimulation of BB1 receptors with an agonist results in inhibition of proceptive behaviour. This inhibitory effect may be prevented by the presence of an antagonist, e.g.
• Compound (1) (nanomolar affinity "mixed" BB ⁇ /BB 2 receptor antagonist) has a dose-dependent stimulatory effect on sexual activity in the female rat, both on proceptivity and receptivity.
• the animals used in that study were ovariectomised, and therefore steroid hormones release can not be expected to occur in response to the compound, there is a possibility that the adrenal glands might secrete steroid hormones in response to Compound (1). If that was the case, the mediation of the stimulatory effects by progesterone would be relevant for rodents, but it would not be the case for primates.
• the bombesin receptor antagonist Compound (I) Oestradiol and pituitary hormones (Luteinising hormone (LH), follicle stimulating hormone (FSH) and prolactin) have also been analysed in the same animals.
• Ovariectomised adult female Sprague Dawley rats (180-200g) were housed in groups of 6 in a reversed lighting system of 12h lightdark (lights off 7.00-19.00h). They were used for the experiments at least two weeks after ovariectomy. Forty eight hours before tests, the animals were primed with oestradiol benzoate (5 ⁇ g/0.1ml in com oil, s.c). Progesterone (0.5mg/0.1ml, in corn oil, s.c.) was administered 4h before blood collection, as a positive control. Compound (1) (3-lOmg/kg, i.p.) was administered in 50% 2-hydroxypropyl- ⁇ -cyclodextrin, lh prior to blood collection. Blood was collected from the trunk, after decapitation. It was immediately centrifuged (3500r.p.m., 4°C, 5min) and the plasmas frozen until assayed for hormonal content,
• progesterone resultsed in a significant increase in the progesterone plasma levels (P ⁇ 0.05), and a significant decrease in LH plasma levels (PO.01), compared to animals injected with vehicle (Kruskal-Wallis followed by Mann- Whitney test).
• Compound (I) (3-lOmg/kg, i.p.) had no effect on the plasma levels of progesterone ( Figure 6, where animals were pre-treated with 5 ⁇ g oestradiol benzoate, s.c, 48h before the test. They were tested lh or 4h post-injection of Compound (1) (3-10mg/kg, p.o.) or progesterone (0.5mg/0.1ml, s.c.) respectively.
• Treatments were administered once a week, with a baseline test in between treatments (4 days intervals between baseline and test day). Treatments were Compound (I) (15mg/kg, dissolved in 50% 2-hydroxypropyl- ⁇ -cyclodextrin in saline), vehicle, or Fluoxetine (20mg/kg dissolved in 100% DMSO). All treatments were administered i.p. in a lml/kg volume, lh before tests.
• Sprague Dawley male rats (Charles River, UK) were kept, 4 rats per cage, in a reversed lighting regime (12:12 hours, lights off at 5.00 h), with free access to food and water.
• the rats were pre-selected by being presented with a receptive female at 4 days intervals, i.e. every third day (having 2 clear days between presentations) until completing 6-7 trials of baseline determination.
• the animals showing consistently vigorous behaviour (ejaculatory latencies ⁇ 300s) were chosen for further experiments. Animals were treated for 3 consecutive days with either vehicle (water) or fluoxetine (20mg/kg, i.p., in a 2ml/kg dosing volume).
• the animals treated with water received vehicle (veh+veh) and the animals treated with fluoxetine received one of the three following treatments: Compound (V) (15mg/kg, dissolved in 50% 2-hydroxypropyl- ⁇ -cyclodextrin in saline), vehicle (cyclodextrine), or yohimbine (2mg/kg dissolved in water). All treatments were administered i.p. in a lml kg volume, lh before tests.
• HPLC A Rt. 11.86 min, 99.8/100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at 1ml min "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC B Rt. 14.32 min, 100/100% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• HPLC A Rt. 17.07 min, 100/100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSID 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC B Rt. 14.35 min, 100/100% purity, 80:20 methanol/Tris buffer at ⁇ H9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• HPLC A Rt. 20.83 min, 98.3/99.6% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 25 min at lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm; HPLC B: Rt. 6.82 min, 100/100% purity, 80:20 methanol Tris buffer at ⁇ H9, lmlmin " Prodigy ODS I 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• HPLC B Rt. 5.72 min, 99.46% purity, 20-90% CH 3 CN/Tris (ImM) over 7 min at 2mlmin " Prodigy Phenyl-Ethyl, 100x4.6mm 5 ⁇ M at 30°C, 200-300 nm.
• Example 13 Washed with brine, saturated NaHCO 3 (x3), brine, dried ( gSO 4 ) and solvent removed under reduced pressure. Residue purified by chromatography using RP silica with 65% MeOH in H O. Pure fractions were evaporated to give Example 13 as a white amorphous solid (320mg, 68%):
• HPLC A Rt. 11.63 min, 97.7/100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSHI 250x4.6mm 5/xM, 215 and 254 nm;
