AU780009B2 - Heterocyclic compounds and methods of use thereof - Google Patents

Heterocyclic compounds and methods of use thereof Download PDF

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AU780009B2
AU780009B2 AU69482/00A AU6948200A AU780009B2 AU 780009 B2 AU780009 B2 AU 780009B2 AU 69482/00 A AU69482/00 A AU 69482/00A AU 6948200 A AU6948200 A AU 6948200A AU 780009 B2 AU780009 B2 AU 780009B2
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pain
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Leo Solomon Bleicher
Nicholas D. P. Cosford
Rowena V. Cube
Stephen D. Hess
Ian A. Mcdonald
Benito Munoz
Edwin J. Schweiger
Mark A. Varney
Jean-Michel Vernier
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Merck Sharp and Dohme LLC
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Merck and Co Inc
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Priority claimed from US09/387,135 external-priority patent/US6956049B1/en
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Description

WO 01/16121 PCT/US00/23923 HETEROCYCLIC COMPOUNDS AND METHODS OF USE THEREOF FIELD OF INVENTION The present invention relates to novel heterocyclic compounds which contain a heterocylic ring bearing at least one substituent attached by a linker containing an acetylenic group, a vinylic group or an azo group. In addition, the present invention relates to therapeutic methods of use of heterocyclic compounds for the treatment and prevention of various disease conditions.
BACKGROUND OF THE INVENTION Unsaturated heterocylic compounds find a wide variety of uses. -For example, compounds of this class find uses as modulators of physiological processes that are mediated by ligand-activated receptors. Receptors that are activated by ligands are located throughout the nervous, cardiac, renal, digestive and bronchial systems, among others. Therefore, in the nervous system, for example, heterocyclic compounds are capable of functioning as agonists or antagonists of receptors for neurotransmitters, neurohormones and neuromodulators. Ligand-activated receptors have been identified in a wide variety of species, including humans, other mammals and vertebrates as well as in invertebrate species. Therefore, compounds of this class are also able to modulate receptor-mediated processes throughout phylogeny and find uses in a wide variety of applications, as pharmaceuticals, insecticides and fungicides.
Receptors activated by excitatory amino acids, such as the amino acid L-glutamic acid (glutamate), are a major excitatory neurotransmitter receptor class in the mammalian central nervous system.
Anatomical, biochemical and electrophysiological analyses suggest that glutamatergic systems are involved in a broad array of neuronal processes, including fast excitatory synaptic transmission, regulation of neurotransmitter release, long-term potentiation, long-term depression, learning and memory, developmental synaptic plasticity, hypoxic-ischemic damage and neuronal cell death, epileptiform seizures, visual processing, as well as the pathogenesis of several neurodegenerative disorders. See generally, Nakanishi et al., Brain Research Reviews 26:230-235 (1998); Monaghan et al., Ann Rev. Pharmacol.
Toxicol. 29:365-402 (1980). This extensive repertoire of functions, especially those related to learning, neurotoxicity, and neuropathology, has stimulated recent attempts to describe and define the mechanisms through which glutamate exerts its effects.
Glutamate has been observed to mediate its effects through receptors that have been categorized into two main groups: ionotropic and metabotropic. Metabotropic glutamate receptors are divided into three groups based on amino acid sequence homology, transduction mechanism and pharmacological properties, namely Group I, Group II and Group III. Each Group of receptors contains one or more types of receptors. For example, Group I includes metabotropic glutamate receptors 1 and 5 (mGluR1 WO 01/16121 PCT/US00/23923 2 and mGluR5), Group II includes metabotropic glutamate receptors 2 and 3 (mGluR2 and mGluR3) and Group III includes metabotropic glutamate receptors 4, 6, 7 and 8 (mGluR4, mGluR6, mGluR7 and mGluR8). Several subtypes of a mGluR type may exist. For example, subtypes of mGluR1 include mGluRla, mGluRlb, mGluRlc and mGluRld.
Anatomical studies demonstrate a broad and selective distribution of metabotropic glutamate receptors in the mammalian nervous system. For example, mGluRI is expressed in the cerebellum, olfactory bulb, hippocampus, lateral septum, thalamus, globus pallidus, entopeduncular nucleus, ventral pallidum and substantia nigra (Petralia el al., (1997) J Chem. Neuroanat., 13:77-93; Shigemoto et al., (1992) J Comp. Neurol., 322:121-135). In contrast, mGluR5 is weakly expressed in the cerebellum, while higher levels of expression are found in the striatum and cortex (Romano et al., (1995) J. Comp.
Neurol., 355:455-46 9 In the hippocampus, mGluR5 appears widely distributed and is diffusely expressed.
Metabotropic glutamate receptors are typically characterized by seven putative transmembrane domains, preceded by a large putative extracellular amino-terminal domain and followed by a large putative intracelluar carboxy-terminal domain. The receptors couple to G-proteins and activate certain second messengers depending on the receptor group. Thus, for example, Group I mGluRs activate phospholipase C. Activation of the receptors results in the hydrolysis of membrane phosphatidylinositol (4,5)-bisphosphate to diacylglycerol, which activates protein kinase C, and inositol trisphosphate, which in turn activates the inositol trisphosphate receptor to promote the release of intracellular calcium.
Ionotropic glutamate receptors are generally divided into two classes: the N-methyl-D-aspartate (NMDA) and non-NMDA receptors. Both classes of receptors are linked to integral cation channels and share some amino acid sequence homology. GluRI-4 are termed AMPA methylisoxazole-4-propionic acid) receptors because AMPA preferentially activates receptors composed of these subunits, while GluR5-7 and KA1-2 are termed kainate receptors as these are preferentially sensitive to kainic acid. Thus, an "AMPA receptor" is a non-NMDA receptor that can be activated by AMPA. AMPA receptors include the GluRI-4 family, which form homo-oligomeric and hetero-oligomeric complexes which display different current-voltage relations and calcium permeability. Polypeptides encoded by GluRI-4 nucleic acid sequences can form functional ligandgated ion channels. An AMPA receptor includes a receptor having a GluRI, GIuR2, GluR3 and/or GluR4 subunit. A NMDA receptor includes a receptor having NMDARI, NMDAR2a, NMDAR2b, NMDAR2c, NMDAR2d and/or NMDAR3 subunits.
Because of the physiological and pathological significance of excitatory amino acid receptors generally and metabotropic glutamate receptors, in particular, there is a need to identify methods of 3 modulating excitatory amino acid receptor-mediated processes, as well as therapeutic methods of treatment and methods for prevention of diseases. Also, there is a continuing need in the art for new members of a compound class that can modulate excitatory amino acid receptors. The present invention satisfies these and related needs.
Brief Description of the Invention Disclosed herein is a novel class of heterocyclic compounds. Compounds disclosed herein may contain a substituted, unsaturated five-, six- or seven-membered heterocyclic ring that includes at least one nitrogen atom and at least one carbon atom. The ring additionally includes three, four or five atoms independently selected from carbon, nitrogen, sulfur and oxygen atoms. The heterocyclic ring has at least one substituent located at a ring position adjacent to a ring nitrogen atom. This mandatory substituent of the ring includes a moiety linked to the heterocyclic ring via a hydrocarbyl or an azo group. Also disclosed are pharmaceutically acceptable salt forms of heterocyclic compounds.
15 According to a first aspect of the invention there is provided a compound having the structure:
.A-L-B,
or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, wherein: 20 A is a thiazolyl ring having the structure:
YX
Z
N
wherein one of X, Y and Z is S; and the remainder ofX, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl; provided that the following compounds are excluded: 3a compounds wherein A is a thiazolyl ring, wherein each R is independently hydrogen, nitro, halogen, Ci-C 4 -alkyl, Ci-C 4 -haloalkyl, Ci-C 4 -alkoxy, Ci-C 4 -haloalkoxy, Ci-C 4 -alkylthio, C 1
-C
4 -haloalkylthio, C 3
-C
6 -alkenyl, or C 3 -Cs-cycloalkyl; L is alkynylene; and B is substituted or unsubstituted aryl, wherein substituents are independently nitro, cyano, CI-C 6 -alkyl, C 1
-C
4 -haloalkyl, Ci-C 4 -alkoxy, C 1
-C
4 haloalkoxy, Cl-C 4 -alkylthio, C 1
-C
4 -haloalkylthio, Ci-C 4 -alkoxycarbonyl, C 3
-C
6 -alkenyl, phenyl, or phenoxy, wherein phenyl and phenoxy may bear further substituents; compounds wherein Y and Z are CR; X is S; R is phenyl; L is unsubstituted alkenylene, and B is unsubstituted phenyl; compounds wherein X and Z are CR, Y is S; R is phenyl; L is unsubstituted alkenylene, and B is unsubstituted phenyl; and compounds wherein A is unsubstituted thiazolyl; L is alkenylene or alkynylene; and B is unsubstituted phenyl.
According to a second aspect of the invention there is provided a pharmaceutical ~15 composition comprising a compound according to the first aspect of the invention, and a pharmaceutically acceptable carrier therefor.
