CA2549965A1 - Alkynes iii - Google Patents

Abstract

The present invention is directed to novel compounds of formula 1, to a process, for their preparation, their use in therapy and pharmaceutical compositions comprising the novel compounds. The novel compounds are useful in therapy, and in particular for the treatment 5 of gastroesophageal reflux disease (GERD).

Description

ALKYNES III
Field of the invention The present invention is directed to novel compounds, to a process for their preparation, their use in therapy and pharmaceutical compositions comprising the novel compounds.
Background of the invention io The metabotropic glutamate receptors (mGluR) are G-protein coupled receptors that are involved in the regulation and activity of many synapses in the central nervous system (CNS). Eight metabotropic glutamate receptor subtypes have been identified and are subdivided into three groups based on sequence similarity. Group I consists of mGluR1 is and mGluRS. These receptors activate phospholipase C and increase neuronal excitability.
Group II, consisting of mGluR2 and mGluR3 as well as group III, consisting of mGluR4, mGluRb, mGluR7 and mGluR8 are capable of inhibiting adenylyl cyclase activity and reduce synaptic transmission. Several of the receptors also exist in various isoforms, occurring by alternative splicing (Chest, C-Y et al., Journal of Physiology (2002), 538.3, zo pp. 773-786; Pig, J P et al., European Jou~hal of Pharmacology (1999), 375, pp. 277-294;
Brau~e~-Osbo~ne, H et al. Journal of Medicinal Chemistr y 0000), 43, pp. 2609-2645;
Schoepp, D.D, Jane D.E. Mohh J.A. Neu~opha~macology (1999), 38, pp. 1431-147.
The lower esophageal sphincter (LES) is prone to relaxing intermittently. As a as consequence, fluid from the stomach can pass into the esophagus since the mechanical barrier is temporarily lost at such times, an event hereinafter referred to as "reflux".
Gastro-esophageal reflux disease (GERD) is the most prevalent upper gastrointestinal tract disease. Current pharmacotherapy aims at reducing gastric acid secretion, or at neutralizing 3o acid in the esophagus. The major mechanism behind reflux has been considered to depend on a hypotonic lower esophageal sphincter. However, e.g. Holloway & Dent (1990) Gastr~oehterol. Cli~c. N. Amer. 19, pp. 517-535, has shown that most reflux episodes occur during transient lower esophageal sphincter relaxations (TLESRs), i.e.
relaxations not triggered by swallows. It has also been shown that gastric acid secretion usually is normal in patients with GERD.
The problem underlying the present invention was to find new compounds useful in the treatment of GERD.
WO 01/16121 A1 discloses a compound A-L-B, where io A is a 5-, 6- or 7-membered heterocycle ~'°X~'i7v c~~Q--~-z\
L is an alkenylene, alkynylene or azo; and B is a hydrocarbyl; cyclohydrocarbyl; heterocycle (optionally containing one or more double bonds); or aryl. These compounds have been described as being useful in inter alia is cerebral ischemia, chronic neurodegeneration, psychiatric disorders, epilepsy and diseases of the pulmonary system as well as the cardiovascular system.
WO 99/02497 A2 discloses compounds of the formula Rz / ~~--N-X Rs 'N
R~
wherein X may be an alkenylene or an alkynylene bonded via vicinal unsaturated carbon atoms, or an azo group; and RS may be an aromatic or heteroaromatic group.
These compounds have been described as being useful in inter alia epilepsy, cerebral ischemia and Alzheimer's disease.

Outline of the invention The present invention is directed to novel compounds according to the general formula I:

R4 Rs R
i R2 ~ Y1 Y2 , Y3 ~I) wherein Rl is selected from hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl, wherein io the aryl or heteroaryl may be substituted by C1-C~ alkyl;
R2 is selected from hydrogen and C1-C4 alkyl;
R3 is selected from hydrogen, C1-C4 alkyl, F, CF3, CHF2 and CHZF;
R4 is selected from hydrogen, F, CF3, CHF2, CH2F and CH3;
Rs is selected from hydrogen and F;
is R6 is selected from hydrogen and F;
Yl is selected from hydrogen; halogen; nitrite; C1-C4 alkoxy; C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom;
b~nzyloxy; nitro in the meta or para position; and C1-C4 alkyl ester;
YZ is selected from hydrogen; halogen; nitrite; C1-C4 alkoxy; C1-C4 alkyl wherein one or ao more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom; and C1-C4 alkyl ester;
Y3 is selected from hydrogen; halogen; nitrite; C1-C4 alkoxy; C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom; and C1-C4 alkyl ester; or Yl and Y2 may form an aromatic or non-aromatic ring, optionally substituted by halogen, nitrite, C1-C4 alkoxy, CI-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted fox a fluorine atom, benzyloxy or C I-C4 alkyl ester;
with the proviso that when YI is hydrogen, Y2 is selected from halogen, nitrite, C1-C4 alkoxy, and C1-C4 alkyl;
as well as pharmaceutically acceptable salts, hydrates, isoforms and/or optical isomers thereof.
The general terms used in the definition of formula I have the following meanings:
io Halogen is chloro, fluoro, bromo or iodo.
C1-C4 alkyl is a straight or branched alkyl group, each independently containing l, 2, 3 or 4 carbon atoms, for example methyl, ethyl, n-propyl, n-butyl ox isopropyl. In one is embodiment, the alkyl groups may contain one or more heteroatoms selected from 4, N
and S. Examples of such groups are methyl-ethylether, methyl-ethylamine and methyl-thiomethyl.
Cycloalkyl is a cyclic alkyl, each independently containing 3, 4, 5 or 6 carbon atoms such zo as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
C1-C4 alkoxy is an alkoxy group containing 1, 2, 3 or 4 carbon atoms, such as methoxy, ethoxy, n-propoxy, n-butoxy or isopropoxy.
as The herein used term aryl means aromatic rings with 6-14 carbon atoms including both single rings and polycyclic compounds, such as phenyl, benzyl or naphtyl.
The term heteroaryl as used herein means aromatic rings with 5-14 carbon atoms, including both single rings and polycyclic compounds, such as imidazopyridine, in which one or several of the ring atoms is either oxygen, nitrogen or sulphur, such as furanyl or thiophenyl.
Within the scope of the invention are also pharmaceutically acceptable salts of the compounds of formula I as well as isomers, hydrates and isoforms thereof:
Pharmaceutically acceptable salts of the compound of formula I are also within the scope of the present invention. Such salts are for example salts formed with mineral acids such as hydrochloric acid; alkali metal salts such as sodium or potassium salts; or alkaline earth io metal salts such as calcium or magnesium salts.
The novel compounds according to the present invention are useful in therapy.
In one aspect of the invention said compounds are useful for the inhibition of transient lower esophageal sphincter relaxations (TLESRs) and thus for treatment or prevention of gastro-is esophageal reflux disorder (GERD). In further embodiments, the compounds according to the present invention are useful for the prevention of reflux, treatment or prevention of regurgitation, treatment or prevention of asthma, treatment or prevention of laryngitis, treatment or prevention of lung disease and for the management of failure to thrive.
ao A further aspect of the invention is the use of a compound according to formula I, for the manufacture of a medicament for the inhibition of transient lower esophageal sphincter relaxations, for the treatment or prevention of GERD, for the prevention of reflux, for the treatment or prevention of regurgitation, treatment or prevention of asthma, treatment or prevention of laryngitis, treatment or prevention of lung disease and for the management of zs failure to thrive.
A further aspect of the invention is the use of a compound according to formula I for the manufacture of a medicament for the treatment or prevention of functional gastrointestinal disorders, such as functional dyspepsia (FD). Yet another aspect of the invention is the use 30 of a compound according to formula I for the manufacture of a medicament for the treatment or prevention of irritable bowel syndrome (IBS), such as constipation predominant IBS, diarrhea predominant IBS or alternating bowel movement predominant IBS.
Still a further aspect of the invention is a method for the treatment of any one of the conditions mentioned above, whereby a pharmaceutically effective amount of a compound according to formula I above, is administered to a subject suffering from said condition(s).
In one aspect of the invention, the compounds of formula I are useful for the treatment io and/or prevention of acute and chronic neurological and psychiatric disorders, anxiety and chronic and acute pain disorders. In a further aspect, said compounds are useful for the prevention and/or treatment of pain related to migraine, inflammatory pain, neuropathic pain disorders such as diabetic neuropathies, arthritis and rheumatoid diseases, low back pain, post-operative pain and pain associated with various conditions including cancer, is angina, renal or billiary colic, menstruation, migraine and gout.
The term "isomers" is herein defined as compounds of formula I, which differ by the position of their functional groups and/or orientation. By "orientation" is meant stereoisomers, diastereoisomers, regioisomers and enantiomers.
ao The term "isoforms" as used herein is defined as compounds of formula I which differ by their crystal lattice, such as crystalline compounds and amorphous compounds.
The wording "TLESR", transient lower esophageal sphincter relaxations, is herein defined zs in accordance with Mittal, R.K., Holloway, R.H., Penagini, R., Blackshaw, L.A., Dent, J., 1995; Transient lower esophageal sphincter ~elaxatioh. Gastroe~terology 109, pp. 601-610.
The wording "reflux" is defined herein as fluid from the stomach being able to pass into so the esophagus, since the mechanical barrier is temporarily lost at such times.

