The present invention relates to ethynyl derivatives of formula I

R ¹ is fluoro or chloro;

or to a pharmaceutically acceptable acid addition salt, to a racemic mixture, or to its

corresponding enantiomer and/or optical isomer and/or stereoisomer thereof.

It has now surprisingly been found that the compounds of general formula I are metabotropic glutamate receptor antagonists (NAM = negative allosteric modulators).

Compounds of formula I are distinguished by having valuable therapeutic properties. They can be used in the treatment or prevention of mGluR5 receptor mediated disorders.

In the central nervous system (CNS) the transmission of stimuli takes place by the interaction of a neurotransmitter, which is sent out by a neuron, with a neuroreceptor.

Glutamate is the major excitatory neurotransmitter in the brain and plays a unique role in a variety of central nervous system (CNS) functions. The glutamate-dependent stimulus receptors are divided into two main groups. The first main group, namely the ionotropic receptors, forms ligand-controlled ion channels. The metabotropic glutamate receptors (mGluR) belong to the second main group and, furthermore, belong to the family of G-protein coupled receptors.

At present, eight different members of these mGluR are known and of these some even have sub-types. According to their sequence homology, signal transduction mechanisms and agonist selectivity, these eight receptors can be sub-divided into three sub-groups:

mGluRl and mGluR5 belong to group I, mGluR2 and mGluR3 belong to group II and mGluR4, mGluR6, mGluR7 and mGluR8 belong to group III. Negative allosteric modulators of metabotropic glutamate receptors, belonging to the first group, can be used for the treatment or prevention of acute and/or chronic neurological disorders such as Parkinson's disease, Fragile-X syndrome, autistic disorders, cognitive disorders and memory deficits, as well as chronic and acute pain and gastroesophageal reflux disease (GERD).

Other treatable indications in this connection are restricted brain function caused by bypass operations or transplants, poor blood supply to the brain, spinal cord injuries, head injuries, hypoxia caused by pregnancy, cardiac arrest and hypoglycaemia. Further treatable indications are ischemia, Huntington's chorea, amyotrophic lateral sclerosis (ALS), dementia caused by AIDS, eye injuries, retinopathy, idiopathic parkinsonism or parkinsonism caused by

medicaments as well as conditions which lead to glutamate-deficiency functions, such as e.g. muscle spasms, convulsions, migraine, urinary incontinence, nicotine addiction, opiate addiction, anxiety, vomiting, dyskinesia and depressions.

Disorders mediated full or in part by mGluR5 are for example acute, traumatic and chronic degenerative processes of the nervous system, such as Alzheimer's disease, senile dementia, Parkinson's disease, Huntington's chorea, amyotrophic lateral sclerosis and multiple sclerosis, psychiatric diseases such as schizophrenia and anxiety, depression, pain and drug dependency (Expert Opin. Then Patents (2002), 12, (12)).

Selective mGluR5 antagonists are especially useful for the treatment of disorders where reduction of mGluR5 receptor activation is desired, such as anxiety and pain, depression, Fragile-X syndrom, autism spectrum disorders, Parkinson's disease, and gastroesophageal reflux disease (GERD).

Objects of the present invention are compounds of formula I and their pharmaceutically acceptable salts, the above-mentioned compounds as pharmaceutically active substances and their production. Further objects of the invention are medicaments based on a compound in accordance with the invention and their manufacture as well as the use of the compounds in the control or prevention of mGluR5 receptor (NAM) mediated disorders, which are anxiety and pain, depression, Fragile-X syndrom, autism spectrum disorders, Parkinson's disease, and gastroesophageal reflux disease (GERD, and, respectively, for the production of corresponding medicaments.

One embodiment of the present invention are compounds of formula I wherein Y is N.

These compounds are

5-(3-fluoro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide 5 - (3 -chloro -phenylethynyl) -pyrimidine-2-c arboxylic acid tert-butyl-ethyl- amide .

One further embodiment of the present invention are compounds of formula I wherein Y is CH.

These compounds are

5-(3-fluoro-phenylethynyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide

5-(3-chloro-phenylethynyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide.