According to a third aspect of the invention there is provided a method of antagonizing the activity of metabotropic glutamate receptors, said method comprising contacting said receptors with at least one compound having the structure A-L-B, or 20 enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, in an amount sufficient to antagonize the activity of said metabotropic glutamate receptor, wherein A is a thiazolyl ring having the structure:
N
wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
3b According to a fourth aspect of the invention there is provided a method for treating a disease condition related to the activity of metabotropic glutamate receptors, said method comprising admiristering to a patient having said disease condition a therapeutically effective amount of at least one compound having the structure A-L-B, or s enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, or administering a pharmaceutical composition comprising said compound together with a pharmaceutically acceptable carrier therefore, wherein A is a thiazolyl ring having the structure:
Y-X
wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, I15 carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or 20 more double bonds, or substituted aryl.
According to a fifth aspect of the invention there is provided a method for preventing a disease condition related to the activity of metabotropic glutamate receptors a subject at risk thereof, said method comprising administering to said subject a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, or administering a pharmaceutical composition comprising said compound together with a pharmaceutically acceptable carrier therefore, wherein A is a thiazolyl ring having the structure: Y-x
N
wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein 3c each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, ainido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
According to a sixth aspect of the invention there is provided the use of a o0 therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for antagonizing the activity of metabotropic glutamate receptors, wherein A is a thiazolyl ring having the structure:
Y-X
15 S. wherein one ofX, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, 20 carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
According to a seventh aspect of the invention there is provided the use of a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for treating a disease condition related to the activity of metabotropic glutamate receptors, wherein A is a thiazolyl ring having the structure: Y-x
Z'N
wherein one of X, Y and Z is S; and the rem'ainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted.
hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
According to an eighth aspect of the invention there is provided the use of a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or 15 pharmaceutically acceptable salts thereof, for the manufacture of a medicament for preventing a disease condition related to the activity of metabotropic glutamate receptors, wherein A is a thiazolyl ring having the structure:
Y-X
Z
N
20 wherein one of X, Y and Z is S; and the remainder ofX, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
According to a ninth aspect of the invention there is provided a pharmaceutically acceptable salt form of a compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, wherein A is a thiazolyl ring having the structure: Y-x
N
wherein one of X, Y and Z is S; and s the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl; and the salt is acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, butyrate, 15 citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, heptanoate, hexanoate, 2-hydroxyethanesulfonate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, tartrate, toluenesulfonate, undecanoate, sulfate, bisulfate, hemisulfate, hydrochloride, hydrobromide, hydroiodide, 20 an ammonium salt, an alkali metal salt, an alkaline earth metal salt, a dicyclohexylamine salt, N-methyl-D-glucamine, phenylethylamine, or an amino acid salt.
Invention compounds are useful for a wide variety of applications. For example heterocyclic compounds of the invention may act to modulate physiological processes by functioning as agonists and antagonists of receptors in the nervous system. Invention compounds may also act as insecticides and as fungicides. Pharmaceutical compositions containing invention compounds also have wide utility.
Also disclosed herein are methods of modulating the activity of excitatory amino acid receptors using a specifically defined class of heterocyclic compounds. One embodiment disclosed herein relates to methods of modulating metabotropic glutamate receptors. Also disclosed herein are methods of treating disease using heterocyclic compounds. Diseases contemplated include cerebral ischemia, chronic neurodegeneration, psychiatric disorders, schizophrenia, mood disorders, emotion disorders, disorders of extrapyramidal motor function, obesity, disorders of respiration, motor control and function, attention deficit disorders, concentration disorders, pain 3f disorders, neurodegenerative disorders, epilepsy, convulsive disorders, eating disorders, sleep disorders, sexual disorders, circadian disorders, drug withdrawal, drug addiction, compulsive disorders, anxiety, panic disorders, depressive disorders, skin disorders, retinal ischemia, retinal degeneration, glaucoma, disorders associated with organ transplantation, asthma, ischemia and astrocytomas. Also disclosed herein are methods of preventing disease conditions related to diseases of the pulmonary system, diseases of the nervous system, diseases of the cardiovascular system, diseases of the gastrointestinal system, diseases of the endocrine system, diseases of the exocrine system, diseases of the skin, cancer and diseases of the ophthalmic system.
:Ioe *i a DETAILED DESCRIPTION OF THE INVENTION Disclosed herein are compounds having the structure:
A-L-.B
or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, wherein: A is a 6- or 7-membered ring having the structure: Y W (R)q zo .wherein at least one ofW, X, Y and Z is (CR)p, wherein p is 0, 1 or 2; the remainder of W, X, Y and Z are each independently O, N or S; and each R is independently halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl or sulfonamide, *wherein q is 0, 1, 2 or3; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted 15 cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted or unsubstituted aryl; provided, that the following compounds are excluded: the compounds wherein A is a 6-membered ring wherein: W, X,Y and Z are wherein p is 1; and R at the W position is hydrogen, lower alkyl, hydroxy, hydroxy-lower alkyl, aminolower alkyl, lower alkylamino-lower alkyl, di-lower alkylamino-lower alkyl, unsubstituted or hydroxysubstituted lower alkyleneamino-lower alkyl, lower alkoxy, lower alkanoyloxy, amino-lower alkoxy, lower alkylamino-lower alkoxy, di-lower alkylamino-lower alkoxy, phthalimido-lower alkoxy, unsubstituted or hydroxy- or 2-oxo-imidazolidin-I-yl-substitued lower alkyleneamino-lower alkoxy, carboxy, esterified or amidated carboxy, carboxy-lower alkoxy or esterified carboxy-lower-alkoxy; R at the X position is hydrogen; R at the Y position is hydrogen, lower alkyl, carboxy, esterified carboxy, amidated carboxy, hydroxy-lower alkyl, hydroxy, lower alkoxy or lower alkanoyloxy; and R at the Z position is hydrogen, lower alkyl, hydroxy-lower alkyl, carboxy, esterified carboxy, amidated carboxy, unsubstituted or lower alkyl-, lower alkoxy-, halo- and/or trifluoromethyl-substituted N-lower alkyl-Nphenylca-bameyl, lower alkoxy, halo-lower alkyl or halo-lower alkoxy; WO 01/16121 PCT/USOO/23923 L is substituted or unsubstituted alkenylene, alkynylene or azo, B is substituted or unsubstituted aryl or heterocycle having two or more double bonds, wherein substituents are independently lower alkyl, lower alkenyl, lower alkynyl, phenyl, phenyl-lower alkynyl, hydroxy, hydroxy-lower alkyl, lower alkoxy, lower alkenyloxy, lower alkylenedioxy, lower alkanoyloxy, phenoxy, phenyl-lower alkoxy, acyl, carboxy, esterified carboxy, amidated carboxy, cyano, nitro, amino, acylamino, N-acyl-N-lower alkylamino, halo and halo-lower alkyl, wherein phenyl, phenyl-lower alkynyl, phenoxy, and phenyl-lower alkoxy may bear further substituents; and the compounds wherein A is a 6-membered ring wherein: W, X, Y and Z are (CR)p wherein p is 1; R at the X position is not hydrogen; and R at the W, Y and Z positions are hydrogen; L is alkenylene or alkynylene; and B is a substituted or unsubstituted aryl or heterocycle containing two or more double bonds; and the compounds wherein A is a 5-membered ring wherein: oneofW, X, Yand Zis (CR) 1 ,and pis0, two ofW, X, Yand Zare (CR)pand p is 1, and the remaining variable ring member is 0 or S; or one of W, X Y and Z is N, one of W, X, Y and Z is (CR)p and p is 1, one of W, X, Y and Z is (CR)p and p is 0, and the remaining variable ring member is 0, S or (CR)p, and p is 1; or two ofW, X, Yand Zare N,one ofW, X, Yand Zis (CR),and pis0, andthe remaining variable ring member is 0 or S or and p is 1; each R is independently hydrogen, nitro, halogen, C 1
-C
4 -alkyl, C 1
-C
4 -haloalkyl, C 1
-C
4 alkoxy, C 1
-C
4 -haloalkoxy, Cj-C 4 -alkylthio, C 1
-C
4 -haloalkylthio, C 3
-C
6 -alkenyl or C 3 -C8cycloalkyl; L is alkynylene; and B is substituted or unsubstituted aryl, wherein substituents are independently nitro, cyano, C 1
-C
6 -alkyl, C 1
-C
4 -haloalkyl, C 1
-C
4 -alkoxy, C 1
-C
4 -haloalkoxy, C 1
-C
4 -alkthio, C 1 -C4haloalkylthio, C 1
-C
4 -alkoxycarbonyl, C 3
-C
6 -alkenyl, phenyl or phenoxy, wherein phenyl and phenoxy may bear furthcr substituents; and the compounds wherein A is a 6-membered ring wherein: W, X, Y and Z are (CR)p, wherein p is I and R is hydrogen, L is alkynylene; and B is unsubstituted I -cyclopenten- I-yl or unsubstituted I -cyclohexen-1I-yl; and the compounds wherein A is a 5-membered ring wherein: WO 01/16121 PCT/US00/23923 6 W is (CR)p, and p is 0, Y and Z are (CR)p, and p is 1, X is N or S; and R is phenyl; or W is (CR)p, and p is 0, X and Z are (CR)p, and p is 1, Y is O, N or S; and R is phenyl; L is unsubstituted alkenylene and B is unsubstituted phenyl; and the compounds wherein A is a 5-membered ring containing two double bonds, wherein one of W, X, Y and Z is and p is 0, and the remaining ring members are and p is 1; and the compounds wherein A is unsubstituted heterocycle containing two or more double bonds; L is alkenylene or alkynylene, and B is unsubstituted phenyl.
