TRIAZINE DERIVATIVES FOR TREATING DISEASES RELATING TO NEUROTROPHINS

Abstract

There is provided a compound of formula I, (I) wherein R.sup.1 and R.sup.2 are as defined herein, which compounds are useful in the treatment of diseases characterised by impaired signalling of neurotrophins and/or other trophic factors, such as Alzheimer's disease and the like.

##STR00001##

Claims

1. A compound of formula I, ##STR00061## wherein: R.sup.1 represents ##STR00062## wherein represents the point of attachment to the nitrogen atom; R.sup.3 represents hydrogen, methyl or methoxymethyl; and R.sup.4 represents hydrogen, methoxy or methoxymethyl; R.sup.2 represents methyl, ethyl, methoxymethyl, methylsulfanyl or -A-R.sup.5; wherein A represents C.sub.1-2 alkylene, —C.sub.1-2alkyleneO—, —OC.sub.1-2alkylene-, which three groups are optionally substituted by one or more groups selected from halo, C.sub.1-2 alkyl and ═O; and R.sup.5 represents oxetanyl or a 4-7-membered nitrogen-containing heterocyclyl group, each of which groups are optionally substituted by one or more groups selected from halo, C.sub.1-2 alkyl and ═O; or a pharmaceutically-acceptable salt thereof, with the proviso that the compound of formula I does not represent ##STR00063##

2. A compound as claimed in claim 1, wherein R.sup.1 represents ##STR00064## wherein , R.sup.3 and R.sup.4 are as defined in claim 1.

3. A compound as claimed in claim 1 or claim 2, wherein at least one of R.sup.3 and R.sup.4 represents hydrogen.

4. A compound as claimed in any one of claims 1 to 3, wherein R.sup.3 represents methyl or methoxymethyl and R.sup.4 represents hydrogen.

5. A compound as claimed in any one of claims 1 to 3, wherein R.sup.3 represents hydrogen and R.sup.4 represents methoxy.

6. A compound as claimed in any one of claims 1 to 3, wherein R.sup.3 and R.sup.4 each represent hydrogen.

7. A compound as claimed in any one of claims 1 to 6, wherein R.sup.2 represents methyl, ethyl or methoxymethyl.

8. A compound as claimed in any one of claims 1 to 7, wherein R.sup.2 represents ethyl or methoxymethyl.

9. A compound as claimed in any one claims 1 to 6, wherein R.sup.2 represents -A-R.sup.5, wherein A and R.sup.5 are as defined in claim 1.

10. A compound as claimed in claim 9, wherein R.sup.5 represents a 4-7-membered nitrogen-containing heterocyclyl group, which heterocyclyl group is optionally substituted by one or more groups selected from halo, C.sub.1-2 alkyl and ═O;

11. A compound as claimed in any one of claims 1 to 6 or 9 to 10, wherein A represents —CH.sub.2— or —OCH.sub.2C(O)—.

12. A compound as claimed in any one of claims 1 to 6 or 9 to 11, wherein R.sup.5 represents a 4-6-membered nitrogen-containing heterocyclyl group, which heterocyclyl group is optionally substituted by one or more groups selected from halo, C.sub.1-2 alkyl and ═O.

13. A compound as claimed in claim 12, wherein R.sup.5 represents a 4-6-membered nitrogen-containing heterocyclyl group selected from: ##STR00065## wherein represents the point of attachment to A, each of which heterocyclyl groups is optionally substituted by a ═O group.

14. A compound as claimed in claim 1, wherein the compound is selected from the group consisting of: ##STR00066## ##STR00067## ##STR00068## ##STR00069## or a pharmaceutically-acceptable salt thereof.

15. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 14, including pharmaceutically-acceptable salts thereof, in combination with one or more pharmaceutically-acceptable excipient.

16. A compound as defined in any one of claims 1 to 14, including pharmaceutically-acceptable salts thereof, or a pharmaceutical composition as defined in claim 15, for use in medicine.

17. A compound as defined in any one of claims 1 to 14, including pharmaceutically-acceptable salts thereof, or a pharmaceutical composition as defined in claim 15, for use in the treatment and/or prevention of a disease characterised by impaired signalling of neurotrophins and/or other trophic factors.

18. A method of treating and/or preventing a disease characterised by impaired signalling of neurotrophins and/or other trophic factors, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound as defined in any one of claims 1 to 14, including pharmaceutically-acceptable salts thereof, or a pharmaceutical composition as defined in claim 15.

19. The use of a compound as defined in any one of claims 1 to 14, including pharmaceutically-acceptable salts thereof, or a pharmaceutical composition as defined in claim 15, for the manufacture of a medicament for the treatment or prevention of a disease characterised by impaired signalling of neurotrophins and/or other trophic factors.

20. The compound for use, method or use as defined in any one of claims 17 to 19, wherein the disease characterised by impaired signalling of neurotrophins and/or other trophic factors is selected from Alzheimer's disease, Lewy body dementia, frontotemporal dementia, cognitive dysfunction, mild cognitive impairment, other dementia disorders, Parkinson's, disease, other Parkinsonian disorders and/or other tauopathies, Huntington's disease, brain injuries, stroke, motor neurone diseases, multiple sclerosis, spinal cord injury, hypoxia, ischemia, hypoxic ischemia injury, coronary artery disease, obesity, diabetes, metabolic syndrome, diabetic neuropathy, Charcot-Marie-Tooth disease and its variants, nerve injury, genetic or acquired or traumatic hearing loss, blindness, posterior eye diseases, anterior eye diseases, dry eye syndrome, neurotrophic keratitis, glaucoma, high intraocular pressure, retinitis pigmentosa, post-traumatic stress disorders, WAGR syndrome, Prader-Willi syndrome, olfactory decline, olfactory dysfunction, fragile X syndrome, congenital hypoventilation syndrome, obsessive-compulsive disorder, anxiety, generalised anxiety disorder, schizophrenia, depression, eating disorders, bipolar disorder, chronic fatigue syndrome, neuromyelitis optica, Rett syndrome, epilepsy, Friedreich's ataxia, obstructive sleep apnea-hypopnea syndrome, pain and constipation.

21. The compound for use, method or use as claimed in claim 20, wherein the disease characterised by impaired signalling of neurotrophins and/or other trophic factors is selected from the group consisting of Alzheimer's disease, Parkinson's disease, other Parkinsonian diseases, other tauopathies, Lewy body dementia, motor neurone disease, Pick's disease, obesity, metabolic syndrome, diabetes, diabetic neuropathy, glaucoma, dry eye syndrome, neurotrophic keratitis, genetic, acquired or traumatic hearing loss and Rett Syndrome.

22. The compound for use, method or use as claimed in claim 21, wherein the disease characterised by impaired signalling of neurotrophins and/or other trophic factors is selected from the group consisting of Alzheimer's disease, Parkinson's disease, cognitive dysfunction, depression, diabetic neuropathy, glaucoma, dry eye syndrome, genetic, acquired or traumatic hearing loss and Rett syndrome.

23. The compound for use, method or use as claimed in claim 22, wherein the disease characterised by impaired signalling of neurotrophins and/or other trophic factors is Alzheimer's disease.

24. A combination product comprising: (I) a compound as defined in any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof; and (II) one or more other therapeutic agent that is useful in the treatment or prevention of a disease characterised by impaired signalling of neurotrophins and/or other trophic factors, wherein each of components (I) and (II) is formulated in admixture, optionally with a pharmaceutically-acceptable excipient, such as a pharmaceutically-acceptable adjuvant diluent or carrier.

25. A kit-of-parts comprising: (a) a pharmaceutical composition comprising a compound as defined in any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, formulated in admixture with a pharmaceutically-acceptable excipient; and (b) a pharmaceutical composition comprising one or more other therapeutic agent that is useful in the treatment or prevention of a disease characterised by impaired signalling of neurotrophins and/or other trophic factors, formulated in admixture with a pharmaceutically-acceptable excipient, which components (a) and (b) are each provided in a form that is suitable for administration in conjunction with the other.

26. A process for the preparation of a compound as defined in any one of claims 1 to 14, including a pharmaceutically-acceptable salt thereof, comprising the step of reacting a compound of formula II, ##STR00070## wherein R.sup.1 and R.sup.2 are as defined in any one of claims 1 to 14, with ethoxycarbonyl isocyanate.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0171] FIG. 1 shows the results of the passive avoidance task performed with the compound of Example 1 described in Biological Example 3. The graph demonstrates that administering the compound of Example 1 to mice treated with scopolamine improves cognitive function, as illustrated by the increased retention latency within the bright area.