The wording "GERD", gastro-esophageal reflux disease, is defined herein in accordance with vav~ Hee~warden, M.A., Smout A.J.P.M., 2000; Diagnosis of reflux disease.
Baillie~e's Clip. Gast~oentef°ol. 14, pp. 759-774.
s Methods of preparation First, a Sonogashira coupling (Tetrahedron Letters 1975, 50, 4467, S. Thorand, N. Krause J. O~g. Chem., 1998, 63, 8551-8553, M. Erdelyi, A. Gogoll, J. Org. Chem., 2001, 66, io 4165-4169) of the aryl bromide A and the alcohol B in the presence of a base such as triethyl amine at room temperature to 60 °C gives the alcohol C which is then converted into the mesylate D with methanesulfonyl chloride in triethyl amine at about 0 to -20 °C.
The mesylate of the primary alcohol is isolated and characterised, while that of the secondary alcohols are made in situ. Finally, the respective mesylate is reacted with the is alcohol. This can either be done by adding the alcohol and a base such as triethyl amine to the mesylate in a solvent such as DCM or by pre-reacting the alcohol with a base such as sodium hydride in a solvent such as THF and subsequently adding the mesylate to this solution to generate product (I) (Scheme 1).

Rs R4 ~ R6 OH (PPf~3)ZPdCl2, Cul, Base + - -R3 N Br R' A B
H OAr MsCI , NEt3 Yz NEt3 O
,.
~S
., R. O

In those cases where the alcohol B is not commercially available with a desired Rl-group, the product (I) is formed by an alternative route (scheme 2): first the aryl bromide A is s coupled with ethynyl(trimethyl)silane F via Sonogashira coupling at 60 °C in triethyl amine to give product G. Deprotection of G at room temperature with potassium carbonate in rnethanol/DCM gives terminal alkyne H, which is deprotonated with lithium bis(trimethylsilyl)amide in THF at - 78 °C. At - 78 °C an aldehyde or a ketone is added and the reaction mixture is allowed to reach room temperature and kept at that temperature io for the appropriate time to form the alcohol I. Having isolated I, the mesylate ~ is formed in situ with methanesulfonyl chloride an'd triethyl amine, either at room temperature or with cooling. The reaction of the alcohol with this mesylate is either performed by adding the alcohol and a base such as triethyl amine to the mesylate in a solvent such as DCM or by pre-reacting the alcohol with a base such as sodium hydride in a solvent such as THF and is subsequently adding the mesylate to this solution to form product (I).

Rs Ra Rs R3 ~ Rs ~ (PPh3)zPdClz, Cul, NEt3 Ra I ~ + - Si- - - Ra N \ /
N Br 60 °C,12 h; then rt, 16h Sip G
A F

MeOH
rt, 2h 1) LiHMDS Rs H THF, -78 °C R3 ~ Rs E
2) R1 RICO Ra N
ooli gh MsCI, NEt3 H
DCM

Yz Rs \ Rs ,O Y3 O~S\ --~,. Ra N ~z O Y1 O
'' R~ ~ ~ Yz (I) Variations of compounds of formula I are investigated by first forming an aryl prop-2-yn-1-yl ether M by reaction of a phenol K with 3-bromoprop-1-yne [= propargyl bromide] L
in the presence of a base such as potassium carbonate in e.g. acetone at temperatures from s room temperature to 60 °C for the appropriate time (Scheme 3). The propargyl ether M is then reacted in a Sonogashira coupling with an aryl bromide in the presence of a base such r as triethyl amine at temperatures from room temperature to 60 °C for the appropriate time.

Y~ ~O Y~
Yz ~ O+ ~Br KzC03 I Yz ' / Acetone, y3 0°C->60°C M
then 60 °C/17h tpph3~2PdCiz, Cul, NEt3 60 °C/ 2h ArX, X=Br or I
Ar ~ O Y~
~Yz ~~,~JY3 In the schemes 1, 2 and 3 above, Rl, R2, R3, R4, R5, R6, yy y2 and y3 are defined as for the compounds of formula I above.
s Experimental details DCM is dried over 3A molecular sieves. THF was distilled from Na/benzophenone just prior to use. All reactions are run under a nitrogen atmosphere. All glassware is dried in at 150 °C for at least two hours prior to its use. Phase separators from International Sorbent io Technology (IST) are used. Purification by chromatography is done either on silica gel 60 (0.040-0.063 mm), or by reverse phase chromatography with a C8 column. All NMR
spectra are measured in chloroform.
2-bromo-6-methylpyridine is commercially available from Aldrich, (PPh3)2PdCl2 from is Avacado, Pd (OAc)2 from Aldrich and CuI from Fluka. If not stated otherwise, the chemicals used are commercially available and are used as such without further purification.

Pharmaceutical formulations For clinical use, the compounds of formula I are in accordance with the present invention suitably formulated into pharmaceutical formulations for oral administration.
Also rectal, parenteral or any other route of administration may be contemplated to the skilled man in the art of formulations. Thus, the compounds of formula I are formulated with at least one pharmaceutically and pharmacologically acceptable carrier or adjuvant. The carrier may be in the form of a solid, semi-solid or liquid diluent.
io In the preparation of oral pharmaceutical formulations in accordance with the invention, the compound of formula I to be formulated is mixed with solid, powdered ingredients such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose derivatives, gelatin, or another suitable ingredient, as well as with disintegrating agents and lubricating agents such as magnesium stearate, calcium stearate, sodium stearyl fuxnarate and is polyethylene glycol waxes. The mixture is then processed into granules or compressed into tablets.
Soft gelatine capsules may be prepared with capsules containing a mixture of the active compound or compounds of the invention, vegetable oil, fat, or other suitable vehicle fox ao soft gelatine capsules. Hard gelatine capsules may contain the active compound in combination with solid powdered ingredients such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives or gelatine.
Dosage units for rectal administration may be prepared (i) in the form of suppositories as which contain the active substances) mixed with a neutral fat base; (ii) in the form of a gelatine rectal capsule which contains the active substance in a mixture with a vegetable oil, paraffin oil, or other suitable vehicle for gelatine rectal capsules;
(iii) in the form of a ready-made micro enema; or (iv) in the form of a dry micro enema formulation to be reconstituted in a suitable solvent just prior to administration.

Liquid preparations for oral administration may be prepared in the form of syrups or suspensions, e.g. solutions or suspensions, containing the active compound and the remainder of the formulation consisting of sugar or sugar alcohols, and a mixture of ethanol, water, glycerol, propylene glycol and polyethylene glycol. If desired, such liquid preparations may contain colouring agents, flavouring agents, saccharine and carboxymethyl cellulose or other thickening agent. Liquid preparations for oral administration may also be prepared in the form of a dry powder to be reconstituted with a suitable solvent prior to use.
io Solutions for parenteral administration may be prepared as a solution of a compound of the invention in a pharmaceutically acceptable solvent. These solutions may also contain stabilizing ingredients and/or buffering ingredients and are dispensed into unit doses in the form of ampoules or vials. Solutions for parenteral administration may also be prepared as a dry preparation to be reconstituted with a suitable solvent extemporaneously before use.
is In one aspect of the present invention, the compounds of formula I may be administered once or twice daily, depending on the severity of the patient's condition.
A typical daily dose of the compounds of formula I is from 0.1-10 mg per kg body ao weight of the subject to be treated, but this will depend on various factors such as the route of administration, the age and weight of the patient as well as of severity of the patient's condition.