As employed herein, "hydrocarbyl" refers to straight or branched chain univalent and bivalent radicals derived from saturated or unsaturated moieties containing only carbon and hydrogen atoms, and having in the range of about I up to 12 carbon atoms. Exemplary hydrocarbyl moieties include alkyl moieties, alkenyl moieties, dialkenyl moieties, trialkenyl moieties, alkynyl moieties, alkadiynal moieties, alkatriynal moieties, alkenyne moieties, alkadienyne moieties, alkenediyne moieties, and the like. The term "substituted hydrocarbyl" refers to hydrocarbyl moieties further bearing substituents as set forth below.
As employed herein, "alkyl" refers to straight or branched chain alkyl radicals having in the range of about I up to 12 carbon atoms; "substituted alkyl" refers to alkyl radicals further bearing one or more substituents such as hydroxy, alkoxy, mercapto, aryl, heterocycle, halogen, trifluoromethyl, pentafluoroethyl, cyano, cyanomethyl, nitro, amino, amide, amidine, amido, carboxyl, carboxamide, carbamate, ester, sulfonyl, sulfonamide, and the like.
As employed herein, "alkenyl" refers to straight or branched chain hydrocarbyl radicals having at least one carbon-carbon double bond, and having in the range of about 2 up to 12 carbon atoms (with radicals having in the range of about 2 up to 6 carbon atoms presently preferred), and "substituted alkenyl" refers to alkenyl radicals further bearing one or more substituents as set forth above.
As employed herein, "alkenylene" refers to straight or branched chain divalent alkenyl moieties having at least one carbon-carbon double bond, and having in the range of about 2 up to 12 carbon atoms (with divalent alkenyl moieties having in the range of about 2 up to 6 carbon atoms presently preferred), and "substituted lower alkenylene" refers to divalent alkenyl radicals further bearing one or more substituents as set forth above.
WO 01/16121 PCT/US00/23923 7 As employed herein, "alkynyl" refers to straight or branched chain hydrocarbyl radicals having at least one carbon-carbon triple bond, and having in the range of about 2 up to 12 carbon atoms (with radicals having in the range of about 2 up to 6 carbon atoms presently being preferred), and "substituted alkynyl" refers to alkynyl radicals further bearing one or more substituents as set forth above.
As employed herein, "alkynylene" refers to straight or branched chain divalent alkynyl moieties having at least one carbon-carbon triple bond, and having in the range of about 2 up to 12 carbon atoms (with divalent alkynyl moieties having two carbon atoms presently being preferred), and "substituted alkynylene" refers to divalent alkynyl radicals further bearing one or more substituents as set forth above.
As employed herein, "cyclohydrocarbyl" refers to cyclic ring-containing) univalent radicals derived from saturated or unsaturated moieties containing only carbon and hydrogen atoms, and having in the range of about 3 up to 20 carbon atoms. Exemplary cyclohydrocarbyl moieties include cycloalkyl moieties, cycloalkenyl moieties, cycloalkadienyl moieties, cycloalkatrienyl moieties, cycloalkynyl moieties, cycloalkadiynyl moieties, spiro hydrocarbon moieties wherein two rings are joined by a single atom which is the only common member of the two rings spiro[3.4]octanyl, and the like), bicyclic hydrocarbon moieties wherein two rings are joined and have two atoms in common bicyclo [3.2.1octane, bicyclo [2.2.1]hept-2-ene, norbomene, decalin, and the like), and the like.
The term "substituted cyclohydrocarbyl" refers to cyclohydrocarbyl moieties further bearing one or more substituents as set forth above.
As employed herein, "cycloalkyl" refers to ring-containing alkyl radicals containing in the range of about 3 up to 20 carbon atoms, and "substituted cycloalkyl" refers to cycloalkyl radicals further bearing one or more substituents as set forth above.
As employed herein, "cycloalkenyl" refers to ring-containing alkenyl radicals having at least one carbon-carbon double bond in the ring, and having in the range of about 3 up to 20 carbon atoms, and "substituted cycloalkenyl" refers to cyclic alkenyl radicals further bearing one or more substituents as set forth above.
As employed herein, "cycloalkynyl" refers to ring-containing alkynyl radicals having at least one carbon-carbon triple bond in the ring, and having in the range of about 3 up to 20 carbon atoms, and "substituted cycloalkynyl" refers to cyclic alkynyl radicals further bearing one or more substituents as set forth above.
As employed herein, "aryl" refers to mononuclear and polynuclear aromatic radicals having in the range of 6 up to 14 carbon atoms, and "substituted aryl" refers to aryl radicals further bearing one or more substituents as set forth above, for example, alkylaryl moieties.
8 As employed herein, "heterocycle" refers to ring-containing radicals having one or more heteroatoms N, O, S) as part of the ring structure, and having in the range of 3 up to 20 atoms in the ring. Heterocyclic moieties may be saturated or unsaturated when optionally containing one or more double bonds, and may contain more than one ring.
Heterocyclic moieties include, for example, monocyclic moieties such as imidazolyl moieties, pyrimidinyl moieties, isothiazolyl moieties, isoxazolyl moieties, and the like, and bicyclic heterocyclic moieties such as azabicycloalkanyl moieties, oxabicycloalkyl moieties, and the like. The term "substituted heterocycle" refers to heterocycles further bearing one or more substituents as set forth above.
As employed herein, "azo" refers to the bivalent moiety wherein each single bond is attached to a different carbon atom.
As employed herein, "halogen" refers to fluoride, chloride, bromide or iodide radicals.
Also disclosed herein are compounds wherein A is a 6- or 7-membered 15 unsaturated heterocyclic moiety, containing a ring having at least one nitrogen atom S. located on the ring in a position adjacent to a carbon atom which bears a linking moiety as a substituent. The ring further contains 3, 4 or 5 independently variable atoms selected from carbon, nitrogen, sulfur and oxygen. Thus, A can be pyridinyl, imidazolyl, pyridazinyl, pyrimidinyl, pyrazolyl, triazolyl, triazinyl, tetrazolyl, tetrazinyl, isoxazolyl, S 20 oxazolyl, oxadiazolyl, oxatriazolyl, oxadiazinyl, isothiazolyl, thiazolyl, dioxazolyl, oxathiazolyl, oxathiazinyl, azepinyl, diazepinyl, and the like. Those of skill in the art will recognize that multiple isomers exist for a single chemical formula; each of the possible isomeric forms of the various empirical formulae set forth herein are contemplated by this disclosure and the invention. When a variable ring atom is carbon, it bears a hydrogen, or is optionally substituted with halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, thiol, nitro, carboxyl, ester, cyano, amine, amide, carboxamide, amidine, amido, sulfonamide, and the like, with presently preferred embodiments having no substituent q is 0) or bearing the following substituents: halogen, alkyl containing one up to four carbon atoms, fluorinated alkyl, containing one up to four carbon atoms, aryl, and amine. Substitution at position Z of the ring is presently preferred.
Also disclosed herein are compounds wherein A is 6- or 7-membered ring containing, as ring members, a nitrogen atom and a sulfur atom. Moieties contemplated for use by this embodiment include those wherein A is isothiazol-3-yl (1,2-thiazol-3-yl), thiazol-4-yl (1,3-thiazol-4-yl), thiazol-2-yl (1,3-thiazol-2-yl), 1,2-thiazin-3-yl, 1,3-thiazin- 9 4-yl, 1,4-thiazin-3-yl, 1,3-thiazin-2-yl, thiazepinyl, and the like. Presently preferred moieties include those wherein A is isothiazol-3-yl (1,2-thiazol-3-yl), thiazol-4-yl (1,3thiazol-4-yl) and thiazol-2-yl (1,3-thiazol-2-yl).
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring s containing, as ring members, a nitrogen atom and an oxygen atom. Moieties contemplated by this embodiment include those wherein A is 1,2-oxazin-3-yl, 1,3-oxazin- 4-yl, 1,4-oxazin-3-yl, 1,3-oxazin-2-yl, oxazol-2-yl, isoxazol-3-yl, oxazol-4-yl, oxazepinyl, and the like. Presently preferred moieties include those wherein A is oxazol-2-yl, isoxazol-3-yl and oxazol-4-yl.
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring containing, as a ring member, a nitrogen atom. Moieties contemplated by this embodiment include those wherein A is 2-pyridinyl and 2-pyrrolyl.
*Also disclosed herein are compounds wherein A is a 6- or 7-membered ring *containing, as ring members, two nitrogen atoms. Moieties contemplated by this embodiment include those wherein A is 3-pyridazinyl (1,2-diazin-3-yl), pyrimidin-4-yl (1,3-diazin-4-yl), pyrazin-3-yl (1,4-diazin-3-yl), pyrimidin-2-yl (1,3-diazin-2-yl), pyrazol- 3-yl (1,2-diazol-3-yl), imidazol-4-yl (1,3-isodiazol-4-yl), imidazol-2-yl (1,3-isodiazol-2yl), diazepinyl, and the like. Presently preferred moieties include those wherein A is 3pyridazinyl (1,2-diazin-3-yl), pyrimidin-4-yl (1,3-diazin-4-yl), pyrazin-3-yl (1,4-diazin-3yl), pyrimidin-2-yl (1,3-diazin-2-yl), 1,3-isodiazol-4-yl and 1,3-isodiazol-2-yl.