A particular embodiment of the invention consists of the following compounds:

5-(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide

5-(3-Chloro-phenylethynyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide

5-(3-Fluoro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

5-(3-Chloro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

Compounds, which are similar to those of the present invention, have been generically described as positive allosteric modulators of the mGluR5 receptor. Surprisingly, it has been found that highly potent mGluR5 antagonists were obtained instead of mGluR5 positive allosteric modulators, which have a completely opposite pharmacology if compared with positive allosteric modulators.

The main difference between positive- and negative allosteric modulators can be seen in figure 1. A mGluR5 positive allosteric modulator (PAM) leads to increased receptor activity (Ca ²⁺ mobilisation) in presence of a fixed concentration of glutamate, whereas an allosteric antagonist (negative allosteric modulator, NAM) leads to a reduction of receptor activation.

Figure 1 shows the general behavior of a NAM and a PAM under the same conditions. The affinity for the receptor in figure 1 is ca. 10 - ^" 7' M for the PAM and between 10 - ^" 7' M and 10 - ^" 8° M for the NAM. These values can also be measured using a binding assay to displace a radioligand (= MPEP), see assay description.

Figure 1: Comparison of an mGluR5 positive allosteric modulator (PAM) and an mGluR5 antagonist (negative allosteric modulator = NAM).

The indications which can be addressed by the compounds are not the same. mGluR5-NAMs are beneficial for indications where a reduction of excessive receptor activity is desired, such as anxiety and pain, depression, Fragile-X syndrom, autism spectrum disorders, Parkinson's disease, and gastroesophageal reflux disease (GERD). mGluR5 PAMs on the other hand are useful in indications where a normalization of decreased receptor activity is desired, such as in psychosis, epilepsy, schizophrenia, Alzheimer's disease and associated cognitive disorders, as well as tuberous sclerosis. This difference can be practically shown for example in an anxiety animal model, such as in the "rat Vogel conflict drinking test", where the compounds of the invention show anxiolytic activity, whereas mGluR-PAMs do not show activity in this animal model.

Biological Assays and Data:

Intracellular Ca ²⁺ mobilization assay

A monoclonal HEK-293 cell line stably transfected with a cDNA encoding for the human mGlu5a receptor was generated; for the work with mGlu5 Positive Allosteric Modulators

(PAMs), a cell line with low receptor expression levels and low constitutive receptor activity was selected to allow the differentiation of agonistic versus PAM activity. Cells were cultured according to standard protocols (Freshney, 2000) in Dulbecco's Modified Eagle Medium with high glucose supplemented with 1 mM glutamine, 10% (vol/vol) heat- inactivated bovine calf serum, Penicillin/Streptomycin, 50 g/ml hygromycin and 15 g/ml blasticidin (all cell culture reagents and antibiotics from Invitrogen, Basel, Switzerland).

About 24 hrs before an experiment, 5xl0 ⁴ cells/well were seeded in poly-D-lysine coated, black/clear-bottomed 96-well plates. The cells were loaded with 2.5 μΜ Fluo-4AM in loading buffer (lxHBSS, 20 mM HEPES) for 1 hr at 37°C and washed five times with loading buffer. The cells were transferred into a Functional Drug Screening System 7000 (Hamamatsu, Paris, France), and 11 half logarithmic serial dilutions of test compound at 37°C were added and the cells were incubated for 10-30 min. with on-line recording of fluorescence. Following this pre- incubation step, the agonist L-glutamate was added to the cells at a concentration corresponding to EC ₂ o (typically around 80 μΜ) with on-line recording of fluorescence; in order to account for day-to-day variations in the responsiveness of cells, the EC ₂ o of glutamate was determined immediately ahead of each experiment by recording of a full dose-response curve of glutamate. Responses were measured as peak increase in fluorescence minus basal (i.e. fluorescence without addition of L-glutamate), normalized to the maximal stimulatory effect obtained with saturating concentrations of L-glutamate. Graphs were plotted with the % maximal stimulatory using XLfit, a curve fitting program that iteratively plots the data using Levenburg Marquardt algorithm. The single site competition analysis equation used was y = A + ((B-A)/(l+((x/C)D))), where y is the % maximal stimulatory effect, A is the minimum y, B is the maximum y, C is the EC ₅₀ , x is the log 10 of the concentration of the competing compound and D is the slope of the curve (the Hill Coefficient). From these curves the EC ₅₀ (concentration at which half maximal stimulation was achieved), the Hill coefficient as well as the maximal response in % of the maximal stimulatory effect obtained with saturating concentrations of L-glutamate were calculated.