Examples Method K
Example 1 s Preparation of 3-(prop-2-yn-1-yloxy~benzene (compound 21~
O
(21 ) Potassium carbonate (1.382 g, 0.1 mol) was added to a solution of phenol (0.941 g, 0.01 io mmol, 1.0 eq.) in acetone (15 mL) at 0 °C. 3-bromoprop-1-yne (1.19 g, 0.89 mL, 0.01 mol, 1.0 eq.) was added. The solution was allowed to reach room temperature and then heated at 60 °C for 17h. After cooling, the solvent was evaporated.
Water (15 mL) was added and the mixture was extracted with EtOAc (3 x 15 mL).
The combined organic phases were washed with water (1 x 15 mL), brine (1 x 15 mL), dried is with magnesium sulphate and evaporated. This gave 1.026 g (78 %) of product.
TLC: Rf (heptane/EtOAc 2:1) = 0.67.
1H NMR (300 MHz): 7.32-7.24 (m, 2H), 7.02-6.92 (m, 3H), 4.64 (d, J = 2.4 Hz, 2H), 2.48 (t, J = 2.4 Hz, 1 H).
isC NMR (75 MHz): 157.6, 129.8, 121.7, 115.1, 78.9, 75.7, 56Ø
Example 2 Preparation of 1-methoxy-3- rop-2~n-1-yloxy)benzene (compound 22'~ according to method K above, with 3-methoxyphenol as starting material.
i 2s (22) Yield: 99 %.

1H NMR (500 MHz): 7.20-7.14 (m 2H), 6.57-6.51 (m, 3H), 4.63 (d, J = 2.3 Hz, 2H), 3.74 (s, 3H), 2.51 (t, J = 2.4 Hz, 1H).
13C NMR (125 MHz): 160.8, 158.8, 129.9, 107.2, 106.9, 101.5, 78.6, 75.6, 55.8, 55.2.
s Example 3 Preparation of 2-methyl-6-(3-~henox3rpro~-1 yn-1-yl)p ridine compound 23):
O
(23) To 2-bromo-6-methylpyridine (1.055 g, 6.13 mmol) was added 3-(prop-2-yn-1-io yloxy)benzene (0.851 g, 6.44 mmol, 1.10 eq.), followed by (PPh3)zPdClz (0.129 g, 0.18 mmol, 0.03 eq.), CuI (0.035 g, 0.18 mmol, 0.03 eq.) and triethylamine (3.50 mL). The mixture was heated under nitrogen at 60 °C fox 2h. Phophate buffer (10 mL, 0.2 M, pH 7), was added and the water phase was extracted with DCM (3 x 10 mL). The combined organic phases were dried with magnesium sulphate, evaporated and then filtered through is a Si-plug, 1 g, while rinsing with diethyl ether/pentane 1:1, ca. 25 mL.
This gave 1.491g after evaporation.
Flash chromatography on Si-gel with heptane/AcOEt, first 9:1, then 3:1, as eluent gave 0.908 g compound.
zo 1H NMR (500 MHz): 7.42 (t, J = 7.8 Hz, 1H), 7.27 (t, J = 7.8, 2H), 7.17 (d, J = 7.7 Hz, 1H), 7.01 (m, 3H), 6.95 (t, J = 7.7 Hz, 1H), 4,88 (s, 2H), 2.94 (s, 3H).
13C NMR (125 MHz): 158.2, 157.1, 140.9, 135.8, 128.9, 123.7, 122.4, 120.8, 114.2, 85.8, 83.1, 55.7, 23.8.
zs Example 4 Preparation of 2-[3-(3-methoxyphenoxy)prop-1-Xn-1-yl]-6-methylp ridine compound 24):
N \ O y O~
(24) s To NaH (0.038 g, 95 % purity, 1.50 mmol, 5.0 eq.) in THF (1 mL) was added 1-methoxy-3-(prop-2-yn-1-yloxy)benzene (0.037 g, 0.30 mmo1,1.0 eq.) in THF (1 mL) at 0 °C under nitrogen. The mixture was stirred for 10 min. at room temperature. Then, 3-(6-methylpyridin-2-yl)prop-2-yn-1-yl methanesulfonate (0.068 g, 0.30 mmol) in THF
(1 mL) was added at 0 °C. The mixture was stirred at room temperature over night (18 h). The ~o mixture was poured onto water (10 mL) and the water phase was extracted with Et20 (2 x 10 mL) and then DCM (2 x 10 mL). The combined organic phases were dried with sodium sulphate and evaporated. This gave 0.041 g crude product which was then purified by reverse phase chromatography. This gave 0.009 g (yield: 12 %) product.
is 1H NMR (300 MHz): 7.52 (t, J = 7.8 Hz, 1H), 7.25 [m (under CDCl3-signal), 1H), 7.20 (t, J
= 8.2 Hz, 1H), 7.10 (d, J = 7.8 Hz, 1H), 6.66-6-53 (m, 3H), 4.91 (s, 2H), 3.79 (s, 3H), 2.55 (s, 3H).
Example 5 2o Pr~aration of 2-[3-~3-methoxyphenoxy)prop-1-yn-1-yl]pyridine (compound 25):
prepared according to example 3 with 2-bromopyridine and 1-methoxy-3-(prop-2-yn-1-yloxy)benzene as starting materials.
w N \ O ~ O~
(25) 2s Example 6, Method A
Preparation of 2-Methyl-6- (3- -p tol loxy-prop-1-ynylypyridine O
i To a stirred mixture of 4-methyl-phenol (l9mg, 0.18mmo1) and K~C03 (3lmg, 0.225mmol) in acetone (0.3m1), methanesulfonic acid 3-(6-methyl-pyridin-2-yl)-prop-2-ynyl ester (34mg, 0. l5mmol) dissolved in acetone (0.2m1) was added at room temperature.
I
io The mixture was stirred at 60°C for 18h. Since all solvent had evaporated but the reaction was not completed, dimethylformamide (0.5m1) was added and the reaction was stirred at 60°C for another 20h. The solid salt was filtered off and the filtrate was purified by HPLC:
Waters FractionLynxsystem with LTV, ELSD and MS. Column: Ace C8 5~. 100 mm x 21,2 mm id, mobile phase A: 95% acetonitrile, mobile phase B: 5% acetonitrile + 95%
0,1 M
is NH4OAc Crradient: From 100% B to 100% A in 10 minutes, Flow: 25 ml/min UV: 254 nm. The title compound, O.OlOg (yield 28%) was isolated. M+H: 238.1 Example 7 ao Preparation of 2-[3-(3-chlorophenoxy)but-1-yn-1-yl]-6-meth~p rid The compound was prepared according to method A using methanesulfonic acid 1-methyl-3-(6-methyl-pyridin-2-yl)-prop-2-ynyl ester and 3-chlorophenol.