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring containing, as ring members, three nitrogen atoms. Moieties contemplated by this embodiment include those wherein A is 1,2,3-triazin-4-yl, 1,2,4-triazin-6-yl, 1,2,4-triazin- 3-yl, 1,2,4-triazin-5-yl, 1,3,5-triazin-2-yl, 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl, triazepinyl, and the like. Presently preferred moieties include those wherein A is 1,2,3-triazin-4-yl, 1,2,4-triazin-6-yl, 1,2,4-triazin-3-yl, 1,2,4-triazin-5-yl, 1,3,5-triazin-2-yl, 1,2,3-triazol-4yl, 1,2,4-triazol-3-yl.
Also disclosed herein are compounds wherein A is a or 7-membered ring containing, as ring members, three nitrogen atoms. Moieties contemplated by this embodiment include those wherein A is tetrazin-2-yl, tetrazin-3-yl, tetrazolyl, tetrazepinyl, and the like. Presently preferred moieties include those wherein A is tetrazolyl.
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring containing, as ring members, one sulfur atom and two nitrogen atoms. Moieties contemplated by this embodiment include those wherein A is 1,2,6-thiadiazin-3-yl, 1,2,5thiadiazin-3-yl, 1,2,4-thiadiazin-3-yl, 1,2,5-thiadiazin-4-yl, 1,2,3-thiadiazin-4-yl, 1,3,4- 1,3,4-thiadiazin-2-yl, 1,2,4-thiadiazin-5-yl, 1,3,5-thiadiazin-4-yl, 1,3,5thiadiazin-2-yl, 1,2,4-thiadiazol-3-yl, 1,2,3-thiadiazol-4-yl, 1,3,4-thiadiazol-2-yl, 1,2,5thiadiazol-3-yl, 1,2,4-thiadiazol-5-yl, thiadiazepinyl, and the like. Presently preferred moieties include those wherein A is 1,2,4-thiadiazol-3-yl, 1,2,3-thiadiazol-4-yi, 1,3,4thiadiazol-2-yl, 1,2,5-thiadiazol-3-yl and 1,2,4-thiadiazol-5-yl.
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring containing, as ring members, one oxygen atom and two nitrogen atoms. Moieties to contemplated by this embodiment include those wherein A is 1,2,6-oxadiazin-3-yl, 1,2,5oxadiazin-3-yl, 1,2,4-oxadiazin-3-yl, 1,2,5-oxadiazin-4-yl, 1,2,3-oxadiazin-4-yl, 1,3,4- 1,3,4-oxadiazin-2-yl, 1,2,4-oxadiazin-5-yl, 1,3,5-oxadiazin-4-yl, 1,3,5oxadiazin-2-yl, 1,2,4-oxadiazol-3-yl, 1,2,3-oxadiazol-4-yl, 1,3,4-oxadiazol-2-yl, 1,2,5oxadiazol-3-yl, 1,2,4-oxadiazol-5-yl, oxadiazepinyl, and the like. Presently preferred S 15s moieties include those wherein A is 1,2,4-oxadiazol-3-yl, 1,2,3-oxadiazol-4-yl, 1,3,4- :oxadiazol-2-yl, 1,2,5-oxadiazol-3-yl and 1,2,4-oxadiazol-5-yl.
Also disclosed herein are compounds wherein A is a 6- or 7-membered ring containing as ring members, one up to six nitrogen atoms, and/or one up to six carbon atoms, and/or zero up to five sulfur atoms, and/or zero up to five oxygen atoms.
20 In accordance with the present invention L is a linking moiety which links moieties A and B. L is selected from substituted or unsubstituted alkenylene moieties, alkynylene moieties or azo moieties. Presently preferred compounds of the invention are those wherein L is alkenylene or alkynylene moieties containing two carbon atoms, with alkynylene most preferred.
Also in accordance with the present invention B is a moiety linked through bridging moiety L to moiety A. Radicals contemplated for use in the invention are those wherein B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, substituted or unsubstituted aryl, and the like.
Presently preferred compounds disclosed herein are those wherein B is a substituted or unsubstituted hydrocarbyl selected from substituted or unsubstituted alkyl moieties, alkenyl moieties, dialkenyl moieties, trialkenyl moieties, alkynyl moieties, alkadiynyl moieties, alkatriynyl moieties, alkenynyl moieties, alkadienynyl moieties, alkenediynyl moieties, and the like.
11 Also disclosed herein are compounds wherein B is a substituted or unsubstituted cyclohydrocarbyl selected from substituted or unsubstituted cycloalkyl moieties, cycloalkefnyl moieties, cycloalkadienyl moieties, cycloalkatrienyl moieties, cycloalkynyl moieties, cycloalkadiynyl moieties, bicyclic hydrocarbon moieties wherein two rings have two atoms in common, and the like. Especially preferred compounds are those wherein B is cycloalkyl and cycloalkenyl having in the range of 4 up to about 8 carbon atoms.
Exemplary compounds include cyclopropanyl, cyclopentenyl and cyclohexenyl. Also especially preferred are bicyclic hydrocarbon moieties wherein two rings have two atoms in common; exemplary compounds include indenyl, dihydroindenyl, naphthalenyl and dihydronaphthalenyl.
Still further preferred compounds disclosed herein are those wherein B is a substituted or unsubstituted heterocycle, optionally containing one or more double bonds.
SExemplary compounds include pyridyl, thiazolyl, furyl, dihydropyranyl, *dihydrothiopyranyl, piperidinyl, isoxazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, and the 15 like. Also preferred are compounds wherein B is substituted or unsubstituted aryl.
Especially preferred compounds are those wherein substituents are aryl and heterocycle, optionally bearing further substituents as described herein, methyl, trifluoromethyl, cyclopropyl, alkoxy, halogen and cyano. Also preferred are compounds wherein B is a bicyclic heterocycle moiety wherein two rings have two atoms in common. Exemplary p 20 compounds include indolyl and isoquinolinyl.
Those of skill in the art recognize that compounds disclosed herein may contain one or more chiral centres, and thus can exist as racemic mixtures. For many applications, it is preferred to carry out stereoselective syntheses and/or to subject the reaction product to appropriate purification steps so as to produce substantially optically pure materials.
Suitable stereoselective synthetic procedures for producing optically pure materials are well known in the art, as are procedures for purifying racemic mixtures into optically pure fractions. Those of skill in the art will further recognize that compounds disclosed herein may exist in polymorphic forms wherein a compound is capable of crystallizing in different forms. Suitable methods for identifying and separating polymorphisms are known in the art.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, esterified carboxy is, for example, lower alkoxycarbonyl, phenyl-lower alkoxycarbonyl or phenyl-lower alkoxycarbonyl substituted in the phenyl moiety by one or more substituents selected from lower alkyl, lower alkoxy, halo and halo-lower alkyl. Esterified carboxy-lower-alkoxy is, for 12 example, lower alkoxycarbonyl-lower-alkoxy. Amidated carboxy is, for example, unsubstituted or aliphatically substituted carbamoyl such as carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl unsubstituted or lower alkyl-, lower alkoxy-, halo- and/or trifluoromethyl-substituted N-phenyl- or N-lower-alkyl-N-phenyls carbamoyl.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, acyl is, for example, lower alkanoyl, lower alkenoyl or unsubstituted or lower alkyl-, lower alkoxy-, halo- and/or trifluoromethyl-substituted benzoyl. Acylamino is, for example, lower alkanoylamino, to and N-acyl-N-lower alkylamino is, for example, N-lower alkanoyl-N-lower-alkylamino or unsubstituted or lower alkyl-, lower alkoxy-, halo- and/or trifluoromethyl-substituted benzoylamino.