• HPLC B Rt. 9.20 min, 100/100% purity, 80:20 methanol/Tris buffer at ⁇ H9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5j- ⁇ M, 215 and 254 nm.
• HPLC B Rt. 17.27 min, 100/100% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• the above compound was synthesized from 2a via 6c as outlined in Scheme 2 using methods analogous to those used for Example 9.
• the acid (6c) (188mg, 0.6mmol), HBTU (228mg, 0.6mmol), and DIPEA (105 ⁇ l, 0.6mmol) were sti ⁇ ed in DMF (10ml) for 5 minutes before adding DIPEA (105jul, O. ⁇ mmol) and [l-(5- methoxy-2-pyridyl)cyclohexyl]-methanamine (see WO 98/07718, 150mg,
• HPLC B Rt. 12.29 min, 100/100% purity, 80:20 methanol/Tris buffer at pH9, 1 mlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC A Rt. 12.30min, 99.4% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm;
• HPLC B Rt. 15.38min, 99.5% purity, 80:20 methanol/Tris buffer at ⁇ H9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm.
• HPLC A Rt. 10.54 min, 100/100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSHI 250x4.6mm 5 M, 215 and 254 nm;
• HPLC B Rt. 10.67 min, 100/100% purity, 80:20 methanol/Tris buffer at ⁇ H9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• Example 17 The method of Example 17 was ' repeated except that 4-bromopyridine hydrochloride (486mg, 2.5mmol) was used.
• step 2 The acid from step 1 (30mg, O.lmmol), HBTU (38mg, O.lmmol), and DIPEA (18j ⁇ tl, O.lmmol) were sti ⁇ ed in DMF (10ml) for 5 minutes before adding DIPEA (18/d, O.lmmol) and [l-(2- pyridyl)cyclohexyl]methylamine (WO 98/07718; 19mg, O.lmmol). After 2 hours at ambient temperature the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate and washed with sodium bicarbonate solution (x2), brine, and dried (MgSO 4 ). The solvent was removed under reduced pressure.
• HPLC A Rt. 7.21 min, 96.1/96.5% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSID 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC B Rt. 6.02 min, 99.1/100% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSID 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• Example 19 was prepared on the same scale and using an analogous method as used for Example 17.
• Example 17 The method of Example 17 was followed except that 4-bromoisoquinoline (520mg, 2.5mmol) was used.
• step 2 The acid from step 1 (40mg, 0.12mmol), HBTU (46mg, 0.12mmol), and DIPEA (21 ⁇ l, 0.12mmol) were sti ⁇ ed in DMF (10ml) for 5 minutes before adding DIPEA (21 ⁇ l, 0.12mmol) and [l-(2- pyridyl)cyclohexyl]methylamine (WO 98/07718; 23mg, 0.12mmol). After 2 hours at ambient temperature the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate and washed with sodium bicarbonate solution (x2) and brine and dried (MgSO 4 ). The solvent was removed under reduced pressure.
• HPLC A Rt. 7.52 min, 100/100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSID 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC B Rt. 8.33 min, 99.7/100% purity, 80:20 methanol Tris buffer at pH9, lmlmin "1 , Prodigy ODSID 250x4.6mm 5 M, 215 and 254 nm;
• step 2 The acid from step 1 (150mg, 0.5mmol), HBTU (190mg, 0.5mmol), and DIPEA (87 ⁇ l, 0.5mmol) were sti ⁇ ed in DMF (10ml) for 5 minutes before adding DIPEA (87 ⁇ l, 0.5mmol) and [l-(2- ⁇ yridyl)cyclohexyl]methylamine (WO 98/07718; 95mg, 0.5mmol). After 2 hours at ambient temperature the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate and washed with sodium bicarbonate solution (x2) and brine and dried (MgSO 4 ). The solvent was removed under reduced pressure.
• the crade product was purified by chromatography using lOg ISCO Redisep cartridge with 90% ethyl acetate in heptane as eluent. Removal of the solvent under reduced pressure gave the desired product as a foam (135mg, 58%):
• HPLC A Rt. 12.65min, 99.65% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm;
• HPLC B Rt. 33.05min, 99.89% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm.
• the above compound was prepared using a one-pot procedure analogous to the method used for Example 16.
• the synthesis was carried out on lmmol scale using l-bromo-3-methyl-benzene (171mg, lmmol).
• the crude product was purified by chromatography using 25g NP silica with 25% ethyl acetate in heptane as eluent. Removal of the solvent under reduced pressure gave the desired compound as a glass (260mg, 54%):
• HPLC A Rt. 11.04 min, 98.3% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm;
• HPLC B Rt. 16.87 min, 99.5% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm.
• the above compound was prepared using a one-pot procedure analogous to the method used for Example 16.
• the synthesis was carried out on 0.4mmol scale using 2-bromo-6-phenyl-pyridine (95mg, 0.4 mmol).
• the crude product was purified by chromatography using 25g NP silica with 55% ethyl acetate in heptane as eluent. Removal of the solvent under reduced pressure gave the desired product as a foam (260mg, 54%):