1H NMR: 7.58 (s, 1H), 7.5 (t, 1H), 7.43 (m, 1H), 7.27 (m, 2H), 7.15 (d, 1H), 7.06 d, 1H), 4.14 (q, 1H), 2.52 (s, 3H), 1.62 (d, 3H) isC NMR: 159.0, 142.4, 136.5, 136.0, 134.6, 132.8, 131.2, 130.0, 128.2, 124.6, 122.9, 89.5, 83.8, 34.1, 24.7, 21.6.
s Example 8 Preparation of 2-[3-(2 3-Dichloro-phenoxy)-pry-1-Vinyl]-6-meth,~pyridine CI
O ~ CI
/
to Using method A starting from 2,3-dichlorophenol (0.029g, 0.18mmol) 0.012g, (yield 27%) of the title compound was isolated. M+H: 292.0 is Example 9 Preparation of 2-[3-(2 3-Dimethyl-phenoxy~rop-1-ynyl]-6-meth l~-,pyridine O
ao Using method A starting from 2,3-dimethylphenol (0.022g, 0.18mmol) 0.010g (yield 26%) of the title compound was isolated. M+H: 252.1 Example 10 Preparation of 2-(3-(2,4-Dichloro-phenoxy)-prod-1-yn~]-6-methyl-p idine N \ O
CI CI
Using method A starting from 2,4-dichlorophenol (0.029g, 0.18mmo1) 0.0098 (yield 20%) of the title compound was isolated. M+H: 292.0 Example 11 io Pret~aration of 2-[3-(2,4-Dimethvl-phenoxy~prop-1-yn~rll-6-methyl-pyridine N ~ O
Using method A starting from 2,4-dimethylphenol (0.0228, 0.18mmo1) 0.0078 (yield 18%) of the title compound was isolated. M+H: 252. I
is Exam-ple 12 Preparation of 2-[3~2 5-Dichloro-phenoxy'~~rop-1-ynyll-6-methyl-pyridine O ~ CI
/

Using method A starting from 2,5-dichlorophenol (0.0298, 0.18mmo1) 0.0248 (yield 54%) of the title compound was isolated. M+H: 292.0 Example 13 Preparation of 2-f3-(2 5-Dimethyl-phenoxy~prop-1-ynyll-6-methyl-pyridine N ~ O
Using method A starting from 2,5-dimethylphenol (0.0228, 0.18mmol) 0.0178 (yield 45%) of the title compound was isolated. M+H: 252.1 io Example 14 Preparation of 2-~~2 6-Dimeth ~~l-phenoxy)-prop-1-ynyl~-6-meth T~l-p riy dine 1\
N \ O
is Using method A starting from 2,6-dimethylphenol (0.0228, 0.18mmol) 0.0198 (yield 50%) of the title compound was isolated. M+H: 252.1 Example 15 ao Preparation of 2-Methyl-6- l3-(2-trifluoromethyl-phenoxy)-prop-1-ynyll-pyridine F F
F
N ~ O \
/

Using method A starting from 2-trifluoromethylphenol (0.029g, 0.18mmol) 0.015g (yield 35%) of the title compound was isolated. M+H: 292.1 Example 16 Preparation of 2-[3-~2-Benzylox~phenoxy~prop-1-ynyll-6-methyl-pyridine /

N ~ O
Using method A starting from benzyl alcohol (0.036g, 0.18mmo1) 0.0168 (yield 32%) of io the title compound was isolated. M+H: 330.2 Example 17 Preparation of 2-f3-(2-Bromo-4 5-dimethyl-phenoxy) -prop-1-~nyl]-6-methyl-pyridine is Using method A starting from 2-bromo-4, 5-dimethylphenol (0.0368, 0.18mmo1) 0.0178 (yield 33%) of the title compound was isolated. M+H: 330.1 zo Example 18 Preparation of 2-f 3-(2-Chloro-4-methox~phenoxy~prop-1-yn~]-6-methyl-pyridine CI
O \
Oi Using method A starting from 2-chloro-4-methoxyphenol (0.029g, 0.18mmol) 0.022g (yield 50%) of the title compound was isolated. M+H: 288.1 Exam lp a 19 io Preparation of 2-f 3-(2-Chloro-5-methyl-phenoxy, -prop-1-yn~]!-6-methyl-pyridine CI
O \
/
Using method A starting from 2-chloro-5-methylphenol (0.026g, 0.18mmo1) 0.021 g (yield is 53%) of the title compound was isolated. M+H: 272.1 Example 20 Preparation of 2-f 3-(2-Chloro-6-methyl-phenoxy)-prop 1-~n~]-6-methyl-p ridine CI
O \

Using method A starting from 2-chloro-6-methylphenol (0.0268, 0.18mmo1) 0.0178 (yield 42%) of the title compound was isolated. M+H: 272.1 s Example 21 Preparation of 2-Methyl-6- (3-o-to~lox -~pro~ 1-Vinyl)-pyridine W
O
Using method A starting from 2-methylphenol (0.0198, 0.18mmo1) 0.0178 (yield 47%) of the title compound was isolated. M+H: 238.1 is Example 22 Preparation of 2-Meths[3-(3 4 5-trimethyl-phenoxy)-drop-1-~nyl]~ ridine zo Using method A starting from 3,4,5-trimethyl phenol (0.0258, 0.18mmo1) 0.0128 (yield 30%) of the title compound was isolated. M+H: 266.1 Exam lp a 23 Preparation of 2-[3-(3,4-Dichloro-~henoxy)-prop-1-ynyll-6-methyl-pyridine O ~ CI
CI
Using method A starting from 3,4-dichloro phenol (0.0298, 0.18mmo1) 0.0218 (yield 49%) of the title compound was isolated. M+H: 292.0 io Example 24 Pr~aration of 2-[3-(3 4-Dimethoxy-phenoxy)-prop-1-ynyll-6-methyl-pyridine N \ O ~ O
O
is Using method A starting from 3,4-dimethoxy phenol (0.0288, 0.18mmo1) 0.0178 (yield 39%) of the title compound was isolated. M+H: 284.1 Example 25 Preparation of 2-~3-(3 4-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine N \ O

Using method A starting from 3,4-dimethyl phenol (0.0228, 0.18mmo1) 0.0128 (yield 32%) of the title compound was isolated. M+H: 252.1 Example 26 Preparation of 2-[3-(3 5-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl pyridine N ~ O \ CI
CI
io Using method A starting from 3,5-dichlorophenol (0.0298, 0.18mmol) 0.0158 (yield 34%) of the title compound was isolated. M+H: 292.0 Example 27 Preparation of 2- f 3-(3 5-Dimethoxy,=phenoxy~prop-1-ynyl] -6-methyl-pyridine is N \ O \ O\
,O
Using method A starting from 3,5-dimethoxyphenol (0.0288, 0.18mmol) 0.0148 (yield 33%) of the title compound was isolated. M+H: 284.1 zo Example 28 Preparation of 2-[3-(3 5-Dimeth ~~l-phenoxy~~rop-1 ~n~]-6-methyl-p rid O
s Using method A starting from 3,5-dimethylphenol (0.0228, 0.18mmol) 0.0188 (yield 47%) of the title compound was isolated. M+H: 252.1 Example 29 io Preparation of 2-[3-(3-Bromo-phenoxy)-prop-1-ynyll-6-methyl-pyridine O ~ Br /
Using method A starting from 3-bromophenol (0.0318, 0.18mmol) 0.0208 (yield 45%) of is the title compound was isolated. M+H: 302.0 Example 30 Preparation of 3- f 3- (6-Methyl-pyridin-2-yl)~rop-2-ynyloxX]-benzonitrile O

Using method A starting from 3-hydroxybenzonitrile (0.021 g, 0.18mmol) 0.01 Og (yield 27%) of the title compound was isolated. M+H: 249.1 s Example 31 Preparation of 2-[3-(3-Ethyl-phenoxy~rprop-lynyl]I-6-methyl-pyridine O
io Using method A starting from 3-ethylphenol (0.0228, 0.18mmol) 0.0308 (yield 79%) of the title compound was isolated. M+H: 252.1 Example 32 is Preparation of 2-methyl-6-[3-(3-methyphenoxylprop-1-yn-1-~]'pyridine O ~ CI
To 3-chlorophenol (0.031 g, 0.24 mmol, 1.2 eq.) in a glass vial was added anhydrous 2o potassium carbonate (0.041 g, 0.30 mmol, 1.5 eq.), followed by 0.5 mL of a 0.4 M solution of 3-(6-methylpyridin-2-yl)prop-2-yn-1-yl methanesulfonate (0.045 g, 0.2 mmol) in acetone. The vial was sealed and heated at 60 °C for 5 h. The material was filtered through Celite and then vacuum centrifuged. Purification was done by reverse phase column chromatography. Yield: 0.023 g (40 %).
2s 1H NMR (500 MHz): 7.53 (t, J = 7.8 Hz, 1H), 7.27-7.20 (m, ca. 2H, overlap with signal), 7.11 (br d, J = 8.0 Hz, 1 H), 7.04-7.02 (m, 1 H), 6.98 (br d, J = 7.8 Hz, 1 H), 6.92 (br d, J = 8.4 Hz, 1H), 4.92 (s, 2H), 2.55 (s, 3H).
MS m/z: 258 (M+1) s Example 33 Preparation of 2-[3-(4-chloro-2-methy~henoxy)prop-1-Xn-1-~l-6-methylp, irk O
CI
to Prepared in analogy to example 32, but with 4-chloro-2-methylphenol as starting material.
Yield: 0.008 g (20 %).
1s MS m/z: 272 (M+1) ao Example 34 Preparation of 2-f3-(4-chloro-3,5-dimethylphenoxy)prop-1-yn-1-yl]'-6-methylpyridine~

