Field of the invention

The present invention relates to novel quinazoline compounds and their use in the treatment, prophylaxis and/or diagnosis of conditions involving a melanin- concentrating hormone.

Background of the invention

Melanin-concentrating hormone (MCH) is a cyclic peptide that originally was isolated from salmoid pituitaries. In the fish, the 17 amino acid peptide causes aggregation of melanin and inhibits the release of ACTH. Mammalian MCH (19 amino acids) is highly conserved between rat, mouse and human exhibiting 100% amino acid identity. In the last decades there has been increasing activity in the research in the physiologic roles of MCH. It has been reported that MCH is involved in the feeding or body weight regulation, in energy balance, in response to stress, in water balance, in energy metabolism, in the general arousal/attention state, memory and cognitive functions and in psychiatric disorders. The biological effects of MCH are believed to be mediated by specific MCH receptors, and the MCH 1 and MCH2 receptors have been described. Antagonists of MCH receptor (e.g. MCH1 receptor) are considered useful as obesity or weight reducing agents, and as antidepressant and/or anxiolytic agents.

Description of the Invention

The present invention provides a compound of formula (I), or a salt, hydrate or solvate thereof:

wherein Ri is:

and R2 is Cl-, CH3-, CF3-, or F3C-O-.

Specific preferred compounds of formula (I) are those of the Examples herein.

Compounds of the invention have a particularly useful combination of drug-like properties, including aqueous solubility and acceptable plasma protein binding.

Pharmaceutical and veterinary compositions comprising a compound of formula (I) or a pharmaceutically or veterinarily acceptable salt, hydrate or solvate thereof, together with a pharmaceutically or veterinarily acceptable carrier, are also included in the invention.

As used herein the term "salt" includes base addition, acid addition and quaternary salts. Compounds of the invention which are acidic can form salts, including pharmaceutically or veterinarily acceptable salts, with bases such as alkali metal hydroxides, e.g. sodium and potassium hydroxides; alkaline earth metal hydroxides e.g. calcium, barium and magnesium hydroxides; with organic bases e.g. Λ/-ethyl piperidine, dibenzylamine and the like. Those compounds (I) which are basic can form salts, including pharmaceutically or veterinarily acceptable salts with inorganic acids, e.g. with hydrohalic acids such as hydrochloric or hydrobromic acids, sulphuric acid, nitric acid or phosphoric acid and the like, and with organic acids e.g. with acetic, tartaric, succinic, fumaric, maleic, malic, salicylic, citric, methanesulphonic and p-toluene sulphonic acids and the like.

The compounds of the invention are highly potent antagonists of the MCH receptor, and are useful for the treatment of MHC -related disorders in mammals, including humans.

MHC-related disorders include overweight, adiposity, obesity and bulimia (e.g. malignant mastocytosis, exogeneous obesity, hyperinsulinar obesity, hyperplasmic obesity, hypophyseal adposity, hypoplasmic obesity, hypophysal adiposity, hypoplasmic obesity, hypothyroid obesity, hypothalamic obesity, symptomatic obesity, infantile obesity, upper body obesity, alimentary obesity, hypogonadal obesity, systemic mastocytosis, simple obesity, central obesity etc.), hyperfagia, emotional disorders, dementia, hormonal disorders, lifestyle diseases such as, e.g., diabetes, diabetic complications (e.g. retinopathy, neuropathy, nephropathy etc.), arteriosclerosis and gonitis..

In the present context the term body mass index or BMI is defined as body weight (kg)/height2 (m2), and the term overweight is intended to indicate a BMI in a range from about 25 to about 29.9, whereas obesity is intended to indicate a BMI, which is at least about 30.

Hence the invention includes (i) the use of a compound of formula (I) above, or a pharmaceutically or veterinarily acceptable salt, hydrate or solvate thereof, in the preparation of a composition for the treatment of an MHC-related disorder and (ii) a method of treating a mammal, including a human, suffering from a MHC- related disorder, comprising administering to the mammal an effective amount of a compound of formula (I) above, or a pharmaceutically or veterinarily acceptable salt, hydrate or solvate thereof.

The MHC-related disorder referred to in these aspects (i) and (ii) of the invention may be, for example, overweight; bulimia; bulimia nervosa; obesity; Syndrome X (metabolic syndrome); a combination of obesity, insulin resistance, dyslipidemia, impaired glucose tolerance and hypertension; Type Il diabetes or Non Insulin Dependent Diabetes Mellitus (NIDDM)1 depression and/or anxiety.