[0172] FIG. 2 shows the results of the passive avoidance task performed with the compound of Example 3 described in Biological Example 3. The graph demonstrates that administering the compound of Example 3 to mice treated with scopolamine improves cognitive function, as illustrated by the increased retention latency within the bright area.

[0173] FIG. 3 shows the results of the Forced Swim Test described in Biological Example 4. The graph demonstrates that administering the compound of Example 1 to mice decreased time spent immobile compared to mice given the vehicle control. Decreased time spent immobile is a model-specific endpoint commonly used as an indicator of anti-depressant effects in pharmaceutical compounds

EXAMPLES

[0174] The present invention will be further described by reference to the following examples, which are not intended to limit the scope of the invention.

Experimental Procedures

[0175] Starting materials and intermediates used in the synthesis of compounds described herein are commercially available or can be prepared by the methods described herein or by methods known in the art.

[0176] Experiments were generally carried out under inert atmosphere (nitrogen or argon), particularly in cases where oxygen- or moisture-sensitive reagents or intermediates were used.

[0177] Mass spectrometry data are reported from liquid chromatography-mass spectrometry (LC-MS) using electrospray ionization. Chemical shifts for NMR data are expressed in parts per million (ppm, δ) referenced to residual peaks from the deuterated solvent used.

[0178] For syntheses referencing general procedures, reaction conditions (such as length of reaction or temperature) may vary. In general, reactions were followed by thin layer chromatography or LC-MS, and subjected to work-up when appropriate. Purifications may vary between experiments: in general, solvents and the solvent ratios used for eluents/gradients were chosen to provide an appropriate R.sub.f and/or retention time.

General Methods

[0179] All solvents were of analytical grade and commercially available anhydrous solvents were routinely used for reactions. Starting materials used were available from commercial sources or prepared according to literature procedures, Room temperature refers to 20-25° C. Solvent mixture compositions are given as volume percentages or volume ratios.

[0180] MW heating was performed in a standard MW reactor producing continuous irradiation at 2450 MHz. It is understood that MWs can be used for the heating of reaction mixtures. Typically, an Anton paar microwave synthesizer 300 was used as a microwave synthesizer.

[0181] Thin layer chromatography (TLC) was performed on Merck TLC-plates (Silica gel 60 F.sub.254) and spots were UV visualized. TLC was generally used to monitor reaction progression and solvents used were for example: ethyl acetate or acetonitrile or DCM with 1-10% of MeOH, ethyl acetate with 0-95% hexane. Straight phase flash column chromatography (“flash chromatography”/“column chromatography”) was manually performed on Merck Silica gel 60 (0.040-0.063 mm) or basic aluminum oxide or neutral aluminum oxide, or automatically using ISCO Combiflash® Companion™ system using RediSep™ normal-phase flash columns (“Combiflash”) using the solvent system indicated.

[0182] NMR spectra was recorded on a 400 MHz NMR spectrometer (Bruker 400 MHz Avance-III) fitted with a probe of suitable configuration. Spectra were recorded at ambient temperature unless otherwise stated. Chemical fields are given in ppm down- and upfield from TMS (0.00 ppm). The following reference signals were used in .sup.1H-NMR: TMS δ 0.00, or residual solvent signal of DMSO-d6 δ 2.49, CDCl.sub.3 δ 7.25 (unless otherwise indicated). Resonance multiplicities are denoted s, d, t, q, m, dd, tt, dt br and app for singlet, doublet, triplet, quartet, doublet of doublet, triplet of triplet, doublet of triplet, multiplet, broad and apparent, respectively. In some cases only diagnostic signals are reported.

[0183] High pressure liquid chromatography (HPLC) was performed on a reversed phase (RP) column. A gradient was applied using for example mobile phase A (5 mM Ammonium acetate+0.1% Formic acid in water) and B (0.1% Formic acid in Acetonitrile) or A (0.1% NH3 in water) and B (0.1% NH3 in acetonitrile) or A (10 mM Ammonium acetate in water) and B (Acetonitrile).

[0184] Reversed phase columns used were for example: BEH C18 (50*2.1 mm), 1.7 μm; X-Bridge C18 (50*4.6 mm), 3.5 μm; X-Bridge/YMCC18 (150*4.6 mm), 5 μm; BEH C18 (50*2.1 mm), 1.7 μm; X-Bridge C8 (250*19) mm, 5 μm. The flowrate used was for example 0.55 ml/min or 1.00 ml/min Mass spectrometry (MS) analysis were performed in positive and/or negative ion mode using electrospray ionization (ESI+/−).

[0185] Preparative HPLC chromatography was run on a Waters e2695 Separation Module with a PDA Detector or on a Shimadzu LC-20AP with an UV detector. Column; X-BRIDGE C18, 150*4.6 mm, 5 μm or X-Bridge C18 (250*19 mm) 5 μm or GEMINI C18 (250*21.2 mm) 5 μm or sunfire c18(150*19) mm, 5 micron or x-bridge c18(150*19) mm, 5 micron or ymc actus triart c18(150*20) mm, 5 micron or kromasil eternity c18 (250*21.2) mm, 5 micron. The flowrate used was for example 10-15 ml/min. The UV spectra were typically recorded at 202 nm & between 214 and 260 nm Lambda max.

[0186] A gradient was applied using for example mobile phase A (0.1% NH.sub.3 in water) and B (0.1% NH3 in acetonitrile); A (0.1% TFA in water) and B (Acetonitrile); A (5 mM ammonium bicarbonate+0.05% ammonia in water) and B (Acetonitrile); A (5 mM ammonium bicarbonate) and B (acetonitrile) for LC-separation at a flow rate 1 ml/min.

[0187] High pressure liquid chromatography (HPLC) was performed on a straight phase column. A linear gradient or isocratic flow was applied using for example phase A (Hexane) and B (XX)

[0188] Compounds have been named using CDD vault from Collaborative Drug Discovery Inc. Burlingame Calif., USA or ChemDoodle 8.1.0/9.02 from iChemLabs LLC, USA or ACD/ChemSketch 2012 (14.01) from Advanced Chemistry Development (ACD/labs) Ontario, Canada. In case of inconsistency between a name of a compound and the structural formula of the same compound, it is the structural formula that is decisive for the molecular structure of the compound.

[0189] In the event that there is a discrepancy between nomenclature and any compounds depicted graphically, then it is the latter that presides (unless contradicted by any experimental details that may be given or unless it is clear from the context).

Intermediate 1

3-(3-methyl-4-phenoxyphenyl)-1-(3-methylphenyl)urea

[0190] ##STR00020##

[0191] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken, 3-methyl-4-phenoxyaniline (3.0 g, 15.1 mmol), NaHCO.sub.3 (3.7 g, 45 mmol) and dichloromethane (30 ml). The mixture was cooled to 0° C. and triphosgene (1.33 g, 4.5 mmol) was added and the reaction mixture was stirred for 1 h. 3-methyl aniline (1.6 g, 15.1 mmol) and NaHCO.sub.3 (3.7 g, 45 mmol) were added and the reaction mixture was allowed to reach room temperature and stirred for 3 h. The reaction mixture was quenched with water (50 ml) and the product was extracted with DCM (3×50 ml). The combined organic layer was washed with brine (50 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product. The crude product was triturated with DCM and Hexane to obtained 4.8 g (95% yield) of the title compound. MS (ES+) m/z 333 [M+H].sup.+

Intermediate 2

1-[3-(methoxymethyl)phenyl]-3-(3-methyl-4-phenoxyphenyl)urea

[0192] ##STR00021##

[0193] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken, 3-methyl-4-phenoxyaniline (commercially available, 1.0 g, 5 mmol) and triethylamine (6.96 ml, 50 mmol) in DCM (10 ml) at room temperature and the mixture was stirred for 10 min. Carbonyldiimidazole (CDI, 4.06 g, 25 mmol) was added and the mixture was stirred for 1 hr. 3-methoxymethyl-phenylisocyanate (0.69 g, 5 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice-water (25 ml) and the product was extracted with DCM (3×25 ml). The combined organic layer was washed with brine (50 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product. The crude product was purified by column chromatography using silica gel (100-200 mesh) and 20% ethyl acetate in hexanes as an eluent to obtain 0.55 g (30% yield) of the title compound. MS (ES+) m/z 363 [M+H].sup.+