s Prepared in analogy to example 32, but in 0.15 mmol scale and with 4-chloro-3,5-dimethylphenol as starting material. Yield: 0.007 g (16 %).
MS '"/z: 286 (M+1) Example 35 Preparation of 2-f 3-(4-chloro-3-meth~phenoxy~prop-1-yn-1-yl]-6-meth,~Tlp, rid O
CI
Prepared in analogy to example 32, but in 0.15 mmol scale and with 4-chloro-3-methyl~henol as starting material. Yield: 0.010 g (25 %).
is MS m/z: 272 (M+1) Example 36 Preparation of 2-[3-(4-chlorophenoxy)prop-1-~n--yl]-6-methylpyridine:
O
W
CI
Prepared in analogy to example 32, but in 0.15 mmol scale and with 4-chlorophenol as starting material. Yield: 0.008 g (21 %).

MS n'/Z: 258 (M+1) Exam lp a 37 s Preparation of 2-[3-(4-methoxyphenoxy~prop-1-~~~-6-methylpyridine:
N \ O \
O~
Prepared in analogy to example 32, but in 0.15 mmol scale and with 4-methoxyphenol as io starting material. Yield: 0.004 g (11 %).
MS m/Z: 254 (M+1) Exam In a 3 8 is Preparation of 2-methyl-6-[3-(4-nitrophenoxy)prop-1-~n-1-~]'pyridine:
N ~ O
\
N
O
Prepared in analogy to example 32, but in 0.15 mmol scale and with 4-nitrophenol as ao starting material. Yield: 0.014 g (35 %).
MS "'/Z: 269 (M+1) Example 39 Preparation of 2-methyl-6-f3-(3-nitrophenoxY)per-1-yn-1-~]pyridine Prepared in analogy to example 32, but in 0.15 mmol scale and with 3-nitrophenol as starting material. Yield: 0.012 g (30 %).
MS n'/z: 269 (M+1) is Example 40 Preparation of 2-methyl-6-[3~3-meth~phenoxy)prop-1-yn-1-~]pyridine' O
zo Prepared in analogy to example 32, but with 3-methylphenol as starting material. Yield:
0.012 (25 %).
IH NMR (500 MHz): 7.52 (t, J = 7.8 Hz, 1H), 7.24 (d, J = 7.8 Hz, 1H), 7.21-7.16 (m, 1H), 7.10 (d, J = 7.8 Hz, 1H), 6.85-6.79 (m, 3H), 4.91 (s, 2H), 2.55 (s, 3H), 2.34 (s, 3H).
2s MS m/z: 238 (M+1) Example 41 Preparation of 2-methyl-6-,[3-(5,6,7,8-tetrah d~ronaphthalen-2-yloxy~prop-1-yn-s yl]p rid N ~ O
Prepared in analogy to example 32, but with 5,6,7,8-tetrahydronaphthalen-2-of as starting io material. Yield: 0.021 g (38 °1°).
1H NMR (300 MHz): 7.52 (t, J = 7.8 Hz, 1H), 7.25 (d, J = ca. 7.8 Hz, 1H, overlap with CDCl3 signal), 7.10 (d, J = 7.8 Hz, 1H), 6.99 (d, J = 8.3 Hz, 1H), 6.85-6.70 (m, 2H), 4.88 (s, 2H), 2.78-2.67 (m, 4H), 2.55 (s, 3H), 1.80-1.74 (m, 4H).
~s MS n'/z: 278 (M+1) Example 42 Preparation of 2-[3-(4-isoprop~phenoxy)prop-1-yn-1-~1-6-meth~p ir' O
Prepared in analogy to example 32, but with 4-isopropylphenol as starting material. Yield:
0.046 (85 %).
2s 1H NMR (500 MHz): 7.52 (t, J = 7.8 Hz, 1H), 7.25 (d, J = ca. 7.7 Hz, 1H, overlap with CDCl3 signal), 7.16 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 7.8 Hz, 1H), 6.96 (d, J
= 8.4 Hz, 2H), 4.90 (s, 2H), 2.90-2.83 (m, J = 7.0 Hz, 1H), 2.55 (s, 3H), 1.23 (d, J = 7.0 Hz, 6H).
MS "'/z: 266 (M+1) s Example 43 Preparation of 2-[3-(4-ter°t-but~phenoxy)prop-1-yn-1- ly 1~6-methylpyridine:
W
O
to Prepared in analogy to example 32, but with 4-tent-butylphenol as starting material. Yield:
0.017 g (30 %).
~s 1H NMR (500 MHz): 7.52 (t, J = 7.8 Hz, 1H), 7.32 (br d, J = 8.8 Hz, 2H), 7.25 (m, partly overlap with CDCI3 signal, 1H(?)), 7.10 (d, J = 7.8 Hz, 1H), 6.96 (br d, J =
8.8 Hz, 2H), 4.90 (s, 2H), 2.55 (s, 3H), 1.30 (s, 9H).
MS m/z: 280 (M+1) zo Example 44 Preparation of 3-(5-fluoro-6-methylpyridin-2-yl)prop-2-yn-1-ol:
6-Bromo-3-fluoro-2-methylpyridine (0.500 g, 2.63 mmol), propargyl alcohol (0.590 g, 10.5 mmol) and bis(triphenylphosphine)palladium(II) chloride (46 mg, 0.065 mmol) were as dissolved in triethylamine (1.0 mL) and finally copper(I) iodide was added.
The mixture was slowly heated to 50 °C and left under stirring overnight. Hereafter KZC03 (1.0 M, 25 mL) was added and the solution was extracted with DCM. The organic phases were pooled, dried (Na2S04), filtrated and evaporated. The resulting crude product was subjected to flash chromatography on silica gel with gradient (heptane/EtOAc 1:0 to 1:4), which gave the pure product 0.332 g (76 %).
F
OH
s 1H-NMR (400 MHz): 7.22 (d, 1H), 7.20 (s, 1H), 4.57 (m, 1H), 4.47 (m, 2H), 2.43 (d, 3H).
13C-NMR (400 MHz): 158.7, 156.1, 147.6 (d), 137.9 (d), 126.4 (d), 122.9 (d), 88.2, 83.4, 50.9, 17.8.
io Example 45 Preparation of 4-(5-fluoro-6-methylpyridin-2-yl)but-3-yn-2-ol:
6-Bromo-3-fluoro-2-methylpyridine (0.500 g, 2.63 mmol), 3-butyn-2-of (0.738 g, 10.5 mmol) and bis(triphenylphosphine)palladium(II) chloride (46 mg, 0.065 mmol) were dissolved in triethylamine (1.0 mL) and finally copper(I) iodide was added.
The mixture is was slowly heated to 50 °C and left under stirring overnight and then heated to 70 °C for additionally 2 h. Hereafter K2C03 (1.0 M, 25 mL) was added and the solution was extracted with DCM. The organic phases were pooled, dried (Na2SO4), filtrated and evaporated. The resulting crude product was subjected to flash chromatography on silica gel with gradient (heptane/EtOAc 1:0 to 1:4), which gave the pure product 0.447 g (95 %).
F
OH
1H-NMR (400 MHz): 7.18 (s, 1H), 7.16 (s, 1H), 4.72 (m, 1H), 4.45 (m, 1H), 2.41 (d, 3H), 1.47 (d, 3H). 13C-NMR (400 MHz): 158.6, 156.0, 147.5 (d), 138.0 (d), 126.3 (d), 122.9 (d), 91.5, 82.1, 58.2, 24.1, 17.8.