Other MHC-related disorders to which aspects (i) and (ii) of the invention apply include a steroid or pituitary hormone disorder, an epinephrine release disorder, a gastrointestinal disorder, a cardiovascular disorder, an electrolyte balance disorder, hypertension, diabetes, a respiratory disorder, asthma, a reproductive function disorder, a muscoskeletal disorder, a neuroendocrine disorder, a cognitive disorder, a memory disorder such as, e.g., Alzheimer's disease, a sensory modulation and transmission disorder, a motor coordination disorder, a sensory integration disorder, a motor integration disorder, a dopaminergic function disorder such as, e.g. Parkinson's disease, a sensory transmission disorder, an olfaction disorder, a sympathetic innervation disorder, an affective disorder such as, e.g. depression, a stress-related disorder, a fluid-balance disorder, a urinary disorder such as, e.g., urinary incontinence, a seizure disorder, pain, psychotic behaviour such as, e.g., schizophrenia, morphine or opioid tolerance, opiate addiction or migraine.

In other specific aspects the invention includes (iii) the use of a compound of formula (I) above, or a pharmaceutically or veterinarily acceptable salt, hydrate or solvate thereof, in the preparation of a composition for (a) modifying the feeding behaviour and/or reducing the body mass of a mammal, including a human and (iv) a method of modifying the feeding behaviour and/or reducing the body mass of a mammal, including a human, comprising administering to the mammal an effective amount of a compound of formula (I) above, or a pharmaceutically or veterinarily acceptable salt, hydrate or solvate thereof.

Pharmaceutical compositions

The compounds with which the invention is concerned are normally presented in the form of a pharmaceutical or a cosmetic composition comprising the specific compound or a physiologically acceptable salt thereof together with one or more physiologically acceptable excipients.

The compounds may be administered to the subject by any convenient administration route such as, e.g., the oral, buccal, nasal, ocular, pulmonary, topical, transdermal, vaginal, rectal, ocular, parenteral (including inter alia subcutaneous, r intramuscular, and intravenous), route in a dose that is effective for the individual purposes. A person skilled in the art will know how to chose a suitable administration route.

The pharmaceutical or cosmetic composition comprising a compound according to the invention may be in the form of a solid, semi-solid or fluid composition.

The solid composition may be in the form of tablets such as, e.g. conventional tablets, effervescent tablets, coated tablets, melt tablets or sublingual tablets, pellets, powders, granules, granulates, particulate material, solid dispersions or solid solutions. A semi-solid form of the composition may be a chewing gum, an ointment, a cream, a liniment, a paste, a gel or a hydrogel.

The fluid form of the composition may be a solution, an emulsion including nano- emulsions, a suspension, a dispersion, a liposomal composition, a spray, a mixture, a syrup or a aerosol.

Fluid compositions, which are sterile solutions or dispersions can be utilized by for example intraveneous, intramuscular, intrathecal, epidural, intraperitoneal or subcutaneous injection of infusion. The compounds may also be prepared as a sterile solid composition, which may be dissolved or dispersed before or at the time of administration using e.g. sterile water, saline or other appropriate sterile injectable medium.

Other suitable dosages forms of the pharmaceutical compositions according to the invention may be vagitories, suppositories, plasters, patches, tablets, capsules, sachets, troches, devices etc.

The dosage form may be designed to release the compound freely or in a controlled manner e.g. with respect to tablets by suitable coatings.

The pharmaceutical composition may comprise a therapeutically effective amount of a compound according to the invention.

The content of a compound of the invention in a pharmaceutical composition of the invention is e.g. from about 0.1 to about 100% w/w of the pharmaceutical composition.

The pharmaceutical or cosmetic compositions may be prepared by any of the method well known to a person skilled in pharmaceutical or cosmetic formulation.

In pharmaceutical or cosmetic compositions, the compounds are normally combined with a pharmaceutical excipient, i.e. a therapeutically inert substance or carrier.

The carrier may take a wide variety of forms depending on the desired dosage form and administration route. The pharmaceutically or cosmetically acceptable excipients may be e.g. fillers, binders, disintegrants, diluents, glidants, solvents, emulsifying agents, suspending agents, stabilizers, enhancers, flavours, colors, pH adjusting agents, retarding agents, wetting agents, surface active agents, preservatives, antioxidants etc. Details can be found in pharmaceutical handbooks such as, e.g., Remington's Pharmaceutical Science or Pharmaceutical Excipient Handbook.