As referred to in reference to compounds excluded herein when referring to *compounds of formula A-L-B disclosed herein, "lower" groups are understood to t I s comprise up to and including seven carbon atoms. N-lower-alkyl-N-phenylcarbamoyl is, for example, N-CI-C 4 alkyl-N-phenylcarbamoyl, such as N-methyl, N-ethyl, N-propyl, N-isopropyl or N-butyl-N-phenylcarbamoyl.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, amino-lower alkyl is, for example, I I S. 5 20 amino-Cl-C 4 alkyl, preferably of the formula -(CH 2 2 in which n is 2 or 3, such as aminomethyl, 2-aminoethyl, 3-aminopropyl or 4-aminobutyl. Hydroxy-lower alkyl is, for C. a example, hydroxy-C 1
-C
4 alkyl, such as hydroxymethyl, 2-hydroxy ethyl, 3-hydroxypropyl, 2-hydroxyisopropyl or 4-hydroxybutyl. Halo-lower alkyl is, for example, polyhalo-Cisee. C 4 alkyl, such as trifluoromethyl.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, lower alkoxy is, for example, C 1
C
7 alkoxy, preferably C 1
-C
4 alkoxy, such as methoxy, ethoxy, propyloxy, isopropyloxy or butyloxy, but may also represent isobutyloxy, sec.butyloxy, tert.-butyloxy or a Cs-
C
7 alkoxy group, such as a pentyloxy, hexyloxy or heptyloxy group. Amino-lower alkoxy is, for example, amino-C 2
-C
4 alkoxy preferably of the formula -O-(CH 2 )n-NRaR, in which n is 2 or 3, such as 2-aminoethoxy, 3-aminopropyloxy or 4-aminobutyloxy. Carboxylower-alkoxy is, for example, carboxy-CI-C4alkoxy, such as carboxymethoxy, 2-carboxyethoxy, 3-carboxypropyloxy or 4-carboxybutyloxy. Lower alkanoyloxy is, for example, C-C 7 alkanoyloxy, such as acetoxy, propionyloxy, butyryloxy, isobutyryloxy, or pivaloyloxy. Halo-lower alkoxy is, for example, halo- or polyhalo-C 1
-C
7 alkoxy, 13 preferably halo- or polyhalo-C 1
-C
4 alkoxy, such as halo- or polyhaloethoxy, halo- or polyhalopropyloxy or butyl-oxy, wherein "poly" refers, for example, to tri- or pentahalo, and "halo" denotes, for example, fluoro or chloro.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, lower alkylamino-lower alkoxy is, for example, Ci-C 4 alkylamino-C 2
-C
4 alkoxy, preferably of the formula -O-(CH 2 )n-NRaRb in which n is 2 or 3 and Ra and Rb, independently of each other, denote lower alkyl groups as defined hereinbefore, such as methyl, ethyl, propyl or butyl. Lower alkylamino-lower alkyl is, for example, Ci-C 4 alkylamino-Ci-C 4 alkyl, preferably of the formula -(CH 2 )n- NRaRb in which n is 2 or 3 and Ra and Rb, independently of each other, denote lower alkyl groups as defined hereinbefore, such as methyl, ethyl, propyl or butyl. Di-lower alkylamino-lower alkyl is, for example, di-Ci-C 4 alkylamino-Ci-C 4 alkyl, preferably of the formula -(CH 2 )n-NRaRb in which n is 2 or 3 and Ra and Rb, independently of each other, denote lower alkyl groups such as methyl, ethyl, propyl or butyl. Di-lower alkylaminolower alkoxy is, for example, di-C 1
-C
4 alkylamino-C 2
-C
4 alkoxy, preferably of the formula
-O-(CH
2 )n-NRaRb in which n is 2 or 3 and Ra and Rb, independently of each other, denote lower alkyl groups such as methyl, ethyl, propyl or butyl.
As used herein, with reference to compounds excluded herein when referring to compounds of formula A-L-B disclosed herein, optionally hydroxy-substituted lower 20 alkyleneamino-lower alkyl is, for example, unsubstituted or hydroxy-substituted 5- to 7membered alkyleneamino-C 1
-C
4 alkyl, preferably of the formula -(CH 2 in which n is S* 2 or 3 and R, is pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino, homopiperidino or hydroxyhomopiperidino. Furthermore, optionally hydroxy-substituted lower alkyleneamino-lower alkoxy is, for example, unsubstituted or hydroxy-substituted 5- to 7-membered alkyleneamine-C 1
-C
4 alkoxy, preferably of the formula -O-(CH 2 )n-R in which n is 2 or 3 and Re is pyrrolidino, hydroxypyrrolidino, piperidino, hydroxypiperidino, homopiperidino or hydroxyhomopiperidino.
Also disclosed herein are pharmaceutical compositions comprising heterocyclic compounds as described above, in combination with pharmaceutically acceptable carriers.
Optionally, compounds disclosed herein can be converted into non-toxic acid addition salts, depending on the substituents thereon. Thus, the above-described compounds (optionally in combination with pharmaceutically acceptable carriers) can be used in the manufacture of medicaments useful for the treatment of a variety of indications.
13a Pharmaceutically acceptable carriers contemplated for use in the practice of the present invention include carriers suitable for oral, sublingual intravenous, subcutaneous, transcutaneous, intramuscular, intracutaneous, intrathecal, epidural, intraocular, intracranial, inhalation, rectal, vaginal, and the like administration. Administration in the s form of creams, lotions, tablets, capsules, pellets, dispersible powders, granules, suppositories, syrups, elixirs, lozenges, injectable solutions, sterile aqueous or nonaqueous solutions, suspensions or emulsions, patches, and the like, is contemplated.
Pharmaceutically acceptable carriers include glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, collidal silica, to potato starch, urea, dextrans, and the like.
Compounds disclosed herein can optionally be converted into non-toxic acid addition salts. Such salts are generally prepared by reacting the compounds disclosed herein with a suitable organic or inorganic acid. Representative salts include hydrochloride, hydrobromide, sulfate, bisulfate, methanesulfonate, acetate, oxalate, adipate, alginate, aspartate, valerate, oleate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate (tosylate), citrate, malate, maleate, fumarate, succinate, tartrate, napsylate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, benzenesulfonate, butyrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, glucoheptanoate, glycerophosphate, heptanoate, hexanoate, undecanoate, 20 2-hydroxyethanesulfonate, ethanesulfonate, and the like. Salts can also be formed with inorganic acids such as sulfate, bisulfate, hemisulfate, hydrochloride, chlorate, perchlorate, hydrobromide, hydroiodide, and the like. Examples of a base salt include ammonium salts; alkali metal salts such as sodium salts, potassium salts, and the like; alkaline earth metal salts such as calcium salts, magnesium salts, and the like; salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, phenylethylamine, and the like; and salts with amino acids such as arginine, lysine, and the like. Such salts can readily be prepared employing methods well known in the art.
Also disclosed herein are methods 14 for the preparation of heterocyclic compounds as described above. For example, many of the heterocyclic compounds described above can be prepared using synthetic chemistry techniques well known in the art (see Comprehensive Heterocyclic Chemistry, Katritzky, A. R. and Rees, C. W. eds., Pergamon Press, Oxford, 1984) from a precursor of the substituted heterocycle of Formula 1 as outlined in Scheme 1.
Scheme 1 (R)q V E B Coupling_ (R)q Y Z N D B B
B
Thus in Scheme 1, a substituted heterocycle precursor (prepared using synthetic chemistry techniques well known in the art) is reacted with an alkyne derivative. In Scheme 1, W, X, Y, Z and B are as defined above and D and E are functional groups which are capable of undergoing a transition metal-catalyzed cross-coupling reaction. For example, D is a group such as hydrogen, 10 halogen, acyloxy, fluorosulfonate, trifluoromethanesulfonate, alkyl- or arylsulfonate, alkyl- or arylsulfinate, alkyl- or arylsulfide, phosphate, phosphinate, and the like, and E is hydrogen or a metallic or metalloid species such as Li, MgX (X is halogen), SnR 3
B(OR)
2 SiR 3 GeR 3 and the like. The coupling may be promoted by a homogeneous catalyst such as PdCI 2 (PPh 3 2 or by a heterogeneous catalyst such as Pd on carbon in a suitable solvent tetrahydrofuran (THF), dimethoxyethane 15 (DME), acetonitrile, dimethylformamide (DMF), etc.). Typically, a co-catalyst such as copper (I) iodide and a base triethylamine, K 2 CO3 etc.) will also be present in the reaction mixture. The coupling reaction typically proceeds by allowing the reaction temperature to warm slowly from about 0° C up to ambient temperature over a period of several hours. The reaction mixture is then maintained at S ambient temperature, or heated to a temperature anywhere between 30° C and 1500 C. The reaction 20 mixture is then maintained at a suitable temperature for a time in the range of about 4 up to 48 hours, with about 12 hours typically being sufficient. The product from the reaction can be isolated and purified employing standard techniques, such as solvent extraction, chromatography, crystallization, distillation, and the like.
Another procedure is illustrated in Scheme 2. A substituted heterocycle precursor is reacted with an alkene derivative in a manner similar to the procedure described for Scheme 1.
Scheme 2 X. oupling (R)q E B Coupling (R)q N' D B The alkene derivative product from Scheme 2 may be converted to an alkyne derivative using the approach outlined in Scheme 3.
Scheme 3 (R)q B Halogenation (R)q G Base (R)qY W z- N j B
N
G B 5 Thus, the alkene derivative may be contacted with a halogenating agent such as chlorine, bromine, iodine, NCS (N-chlorosuccinimide), NBS (N-bromosuccinimide), NIS (N-iodosuccinimide), iodine monochloride, etc. in a suitable solvent (CC4, CHCI 3
CH
2 C12, acetic acid, and the like). The resulting halogenated derivative (G halogen) is then treated with a suitable base such as NaOH, KOH, S: DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), DBN (diazabicyclononene), DABCO (1,4-diazabicyclo[2.2.2]octane), and the like, which promotes a double elimination reaction to afford S the alkyne. The reaction is carried out in a suitable solvent such as ethanol, acetonitrile, toluene, etc. at an appropriate temperature, usually between about 0° C and 150° C.
e In accordance with another procedure disclosed herein, a substituted heterocyclic derivative is reacted with an aldehyde or ketone to provide a substituted alkene. (See Scheme 4).