Example 46 Preparation of 3-(5-fluoro-6-methylpyridin-_ 2-yl)pro~2-yn-1-yl methanesulfonate~
3-(5-fluoro-6-methylpyridin-2-yl)prop-2-yn-1-of (40.0 mg, 0.24 mmol) and diisopropylethylamine (125 mg, 0.97 mmol) were dissolved in DCM (1.0 mL) and mesyl chloride was added dropwise. The mixture was stirred for 2 h at room temperature. The reaction was quenched with water (25 mL) and extracted with DCM. The organic phases were pooled, dried (Na2S04), filtrated and evaporated. The crude material was sufficiently pure (LC-MS) to be used for further synthesis.
F
N ~ O~ /O
/j \
to O
Example 47 is Pret~aration of 3-(5-fluoro-6-meth~pyridin-2-yl -1-meth~prop-2-yn-1-Xl methanesulfonate:
4-(5-fluoro-6-methylpyridin-2-yl)but-3-yn-2-of (43.4 mg, 0.24 mmol) and diisopropylethylamine (125 mg, 0.97 mmol) were dissolved in DCM (1.0 mL) and mesyl chloride (55.4 mg, 0.48 mmol) was added dropwise. The mixture was stirred for 2 h at zo room temperature. The reaction was quenched with water (25 mL) and extracted with DCM. The organic phases were pooled, dried (Na2S04), filtrated and evaporated.
The crude material was sufficiently pure (LC-MS) to be used for further synthesis.
F
N ~ O~ /O
lj \
O

Example 48 Preparation of 6-f3-(3 4-dimethylphenoxy~pro~ 1-ynl-yl]-3-fluoro-2-methylpyridine:
3-(5-fluoro-6-methylpyridin-2-yl)prop-2-yn-1-yl methanesulfonate (10.0 mg, 0.041 mmol) and 3,4-dimethylphenol (7.5 mg, 0.061 mmol) were dissolved in acetone/DMF
(2:1, 3 mL) and K2C03 (11.4 mg, 0.082 mmol) was added. The mixture was heated to 60 °C overnight.
Hereafter K2C03 (1.0 M, 25 mL) was added and the solution was extracted with DCM.
The organic phases were pooled, dried (Na2S04), filtrated and evaporated. The resulting crude product was purified by preparative HPLC (C8 kromasil), which afforded the pure io product 2.0 mg (9.0 %).
F
N ~O \
1H-NMR (400 MHz): 7.26 (m, 2H), 7.04 (d, 1H), 6.76 (dd, 2H), 4.87 (s, 2H), 2.51 (d, 3H), 2.24 (s, 3H), 2.19 (s, 3H).
is Example 49 Preparation of 6-L-(3 4-dimethy~henoxy but-1-yn-1-yll-3-fluoro-2-methylpyridine:
3-(5-fluoro-6-methylpyridin-2-yl)-1-methylprop-2-yn-1-yl methanesulfonate (62.3 mg, zo 0.242 mmol) and 3,4-dimethylphenol (44.4 mg, 0.363 mmol) were dissolved in acetone/DMF (2:1, 3 mL) and K2CO3 (66.9 mg, 0.484 mmol) was added. The mixture was heated to 60 °C overnight. Hereafter KZC03 (1.0 M, 25 mL) was added and the solution was extracted with DCM. The organic phases were pooled, dried (NaZS04), filtrated and evaporated. The resulting crude product was purified by preparative HPLC (C8 kromasil), as which afforded the pure product 22.0 mg (32 %).

F
i O
1H-NMR (400 MHz): 7.22 (m, 2H), 7.03 (d, 1H), 6.82 (m, 2H), 5.04 (q, 1H), 2.50 (d, 3H), 2.23 (s, 3H), 2.19 (s, 3H), 1.71 (d, 3H) s Example 50 Preparation of 4-(6-methvlpyridin-2-Xl)but-3-yn-2-ol:
2-bromo-6-methylpyridine (0.258 g, 1.5 mmol) was mixed with but-3-yn-2-of (0.116 g, 1.65 mmol, 1.1 eq.) and (PPh3)2PdCl2 (0.032 g, 0.045 mmol, 0.03 eq.). At 0 °C
io triethylamine (0.61 g, 0.84 mL, 6.0 mmol, 4.0 eq.) was added. The mixture was stirred at 0 °C for 10 min and CuI (0.006 g, 0.03 mmol, 0.02 eq.) was added. The mixture was allowed to reach room temperature and was finally heated at 60 °C for 4h.
Phosphate buffer (10 mL, 0.2 M, pH 7) was added and the water phase was extracted with DCM (3x10 mL) by using a phase separator. The combined organic phases were dried with is sodium sulphate and evaporated. This gave 0.286 g crude product.
After flash chromatography on Si with pentane/EtOAc fractions (first 1:1, then 3:2 and finally 1:2) as eluent 0.163 g (Yield: 67 %) pure product was isolated as a yellow oil.
TLC: Rf (pentane/EtOAc l :l) = 0.20.
OH
1H NMR (300 MHz): 7.40 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.8 Hz, 1H), 6.96 (d, J = 7.8 Hz, 1H), 4.90 (b, 1H), 4.76 (q, J = 6.8 Hz, 1H), 2.43 (s, 3H), 1.49 (d, J = 6.7 Hz, 3H).
isC NMR (75 MHz): 158.2, 141.7, 136.2, 123.9, 122.4, 91.7, 82.3, 57.6, 23.9, 23.8.
2s Example 51 Preparation of Methanesulfonic acid 1-methyl-3-(6-metl~l-pyridin-2-yl)-prop-2-yn 1 ester The compound was prepared according to the method in example 46 using 4-(6-methylpyridin-2-yl)but-3-yn-2-of as starting material.
i ov N ~ O~S O
MS "'/Z: 240 (M+1) Example 52 io Preparation of 3-(6-methylpyridin-2-yl)prop-2;yn-1-ol~
To 2-bromo-6-methylpyridine (1.72 g, 0.01 mol) was added (PPh3)2PdC12 (0.116 g, 0.2 mmol, 0.02 eq.) and CuI (0.063 g, 0.3 mmol, 0.03 eq.) at 0 °C under nitrogen, followed by ro -2- n-1-of 2.24 2.33 mL 0.4 mol 4.0 a ~ and trieth lamine 1 50 mL The p p Y ( g> > > q~) Y ( ~ ).
reaction mixture was allowed to reach room temperature and then heated at 60 °C for 3.5 h.
is Then the reaction mixture was added to water (10 mL) and the pH was adjusted to 6-7 with 2 M HCI. The water phase was extracted with DCM (3 x 10 mL) and the combined organic phases were dried with sodium sulphate and evaporated. This gave 1.719 g crude product.
1.098 g hereof was subjected to flash chromatography on silica gel with pentane/EtOAc, first 1:1, then 1:2, finally 1:3, as eluent. This gave 0.578 g product.
N ~ OH
13C NMR (75 MHz): 157.7, 141.1, 136.2, 123.7, 122.3, 88.4, 83.0, 49.9, 23.5.
2s Example 53 Preparation of 3-(6-meth~pyridin-2-yl)prop-2-yn-1-yl methanesulfonate:
3-(6-methylpyridin-2-yl)prop-2-yn-1-of (0.300 g, 2.04 mmol) was dissolved in DCM (10 mL) under nitrogen over 5-10 min. The solution was cooled to -20 °C
(cooling bath:
s acetone + pieces of dry ice). Triethylamine (0.268 g, 0.37 mL, 0.27 mmol, 1.30 eq.) was added. Methanesulfonyl chloride (0.280 g, 0.19 mL, 0.24 mmol, 1.2 eq.) in DCM
(1.5 mL) was added over 3 min. The reaction mixture was stirred at -18 to -22 °C
for lh after which time LC/MS only showed product. Water (10 mL) was added. The organic phase was separated and the water phase was extracted with DCM (3 x 10 mL). The organic phases io were pooled, dried with magnesium sulphate and evaporated. This gave 0.450 g (yield: 98 %) as a yellow oil.
N \ O. ,O
,S~
O
1H NMR (300 MHz): 7.61 (t, J = 7.7 Hz, 1H), 7.31 (d, J =7.7 Hz,lH), 7.19 (d, J
= 7.7 Hz, is 1H), 5.10 (s, 2H), 3.18 (s, 3H), 2.58 (s, 3H).
13C NMR (75 MHz): 158.9, 140.2, 136.8, 124.5, 123.8, 87.8, 80.7, 57.7, 38.9, 24.2.
Bioloe~ical evaluation zo Functional assessment of mGluRS antagonism in cell lines expressing mGluRSd The properties of the compounds of the invention can be analyzed using standard assays for pharmacological activity. Examples of glutamate receptor assays are well known in the art as described in for example Aramori et al., Neuron 8:757 (1992), Tanabe et al., Neu~oh zs 8:169 (1992), Miller et al., J. Neuroscience 15: 6103 (1995), Balazs, et al., J.
Neurochemistry 69:151 (1997). The methodology described in these publications is incorporated herein by reference. Conveniently, the compounds of the invention can be studied by means of an assay (FLIPR) that measures the mobilization of intracellular calcium, [Ca2~]; in cells expressing mGluRS or another assay (IP3) that measures inositol phosphate turnover.
FLIPR Assay s Cells expressing human mGluRSd as described in W097/05252 are seeded at a density of 100,000 cells per well on collagen coated clear bottom 96-well plates with black sides and experiments are done 24 h following seeding. All assays are done in a buffer containing 127 mM NaGI, 5 mM KCI, 2 mM MgGl2, 0.7 mM NaHZP04, 2 mM CaCl2, 0.422 mg/ml NaHC03, 2.4 mg/ml HEPES, 1.8 mg/ml glucose and 1 mg/ml BSA Fraction IV (pH
7.4).
io Cell cultures in the 96-well plates are loaded for 60 minutes in the above mentioned buffer containing 4 ~M of the acetoxymethyl ester form of the fluorescent calcium indicator fluo-3 (Molecular Probes, Eugene, Oregon) in 0.01% pluronic acid (a proprietary, non-ionic surfactant polyol - CAS Number 9003-11-6). Following the loading period the fluo-3 buffer is removed and replaced with fresh assay buffer. FLIPR experiments are done using is a laser setting of 0.800 W and a 0.4 second CCD camera shutter speed with excitation and emission wavelengths of 488 nm and 562 nm, respectively. Each experiment is initiated with 160 q.l of buffer present in each well of the cell plate. A 40 ~,1 addition from the antagonist plate was followed by a 50 ~L addition from the agonist plate. A 90 second interval separates the antagonist and agonist additions. The fluorescence signal is sampled ao 50 times at 1 second intervals followed by 3 samples at 5 second intervals immediately after each of the two additions. Responses are measured as the difference between the peak height of the response to agonist, less the background fluorescence within the sample period. ICso determinations are made using a linear least squares fitting program.
zs IP3 Assay An additional functional assay for mGluRSd is described in W097/05252 and is based on phosphatidylinositol turnover. Receptor activation stimulates phospholipase C
activity and leads to increased formation of inositol 1,4,S,triphosphate (IP3).
so GHEK stably expressing the human mGluRSd are seeded onto 24 well poly-L-lysine coated plates at 40 x 104 cells /well in media containing 1 ~,Ci/well [3H] myo-inositol.