Intermediate 3

2-(methoxymethyl)-4-nitro-1-phenoxybenzene

[0194] ##STR00022##

[0195] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken, Phenol (1.14 g, 12.2 mmol) and K.sub.2CO.sub.3 (3.36 g, 24.3 mmol) in N-Methyl-2-pyrrolidone (15 ml) and stirred for 1 h. 1-Fluoro-2-(methoxymethyl)-4-nitrobenzene (1.5 g, 8.1 mmol) was added in to the mixture and the reaction mixture was heated to 100° C. and stirred for 16 h. The reaction mixture was quenched with water (50 ml) and product was extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (50 ml), dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was purified by column chromatography using silica gel (100-200 mesh) and 5% ethyl acetate in hexanes as an eluent to obtain 1.6 g (76% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.29 (d, J=2.40 Hz, 1H), 8.15 (dd, J=2.80, 9.20 Hz, 1H), 7.49 (app t, 2H), 7.30 (t, J=7.20 Hz, 1H), 7.16 (app d, 2H), 6.85 (d, J=9.20 Hz, 1H), 4.62 (s, 2H), 3.43 (s, 3H). MS (ES+) m/z 260 [M+H].sup.+

Intermediate 4

3-(methoxymethyl)-4-phenoxyaniline

[0196] ##STR00023##

[0197] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was added 2-(methoxymethyl)-4-nitro-1-phenoxybenzene (Intermediate 3, 1.6 g, 8.0 mmol) in Methanol (16 ml). 10% Pd/C (50% wet, 0.32 g) was added under N.sub.2 atmosphere. The suspension was stirred under H.sub.2 (gas) bubbling. The completion of reaction was confirmed by the TLC using Hexane:EtOAc (7:3) as mobile phase. The TLC was visualized using UV light. After completion of the reaction, reaction mixture was filtered through celite bed washed with methanol and solvent was removed under reduced pressure to obtain 1.4 g (97% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 7.28 (app t, 2H), 6.97 (t, J=6.80 Hz, 1H), 6.79 (app d, 2H), 6.68 (s, 2H), 6.51 (d, J=8.40 Hz, 1H), 5.05 (s, 2H), 4.20 (s, 2H), 3.20 (s, 3H). MS (ES+) m/z 230 [M+H].sup.+

Intermediate 5

3-[3-(methoxymethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea

[0198] ##STR00024##

[0199] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(methoxymethyl)-4-phenoxyaniline, (Intermediate 4, 0.35 g, 1.5 mmol) and NaHCO.sub.3 (0.38 g, 4.5 mmol) in DMF (3.5 ml) and the mixture was cooled to 0° C. 3-methyl-phenylisocyanate (0.203 g, 1.5 mmol) was added and the resulting reaction mixture was allowed to reach 25° C. and stirred for 3 h. The reaction mixture was quenched with ice-water (50 ml) and the solid precipitate obtained was collected by filtration, washed with water (50 ml) and dried under vacuum to yield 0.4 g (72% yield) of the title compound, that was used in the next step without further purification. .sup.1H NMR (400 MHz, DMSO-d6): δ 8.74 (s, 1H), 8.55 (s, 1H), 7.62 (s, 1H), 7.32-7.34 (m, 4H), 7.22 (d, J=7.60 Hz, 1H), 7.15 (t, J=7.60 Hz, 1H), 7.06 (t, J=7.20 Hz, 1H), 6.88 (app d, 3H), 6.78 (d, J=7.2 Hz, 1H), 4.35 (s, 2H), 3.27 (s, 3H), 2.27 (s, 3H). MS (ES+) m/z 363 [M+H].sup.+

Intermediate 6

Phenyl N-(2-methyl-1-benzofuran-7-yl)carbamate

[0200] ##STR00025##

[0201] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-Methyl-1-benzofuran-7-ylamine (commercially available, 0.8 g, 5.3 mmol) and K.sub.2CO.sub.3 (2.19 g, 16 mmol) in THE (16 ml) at 25° C. and the mixture was stirred for 30 min. Phenyl chloroformate (1.25 g, 8 mmol) was added drop wise in to the reaction mixture and it was stirred at 25° C. for 16 h. The reaction mixture was quenched with ice-water (30 ml) and the product was extracted with Ethyl Acetate (3×20 ml). The combined organic layer was washed with brine (30 ml). The organic layer was dried over sodium sulphate and the solvent removed under reduced pressure to obtain 1.44 g (99%) of the title compound. MS (ES+) m/z 268 [M+H].sup.+

Intermediate 7

3-(2-methyl-1-benzofuran-7-yl)-1-(3-methyl-4-phenoxyphenyl)urea

[0202] ##STR00026##

[0203] In a seal tube previously equipped with a magnetic stirrer was taken Phenyl N-(2-methyl-1-benzofuran-7-yl)carbamate (Intermediate 6, 1.00 g 3.7 mmol) and 3-methyl-4-phenoxyaniline (commercially available, 0.73 g, 3.7 mmol) in DMF (10 ml) at 25° C. Triethylamine (0.74 g, 7.4 mmol) was added and the reaction mixture was heated to 100° C. and stirred for 16 h. The reaction mixture was quenched with ice-water (40 ml) and the product was extracted with ethyl acetate (3×25 ml). The combined organic layer was washed with brine (30 ml). The organic layer was dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was purified by column chromatography using 20% Ethyl acetate in Hexane as a mobile phase and 60-120 silica on column chromatography to yield 0.26 g (18% yield) of the title compound. MS (ES+) m/z 373 [M+H].sup.+

Intermediate 8

methyl 5-{[(3-methylphenyl)carbamoyl]amino}-2-phenoxybenzoate

[0204] ##STR00027##

[0205] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was added, 3-methylaniline (1.35 g, 12.6 mmol), NaHCO.sub.3 (3.17 g, 37.8 mmol) and DCM (17 ml). The mixture was cooled to 0° C., and triphosgene (1.23 g, 4.1 mmol) was added and reaction mixture was stirred for 2 h at 0° C. Methyl 5-amino-2-phenoxybenzoate (commercially available, 3.08 g, 12.6 mmol) and NaHCO.sub.3 (3.17 g, 37.8 mmol) were added to the reaction mixture. The reaction mixture was allowed to reach to 25° C. and stirred for 2 h. The reaction mixture was quenched with water (50 ml) and the product was extracted with DCM (3×50 ml). The combined organic layer was washed with brine (50 ml), dried over sodium sulphate and the solvent removed under reduced pressure and the crude product was purified by column chromatography using silica gel (60-120 mesh) and 30% ethyl acetate in hexanes as an eluent to obtain 4.5 g (94% yield) of the title compound. MS (ES+) m/z 377 [M+H].sup.+

Intermediate 9

3-[3-(hydroxymethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea

[0206] ##STR00028##

[0207] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was added, methyl 5-{[(3-methylphenyl)carbamoyl]amino}-2-phenoxybenzoate (Intermediate 8, 4.50 g, 11.9 mmol) in THE (45 ml) and the mixture was cooled to 0° C. LiBH.sub.4 (1.56 g, 71.7 mmol) was added portion wise at 0° C. The reaction mixture was allowed to reach room temperature and stirred for 4 h. The reaction mixture was quenched with water (50 ml) and the product was extracted with ethylacetate (3×50 ml), dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was triturated by DCM (2×15 ml) and the solid product was collected by filtration to yield 3.9 g (93%) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.76 (s, 1H), 8.57 (s, 1H), 7.64 (s, 1H), 7.35-7.40 (m, 4H), 7.33-7.15 (m, 2H), 7.05-7.06 (m, 1H), 6.81-6.79 (m, 4H), 5.21 (d, J=4.80 Hz, 1H), 4.45 (d, J=4.80 Hz, 2H), 2.30 (s, 3H). MS (ES+) m/z 349 [M+H].sup.+

Intermediate 10

3-[3-(chloromethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea

[0208] ##STR00029##

[0209] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was added 3-[3-(hydroxymethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea (Intermediate 9, 3.80 g, 10.9 mmol) in dichloromethane (38 ml). A catalytic amount of DMF (0.5 ml) was added and the mixture was cooled to 0° C. and stirred for 10 min at 0° C. Thionyl chloride (2.59 g, 21.8 mmol) was added drop wise and the resulting reaction mixture was allowed to reach room temperature and stirred for 2 h. The reaction mixture was quenched with water (30 ml) and the aqueous layer was extracted with dichloromethane (3×30 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was purified by column chromatography using silica gel (100-200 mesh) and neat hexanes as an eluent to obtain 3.0 g (74% yield) of the title compound. MS (ES+) m/z 367 [M+H].sup.+

Intermediate 11

1-[3-(chloromethyl)-4-phenoxyphenyl]-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione

[0210] ##STR00030##

[0211] To a microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was added 3-[3-(chloromethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea (Intermediate 10, 1.00 g, 2.3 mmol) in bromobenzene (10 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (0.53 g, 4.5 mmol) was added and resulting reaction mixture was allowed to reach 25° C. and heated at 150° C. for 3 h in an Anton par microwave synthesizer-300. The reaction mixture was quenched with water (10 ml) and aqueous layer was extracted with ethyl acetate (2×30 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was purified by Combi-flash chromatography and 20% ethyl acetate in hexanes as an eluent to obtain 0.53 g of the crude title compound that was used in the next step without further purification. MS (ES−) m/z 434 [M−H].sup.−