Positive signals obtained during the pre-incubation with the PAM test compounds (i.e. before application of an EC ₂₀ concentration of L-glutamate) were indicative of an agonistic activity, the absence of such signals were demonstrating the lack of agonistic activities. A depression of the signal observed after addition of the EC ₂ o concentration of L-glutamate was indicative of an inhibitory activity of the test compound.

In the list of examples below are shown the corresponding results for compounds, which all have EC ₅₀ values less or equal 100 nM.

ΜΡΕΡ binding assay:

For binding experiments, cDNA encoding human mGlu 5a receptor was transiently transfected into EBNA cells using a procedure described by Schlaeger and Christensen

[Cytotechnology 15: 1-13 (1998)]. Cell membrane homogenates were stored at -80°C until the day of assay where upon they were thawed and resuspended and polytronised in 15 mM Tris- HC1, 120 mM NaCl, 100 mM KC1, 25 mM CaCl ₂ , 25 mM MgCl ₂ binding buffer at pH 7.4 to a final assay concentration of 20 μg protein/ well.

Saturation isotherms were determined by addition of twelve [ H]MPEP concentrations (0.04-100 nM) to these membranes (in a total volume of 200 μΐ) for 1 h at 4°C. Competition experiments were performed with a fixed concentration of [ H]MPEP (2nM) and IC ₅₀ values of test compounds evaluated using 11 concentrations (0.3-10,000nM). Incubations were performed for 1 h at 4° C.

At the end of the incubation, membranes were filtered onto unifilter (96-well white microplate with bonded GF/C filter preincubated 1 h in 0.1% PEI in wash buffer, Packard Bioscience,

Meriden, CT) with a Filtermate 96 harvester (Packard Bioscience) and washed 3 times with cold 50 mM Tris-HCl, pH 7.4 buffer. Nonspecific binding was measured in the presence of 10 μΜ MPEP. The radioactivity on the filter was counted (3 min) on a Packard Top-count microplate scintillation counter with quenching correction after addition of 45 μΐ of microscint 40 (Canberra Packard S.A., Ziirich, Switzerland) and shaking for 20 min.

In the list of examples below are shown the corresponding results for compounds which all have EC ₅₀ values less or equal to 100 nM.

Comparison of compounds of the invention versus the reference compounds 1 and 2

As can be seen in the table below, the compounds of the invention (NAM) show a clearly different profile compared to structurally similar reference compounds 1 and 2 (PAM).

ECso (nM) Ki (nM)

Activity

Examples Structure mGlu5 PAM MPEP

profile assay binding

Reference

16 29 PAM compound 1

Reference

23 51 PAM compound 2

The compounds of formula I can be manufactured by the methods given below, by the methods given in the examples or by analogous methods. Appropriate reaction conditions for the individual reaction steps are known to a person skilled in the art. The reaction sequence is not limited to the one displayed in the schemes, however, depending on the starting materials and their respective reactivity the sequence of reaction steps can be freely altered. Starting materials are either commercially available or can be prepared by methods analogous to the methods given below, by methods described in references cited in the description or in the examples, or by methods known in the art.

The present compounds of formula I and their pharmaceutically acceptable salts may be prepared by methods, known in the art, for example by the process variants described below, which process comprises

reacting a compound of formula

with a compound of formula

to form a compound of formu

wherein the substituents are described above.

The preparation of compounds of formula I is further described in more detail in scheme 1 and in examples 1 - 4.