• the crade product was purified by chromatography using 50% ethyl acetate in heptane and then RP C18 silica with 70% methanol in water as eluent. Removal of the solvent under reduced pressure gave the desired product as a white amorphous solid (0.15g, 22%):
• HPLC A Rt. 4.51 min, 100% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 10 min at 1.5 mlmin "1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 200-300 nm;
• HPLC B Rt. 13.15 min, 99.14% purity, 80:20 methanoi/Tris buffer at pH9, lmlmin "1 , Prodigy ODS T 250x4.6mm 5 ⁇ M/200-300 nm.
• Boc-(S)-oMeTrp-OH as an orange oil (14.5g, 99%).
• HBTU 8.0g, 22mmol
• triethylamine 5ml, 35mmol
• [l-(2- ⁇ yridyl)cyclohexyl]methylamine WO 98/07718; 4.2g, 22mmol.
• HPLC A Rt. 8.52 min, 99.0/98.6% purity, 20-100% CH 3 CN in H 2 O (+0.1%TFA) over 15 min at lmlmin 1 , Prodigy ODSDI 250x4.6mm 5 ⁇ M, 215 and 254 nm;
• HPLC B Rt. 23.84 min, 99.6/100% purity, 80:20 methanol/Tris buffer at pH9, lmlmin "1 , Prodigy ODSID 250x4.6mm 5 ⁇ M, 215 and 254 nm.
• BBj and BB2 binding were as follows. CHO-K1 cells stably expressing cloned human NMB (for (BB ⁇ assay) and GRP receptors (for BB2 assay) were routinely grown in Ham's F12 culture medium supplemented with 10% foetal calf serum and 2 mM glutamine. For binding experiments, cells were harvested by trypsinization, and stored frozen at -70°C in Ham's F12 culture medium containing 5% DMSO until required. On the day of use, cells were thawed rapidly, diluted with an excess of culture medium, and centrifuged for 5 minutes at 2000 g.
• Cells were resuspended in 50 mM Tris-HCl assay buffer (pH 7.4 at 21°C, containing 0.02% BSA, 40 ⁇ g/mL bacitracin, 2 ⁇ g/mL chymostatin, 4 ⁇ g/mL leupeptin, and 2 ⁇ M phosphoramidon), counted, and polytronned (setting 5, 10 sec) before centrifuging for 10 minutes at 28,000 g. The final pellet was resuspended in assay buffer to a final cell concentration of 1.5 x lONmL.
• Tests were carried out in a circular arena of 90cm diameter, su ⁇ ounded by a 30cm high wall.
• Two small cages with wire-mesh front (15x15cm) are fixed into the wall such that the front of the cage is "flush” with the wall and the 2 cages are opposite each other.
• They contain two stimuli animals: an intact sexually experienced male and a receptive female (ovariectomised, primed with 5 ⁇ g oestradiol benzoate dissolved in com oil and injected subcutaneously 48 hours before the test and with 0.5 mg of progesterone four hours before the test).
• sexually naive test and control animals were used.
• test and control ammals were primed with 5 ⁇ g oestradiol benzoate.
• Test animals were treated with the above compound (30-lOOmg kg) which was dissolved in PEG 200 vehicle and administered orally in a lml/kg volume lh before each test.
• progesterone 0.5mg/0.1ml
• s.c subcutaneously
• Test and control animals were introduced one at a time for 10 minute periods into the arena. During the lOmin test, the time that the test or positive control animal spent investigating each stimulus animal was noted. The arena was thoroughly cleaned between animals. The position of the male/female stimuli boxes was randomised between animals, in order to avoid place preference. The difference in the percentage of time spent investigating male minus female was calculated, out of the total time spent investigating stimuli animals.
• Example 31 was repeated except that the above compound (3-30 mg kg) was dissolved in 0.5% methyl cellulose and was administered p.o. in a dosing volume of 3ml kg lh before tests.
• Progesterone, (0.5mg/0.1ml) was dissolved in com oil and administered s.c, 4h before test, as a positive control.
• Ovariectomised adult female Sprague Dawley rats (180-200g, from Charles River) were housed in groups of 6 in a reversed lighting system of 12h light:dark (lights off 7.00-19.00h). Two weeks after ovariectomy they were used for sexual activity tests. The experiments started at least 5h into the dark period.
• Intermediates DI-6 and flI-7 are made by (i) protecting the amino group of the starting amino acid a with di-t-butyl carbonate and potassium carbonate in dioxane/water, (ii) forming an amide by reaction of the N- protected amino acid with an amine bl or b_2 in dimethylformamide in the presence of O-benzotriazol-l-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) and NN-diisopro ⁇ yl-ethylamine (DIPEA), and (iii) deprotecting the amino group of the product cl or c2 by reaction with trifluoroacetic acid in dichloromethane.
• HBTU O-benzotriazol-l-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate
• DIPEA NN-diisopro ⁇ y
• IA IC50 > 10000 nM
• R2 represents the rest of the intermediate e.
• HPLC column: Phenomenex primesphere 10 ⁇ C18-HC 110A, 100x21.20 mm; mobile phase: methanol/water 10 to 100% gradient.
• the products were characterised and analysed by LCMS (column: 50x4.6 mm Prodigy ODSDI (5 ⁇ ) column; mobile phase: acetonitrile/water (0.1% formic acid) 5 to 100% gradient over 2 min, held at 100% acetonitrile for 1 min; flow rate 4 mL/min; UN detection at 215 nm; mass spec: 150-900 Da full scan APCI+ centroid data).