Scheme 4 Y X. W 0 B Cty Y'.W OR
W
(R)q O B Catalyst (R)q W (R)qiOR N K B N-BB
J
Thus, in Scheme 4, J is hydrogen, PR 3
P(O)(OR)
2
SO
2 R, SiR 3 and the like, K is hydrogen, alkyl or aryl (as defined previously) and R is hydrogen, acetyl, and the like. Suitable catalysts for this reaction include bases such as NaH, n-buytllithium, lithium diisopropylamide, lithium hexamethyl disilazide, H 2 NR, HNR 2
NR
3 etc., or electropositive reagents such as Ac 2 0, ZnCl 2 and the like. The reaction is carried out in a suitable solvent (THF, acetonitrile, etc.) at an appropriate temperature, usually between about 0* C and 1500 C. Sometimes an intermediate is isolated and purified or partially purified before continuing through to the alkene product.
16 In another procedure disclosed herein, a substituted heterocyclic aldehyde or ketone is reacted with an activated methylene-containing compound to provide a substituted alkene. (See Scheme Scheme
X
(R)q 0
K
rB Catalyst
J
(R)q.JY W 10 10 Thus, in Scheme 5, J, K, R, the-catalyst and reaction conditions are as described for Scheme 4.
Again, as in Scheme 4, sometimes an intermediate is isolated and purified or partially purified before continuing through to the alkene product.
The alkene products from the reactions in Scheme 4 and Scheme 5 may be converted to an alkyne derivative using reagents and conditions as described for Scheme 3.
Another method for the preparation of heterocyclic compounds of Formula I is depicted in Scheme Scheme 6 see .00: .00.
0 (R)q NH )q 0 L-B Heat JR)q (R)q N L BB Y= O S In scheme 6, Y is O or S and G is halogen or a similar leaving group, and L and B are as defined previously. The reagents are contacted in a suitable solvent such as ethanol, DMF, and the like and stirred until the product forms. Typically reaction temperatures will be in the range of ambient through to about 1500 C, and reaction times will be from about 1 h to about 48 h, with about 700 C and 4 h being presently preferred. The heterocycle product can be isolated and purified employing standard techniques, such as solvent extraction, chromatography, crystallization, distillation, and the like. Often, the product will be isolated as the hydrochloride or hydrobromide salt, and this material may be carried onto the next step with or without purification.
Yet another method for the preparation of heterocyclic compounds of Formula I is depicted in Scheme 7.
17 Scheme 7 (R)q G W Heat (R)q S HN -B W O, S (R)q O H (R)q N L-B In Scheme 7, W may be O or S, G is halogen or a similar leaving group, and L and B are as defined previously. The reaction conditions and purification procedures are as described for Scheme 6.
In another procedure disclosed herein, depicted in Scheme 8, an alkynyl-substituted heterocycle precursor (prepared using synthetic chemistry techniques well known in the art) is reacted with a species B, bearing a reactive functional group D (See Scheme 8).
Scheme 8 N+ D B N Coung
**B
•In Scheme 8, W, X, Y,Z and B are as defined above and D and E are functional groups which are capable of undergoing a transition metal-catalyzed cross-coupling reaction. For example, D is a group such as hydrogen, halogen, acyloxy, fluorosulfonate, trifluoromethanesulfonate, alkyl- or S arylsulfonate, alkyl- or arylsulfinate, alkyl- or arylsulfide, phosphate, phosphinate, and the like, and E is S: hydrogen or a metallic or metalloid species such as Li, MgX (X is halogen), SnR 3
B(OR)
2 SiR 3 GeR 3 and the like. The coupling may be promoted by a homogeneous catalyst such as PdCI 2 (PPh 3 2 or by a heterogeneous catalyst such as Pd on carbon in a suitable solvent tetrahydrofuran (THF), dimethoxyethane (DME), acetonitrile, dimethylformamide (DMF), etc.). Typically a co-catalyst such as copper iodide and the like and a base triethylamine, K 2 CO3. etc.) will also be present in the reaction mixture. The coupling reaction is typically allowed to proceed by allowing the reaction temperature to warm slowly from about 0* C up to ambient temperature over a period of several hours.
The reaction mixture is then maintained at ambient temperature, or heated to a temperature anywhere between about 300 C up to about 1500 C. The reaction mixture is then maintained at a suitable temperature for a time in the range of about 4 up to about 48 hours, with about 12 hours typically being sufficient The product from the reaction can be isolated and purified employing standard techniques, such as solvent extraction, chromatography, crystallization, distillation, and the like.
Another procedure is illustrated in Scheme 9.
18 Scheme 9 (R)q Y Coupling (R)Y W Z-I E D (R)q zB An alkenyl-substituted heterocycle precursor is reacted with an alkene derivative in a manner similar to the procedure described for Scheme 8. The product alkene derivative from Scheme 9 may be converted to an alkyne derivative using the approach outlined previously in Scheme 3 above.
In yet another procedure disclosed herein, depicted in Scheme 10, an alkynyl-substituted heterocycle precursor is reacted with a species composed of a carbonyl group bearing substituents R' and CHR"R"'.
Scheme O Catal (R)q R"' z'N R 'R N OR
N
E R' RR" R" R'
R
Thus in Scheme 10, R" and may be hydrogen or other substituents as described 10 previously, or may optionally combine to form a ring (this portion of the molecule constitutes B in the final compound). E is hydrogen or a metallic or metalloid species such as Li, MgX, wherein X is halogen, SnR 3
B(OR)
2 SiR 3 GeR 3 and the like. Suitable catalysts for this reaction include bases such as NaH, n-butyllithium, lithium diisopropylamide, lithium hexamethylsilazide, H 2 NR, HNR 2
NR
3 nBu4NF, ethylmagnesium halide, etc. R in Scheme 10 may be hydrogen, Ac, and the like. Typically 15 the reaction is carried out in a suitable solvent such as diethylether, THF, DME, toluene, and the like, and at an appropriate temperature, usually between -100° C and 25* C. The reaction is allowed to proceed for an appropriate length of time, usually from about 15 minutes to about 24 hours. The intermediate bearing the -OR group may be isolated and purified as described above, partially purified or carried on to the next step without purification. Elimination of the -OR group to provide the alkene derivative may be accomplished using a variety of methods well known to those skilled in the art. For example, the intermediate may be contacted with POCI3 in a solvent such as pyridine and stirred at a suitable temperature, typically between about 00 C and about 150* C, for an appropriate amount of time, usually between about 1 h and about 48 h. The product from the reaction can be isolated and purified employing standard techniques, such as solvent extraction, chromatography, crystallization, distillation, and the like.
19 Also disclosed herein are methods of modulating the activity of excitatory amino acid receptors, said method comprising contacting said receptors with at least one comipound as described above, as well as additional compounds of the same basic structure but having substitution patterns which may have been previously disclosed but never contmplated for use for such purposes. Thus, compounds contemplated for use in accordance with methods disclosed herein include those having the structure A-L-B or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, in an amount sufficient to modulate the activity of said excitatory amino acid receptor, wherein: A is a 6- or 7-membered ring having the structure:
V
wherein at least one of V, W, X, Y and Z is (CR)p, whereifp is 0, 1 or 2; at least one of V, W, X, Y and Z is O, N or S; the remainder of V, W, X, Y and Z are each independently O, N or S; and each R is independently halogen, substituted or unsubstituted 15 hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl or sulfonamide, wherein q is 0, 1, 2 or 3; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted .20 cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted or unsubstituted aryl; provided, that the following compounds are excluded: the compounds wherein A is a 6-membered ring wherein: V, W, X and Y are (CR)p, wherein p is 1, ZisN; R at the V position is hydrogen, lower alkyl, hydroxy, hydroxy-lower alkyl, amino-lower alkyl, lower alkylamino-lower alkyl, di-lower alkylamino-lower alkyl, unsubstituted or hydroxy-substituted lower alkyleneamino-lower alkyl, lower alkoxy, lower alkanoyloxy, amino-lower alkoxy, lower alkylamino-lower alkoxy, di-lower alkylamino-lower alkoxy, phthalimido-lower alkoxy, unsubstituted or hydroxy- or 2-oxo-imidazolidin-l-yl-substitued lower alkyleneamino-lower alkoxy, carboxy, esterified or amidated carboxy, carboxy-lower alkoxy or esterified carboxy-lower-alkoxy; R at WO 01/16121 PCT/US00/23923 the W position is hydrogen; R at the X position is hydrogen, lower alkyl, carboxy, esterified carboxy, amidated carboxy, hydroxy-lower alkyl, hydroxy, lower alkoxy or lower alkanoyloxy; and R at the Y position is hydrogen, lower alkyl, hydroxy-lower alkyl, carboxy, esterified carboxy, amidated carboxy, unsubstituted or lower alkyl-, lower alkoxy-, halo- and/or trifluoromethyl-substituted N-lower alkyl-Nphenylcarbamoyl, lower alkoxy, halo-lower alkyl or halo-lower alkoxy; L is substituted or unsubstituted alkenylene, alkynylene or azo, and B is substituted or unsubstituted aryl or heterocycle having two or more double bonds, wherein substituents are independently lower alkyl, lower alkenyl, lower alkynyl, phenyl, phenyl-lower alkynyl, hydroxy, hydroxy-lower alkyl, lower alkoxy, lower alkenyloxy, lower alkylenedioxy, lower alkanoyloxy, phenoxy, phenyl-lower alkoxy, acyl, carboxy, esterified carboxy, amidated carboxy, cyano, nitro, amino, acylamino, N-acyl-N-lower alkylamino, halo and halo-lower alkyl, wherein phenyl, phenyl-lower alkynyl, phenoxy, and phenyl-lower alkoxy may bear further substituents.