Optimal dosages to be administered may be determined by those skilled in the art, and will vary with the particular compound in use, the strength of the composition, the route of administration, the frequency of administration, the age, weight, gender, diet and condition of the subject to be treated and the condition being treated and the advancement of the disease condition etc.

Suitable dosages may be from about 0.001 mg to about 1 g such as, e.g. from about 0.005 to about 750 mg, from about 0.01 to about 500 mg, from about 0.05 to about 500 mg, from about 0.1 to about 250 mg, from about 0.1 to about 100 mg or from about 0.5 to about 50 mg.

The amounts can be divided into one or several doses for administration daily, every second day, weekly, every two weeks, monthly or with any other suitable frequency. Normally, the administration is daily.

A compound or a pharmaceutical composition according to the invention may be used in combination with other drug substances such as agents for treating disorders like e.g. diabetes, diabetes complications, obesity, hypertension, hyperlipidemia, arteriosclerosis, arthritis, anxiety, and/or depression etc.

The following examples describe the preparation of specific compounds of the invention. Other compounds within the scope of formula (I) above may be synthesised by methods directly analogous to those of the examples

Examples

General comments; 1H-NMR data are given either in full detail or with selected characteristic peaks. LC/MS was performed on an Agilent 1100-series instrument with the column, Waters XTerra MS C18 (2.1x5 mm, 5 μ). The method in use; Flow: 1.0 mL/min; Gradient: 0-5 min: 10-100% MeCN, 5-7.5 min: 100% MeCN; MS-ionization mode API-ES (pos.).

Example 1 Step i

2-Chloro-4-methyl-quinazoline. The title compound was synthesized by adding dimethyl aniline (1.5 mL) to a suspension of 4-Methyl-1 H-quinazolin-2-one (3.3 g, 20 mmol) in Phosphorus oxychloride (30 mL) (ref. Shigeho lnaba et al, 1975, US3859237, Quinazoline Derivatives). The reaction mixture was heated to 120 ° C until the reactants came into solution. The reaction mixture was cooled and poured onto ice water followed by extraction with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and evaporated. The crude product was chromatographed (Silica, EtOAc/Heptane, 1 :1) yielding 2.2 g of the product. LC/MS : Rt = 2.85 min, m/z 178.8 [M+]. 1H-NMR(300MHz,CDCI3): δ = 3.0 (s, 3H), 7.65 (t, 1 H), 7.95 (m, 2H), 8.15 (d, 1 H)

Step 2

4-Methyl-2-(4-pyrrolidin-1-yl-piperidin-1-yl)-quinazoline . 2-Chloro-4-methyl- quinazoline (0.8 g, 4.5 mmol) and 4-(1-pyrrolidinyl)-piperidine (1.0 g, 6.5 mmol) were mixed and subjected to microwave heating (150 ° C, 8 min, fixed hold time, 10 sec prestirring). The mixture was dissolved in EtOAc/water, neutralized with NaOH, and extracted with EtOAc. The combined organic phases were washed twice with 4 M HCI. The water phase was neutralized (pH = 8-9) followed by extraction with EtOAc. The organic phases were dried over Na2SO4, filtered, and evaporated, yielding 1.3 g of the title product. 1H-NMR(300MHz,CDCI3): δ = 1.6 - 2.9 (m + s, 13H + 3H), 3.0 (t, 2H), 5.05 (d, 2H), 7.2 (t, 1 H), 7.6 (m, 2H), 7.85 (d, 1 H) Step 3

4-Methyl-2-(4-pyrrolidin-1 -yl-piperidin-1 -yl)-quinazolin-6-ylamine. 4-Methyl-2-(4- pyrrolidin-1 -yl-piperidin-1 -yl)-quinazoline (0.50 g /1.5 mmol) was added portionwise to HNO3 (90%, 25 mL) at -35 ° C and the reaction mixture was stirred for 2h at -30 ° C. The reaction was poured onto ice before NaOH (8M) was added (pH = 8) and a precipitate were formed. The solid was collected, washed with water, and dried in vacuo, yielding 0.38 g of 4-Methyl-6-nitro-2-(4-pyrrolidin-1 -yl-piperidin-1 -yl)- quinazoline. LC/MS; Rt = 4.88 min, m/z 342.2 [MH+]. 4-Methyl-6-nitro-2-(4-pyrrolidin-1 -yl-piperidin-1 -yl)-quinazoline (0.3 g, 0.88 mmol) was dissolved in MeOH (20 mL) and 10 % Pd/C (60 mg) was added. The reaction mixture was heated to 60 ° C under hydrogen atmosphere and stirred for 3h, thereafter was the mixture filtered and the solvent removed in vacuo. The crude product was chromatographed (Silica, CH2CI2/Me0H/NH3, 100:10:1) yielding 0.31 g of the product. LC/MS: Rt = 1.77 min, m/z 311.8 [MH+].