Scheme 1

An ethynyl-pyridine or ethynyl-pyrimidine compound of formula I can be obtained for example by Sonogashira coupling of 5-bromo-pyridine-2-carboxylic acid methyl ester or 5- bromo-pyrimidine-2-carboxylic acid methyl ester 1 with an appropriately substituted arylacetylene 2 followed by saponification with a base such as LiOH to yield the corresponding acid 3 or by Sonogashira coupling of 5-bromo-pyridine-2-carboxylic acid or 5-bromo- pyrimidine-2-carboxylic acid 1 with an appropriately substituted arylacetylene 2 to yield directly the corresponding acid 3. Reacting the corresponding acid 3 with tert.-butylethylamine 4 in the presence of a base such as Hunig's Base and a peptide coupling reagent such as TBTU in a solvent such as dioxane or by preparing in-situ the corresponding acid chloride with oxalyl chloride and DMF (cat.) in a solvent such as dichloromethane followed by reaction with tert.- butylethylamine 4 in the presence of a base such as pyridine yield the desired ethynyl

compounds of general formula I (scheme 1).

Pharmaceutically acceptable salts of compounds of formula I can be manufactured readily according to methods known per se and taking into consideration the nature of the compound to be converted into a salt. Inorganic or organic acids such as, for example, hydrochloric acid, hydrobromic acid, sulphuric acid, nitric acid, phosphoric acid or citric acid, formic acid, fumaric acid, maleic acid, acetic acid, succinic acid, tartaric acid, methanesulphonic acid, p-toluenesulphonic acid and the like are suitable for the formation of pharmaceutically acceptable salts of basic compounds of formula I. Compounds which contain the alkali metals or alkaline earth metals, for example sodium, potassium, calcium, magnesium or the like, basic amines or basic amino acids are suitable for the formation of pharmaceutically acceptable salts of acidic compounds.

Moreover the invention relates also to medicaments containing one or more compounds of the present invention and pharmaceutically acceptable excipients for the treatment and prevention of mGluR5 receptor mediated disorders (NAM), such as anxiety and pain, depression, Fragile -X syndrom, autism spectrum disorders, Parkinson's disease, and gastroesophageal reflux disease (GERD). The invention also relates to the use of a compound in accordance with the present invention as well as its pharmaceutically acceptable salt for the manufacture of medicaments for the treatment and prevention of mGluR5 receptor mediated disorders (NAM) as outlined above.

The pharmacological activity of the compounds was tested using the following method: cDNA encoding rat mGlu 5a receptor was transiently transfected into EBNA cells using a procedure described by E.-J. Schlaeger and K. Christensen (Cytotechnology 1998, 15, 1-13). [Ca ²⁺ ]i measurements were performed on mGlu 5a transfected EBNA cells after incubation of -l ithe cells with Fluo 3-AM (obtainable by FLUKA, 0.5 μΜ final concentration) for 1 hour at 37°C followed by 4 washes with assay buffer (DMEM supplemented with Hank's salt and 20 mM HEPES. [Ca ²⁺ ]i measurements were done using a fluorometric imaging plate reader (FLIPR, Molecular Devices Corporation, La Jolla, CA, USA). When compounds were evaluated as antagonists they were tested against 10 μΜ glutamate as agonist.

The inhibition (antagonists) curves were fitted with a four parameter logistic equation giving IC ₅₀ , and Hill coefficient using the iterative non linear curve fitting software Origin (Microcal Software Inc., Northampton, MA, USA).

The Ki values of the compounds tested are given. The Ki value is defined by the following formula:

1 ₊ JiJ

^EC 50

in which the IC50 values are those concentrations of the compounds tested in μΜ by which 50 % of the effect of compounds are antagonised. [L] is the concentration and the EC50 value is the concentration of the compounds in μΜ which brings about 50 % stimulation.

The compounds of the present invention are mGluR5a receptor antagonists. The activities of compounds of formula I as measured in the assay described above are in the range of Ki < 100 μΜ.

The compounds of formula I and pharmaceutically acceptable salts thereof can be used as medicaments, e.g. in the form of pharmaceutical preparations. The pharmaceutical preparations can be administered orally, e.g. in the form of tablets, coated tablets, dragees, hard and soft gelatine capsules, solutions, emulsions or suspensions. However, the administration can also be effected rectally, e.g. in the form of suppositories, or parenterally, e.g. in the form of injection solutions.