Applications Claiming Priority (1)

Publications (1)

Family

ID=9886325

Family Applications (1)

Country Status (13)

Families Citing this family (3)

Family Cites Families (6)

• 2000
  • 2000-11-17 IL IL15581400A patent/IL155814A0/xx unknown
  • 2000-11-17 AU AU2001214046A patent/AU2001214046A1/en not_active Abandoned
  • 2000-11-17 PL PL00366002A patent/PL366002A1/xx not_active Application Discontinuation
  • 2000-11-17 CN CNA008201218A patent/CN1479618A/zh active Pending
  • 2000-11-17 CA CA002426521A patent/CA2426521A1/en not_active Abandoned
  • 2000-11-17 BR BR0017374-6A patent/BR0017374A/pt not_active IP Right Cessation
  • 2000-11-17 WO PCT/GB2000/004380 patent/WO2002040022A1/en not_active Application Discontinuation
  • 2000-11-17 KR KR10-2003-7006676A patent/KR20040062416A/ko active IP Right Grant
  • 2000-11-17 HU HU0303500A patent/HUP0303500A2/hu unknown
  • 2000-11-17 EP EP00976165A patent/EP1333829A1/de not_active Withdrawn
  • 2000-11-17 MX MXPA03003481A patent/MXPA03003481A/es unknown
  • 2000-11-17 JP JP2002542395A patent/JP2004525864A/ja active Pending
• 2003
  • 2003-04-25 ZA ZA200303250A patent/ZA200303250B/en unknown

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events