Step 4

Λ/-[4-Methyl-2-(4-pyrrolidin-1 -yl-piperidin-1 -yl)-quinazolin-6-yl]-2-(4- trifluoromethoxy-phenoxy)-acetamide. p-Trifluoromethoxy-phenol (0.78 mL, 6.0 mmol) and bromoethylacetat (0.7 mL, 6.0 mmol) were dissolved in acetone (10 mL), followed by addition of potassium carbonate (0.91 g, 6.6 mmol). The reaction mixture was heated to 150 ° C for 10 min and then cooled to room temperature whereupon water was added. The reaction mixture was extracted with EtOAc. The combined organic phases were dried over MgSO4, filtered, and evaporated. The crude product was chromatographed (Silica, CH2CI2ZMeOHZNH3, 100:10:1) yielding 1.0 g of (4- trifluoromethoxy-phenoxy)-acetic acid ethyl ester. To this ester were methanol and lithium hydroxid monohydrate added and the suspension was stirred in room temperature for 2 h followed by stirring at 60 ° C for 2h. The reaction mixture was cooled to room temperature before the solvent was removed in vacuo, yielding 1.21 g of the lithium salt of the hydrolyzed ester. To lithium (4-trifluoromethoxy-phenoxy)-acetic carboxylate (0.09 g, 0.35 mmol) dissolved in CH2CI2 (6 mL) was hydroxybenzotriazole (0.07 g, 0.48 mmol) added. The solution was cooled to 0 ° C, whereupon 1-ethyl-3(-3- dimethylaminopropyl)carbodiimid hydrochlorid (0.08 g, 0.42 mmol) and diisopropyl ethyl amine (0.02g, 0.16 mmol) were added. The reaction mixture was stirred at room temperature for 12 h before some NaHCO3-solution was added, followed by extraction with EtOAc. The combined organic phases was dried over MgSO4, filtered and evaporated. The crude product was chromatographed (silica, CH- 2CI2ZMeOHZNH3, 100:10:1) yielding the title product. LCZMS: Rt = 5.22 min, mZz 530.2 [MH+]. 1H-NMR(300MHz,CDCI3): δ = 8.29 (s, 1 H), 7.53-7.64 (m, 2H), 7.23 (d, 2H), 7.03 (d, 2H), 4.92 (d, 2H), 4.64 (s, 2H), 2.95-3.04 (m, 2H), 2.77 (s, 3H), 2.50-2.67 (m, 6H), 2.25-2.33 (m, 2H), 2.00-2.04 (m, 3H), 1.51-1.63 (m, 3H).

Example 2

2-(4-Chloro-phenoxy)-Λ/-[4-methyl-2-(4-pyrrolidin-1-yl-p iperidin-1-yl)-quinazolin- 6-yl]-acetamide. To a solution of 4-Methyl-2-(4-pyrrolidin-1 -yl-piperidin-1 -yl)- quinazolin-6-ylamine (1.5 g, 4.8 mmol) in CH2CI2 was added (4-Chloro-phenoxy)- acetyl chloride dropwise at 0 ° C. The reaction mixture was stirred under N2 atmosphere until complete. The product was precipitated by the addition of HCI in ether (2 M, 2 ml_). The crude product was recrystallised and triturated from hot methanol giving 1.2 g of the title product. LC/MS; Rt = 5.85, m/z 479.6 [MH+]. 1H- NMR (300MHz, D2O): δ = 8.18 (s, 1 H), 7.84 (d, 1 H), 7.56 (d, 1 H), 7.12 (d, 2H), 6.71 (d, 2H), 4.70 (2H, obscured by solvent signal), 4.38 (s, 2H), 3.52-3.63 (m 3H), 3.11- 3.28 (m, 5H), 2.75 (s, 3H), 2.35 (d, 2H), 2.05-2.19 (m, 2H), 1.91-1.96 (m, 2H), 1.73- 1.79 (m, 2H).