The compounds of formula I and pharmaceutically acceptable salts thereof can be processed with pharmaceutically inert, inorganic or organic carriers for the production of pharmaceutical preparations. Lactose, corn starch or derivatives thereof, talc, stearic acid or its salts and the like can be used, for example, as such carriers for tablets, coated tablets, dragees and hard gelatine capsules. Suitable carriers for soft gelatine capsules are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols and the like; depending on the nature of the active substance no carriers are, however, usually required in the case of soft gelatine capsules. Suitable carriers for the production of solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose and the like. Adjuvants, such as alcohols, polyols, glycerol, vegetable oils and the like, can be used for aqueous injection solutions of water-soluble salts of compounds of formula I, but as a rule are not necessary. Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

In addition, the pharmaceutical preparations can contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They can also contain still other therapeutically valuable substances.

As mentioned earlier, medicaments containing a compound of formula I or

pharmaceutically acceptable salts thereof and a therapeutically inert excipient are also an object of the present invention, as is a process for the production of such medicaments which comprises bringing one or more compounds of formula I or pharmaceutically acceptable salts thereof and, if desired, one or more other therapeutically valuable substances into a galenical dosage form together with one or more therapeutically inert carriers.

The dosage can vary within wide limits and will, of course, be fitted to the individual requirements in each particular case. In general, the effective dosage for oral or parenteral administration is between 0.01-20 mg/kg/day, with a dosage of 0.1-10 mg/ kg/day being preferred for all of the indications described. The daily dosage for an adult human being weighing 70 kg accordingly lies between 0.7-1400 mg per day, preferably between 7 and 700 mg per day.

The following examples are provided to further elucidate the invention:

Example 1

5-(3-Fluoro-phenylethynyl -pyridine-2-carboxylic acid tert-butyl-ethyl-amide

Step 1: 5-(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid methyl ester

Bis-(triphenylphosphine)-palladium(II)dichloride (406 mg, 580 μιηοΐ, 0.05 equiv.) was dissolved in 25 ml DMF. (2.5 g, 11.6 mmol) 5-Bromo-pyridine-2-carboxylic acid methyl ester and 3- fluorophenylacetylene (2.22 g, 18.5 mmol, 1.6 equiv.) were added at room temperature.

Triethylamine (3.5 g, 4.84 ml, 34.7 mmol, 3 equiv.), triphenylphosphine (91 mg, 347 μιηοΐ, 0.03 equiv.) and copper(I)iodide (66 mg, 347 μιηοΐ, 0.03 equiv.) were added and the mixture was stirred for 20 hours at 80°C. The reaction mixture was cooled and evaporated to dryness with Isolute® sorbent. The crude product was purified by flash chromatography on silica gel (70 g) eluting with an ethyl acetate:heptane gradient 0: 100 to 80:20. The desired 5-(3-fluoro- phenylethynyl)-pyridine-2-carboxylic acid methyl ester (1.95 g, 66 % yield) was obtained as a light yellow solid, MS: m/e = 256.3 (M+H ⁺ ).

Step 2: 5-(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid

(1.9 g, 7.44 mmol) 5-(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid methyl ester

(Example 1, step 1 ) was dissolved in THF (30 ml) and water (30 ml) and LiOH (357 mg, 24.9 mmol, 2 equiv.) was added at room temperature. The mixture was stirred for 16 hours at room temperature. The reaction mixture was acidified with 4N HCl to pH 2.5 and THF was evaporated to form a yellow suspension. The suspension was cooled to 0-5°C and filtered. The crystals were washed with cold water and evaporated to dryness. The desired 5-(3-fluoro-phenylethynyl)- pyridine-2-carboxylic acid (1.71 g, 95 % yield) was obtained as a light yellow solid, MS: m/e = 239.9 (M+H ⁺ ). -(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide

(100 mg, 0.41 mmol) 5-(3-Fluoro-phenylethynyl)-pyridine-2-carboxylic acid (Example 1, step 2) was dissolved in dioxane (1 ml) and Hunig's Base (217 μΐ, 1.24 mmol, 3 equiv.), tert.- butylethylamine (63 mg, 0.62 mmol, 1.5 equiv.) and TBTU (146 mg, 0.45 mmol, 1.1 equiv.) were added at room temperature. The mixture was stirred for 16 hours at room temperature. The reaction mixture was evaporated and extracted saturated NaHC0 ₃ solution and two times with a small volume of dichloromethane. The crude product was purified by flash chromatography by directly loading the dichloromethane layers onto a silica gel column and eluting with an ethyl acetate:heptane gradient 0: 100 to 0: 100. The desired 5-(3-fluoro-phenylethynyl)-pyridine-2- carboxylic acid tert-butyl-ethyl-amide (102 mg, 76 % yield) was obtained as a light yellow oil, MS: m/e = 325.3 (M+H ⁺ ).

Example 2

5-(3-Chloro-phenylethyny -pyridine-2-carboxylic acid tert-butyl-ethyl-amide

Step 1: 5-(3-Chloro-phenylethynyl)-pyridine-2-carboxylic acid

The title compound was obtained as a white solid, MS: m/e = 258.4/260.4 (M+H ⁺ ), using chemistry similar to that described in Example 1, step 1 from 5-bromo-pyridine-2-carboxylic acid and 3-chlorophenylacetylene. -(3-Chloro-phenylethvnyl)-pyridine-2-carboxylic acid tert-butyl-ethyl-amide

The title compound was obtained as a light yellow oil, MS: m/e = 341.5/343.5 (M+H ⁺ ), using chemistry similar to that described in Example 1, step 3 from 5-(3-chloro-phenylethynyl)- pyridine-2-carboxylic acid (Example 2, step 1 ) and tert.-butylethylamine. Example 3

5-(3-Fluoro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

Step 1: 5-(3-Fluoro-phenylethynyl)-pyrimidine-2-carboxylic acid

The title compound was obtained as a light yellow solid, MS: m/e = 243.4 (M+H ⁺ ), using chemistry similar to that described in Example 1, step 1 from 5-bromo-pyrimidine-2-carboxylic acid and 3-fluorophenylacetylene. -(3-Fluoro-phenylethvnyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

(100 mg, 0.41 mmol) 5-(3-Fluoro-phenylethynyl)-pyrimidine-2-carboxylic acid (Example 3, step 1) was suspended in dichloromethane (1 ml) and DMF (10 μΐ). Oxalyl chloride (40 μΐ, 0.45 mmol, 1.1 equiv.) was added drop wise at room temperature and the mixture was stirred for 1 hour at reflux. The reaction mixture was then added to a mixture of diisopropylethylamine (235 μΐ, 1.34 mmol, 3.3 equiv.) and tert-butylethylamine (43 mg, 0.41 mmol, 1 equiv.) in THF (2 ml). The mixture was stirred for 16 hours at room temperature and evaporated in presence of Isolute® sorbent to dryness. The crude product was purified by flash chromatography with a 20 g silica gel column eluting with heptane:ethyl acetate 100:0 -> 0: 100. The desired 5-(3-fluoro- phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide (97 mg, 78% yield) was obtained as a white solid, MS: m/e = 326.5 (M+H ⁺ ).

Example 4

5-(3-Chloro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

Step 1: 5-(3-Chloro-phenylethvnyl)-pyrimidine-2-carboxylic acid

The title compound was obtained as a white solid, MS: m/e = 259.4/261.4 (M+H ⁺ ), using chemistry similar to that described in Example 1, step 1 from 5-bromo-pyrimidine-2-carboxylic acid and 3-chlorophenylacetylene. -(3-Chloro-phenylethynyl)-pyrimidine-2-carboxylic acid tert-butyl-ethyl-amide

The title compound was obtained as a white solid, MS: m/e = 342.6/344.6 (M+H ⁺ ), using chemistry similar to that described in Example 3, step 2 from 5-(3-chloro-phenylethynyl)- pyrimidine-2-carboxylic acid (Example 4, step 1 ) and tert-butylethylamine.