As employed herein, "excitatory amino acid receptors" refers to a class of cell-surface receptors which are the major class of excitatory neurotransmitter receptors in the central nervous system. In addition, receptors of this class also mediate inhibitory responses. Excitatory amino acid receptors are membrane spanning proteins that mediate the stimulatory actions of the amino acid glutamate and possibly other endogenous acidic amino acids. Excitatory amino acids are crucial for fast and slow neurotransmission and they have been implicated in a variety of diseases including Alzheimer's disease, stroke, schizophrenia, head trauma, epilepsy, and the like. In addition, excitatory amino acids are integral to the processes of long-term potentiation and depression which are synaptic mechanisms underlying learning and memory. There are three main subtypes of excitatory amino acid receptors: (1) the metabotropic receptors; the ionotropic NMDA receptors; and the non-NMDA receptors, which include the AMPA receptors and kainate receptors.
As employed herein, the phrase "modulating the activity of" refers to altered levels of activity so that the activity is different with the use of the invention method when compared to the activity without the use of the invention method. Modulating the activity of excitatory amino acid receptors includes the suppression or augmentation of the activity of receptors. Suppression of receptor activity may be accomplished by a variety of means, including blocking of a ligand binding site, biochemical and/or physico-chemical modification of a ligand binding site, binding of agonist recognition domains, preventing ligand-activated conformational changes in the receptor, preventing the activated receptor from stimulating second messengers such as G-proteins, and the like. Augmentation of receptor activity may be accomplished by a variety of means including, stabilization of a ligand binding site, biochemical and/or physico-chemical modification of a ligand binding site, binding of agonist recognition domains, promoting ligand-activated conformational changes in the receptor, and the like.
WO 01/16121 PCT/US00/23923 21 Excitatory amino acid receptor activity can be involved in numerous disease states. Therefore modulating the activity of receptors also refers to a variety of therapeutic applications, such as the treatment of cerebral ischemia, chronic neurodegeneration, psychiatric disorders, schizophrenia, mood disorders, emotion disorders, disorders of extrapyramidal motor function, obesity, disorders of respiration, motor control and function, attention deficit disorders, concentration disorders, pain disorders, neurodegenerative disorders, epilepsy, convulsive disorders, eating disorders, sleep disorders, sexual disorders, circadian disorders, drug withdrawal, drug addiction, compulsive disorders, anxiety, panic disorders, depressive disorders, skin disorders, retinal ischemia, retinal degeneration, glaucoma, disorders associated with organ transplantation, asthma, ischemia or astroytomas, and the like.
The compounds contemplated for use in accordance with of invention modulatory methods are especially useful for the treatment of mood disorders such as anxiety, depression, psychosis, drug withdrawal, tobacco withdrawal, memory loss, cognitive impairment, dementia, Alzheimer's disease, and the like; disorders of extrapyramidal motor function such as Parkinson's disease, progressive supramuscular palsy, Huntington's disease, Gilles de la Tourette syndrome, tardive dyskinesia, and the like.
Compounds contemplated for use in accordance with invention modulatory methods are also especially useful for the treatment of pain disorders such as neuropathic pain, chronic pain, acute pain, painful diabetic neuropathy, post-herpetic neuralgia, cancer-associated pain, pain associated with chemotherapy, pain associated with spinal cord injury, pain associated with multiple sclerosis, causalgia and reflex sympathetic dystrophy, phantom pain, post-stroke (central) pain, pain associated with HIV or AIDS, trigeminal neuralgia, lower back pain, myofacial disorders, migraine, osteoarthritic pain, postoperative pain, dental pain, post-bum pain, pain associated with systemic lupus, entrapment neuropathies, painful polyneuropathies, ocular pain, pain associated with inflammation, pain due to tissue injury, and the like.
"Contacting" may include contacting in solution or in solid phase.
"Pharmaceutically acceptable salt" refers to a salt of the compound used for treatment which possesses the desired pharmacological activity and which is physiologically suitable. The salt can be formed with organic acids such as acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, heptanoate, hexanoate, 2-hydroxyethanesulfonate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, tartrate, toluenesulfonate, undecanoate, and the like. The salt can also be formed with inorganic acids such as sulfate, bisulfate, chlorate, perchlorate, hemisulfate, hydrochloride, hydrobromide, hydroiodide, and the like. In addition, the salt can be formed with a base salt, including ammonium salts, alkali metal salts such as sodium salts, potassium salts, and the like; alkaline earth metal salts such as calcium salts, magnesium salts, and the like; salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, phenylethylamine, and the like; and sals with amino acids such as arginine, lysine, and the like.
Salt forms of compounds herein find several advantages. Certain pharmaceutically acceptable salt forms of heterocyclic compounds described herein, achieve higher solubility as compared with nonsalt forms. In addition, certain salt forms are more compatible with pharmaceutical uses. For example, the hydrochloric acid salt of 2-(phenylethynl)-l,3-thiazole is an oil while the toluene sulfonic acid salt form of 2-(phenylethynl)-l,3-thiazole is a solid that is soluble in aqueous medium. (See Example 187.) Characteristics of salt forms of compounds depend on the characteristics of the compound so treated, and on the particular salt employed.
Also disclosed herein are methods of S modulating the activity of metabotropic glutamate receptors, said method comprising contacting 0.
metabotropic glutamate receptors with a concentration of a heterocylic compound as described above in accordance with invention methods for modulating the acitivity of excitatory amino acid receptors, o sufficient to modulate the activity of said metabotropic glutamate receptors.
As used herein, the phrase "metabotropic glutamate receptor" refers to a class of cell-surface receptors which participates in the G-protein-coupled response of cells to glutamatergic ligands. Three .0 groups of metabotropic glutamate receptors, identified on the basis of amino acid sequence homology, transduction mechanism and binding selectivity are presently known and each group contains one or more types of receptors. For example, Group I includes metabotropic glutamate receptors 1 and (mGluRI and mGluR5), Group II includes metabotropic glutamate receptors 2 and 3 (mGluR2 and mGluR3) and Group 1Il includes metabotropic glutamate receptors 4, 6, 7 and 8 (mGluR4, mGluR6, mGluR7 and mGluR8). Several subtypes of each mGluR type may be found; for example, subtypes of mGluRl include mGluRla, mGluRlb and mGluRlc.
Also disclosed herein are methods of treating a wide variety of disease conditions, said method comprising administering to a patient having a disease condition a therapeutically effective amount of at least one of the heterocyclic compounds described above in accordance with invention methods for modulating the activity of excitatory amino acid receptors.
As used herein, "treating" refers to inhibiting or arresting the development of a disease, disorder or condition and/or causing the reduction, remission, or regression of a disease, disorder or condition. Those of skill in the art will understand that various methodologies and assays may be used to assess the development of a disease,.disorder or condition, and similarly, various methodologies and assays may be used to assess the reduction, remission or regression of a disease, disorder or condition.
WO 01/16121 PCT/US00/23923 23 Disease conditions contemplated for treatment in accordance with the invention include cerebral ischemia, chronic neurodegeneration, psychiatric disorders, schizophrenia, mood disorders, emotion disorders, disorders of extrapyramidal motor function, obesity, disorders of respiration, motor control and function, attention deficit disorders, concentration disorders, pain disorders, neurodegenerative disorders, epilepsy, convulsive disorders, eating disorders, sleep disorders, sexual disorders, circadian disorders, drug withdrawal, drug addiction, compulsive disorders, anxiety, panic disorders, depressive disorders, skin disorders, retinal ischemia, retinal degeneration, glaucoma, disorders associated with organ transplantation, asthma, ischemia, astrocytomas, and the like.
Disease conditions contemplated for treatment in accordance with the present invention further include diseases of the pulmonary system, diseases of the nervous system, diseases of the cardiovascular system, diseases of the gastrointestinal system, diseases of the endocrine system, diseases of the exocrine system, diseases of the skin, cancer, diseases of the ophthalmic system, and the like.
As used herein, "administering" refers to means for providing heterocyclic compounds and/or salts thereof, as described herein, to a patient, using oral, sublingual intravenous, subcutaneous, transcutaneous, intramuscular, intracutaneous, intrathecal, epidural, intraoccular, intracranial, inhalation, rectal, vaginal, and the like administration. Administration in the form of creams, lotions, tablets, capsules, pellets, dispersible powders, granules, suppositoiries, syrups, elixirs, lozenges, injectable solutions, sterile aqueous or non-aqueous solutions, suspensions or emulsions, patches, and the like, is also contemplated. The active ingredients may be compounded with non-toxic, pharmaceutically acceptable carriers including, glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, dextrans, and the like.
For purposes of oral administration, tablets, capsules, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, elixirs and lozenges containing various excipients such as calcium carbonae, lactose, calcium phosphate, sodium phosphate, and the like may be employed along with various granulating and disintegrating agents such as corn starch, potato starch, alginic acid, and the like, together with binding agents such as gum tragacanth, corn starch, gelatin, acacia, and the like. Lubricating agents such as magnesium stearate, stearic acid, talc, and the like may also be added. Preparations intended for oral use may be prepared according to any methods known to the art for the manufacture of pharmaceutical preparations and such preparations may contain one or more agents selected from the group consisting of a sweetening agent such as sucrose, lactose, saccharin, and the lake, flavoring agents such as peppermint, oil of wintergreen, and the like, coloring agents and preserving agents in order to provide pharmaceutically palatable preparations.