Intermediate 12

1-(5-nitro-2-phenoxyphenyl)ethan-1-one

[0212] ##STR00031##

[0213] In a sealed tube, previously equipped with a magnetic stirrer and nitrogen balloon, was taken phenol (7.39 g, 78.6 mmol) in o-Xylene (150 ml) at room temperature and K.sub.2CO.sub.3 (12.07 g, 87.3 mmol) was added. The mixture was stirred for 1 h at room temperature and 1-(2-fluoro-5-nitrophenyl)ethan-1-one (commercially available, 8.0 g, 43.6 mmol) in o-Xylene (42 ml) was added and the reaction mixture was stirred at 140° C. for 16 h. The reaction mixture was quenched with ice-water (300 ml) and the product was extracted with Ethyl Acetate (3×200 ml). The combined organic layer was washed with brine (300 ml), dried over sodium sulphate and the solvent removed under reduced pressure. The obtained crude product was triturated by dichloromethane:Hexane (1:9; 100 ml) to obtain 7.92 g (70% yield) of the title compound. 1H NMR (400 MHz, DMSO-d6): δ 8.49 (s, 1H), 6.33-7.31 (m, 1H), 7.52 (s, 2H), 7.33-7.28 (m, 3H), 6.98 (d, J=8.0 Hz, 1H), 2.67 (s, 3H).

Intermediate 13

2-ethyl-4-nitro-1-phenoxybenzene

[0214] ##STR00032##

[0215] In a RBF previously equipped with a magnetic stirrer was taken 1-(5-nitro-2-phenoxyphenyl)ethan-1-one (Intermediate 12, 7.92 g, 30.8 mmol) in TFA (159 ml) and the mixture was purged with nitrogen gas for 10 minutes. Et.sub.3SiH (14.33 g, 123.2 mmol) and LiClO.sub.4 (0.034 g, 0.31 mmol) were added under N.sub.2 atmosphere. The reaction mixture was stirred at RT for 6 h. The solvent was removed under reduced pressure to obtain 12.0 g of the crude title compound that was used without further purification in the next step. MS (ES+) m/z 244 [M+H].sup.+

Intermediate 14

3-ethyl-4-phenoxyaniline

[0216] ##STR00033##

[0217] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-ethyl-4-nitro-1-phenoxybenzene (Intermediate 13, 12 g, 49.3 mmol) in methanol (150 ml). 10% Pd/C (3.0 g, 50% wet) was added under N.sub.2 atmosphere. The suspension was stirred for 6 h under H.sub.2 (gas) bubbling. The reaction mixture was filtered through a celite pad and the solvent removed under reduced pressure. The obtained crude was purified by column chromatography (30% Ethyl acetate in hexanes) to obtain 4.5 g (68%) of the title compound. 1H NMR (400 MHz, DMSO-d6): δ 7.26 (app t, J=8.00 Hz, 2H), 6.95 (t, J=7.20 Hz, 1H), 6.78 (app d, J=8.00 Hz, 2H), 6.66 (d, J=8.40 Hz, 1H), 6.52 (d, J=2.00 Hz, 1H), 6.43-6.45 (m, 1H), 4.95 (s, 2H), 2.34 (q, J=7.20 Hz, 2H), 1.03 (t, J=7.20 Hz, 3H). MS (ES+) m/z 214 [M+H].sup.+

Intermediate 15

3-(3-ethyl-4-phenoxyphenyl)-1-phenylurea

[0218] ##STR00034##

[0219] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-ethyl-4-phenoxyaniline (Intermediate 14, 1.20 g, 5.6 mmol) in DMF (12.0 ml) and the mixture was cooled to 0° C. NaHCO.sub.3 (1.418 g, 16.4 mmol) was added and the resulting mixture was stirred for 20 min at 0° C. Phenyl isocyanate (0.804 g, 6.7 mmol) was added and the reaction mixture was stirred at RT for 4 h. The reaction mixture was quenched with water (30 ml) and the product was extracted with EtOAc (3×30 ml). The combined organic layer was washed with brine (20 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain 1.21 g (64%) of the title compounds that was used in the next step without further purification. MS (ES+) m/z 333 [M+H].sup.+

Intermediate 16

3-(3-ethyl-4-phenoxyphenyl)-1-(3-methylphenyl)urea

[0220] ##STR00035##

[0221] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-ethyl-4-phenoxyaniline (Intermediate 14, 1.20 g, 5.6 mmol) in DMF (12.0 ml) and the mixture was cooled to 0° C. NaHCO.sub.3 (1.418 g, 16.8 mmol) was added and the resulting mixture was stirred for 20 min at 0° C. 3-Methyl-phenyl isocyanate (0.898 g, 6.7 mmol) was added and the reaction mixture stirred at RT for 4 h. The reaction mixture was quenched with water (30 ml) and the product was extracted with EtOAc (3×30 ml). The combined organic layer was washed with brine (20 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain 1.22 g (62%) of the title compound that was used in the next step without further purification. MS (ES+) m/z 347 [M+H].sup.+

Intermediate 17

N-cyano-N′-(3-methyl-4-phenoxyphenyl)(methylsulfanyl)methanimidamide

[0222] ##STR00036##

[0223] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-methyl-4-phenoxyaniline (commercially available, 2.0 g, 10 mmol) in Ethanol (30 ml). To this solution dimethyl N-cyanodithioiminocarbonate (1.47 g, 10 mM) was added at room temperature. The reaction mixture was heated to 80° C. and stirred for 72 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude product was purified by Combi-flash chromatography using 50% ethyl acetate in hexanes as an eluent to obtain 2.2 g (73% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 10.14 (s, 1H), 7.40-7.36 (m, 3H), 7.29 (d, J=8.4 Hz, 1H), 7.11 (t, J=7.2 Hz, 1H), 6.92 (app d, J=8.4 Hz, 3H), 2.70 (s, 3H), 2.18 (s, 3H). MS (ES+) m/z 298 [M+H].sup.+

Intermediate 18

1-(4-methoxyphenyl)-3-[1-(4-methoxyphenyl)-5-(3-methyl-4-phenoxyphenyl)-4-(methylsulfanyl)-6-oxo-1,2,5,6-tetrahydro-1,3,5-triazin-2-ylidene]urea

[0224] ##STR00037##

[0225] In a 25 ml RBF previously equipped with a magnetic stirrer was taken N-cyano-N′-(3-methyl-4-phenoxyphenyl)(methylsulfanyl)methanimidamide (Intermediate 17, 0.40 g, 1.3 mmol) and 4-methoxy-phenyl isocyanate (0.34 g, 2.3 mmol) in DCM (8.0 ml). To the mixture, triethylamine (1.90 ml, 13.4 mmol) was added followed by portion wise (3-4 portions) addition of 1,1′-Carbonyldiimidazole (1.10 g, 6.7 mmol). The reaction mixture was stirred at room temperature for 6 h. The reaction mixture was quenched with water (30 ml) and both the layers were separated. Aqueous layer was extracted with DCM (2×30 ml). The combined organic layer was washed with brine (30 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by Combi-flash chromatography using 50% ethyl acetate in hexanes as an eluent to obtain 0.45 g of the title compound that was used in the next step without further purification. MS (ES+) m/z 596 [M+H].sup.+

Intermediate 19

3-(3-hydroxy-4-phenoxyphenyl)-1-(3-methylphenyl)urea

[0226] ##STR00038##

[0227] To a RBF previously equipped with a magnetic stirrer and nitrogen balloon was added 5-Amino-2-phenoxyphenol (commercially available, 2.0 g, 9.9 mmol) in DMF (20 ml) and the mixture was cooled to 0° C. NaHCO.sub.3 (2.49 g, 29.7 mmol) was added and the resulting mixture was stirred for 20. 3-Methyl-phenylisocyanate (1.32 g, 9.9 mmol) was added and reaction mixture was stirred at room temperature for 4 h. The reaction mixture was quenched with water (500 ml) and product was extracted with EtOAc (3×100 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was triturated with dichloromethane and hexane to obtain 3.5 g of the title compound. MS (ES+) m/z 335 [M+H].sup.+