Cells were incubated overnight (16 h), then washed three times and incubated for 1 h at 37°C in HEPES buffered saline (146 mM NaCI, 4.2 mM KCI, 0.5 mM MgCl2, O.I%
glucose, 20 mM HEPES, pH 7.4) supplemented with 1 unit/ml glutamate pyruvate transaminase and 2 mM pyruvate. Cells are washed once in HEPES buffered saline and s pre-incubated for 10 min in HEPES buffered saline containing IO mM LiCI.
Compounds are incubated in duplicate at 37°C for I S min, then either glutamate (80 ~M) or DHPG (30 ~M) is added and incubated for an additional 30 min. The reaction is terminated by the addition of 0.5 ml perchloric acid (5%) on ice, with incubation at 4°C
for at least 30 min.
Samples are collected in I S ml polyproplylene tubes and inositol phosphates are separated io using ion-exchange resin (Dower AG1-X8 formate form, 200-400 mesh, BIORAD) columns. Inositol phosphate separation was done by first eluting glycero phosphatidyl inositol with 8 ml 30 mM ammonium formate. Next, total inositol phosphates is eluted with 8 ml 700 mM ammonium formate / 100 mM formic acid and collected in scintillation vials. This eluate is then mixed with 8 ml of scintillant and [3H] inositol incorporation is is determined by scintillation counting. The dpm counts from the duplicate samples are plotted and ICso determinations are generated using a linear least squares fitting program.
Abbreviatio~zs Zo BSA Bovine Serum Albumin CCD Charge Coupled Device CRC Concentration Response Curve DHPG 3,5-dihydroxyphenylglycine DPM Disintegrations per Minute as EDTA Ethylene Diamine Tetraacetic Acid FLIPR Fluorometric Imaging Plate reader GHEK GLAST-containing Human Embrionic Kidney GLAST glutamatelaspartate transporter HEPES 4-(2-hydroxyethyl)-I-piperazineethanesulfonic acid (buffer) 3o IP3 inositol triphosphate Generally, the compounds are active in the assay above with ICSO values less than 10 000 nM. In one aspect of the invention, the ICSO value is less than 1 ~.M. In a further aspect of the invention, the ICso value is less than 100 nM.
Examples of ICso values for individual compounds is given below:
Compound.-__ _ FLIPR ICso 3- [3- (6-Methyl-pyridin-2-yl)-prop-2-545 ynyloxy]-benzonitrile 2-methyl-6-[3-(4-nitrophenoxy)prop-1-yn-268 1- 1 yridine 6-[3-(3,4-dimethylphenoxy)prop-1-yn-1-yl]-177 3-fluoro-2-methyl yridine io Screeuiv~g for compounds active against TLESR
Adult Labrador retrievers of both genders, trained to stand in a Pavlov sling, are used.
Mucosa-to-skin esophagostomies are formed and the dogs are allowed to recover is completely before any experiments are done.
Motility measurement In brief, after fasting for approximately 17 h with free supply of water, a multilumen zo sleeve/sidehole assembly (Dentsleeve, Adelaide, South Australia) is introduced through the esophagostomy to measure gastric, lower esophageal sphincter (LES) and esophageal pressures. The assembly is perfused with water using a low-compliance manometric perfusion pump (Dentsleeve, Adelaide, South Australia). An air-perfused tube is passed in the oral direction to measure swallows, and an antimony electrode monitored pH, 3 cm above the LES. All signals are amplified and acquired on a personal computer at 10 Hz.
When a baseline measurement free from fasting gastric/LES phase III motor activity has s been obtained, placebo (0.9% NaCl) or test compound is administered intravenously (i.v., 0.5 ml/kg) in a foreleg vein. Ten min after i.v. administration, a nutrient meal (10%
peptone, 5% D-glucose, 5% Intralipid, pH 3.0) is infused into the stomach through the central lumen of the assembly at 100 ml/min to a final volume of 30 mlfkg. The infusion of the nutrient meal is followed by air infusion at a rate of 500 ml/min until an intragastric to pressure of 10~1 mmHg is obtained, The pressure is then maintained at this level throughout the experiment using the infusion pump for further air infusion or for venting air from the stomach. The experimental time from start of nutrient infusion to end of air insufflation is 45 min. The procedure has been validated as a reliable means of triggering TLESRs.
is TLESRs is defined as a decrease in lower esophageal sphincter pressure (with reference to intragastric pressure) at a rate of >1 mmHg/s. The relaxation should not be preceded by a pharyngeal signal <2s before its onset in which case the relaxation is classified as swallow-induced. The pressure difference between the LES and the stomach should be less than zo 2 mmHg, and the duration of the complete relaxation longer than 1 s.