Example 3

Λ/-[2-(4-Amino-piperidin-1-yl)-4-methyl-quinazolin-6-yl] -2-(4-trifluoromethoxy- phenoxy)-acetamide. The title compound was synthesized according to the method of Example 1 utilizing 4-BOC-amino piperidine in the coupling with2-Chloro-4-methyl- quinazoline. LC/MS; Rt = 3.47 min, m/z 475.6 [MH+].1H-NMR (300 MHz, CDCI3): δ = 8.29-8.32 (m, 2H), 7.54-7.64 (m, 2H), 7.24 (d, 2H), 7.05 (d, 2H), 4.92 (d, 2H), 4.66 (s, 2H), 2.94-3.10 (m, 3H), 2.80 (s, 3H), 1.93-1.98 (m, 2H) and 1.36-1.56 (m, 4H).

Example 4

Λ/-[2-(4-Amino-piperidin-1-yl)-4-methyl-quinazolin-6-yl] -2-(4-chloro-phenoxy)- acetamide. The title compound was synthesized according to the method of Example 2 utilizing 4-BOC-amino piperidine in the coupling with 2-Chloro-4-methyl- quinazoline. LC/MS Rt = 3.26 min, m/z 425.6 [MH+].

Biological assays

Materials and methods

Transfections and Tissue Culture - The cDNA encoding the human MCH-1 receptor was cloned from a human brain cDNA library and cloned into the eukaryotic expression vector pcDNA3.1 (Invitrogen). Assays were performed on transiently transfected COS-7 cells or stably transfected CHO (Chinese Hamster Ovary) cells, expressing the human MCH-1 receptor in pcDNA3.1. Stable MCH-1 receptor transfectants of CHO cells were obtained using 5 μg plasmid cDNA and a standard calcium phosphate transfection method (Johansen et al., 1990; Gether ef a/., 1992) with subsequent selection in 1 mg/ml G418 (Life Technology). Clones were screened by a MCH receptor radioligand binding assay (as described below). Stably transfected CHO cells were maintained in RPMI 1640 culture medium (Invitrogen), supplemented with 10 % fetal calf serum (Invitrogen), 100 U/ml penicillin, 100 μg/ml streptomycin (Life Technology), and 500 μg/ml G418 (Life Technology). COS-7 cells were grown in Dulbecco's modified Eagle's medium (DMEM) 1885 (Invitrogen) supplemented with 10 % fetal calf serum, 100 U/ml penicillin, 100 μg/ml streptomycin, and were transiently transfected by a standard calcium phosphate transfection method (Johansen et al., 1990; Gether et al., 1992) two days before assay.

Radioligand Binding Assay -Transiently transfected COS-7 cells or stably transfected CHO cells, expressing human MCH-1 receptor were seeded in multi-well culture plates one day before the assay. The number of cells per well was determined by the apparent expression efficiency of the cell line aiming at 5 - 10 % binding of the added radioligand. Cells were assayed by competition binding for 3 hours at room temperature using 15 pM [125I]-MCH (Amersham Pharmacia Biotech) plus variable amounts of unlabeled ligand in 0.5 ml of a 25 mM Hepes buffer, pH 7.4, supplemented with 10 mM MgCI2, 5 mM MnCI2, 10 mM NaCI, 0.1 % (w/v) bovine serum albumin (BSA), 100 μg/ml bacitracin. The assay was performed in duplicate. Nonspecific binding was determined as the binding in the presence of 1 μM MCH (Bachem). Binding data were analyzed and IC50 values determined by non-linear regression using the Prism software (GraphPad software, San Diego). The compounds of the examples had IC50S in the above assay of 25nM or less.

References: Gether, U., Marray, T., Schwartz, T.W., and Johansen, T.E. (1992). Stable expression of high affinity NKi (substance P) and NK2 (neurokinin A) receptors but low affinity NK3 (neurokinin B) receptors in transfected CHO cells. FEBS Lett., 296, 241-244.

Johansen, T.E., Schøller, M.S., Tolstoy, S. and Schwartz, T.W. (1990). Biosynthesis of peptide precursors and protease inhibitors using new constitutive and inducible eukaryotic expressions vectors. FEBS Lett., 267, 289-294.