Intermediate 20

ethyl 2-(5-{[(3-methylphenyl)carbamoyl]amino}-2-phenoxyphenoxy)acetate

[0228] ##STR00039##

[0229] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(3-hydroxy-4-phenoxyphenyl)-1-(3-methylphenyl)urea (Intermediate 19, 3.00 g, 8.97 mmol) and K.sub.2CO.sub.3 (1.85 g, 13.46 mmol) in DMF (30 ml) and the mixture was cooled to 0° C. To the reaction mixture, BrCH.sub.2COOEt (2.24 g, 13.46 mmol) was added and the resulting reaction mixture was stirred for 16 h at room temperature. The reaction mixture was diluted with ice cold water (100 ml) and extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was purified by column chromatography using silica gel (60-120 mesh) and 20% ethyl acetate in hexane as an eluent to obtain 3.5 g of the title compound. MS (ES+) m/z 421 [M+H].sup.+

Intermediate 21

ethyl 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetate

[0230] ##STR00040##

[0231] In a sealed tube previously equipped with a magnetic stirrer and nitrogen balloon was taken ethyl 2-(5-{[(3-methylphenyl)carbamoyl]amino}-2-phenoxyphenoxy)acetate (Intermediate 20, 3.50 g, 8.32 mmol) in Chlorobenzene (35 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (3.83 g, 33.3 mmol) was added drop-wise and the resulting reaction mixture was allowed to reach room temperature and heated at 150° C. for 16 h. The solvent was removed under reduced pressure and the crude product was purified by column chromatography using silica gel (100-200 mesh) and 70% ethyl acetate in hexanes as an eluent to obtain 1.071 g of the crude title compound that was used in the next step without further purification. MS (ES−) m/z 488 [M−H].sup.−.

Intermediate 22

2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetic acid

[0232] ##STR00041##

[0233] In a RBF previously equipped with a magnetic stirrer was taken ethyl 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetate (Intermediate 21, 1.0 g) in THF:H.sub.2O (8 ml: 2 ml). LiOH (1.85 g, 13.4 mmol) was added and the resulting reaction mixture was stirred for 1 h at RT. The reaction mixture was concentrated under reduced pressure and the crude mass was diluted with water (5 ml) and extracted with EtOAc (2×10 ml). The aqueous layer was acidified using 1M HCl to pH-3-4 and then again extracted with EtOAc (3×20 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure. The crude product was triturated with DCM and Hexane to obtain the 0.8 g of the title compound. MS (ES−) m/z 460 [M−H].sup.−.

Intermediate 23

2-(methylsulfanyl)-4-nitro-1-phenoxybenzene

[0234] ##STR00042##

[0235] In a seal tube previously equipped with a magnetic stirrer and nitrogen balloon was taken Phenol (0.84 g, 8.97 mmol) and Cs2CO.sub.3 (2.92 g, 8.97 mmol) in DMSO (84 ml) and the mixture was cooled to 0° C. To the reaction mixture, 1-fluoro-2-(methylsulfanyl)-4-nitrobenzene (commercially available, 1.4 g, 7.48 mmol) was added and the resulting reaction mixture was allowed to reach room temperature and then stirred at 110° C. for 5 h. The reaction mixture was quenched with water (50 ml) and product was extracted with EtOAc (3×40 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by column chromatography using silica gel (60-120 mesh) and 10% ethyl acetate in hexane as an eluent to obtain 1.0 g (51% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.06-8.01 (m, 2H), 7.50 (app t, 2H), 7.30 (t, J=7.20 Hz, 1H), 7.15 (app d, 2H), 6.89 (d, J=9.20 Hz, 1H), 2.60 (s, 3H).

Intermediate 24

3-(methylsulfanyl)-4-phenoxyaniline

[0236] ##STR00043##

[0237] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(methylsulfanyl)-4-phenoxyaniline (Intermediate 23, 0.50 g, 1.91 mmol) in Methanol (5.0 ml). 10% Pd/C (50% wet, 0.101 g, 0.956 mmol) was added under N.sub.2 atmosphere. The mixture was stirred under H.sub.2 (gas) bubbling. The completion of reaction was confirmed by the TLC using Hexane:EtOAc (8:2) as mobile phase and monitored using TLC (visualized using UV light). After completion, the reaction mixture was filtered through a celite bed, washed with methanol and the solvent was removed under reduced pressure to obtain 0.400 g (90%) of the title compound. MS (ES+) m/z 232 [M+H].sup.+

Intermediate 25

3-[3-(methylsulfanyl)-4-phenoxyphenyl]-1-phenylurea

[0238] ##STR00044##

[0239] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(methylsulfanyl)-4-phenoxyaniline (Intermediate 24, 0.400 g, 1.729 mmol) in DMF (4.0 ml) and the mixture was cooled to 0° C. To the reaction mixture, NaHCO.sub.3 (0.435 g, 5.187 mmol) was added and the resulting mixture was stirred for 20 min at 0° C. Phenyl isocyanate (0.205 g, 1.729 mmol) was added and the reaction stirred at RT for 3 h. The reaction mixture was quenched with water (30 ml) and the product was extracted with EtOAc (3×30 ml). The combined organic layer was washed with brine (20 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain the crude product that was purified by column chromatography using Hexanes:ethyl acetate as an eluent using 60-120 mesh silica gel to yield 0.280 g (46%) of the title compound. MS (ES+) m/z 351 [M+H].sup.+

Intermediate 26

1-(3-methylphenyl)-3-[3-(methylsulfanyl)-4-phenoxyphenyl]urea

[0240] ##STR00045##

[0241] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken, 3-methyl-aniline (0.185 g, 1.729 mmol), NaHCO.sub.3 (0.435 g, 5.187 mmol) and DCM (12 ml). The mixture was cooled to 0° C. and Triphosgene (0.168 g, 0.570 mmol) was added and the reaction mixture was stirred for 1 h. After that, 3-(methylsulfanyl)-4-phenoxyaniline (Intermediate 24, 0.400 g, 1.729 mmol) and NaHCO.sub.3 (0.435 g, 5.187 mmol) were added and the reaction mixture was allowed to reach room temperature. The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was quenched with water (50 ml) and the product was extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (50 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product. The crude product was purified by column chromatography using silica gel (60-120 mesh) and 2% methanol in Dichloromethane as an eluent to obtain 0.320 g (50% yield) of the title compound. MS (ES+) m/z 365 [M+H].sup.+

Example 1

1-(3-methyl-4-phenoxyphenyl)-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione

[0242] ##STR00046##

[0243] In a microwave tube previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(3-methyl-4-phenoxyphenyl)-1-(3-methylphenyl)urea (Intermediate 1, 2.8 g, 8.4 mmol) in Bromobenzene (28 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (3.88 g, 33.7 mmol) was added and resulting reaction mixture was allowed to come to 25° C. and then heated at 150° C. for 16 h. The reaction mixture was quenched with water (10 ml) and the aqueous layer was extracted with ethyl acetate (2×30 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure to obtain the crude product that was purified by preparative RP-HPLC (acetonitrile 25-100% in water [5 mM Ammonium bicarbonate+0.1% NH3]) to yield 89 mg, (26% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.03 (s, 1H), 7.43-7.33 (m, 4H), 7.25-7.13 (m, 5H), 6.99 (s, 1H), 6.97 (s, 1H), 6.93 (d, J=8.4 Hz, 1H), 2.35 (s, 3H), 2.22 (s, 3H). MS (ES−) m/z 400 [M−H].sup.−

Example 2

1-[3-(methoxymethyl)phenyl]-3-(3-methyl-4-phenoxyphenyl)-1,3,5-triazinane-2,4,6-trione

[0244] ##STR00047##

[0245] In a microwave tube previously equipped with a magnetic stirrer and nitrogen balloon was taken 1-[3-(methoxymethyl)phenyl]-3-(3-methyl-4-phenoxyphenyl)urea (Intermediate 2, 0.5 g, 1.4 mmol) in Chlorobenzene (5 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (0.644 g, 5.6 mmol) was added and resulting reaction mixture was allowed to reach room temperature and then heated at 150° C. for 3 h in Anton paar microwave synthesizer-300. The solvent was removed under reduced pressure to obtain that crude product, that was purified by preparative RP-HPLC (acetonitrile 20-100% in water [5 mM Ammonium bicarbonate+0.1% NH3]) to yield 0.055 g (9% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1H), 7.46-7.27 (m, 7H), 7.2 (d, J=8.0, 1H), 7.13 (t, J=7.2 Hz, 1H), 6.89-7.01 (m, 3H), 4.45 (s, 2H), 3.49 (s, 3H), 2.21 (s, 3H). MS (ES−) m/z 430 [M−H].sup.−

Example 3

1-[3-(methoxymethyl)-4-phenoxyphenyl]-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione

[0246] ##STR00048##

[0247] In a microwave tube previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-[3-(methoxymethyl)-4-phenoxyphenyl]-1-(3-methylphenyl)urea (Intermediate 5, 0.2 g, 0.6 mmol) in Chlorobenzene (2 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (0.191 g, 1.7 mmol) was added and the resulting reaction mixture was allowed to reach room temperature and heated at 150° C. for 3 h in an Anton par microwave synthesizer-300. The solvent was removed under reduced pressure and the crude product was purified by preparative RP-HPLC (acetonitrile 35-100% in water [0.1% formic acid]) to yield 0.022 g (9% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.98 (s, 1H), 7.49 (s, 1H), 7.42 (t, J=7.60 Hz, 2H), 7.35 (t, J=7.60 Hz, 1H), 7.27 (dd, J=2.00, 8.60 Hz, 1H), 7.23 (d, J=7.60 Hz, 1H), 7.15-7.18 (m, 3H), 7.02 (d, J=7.60 Hz, 2H), 6.89 (d, J=8.40 Hz, 1H), 4.49 (s, 2H), 3.34 (s, 3H), 2.34 (s, 3H). MS (ES−) m/z 430 [M−H].sup.−

Example 4

1-(2-methyl-1-benzofuran-7-yl)-3-(3-methyl-4-phenoxyphenyl)-1,3,5-triazinane-2,4,6-trione

[0248] ##STR00049##

[0249] In a 10 ml microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(2-methyl-1-benzofuran-7-yl)-1-(3-methyl-4-phenoxyphenyl)urea (Intermediate 7, 0.210 g, 0.5 mmol) in Bromobenzene (2.1 ml) and the mixture was cooled to 0° C. Ethoxy carbonyl isocyanate (0.227 g, 1.9 mmol) was added drop wise and resulting reaction mixture was allowed to reach room temperature and heated at 150° C. for 3 h in Anton paar microwave synthesizer-300. The solvent was removed under reduced pressure to obtain crude product. The crude product was purified by preparative RP-HPLC (acetonitrile 55-100% in water [0.1% formic acid]) to yield 30 mg (12% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d6): δ 12.22 (s, 1H), 7.61-7.58 (m, 1H), 7.41-7.37 (m, 3H), 7.28-7.26 (m, 3H), 7.13 (t, J=7.20 Hz, 1H), 6.99 (m, 3H), 6.68 (s, 1H), 2.45 (s, 3H), 2.22 (s, 3H). MS (ES−) m/z 440 [M−H].sup.−

Example 5

1-(3-methylphenyl)-3-{3-[(2-oxoimidazolidin-1-yl)methyl]-4-phenoxyphenyl}-1,3,5-triazinane-2,4,6-trione

[0250] ##STR00050##

[0251] To a microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was added 2-Imidazolidinone (0.039 g, 0.458 mmol) in THE (1.0 ml) and the mixture was cooled to 0° C. Sodium hydride (60%, 0.018 g, 0.458 mmol) was added at 0° C. and reaction mixture was stirred for 20 min at 0° C. 1-[3-(chloromethyl)-4-phenoxyphenyl]-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione (Intermediate 11, 0.100 g, 0.229 mmol) was added and resulting reaction mixture was stirred room temperature for 12 h. The completion of reaction was confirmed by the TLC using DCM:MeOH (9:1) as mobile phase. The TLC was visualized using UV light. After completion of the reaction, reaction mixture was quenched with water (10 ml) and product was extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (35 ml), dried over sodium sulphate and the solvent removed under reduced. The crude product was purified by preparative HPLC (acetonitrile 20-100% in water [5 mM Ammonium bicarbonate+0.1% NH.sub.3]) to yield 0.006 g (5% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.96 (s, 1H), 7.44-7.42 (m, 2H), 7.40-7.35 (m, 2H), 7.26-7.16 (m, 5H), 7.08-6.98 (m, 2H), 6.90 (d, J=8.8 Hz, 1H), 6.47 (s, 1H), 4.28 (s, 2H), 3.25-3.26 (m, 4H), 2.33 (s, 3H); MS (ES+) m/z 486 [M+H].sup.+

Example 6

1-(3-ethyl-4-phenoxyphenyl)-3-phenyl-1,3,5-triazinane-2,4,6-trione

[0252] ##STR00051##

[0253] In a microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(3-ethyl-4-phenoxyphenyl)-1-phenylurea (Intermediate 15, 1.20 g, 3.6 mmol) in Bromobenzene (14.0 ml). Ethoxy carbonyl isocyanate (1.246 g, 10 mmol) was added and the resulting reaction mixture heated at 150° C. for 4 h in an Anton paar microwave synthesizer-300. The solvent was removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 55-100% in water [0.1% formic acid]) to yield 130 mg (8.9% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.05 (s, 1H), 7.51-7.33 (m, 8H), 7.21 (d, J=7.6 Hz, 1H), 7.13 (t, J=7.2 Hz, 1H), 7.01-6.95 (m, 2H), 6.90 (d, J=8.4 Hz, 1H), 2.60 (q, J=7.20 Hz, 2H), 1.15 (t, J=7.60 Hz, 3H). MS (ES−) m/z 400 [M−H].sup.−

Example 7

1-(3-ethyl-4-phenoxyphenyl)-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione

[0254] ##STR00052##

[0255] In a 30 ml microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-(3-ethyl-4-phenoxyphenyl)-1-(3-methylphenyl)urea (Intermediate 16, 1.20 g, 3.4 mmol) in Bromobenzene (12 ml). Ethoxy carbonyl isocyanate (1.19 g, 10.4 mmol) was added and the resulting reaction mixture was heated to 150° C. for 4 h in an Anton paar microwave synthesizer-300. The solvent was removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 60-100% in water [0.1% formic acid]) to yield 84 mg (5%) of the title compound. 1H NMR (400 MHz, DMSO-d6): δ 11.99 (s, 1H), 7.43-7.32 (m, 4H), 7.26-7.11 (m, 5H), 7.01-6.95 (m, 2H), 6.90 (d, J=8.4 Hz, 1H), 2.60 (m, 2H), 2.34 (s, 3H) 1.16 (t, J=7.20 Hz, 3H). MS (ES−) m/z 416 [M−H].sup.−

Example 8

1-(4-methoxyphenyl)-3-(3-methyl-4-phenoxyphenyl)-1,3,5-triazinane-2,4,6-trione

[0256] ##STR00053##

[0257] In a RBF previously equipped with a magnetic stirrer and nitrogen balloon was taken 1-(4-methoxyphenyl)-3-[1-(4-methoxyphenyl)-5-(3-methyl-4-phenoxyphenyl)-4-(methylsulfanyl)-6-oxo-1,2,5,6-tetrahydro-1,3,5-triazin-2-ylidene]urea (Intermediate 18, 0.45 g, 0.7 mmol) in 1,4-Dioxane (4.5 ml). To the solution 2M aq. HCl (6.75 ml) was added and the reaction mixture was heated to 100° C. and stirred at same temperature for 2 h. The reaction mixture was allowed to reach room temperature and quenched with ice cold water (20 ml) and stirred for 10 min. The aqueous layer was extracted with Ethyl Acetate (2×30 ml) and the combined organic layer was washed with ice cold water (20 ml) followed by brine (20 ml), dried over sodium sulphate and the solvent removed under reduce pressure to obtain crude product. The crude product was purified by column chromatography using 25% ethyl acetate in hexanes as an eluent to yield 0.085 g (13%) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6: δ 11.96 (s, 1H), 7.46-7.39 (m, 2H), 7.33-7.28 (m, 3H), 7.22 (dd, J=8.4 Hz, 2.0 Hz, 1H), 7.15 (t, J=7.2 Hz, 1H), 7.03-6.97 (m, 4H), 6.92 (d, J=8.8 Hz, 1H), 3.80 (s, 3H), 2.23 (s, 3H). MS (ES+) m/z 418 [M+H].sup.+

Example 9

1-(3-methylphenyl)-3-{3-[2-oxo-2-(pyrrolidin-1-yl)ethoxy]-4-phenoxyphenyl}-1,3,5-triazinane-2,4,6-trione

[0258] ##STR00054##

[0259] In a vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetic acid (Intermediate 22, 0.100 g, 0.216 mmol) in DMF (1.0 ml). To it HATU (0.247 g, 0.650 mmol) was added and resulting reaction mixture was stirred for 1 h at 0° C. After that, pyrrolidine (0.023 g, 0.325 mmol) and N,N-Diisopropylethylamine (0.15 ml, 0.866 mmol) were added and the reaction mixture was allowed to reach room temperature and then stirred for 2 h. The reaction mixture was diluted with ice cold water (100 ml) and extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified using preparative RP-HPLC (acetonitrile 10-100% in water [5 mM ammonium bicarbonate+0.1% NH.sub.3]) to yield 42 mg (37% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.85 (s, 1H), 7.39-7.31 (m, 3H), 7.23 (d, J=7.2 Hz, 1H), 7.16 (app s, 3H), 7.11-7.04 (m, 2H), 7.02-6.92 (m, 3H), 4.67 (s, 2H), 3.32-3.23 (m, 4H), 2.34 (s, 3H), 1.85-1.67 (m, 4H). MS (ES−) m/z 513 [M−H].sup.−