Claims (13)

1. A compound of formula I
wherein R1 is selected from hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl, wherein the aryl or heteroaryl may be substituted by C1-C4 alkyl;
R2 is selected from hydrogen and C1-C4 alkyl;
R3 is selected from hydrogen, C1-C4 alkyl, F, CF3, CHF2 and CH2F;
R4 is selected from hydrogen, F, CF3, CHF2, CH2F and CH3;
R5 is selected from hydrogen and F;
R6 is selected from hydrogen and F;
Y1 is selected from hydrogen; halogen; nitrile; C1-C4 alkoxy; C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom; benzyloxy; nitro in the meta or para position; and C1-C4 alkyl ester;
Y2 is selected from hydrogen; halogen; nitrile; C1-C4 alkoxy; C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom; and C1-C4 alkyl ester;
Y3 is selected from hydrogen; halogen; nitrile; C1-C4 alkoxy; C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom; and C1-C4 alkyl ester; or Y1 and Y2 may form an aromatic or non-aromatic ring, optionally substituted by halogen, nitrite, C1-C4 alkoxy, C1-C4 alkyl wherein one or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom, benzyloxy or alkyl ester;
with the proviso that when Y1 is hydrogen, Y2 is selected from halogen, nitrile, C1-C4 alkoxy, and C1-C4 alkyl;
as well as pharmaceutically acceptable salts, hydrates, isoforms and/or optical isomers thereof.

2. A compound of formula I
wherein R1 is selected from hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl, wherein the aryl or heteroaryl may be substituted by C1-C4 alkyl;
R2 is selected from hydrogen and C1-C4 alkyl;
R3 is selected from hydrogen, C1-C4 alkyl, F, CF3, CHF2 and CH2F;
R4 is selected from hydrogen, F, CF3, CHF2, CH2F and CH3;
R5 is selected from hydrogen and F;
R6 is selected from hydrogen and F;
Y1 is selected from hydrogen, halogen, nitrile, C1-C4 alkoxy, and C1-C4 alkyl;
Y2 is selected from hydrogen, halogen, nitrile, C1-C4 alkoxy, and C1-C4 alkyl;
Y3 is selected from hydrogen, halogen, nitrile, C1-C4 alkoxy, and C1-C4 alkyl;
with the proviso that when Y1 is hydrogen, Y2 is selected from halogen, nitrite, C1-C4 alkoxy, and C1-C4 alkyl;

as well as pharmaceutically acceptable salts, hydrates, isoforms and/or optical isomers thereof.

3. A compound according to formula I of claim 1 or 2, wherein R1 is hydrogen or C1-C3 alkyl;
R2 is hydrogen;
R3 is selected from hydrogen and C1-C2 alkyl;
R4 is hydrogen;
R5 is hydrogen;
R6 is hydrogen;
Y1 is selected from hydrogen, chloro, C1-C2 alkoxy, and C1-C2 alkyl; and Y2 is selected from hydrogen, chloro, C1-C2 alkoxy, and C1-C2 alkyl;
with the proviso that when Y1 is hydrogen, Y2 is selected from chloro, C1-C2 alkoxy, and C1-C2 alkyl; and Y3 is hydrogen.

4. A compound according to claim 1 selected from;
2-[3-(3-methoxyphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(3-methoxyphenoxy)prop-1-yn-1-yl]pyridine;
2-[3-(3-chlorophenoxy)but-1-yn-1-yl]-6-methylpyridine;
2-Methyl-6-(3-p-tolyloxy-prop-1-ynyl)-pyridine;
2-[3-(2,3-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,3-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,3-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,4-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,4-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,5-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,5-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2,6-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-Methyl-6-[3-(2-trifluoromethyl-phenoxy)-prop-1-ynyl]-pyridine;

2-[3-(2-Benzyloxy-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2-Bromo-4,5-dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2-Chloro-4-methoxy-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2-Chloro-5-methyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(2-Chloro-6-methyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-Methyl-6-(3-o-tolyloxy-prop-1-ynyl)-pyridine;
2-Methyl-6-[3-(3,4,5-trimethyl-phenoxy)-prop-1-ynyl]-pyridine;
2-[3-(3,4-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3,4-Dimethoxy-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3,4-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3,5-Dichloro-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3,5-Dimethoxy-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3,5-Dimethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-[3-(3-Bromo-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
3-[3-(6-Methyl-pyridin-2-yl)-prop-2-ynyloxy]-benzonitrile;
2-[3-(3-Ethyl-phenoxy)-prop-1-ynyl]-6-methyl-pyridine;
2-methyl-6-[3-(3-methylphenoxy)prop-1-yn-1-yl] pyridine;
2-[3-(4-chloro-2-methylphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(4-chloro-3,5-dimethylphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(4-chloro-3-methylphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(4-chlorophenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(4-methoxyphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-methyl-6-[3-(4-nitrophenoxy)prop-1-yn-1-yl]pyridine;
2-methyl-6-[3-(3-nitrophenoxy)prop-1-yn-1-yl]pyridine;
2-methyl-6-[3-(3-methylphenoxy)prop-1-yn-1-yl]pyridine;
2-methyl-6-[3-(5,6,7,8-tetrahydronaphthalen-2-yloxy)prop-1-yn-1-yl]pyridine;
2-[3-(4-isopropylphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
2-[3-(4-tert-butylphenoxy)prop-1-yn-1-yl]-6-methylpyridine;
6-[3-(3,4-dimethylphenoxy)prop-1-yn-1-yl]-3-fluoro-2-methylpyridine; and 6-[3-(3,4-dimethylphenoxy)but-1-yn-1-yl]-3-fluoro-2-methylpyridine.

5. A compound according to any one of claims 1-4 for use in therapy.

6. A compound according to claim 5, wherein the therapy is treatment or prevention of gastroesophageal reflux disease.

7. Use of a compound according to formula I of claim 1 or 2, or a pharmaceutically acceptable salt or an optical isomer thereof, for the manufacture of a medicament for the inhibition of transient lower esophageal sphincter relaxations.

8. Use of a compound according to formula I of claim 1 or 2, or a pharmaceutically acceptable salt or an optical isomer thereof, for the manufacture of a medicament for treatment or prevention of gastroesophageal reflux disease.

9. A pharmaceutical composition comprising a compound of formula I of claim 1 or 2 as an active ingredient, together with a pharmacologically and pharmaceutically acceptable carrier.

10. A process for the preparation of a compound of formula I, whereby a coupling reaction of the aryl bromide A

is performed in the presence of a base such as triethyl amine to give the alcohol C

which is then converted into the mesylate D

and reacted with an alcohol, and wherein R1 is selected from hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl, wherein the aryl or heteroaryl may be substituted by C1-C4 alkyl;
R2 is selected from hydrogen and C1-C4 alkyl;
R3 is selected from hydrogen, C1-C4 alkyl, F, CF3, CHF2 and CH2F;
R4 is selected from hydrogen, F, CF3, CHF2, CH2F and CH3;
R5 is selected from hydrogen and F;
R6 is selected from hydrogen and F.

11. A compound selected from 3-(5-fluoro-6-methylpyridin-2-yl)prop-2-yn-1-ol;
4-(5-fluoro-6-methylpyridin-2-yl)but-3-yn-2-ol; 3-(5-fluoro-6-methylpyridin-2-yl)prop-2-yn-1-yl methanesulfonate; 3-(5-fluoro-6-methylpyridin-2-yl)-1-methylprop-2-yn-1-yl methanesulfonate; 4-(6-methylpyridin-2-yl)but-3-yn-2-ol; and Methanesulfonic acid 1-methyl-3-(6-methyl-pyridin-2-yl)-prop-2-ynyl ester.

12. A method for the inhibition of transient lower esophageal sphincter relaxations whereby an effective amount of a compound of formula I of claim 1 or 2 is administered to a subject in need of such inhibition.

13. A method for the treatment or prevention of gastroesophageal reflux disease, whereby an effective amount of a compound of formula I of claim 1 or 2 is administered to a subject in need of such treatment or prevention.