Example 10

1-(3-methylphenyl)-3-{3-[2-oxo-2-(piperidin-1-yl)ethoxy]-4-phenoxyphenyl}-1,3,5-triazinane-2,4,6-trione

[0260] ##STR00055##

[0261] In a vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetic acid (Intermediate 22, 0.100 g, 0.216 mmol) in DMF (1.0 ml). To it HATU (0.247 g, 0.650 mmol) was added and resulting reaction mixture was stirred for 1 h at 0° C. After that, piperidine (0.027 g, 0.325 mmol) and N,N-Diisopropylethylamine (0.15 ml, 0.866 mmol) were added and the reaction mixture was allowed to reach room temperature and then stirred for 2 h at room temperature. The reaction mixture was diluted with ice cold water (100 ml) and extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 20-100% in water [5 mM ammonium bicarbonate+0.1% NH.sub.3]) to yield 30 mg (26% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.58 (s, 1H), 7.38-7.32 (m, 3H), 7.23 (d, J=7.2 Hz, 1H), 7.18-7.12 (m, 3H), 7.10-7.04 (m, 2H), 7.01-6.91 (m, 3H), 4.75 (s, 2H), 3.42-3.35 (m, 2H), 3.26 (br s, 2H), 2.34 (s, 3H), 1.51 (br s, 2H), 1.40 (br s, 4H). MS (ES−) m/z 527 [M−H].sup.−

Example 11

1-(3-methylphenyl)-3-{3-[2-(morpholin-4-yl)-2-oxoethoxy]-4-phenoxyphenyl}-1,3,5-triazinane-2,4,6-trione

[0262] ##STR00056##

[0263] In a vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetic acid (Intermediate 22, 0.100 g, 0.216 mmol) in DMF (1.0 ml). To it HATU (0.247 g, 0.650 mmol) was added and resulting reaction mixture was stirred for 1 h at 0° C. After that, morpholine (0.028 g, 0.325 mmol) and N,N-Diisopropylethylamine (0.15 ml, 0.866 mmol) were added and the reaction mixture was allowed to reach room temperature and then stirred for 2 h at room temperature. The reaction mixture was diluted with ice cold water (100 ml) and extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 10-100% in water [5 mM ammonium bicarbonate+0.1% NH.sub.3]) to yield 17 mg (14% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.99 (s, 1H), 7.42-7.30 (m, 3H), 7.24 (d, J=8 Hz, 1H), 7.22-7.13 (m, 3H), 7.12-7.03 (m, 2H), 7.01 (d, J=7.2 Hz, 1H), 6.98-6.89 (d, 2H), 4.79 (s, 2H), 3.56-3.45 (m, 4H), 3.45-3.36 (m, 4H), 2.34 (s, 3H). MS (ES−) m/z 529 [M−H].sup.−

Example 12

1-{3-[2-(azetidin-1-yl)-2-oxoethoxy]-4-phenoxyphenyl}-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione

[0264] ##STR00057##

[0265] In a vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 2-{5-[3-(3-methylphenyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]-2-phenoxyphenoxy}acetic acid (Intermediate 22, 0.075 g, 0.162 mmol) in DMF (0.75 ml). To it HATU (0.185 g, 0.487 mmol) was added and the resulting mixture was stirred for 1 h at 0° C. After that, azetidine (0.014 g, 0.243 mmol) and N,N-Diisopropylethylamine (0.11 ml, 0.650 mmol) were added and the reaction mixture was allowed to reach room temperature and then stirred for 2 h at room temperature. The reaction mixture was diluted with ice cold water (100 ml) and extracted with EtOAc (3×50 ml). The combined organic layer was washed with brine (100 ml), dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 40-100% in water [0.1% formic acid]) to yield 9 mg (11% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 12.02 (s, 1H), 7.41-7.32 (m, 3H), 7.23 (d, J=7.6 Hz, 1H), 7.16 (app s, 3H), 7.12-7.05 (m, 2H), 7.04-6.99 (m, 1H), 6.98-6.92 (m, 2H), 4.53 (s, 2H), 4.07 (t, J=7.2 Hz, 2H), 3.85 (t, J=7.6 Hz, 2H), 2.34 (s, 3H), 2.18-2.10 (m, 2H). MS (ES−) m/z 499 [M−H].sup.−

Example 13

1-[3-(methylsulfanyl)-4-phenoxyphenyl]-3-phenyl-1,3,5-triazinane-2,4,6-trione

[0266] ##STR00058##

[0267] In a microwave vial previously equipped with a magnetic stirrer and nitrogen balloon was taken 3-[3-(methylsulfanyl)-4-phenoxyphenyl]-1-phenylurea (Intermediate 25, 0.150 g, 0.428 mmol) in Chlorobenzene (1.5 ml). The solution was cooled to 0° C. and Ethoxy carbonyl isocyanate (0.197 g, 1.712 mmol) was added and resulting reaction mixture heated to 150° C. for 4 h in an Anton paar microwave synthesizer-300. After completion of the reaction, solvent was evaporated under reduced pressure to obtain crude product. The crude product was purified by preparative RP-HPLC (acetonitrile 25-100% in water [0.1% formic acid]) to yield 0.023 g (12% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6: δ 12.07 (s, 1H), 7.54-7.37 (m, 8H), 7.21-7.14 (m, 2H), 7.03-6.94 (m, 3H), 2.43 (s, 3H). MS (ES−) m/z 418 [M−H].sup.−

Example 14

1-(3-methylphenyl)-3-[3-(methylsulfanyl)-4-phenoxyphenyl]-1,3,5-triazinane-2,4,6-trione

[0268] ##STR00059##

[0269] In a microwave tube previously equipped with a magnetic stirrer and nitrogen balloon was taken 1-(3-methylphenyl)-3-[3-(methylsulfanyl)-4-phenoxyphenyl]urea (Intermediate 26, 0.150 g, 0.411 mmol) in Toluene (1.5 ml). The solution was cooled to 0° C. and Ethoxy carbonyl isocyanate (0.189 g, 1.646 mmol) was added and resulting reaction mixture was allowed to reach room temperature and then heated at 150° C. for 3 h in an Anton paar microwave synthesizer-300. After completion of the reaction, solvent was evaporated under reduced pressure to obtain the crude product that was purified by preparative RP-HPLC (acetonitrile 50-100% in water [0.1% formic acid]) to yield 0.011 g (6% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6: δ 12.05 (s, 1H), 7.44-7.32 (m, 4H), 7.24 (d, J=7.20 Hz, 1H), 7.19-7.12 (m, 4H), 7.01-6.92 (m, 3H), 2.41 (s, 3H), 2.34 (s, 3H). MS (ES−) m/z 432 [M−H].sup.−

Example 15

1-(3-methylphenyl)-3-{3-[(2-oxopyrrolidin-1-yl)methyl]-4-phenoxyphenyl}-1,3,5-triazinane-2,4,6-trione

[0270] ##STR00060##

[0271] In a sealed tube previously equipped with a magnetic stirrer and nitrogen balloon was taken 1-[3-(chloromethyl)-4-phenoxyphenyl]-3-(3-methylphenyl)-1,3,5-triazinane-2,4,6-trione (Intermediate 11, 0.10 g, 0.2 mmol) in THE (1.0 ml) and to it NaH (60%, 0.018 g, 0.4 mmol) was added and the mixture was stirred for 10 min at room temperature. 2-Pyrrolidone (0.039 g, 0.4 mmol) was added and the reaction mixture was heated at 60° C. for 1 h. The reaction mixture was quenched with water (10 ml) and the aqueous layer was extracted with ethyl acetate (3×10 ml). The combined organic layer was dried over sodium sulphate and the solvent removed under reduced pressure to obtain crude product that was purified by preparative RP-HPLC (acetonitrile 15-100% in water [5 mM Ammonium bicarbonate+0.1% NH.sub.3]) to yield 15 mg (13% yield) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.97 (s, 1H), 7.43-7.17 (m, 9H), 7.03-6.92 (m, 3H), 4.42 (s, 2H), 3.27 (br s, 2H), 2.35 (s, 3H), 2.22 (br s, 2H), 1.90 (br s, 2H). MS (ES−) m/z 483 [M−H].sup.−

BIOLOGICAL EXAMPLES

Biological Example 1

In Vitro Trk Receptor Modulation Assay

[0272] A high throughput cell-based screen was used to identify positive modulators of TrkA and TrkB. The screen involves the use of cell-based assay overexpressing TrkA, or TrkB. The purpose of the assay is to identify compounds that modulate neurotrophin signalling (Forsell et al 2012). The assay can be used in inhibitor mode using a high concentration of ligand, in modulator mode using an intermediate concentration and in agonist mode using a low concentration of ligand.

[0273] The assay uses Enzyme Fragment Complementation (EFC) technique, which is a proximity-based assay. Briefly, cells used in this assay over-express two fusion proteins, i.e. the receptor, which can be one of TrkA or TrkB fused to a small peptide of beta-galactosidase and an adaptor protein, i.e. SHC1 (or any other Trk-adaptor protein) fused to the major part of beta-galactosidase. Ligand binding to the receptor induces phosphorylation of the intracellular domain and hence, recruitment of the adaptor protein to the receptor. The proximity between the small activating peptide on the receptor and the major part of beta-galactosidase on the adaptor protein leads to an active beta-galactosidase enzyme. The activation of the receptor is quantified by measuring the amount of active beta-galactosidase by its conversion of a non-luminescent substrate into a luminescent product.

[0274] U2OS-cells, over-expressing TrkA or TrkB, were plated in 96- or 384-well plates and incubated overnight. On the following day, test compound was pre-mixed with ligand (NGF) and the ligand-compound mixture is then added to the cells to yield a final ligand concentration of 10 ng/mL. After 3 hours of incubation at room temperature, the incubation is stopped by the addition of a beta-galactosidase substrate mixture containing detergents. The substrate mixture is incubated for 60 minutes at ambient temperature. The luminescence is thereafter read by the use of a plate reader.

Results

[0275]

TABLE-US-00001 TABLE 1 Data from these assays for representative compounds is shown in the table below. The potency is expressed as EC50 (μM) for the individual receptors. The data indicate that the compounds of the invention are highly potent modulators of TrkA and TrkB receptor signalling and are therefore expected to possess useful therapeutic properties. Example TrkA TrkB 1 0.12 0.15 2 0.43 0.26 3 0.33 0.31 4 0.30 0.27 5 0.30 0.12 6 0.21 0.22 7 0.13 0.09 8 0.35 0.23 9 0.21 0.15 10 0.28 0.32 11 0.27 0.18 12 0.41 0.22 13 0.64 0.38 14 0.65 0.30 15 0.04 0.11

Biological Example 2

Human Liver Microsomal Stability

[0276] A 10 mM stock solution (in DMSO) was prepared for each compound. From the intermediate stock solution of 2 mM, a working solution of 0.5 mM was prepared by diluting the compound in Acetonitrile:water (50:50). The compound (1.8 μL of working solution) was spiked in 0.1 M potassium phosphate buffer (260.7 μL), pH 7.4 at a concentration of 3 μM (0.1% DMSO). Following this, human liver microsomes (pooled, Invitrogen, #HMMCPL) (7.5 μL; final protein conc. 0.5 mg/mL) were added. The aforementioned sample was incubated at 37° C. for 5 min. Subsequently, 30 μL of 10 mM NADPH prepared in 0.1 M potassium phosphate buffer was added (as a co-factor) to initiate the reaction. The samples were then incubated at 37° C. for 0, 20, 40, 60 and 120 min.

[0277] At each time point (0, 20, 40, 60 and 120 min), 40 μL of the samples were withdrawn and reactions were stopped using 360 μL chilled acetonitrile or methanol containing suitable internal standard (Carbamazepine). The samples were centrifuged and the supernatants was analyzed in duplicate by LC-MS/MS. The percent compound remaining at each time point was calculated with respect to that of the 0 min sample. Control samples were run without NADPH for initial and final time point and blank samples was prepared using DMSO (without the test compound).

[0278] For comparison, the experiment was also performed with known Trk receptor signalling modulator 1-methyl-3-[3-methyl-4-(4-trifluoromethanesulfonylphenoxy)phenyl]-1,3,5-triazinane-2,4,6-trione) (ponazuril). Ponazuril is metabolized very slowly in vivo and this prolonged exposure creates an increased risk of side effects if the compound is administered systemically. This causes significant challenges to the development of the compound as a systemic drug.

Results

[0279]

TABLE-US-00002 TABLE 2 % metabolism of representative compounds in human liver microsomes after incubation for 120 min. % of parent compound metabolized Example after 120 min 1 26.8 2 13.3 3 82.0 4 37.7 5 15.4 6 NT 7 48.8 8 NT 9 82.8 10 99.8 11 57.3 12 NT 13 4.7 14 53.4 15 29.6 ponazuril 3.1

[0280] These data indicate that the compounds are metabolised more quickly than ponazuril and are therefore likely to have improved metabolic profiles for pharmaceutical development as systemic drugs and/or may offer other metabolic advantages for pharmaceutical development.

Biological Example 3

Passive Avoidance Task

[0281] The passive avoidance (PA) is an aversive learning task based on classical (Pavlovian) fear conditioning that allows for analysis of both facilitation and impairment of memory function by adjusting the unconditioned stimulus, i.e. the electrical foot shock. Commonly a cognitive-impairing agent is administered to the animals to mimic the neurochemical disturbances present in various cognitive disorders e.g. cholinergic (scopolamine) and glutamatergic (MK-801) deficits.

[0282] Prior to testing, the animals are brought to the experimental room where they were allowed to habituate for 60 min. The test is conducted using a modified shuttle box with two communicating compartments of equal size with a small sliding door built into the separating wall and a stainless-steel bar floor. One of the compartments is not illuminated and thus black whereas the other compartment (the light one) is illuminated by an electric bulb, installed on the top of a plexiglass cover. The PA training is conducted in a single session. The animals are allowed to explore the compartment for 60 sec, after which the sliding door is automatically opened and the mouse is allowed to cross over into the dark compartment. Once the mouse has entered the dark chamber with all four feet, the sliding door is automatically closed and a scrambled electrical current is delivered through the grid floor. Latency to cross over into the dark compartment (training latency) is recorded. The memory test is conducted on day 2 (24 hrs later) whereby the animals are placed in the light compartment and allowed to explore for 15 sec, then the sliding door is opened allowing free access to the dark compartment for a period of 300 sec. The latency to cross over into the dark compartment with all four feet is measured (retention latency) as well as time in bright compartment and a number of other relevant parameters (e.g. number of visits in the dark compartment).

[0283] Vehicle (20% DMSO in 0.1M PBS) or different doses of the compounds of Example 1 and Example 3 were administered to C57/Bl6 mice as a single s.c. administration 60 min prior to PA training. PA training was then performed according to the procedure described above. On the day of PA training, scopolamine at 0.3 mg/kg, or vehicle, was administered subcutaneously 30 min prior to training.

[0284] Data on time spend in the bright compartment for mice treated with different doses of the compound of Example 1, the vehicle control group and scopolamine alone are shown in FIG. 1, and for mice treated with different doses of the compound of Example 3, the vehicle control group and scopolamine alone are shown in FIG. 2.

Biological Example 4

Forced Swim Test

[0285] The Forced Swim Test (FST) is a commonly used test in rodents to assess depression-like behaviour (Porsolt et al., Nature, 1977, 266, 730-732). The model measures a learned helplessness response and has shown to have predictive validity for clinical antidepressants, e.g. SSRIs.

[0286] The mouse version consists of a vertical glass cylinder (25 cm high, 13 cm in diameter) filled with tap water up to 16 cm in height (25±0.5° C.). This means that the animal cannot escape (climb out) nor stand on its tail or hindlimbs. After a certain time the animal realize that it cannot get out and will stop struggling to get out, and instead become immobile and float.

[0287] The animal is placed inside the cylinder and allowed two swimming sessions: a 10 min pre-test (on day 1), 24 h later (day 2) a second 6 min test is conducted. The total duration of immobility and latency to the first immobility are recorded during the second 6 min test. Immobility is defined as floating passively in an upright position in the water, with only small movements necessary to keep the head above the water surface. Animals are then gently removed from water and dried.

[0288] Vehicle (20% DMSO in 0.1M PBS) or 1 mg/kg of the compound of Example 1 were administered to C57/Bl6 mice as a single s.c. administration 30 min prior to FST on day 2. As comparator, fluoxetine is commonly used a dose of 20 mg/kg. Data on the total time of immobility for mice treated with the compound of Example 1 or the vehicle control are shown in FIG. 3.