Preparations for oral use may also contain suitable carriers include emulsions, solutions, suspensions, syrups, and the like, optionally containing additives such as wetting agents, emulsifying and suspending WO 01/16121 PCT/US00/23923 24 agents, sweetening, flavoring and perfuming agents, and the like. Tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period of time.
For the preparation of fluids for parentcral administration, suitable carriers include sterile aqueous or non-aqueous solutions, suspensions, or emulsions. For parenteral administration, solutions for the practice of the invention may comprise sterile aqueous saline solutions, or the corresponding water soluble pharmaceutically acceptable metal salts, as previously described. For parenteral administration, solutions of the compounds used in the practice of the invention may also comprise non-aqueous solutions, suspensions, emulsions, and the like. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Such dosage forms may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. They may be sterilized, for example, by filtration through a bacteria-retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. They can also be manufactured in the form of sterile water, or some other sterile injectable medium immediately before use.
Aqueous solutions may also be suitable for intravenous, intramuscular, intrathecal, subcutaneous, and intraperitoneal injection. The sterile aqueous media employed are all readily obtainable by standard techniques well known to those skilled in the art. They may be sterilized, for example, by filtration through a bacteria-retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, by heating the compositions, and the like. They can also be manufactured in the form of sterile water, or some other sterile medium capable of injection immediately before use.
Compounds contemplated for use in accordance with the present invention may also be administered in the form of suppositories for rectal or vaginal administration. These compositions may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters of polyethylene glycols, and the like, such materials being solid at ambient temperatures but liquify and/or dissolve in internal cavities to release the drug.
The preferred therapeutic compositions for inocula and dosage will vary with the clinical indication. Some variation in dosage will necessarily occur depending upon the condition of the patient being treated, and the physician will, in any event, determine the appropriate dose for the individual patient. The effective amount of compound per unit dose depends, among other things, on the body weight, physiology, and chosen inoculation regimen. A unit dose of compound refers to the weight of compound without the weight of carrier (when carrier is used).
The route of delivery of compounds and compositions used for the practice of the invention is determined by the disease and the site where treatment is required. Since the pharmacokinetics and pharmacodynamics of compounds and compositions described herein will vary somewhat, the most preferred method for achieving a therapeutic concentration in a tissue is to gradually escalate the dosage and monitor the clinical effects. The initial dose, for such an escalating dosage regimen of therapy, will depend upon the route of administration.
In accordance with methods disclosed herein, the medicinal preparation can be introduced parenterally, by dermal application, and the like, in any medicinal form or composition. It is used as a solitary agent of medication or in combination with other medicinal preparations. Single and multiple therapeutic dosage regimens may prove useful in therapeutic protocols.
As employed herein, the phrase "a therapeutically effective amount", when used in reference to invention methods employing heterocyclic compounds and pharmaceutically acceptable salts thereof, refers to a dose of compound sufficient to provide circulating concentrations high enough to impart a beneficial effect on the recipient thereof. The specific therapeutically effective dose level for any 4, 15 particular patient will depend upon a variety of factors including the disorder being treated, the severity o* of the disorder, the activity of the specific compound used, the route of administration, the rate of clearance of the specific compound, the duration of treatment, the drugs used in combination or coincident with the specific compound, the age, body weight, sex, diet and general health of the patient, es- and like factors well known in the medical arts and sciences. Dosage levels typically fall in the range of about 0.001 up to 100 mg/kg/day; with levels in the range of about 0.05 up to 10 mg/kg/day being *b*4 d preferred.
Also disclosed herein are methods for preventing disease ,t conditions in a subject at risk thereof, said method comprising administering to said subject a therapeutically effective amount of at least one of the heterocyclic compounds described above in accordance with invention methods for modulating the activity of excitatory amino acid receptors.
As used herein, the phrase "preventing disease conditions" refers to preventing a disease, disorder or condition from occurring in a subject who may be at risk for the disease, but has not yet been diagnosed as having the disease. Those of skill in the art will understand that a variety of methods may be used to determine a subject at risk for a disease, and that whether a subject is at risk for a disease will depend on a variety of factors known to those of skill in the art, including genetic make-up of the subject, age, body weight, sex, diet, general health, occupation, exposure to environmental conditions, marital status, and the like, of the subject.
Those of skill in the art can readily identify a variety of assays that can be used to assess the activity of excitatory amino acid receptors. For receptor species that activate a second messenger WO 01/16121 PCT/US00/23923 26 pathway, assays that measure receptor-activated changes in intracellular second messengers can be employed to monitor receptor activity. For example, inhibition of G-protein-coupled metabotropic glutamate receptors by antagonists can lead to inhibition of the glutamate-evoked increase in phosphatidylinositol (PI) hydrolysis, which can be assessed by measuring decreases in glutamatestimulated products of PI hydrolysis. (See Berridge el al, (1982) Biochem. J. 206:587-5950; and Nakajima el al., J Biol. Chem. 267:2437-2442 (1992) and Example 23.) Similarly, activation of excitatory amino acid receptors that leads to the release of intracellular calcium or changes in intracellular calcium concentration can also be used to assess excitatory amino acid receptor activity.
Methods of detection of transient increases in intracellular calcium concentration are well known in the art. (See Ito et al., J. Neurochem. 56:531-540 (1991) and Example 22). Furthermore, for receptor species that mediate analgesia, assays that measure analgesic efficacy can be employed to monitor receptor activity (See Example 24).
The following examples are intended to illustrate but not to limit the invention in any manner, shape, or form, either explicitly or implicitly. While they are typical of those that might be used, other procedures, methodologies, or techniques known to those skill in the art may alternatively be used.
Example 1 Synthesis of 2-(l-Cyclohexen-l-vlethynvy)-1,3-thiazole Triphenylphosphine (570 mg, 2.0 mmol) was dissolved in tetrahydrofuran (THF) (20 mL), then argon was bubbled through the solution for several minutes to deoxygenate it Palladium(ll) acetate (120 mg, 0.54 mmol) was added, and the reaction mixture was heated to 60 0 C for 0.5 h, and then cooled to ambient temperature. Cul (308 mg, 1.6 mmol), 2-bromo-l,3-thiazole (3.0 g, 18 mmol), 1ethynylcyclohexene (2.4 g, 20 mmol), potassium carbonate (6 g, 45 mmol) and water (1.0 mL, 58 mmol) were dissolved in 50 mL dimethoxyether (DME) and argon was bubbled through the solution for several minutes to deoxygenate the mixture. The catalyst solution of triphenylphosphine and palladium (nI) acetate in THF was added to the reaction flask which was heated to 75°C for 2h. After 2 h, heating was discontinued and the reaction was allowed to cool to ambient temperature. After stirring for 16 h, gas chromatography/mass spectrometry (GC/MS) analysis showed the reaction to be complete. The mixture was filtered through Celite T M the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL) and washed with water (200 mL), brine (200 mL), dried over Na 2 SO4. filtered, and concentrated in vacuo.
The residue was purified by column chromatography eluting with hexane then 97:3 hexane:ethyl acetate to afford 2-(l-cyclohexen-l-ylethynyl)-l,3-thiazole (2.56 g, 74% yield) as a brown oil. 'H NMR
(CDCI
3 300 MHz) 8 7.79 J=3.0 Hz, 1H), 7.31 J=3.0 Hz, 1H), 6.37-6.35 1H), 2.23-2.14 (m, 4H), 1.71-1.57 4H). MS (ESI) 190.0 WO 01/16121 PCT/US00/23923 27 Example 2 Synthesis of 2-Methyl-4-1 3-thiazol-2-vI)-3-butvn-2-ol 2-Bromo-l,3-thiazole (6.0 g, 37 mmol) and Cul (1.3 g, 7.3 mmol) were combined in DME (150 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture.
Triethylamine (25 mL, 180 mmol) and PdCI 2 (PPh 3 2 (2.5 g, 3.7 mmol) were added and 2-methyl-3butyne-2-ol (4.6 g, 55 mmol) was added dropwise. After stirring at ambient temperature for 16 h, GC/MS showed the reaction was not complete. The reaction was heated to reflux for 2 h. The mixture was filtered through Celite T M the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (600 mL), washed with water (600 mL), brine (600 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo.
The residue was purified by column chromatography eluting with hexane then 7:3 hexane:ethyl acetate to afford 4-(2-thiazolyl)-2-methyl-3-butyn-2-ol contaminated with 2,7-dimethyl-but-3,5-diyne-2,7-diol (the dimer of 2-methyl-3-butyne-2-ol). The product was crystallized from boiling hexane to afford 2methyl-4-(l,3-thiazol-2-yl)-3-butyn- 2 -ol (2.18 g, 36% yield) as off white crystals that were contaminated with a small amount of 2,7-dimethyl-but-3,5-diyne-2,7-diol. M.p. 69-70 0 C. 'H NMR
(CDCI
3 300 MHz) 8 7.80 J=3.0 Hz, 1H), 7.34 J=3.0 Hz, 1H), 4.40 1H), 1.65 6H). MS (ESI) 168.1 Example 3 Synthesis of 5-Chloro-3-pyridinvl trifluoromethanesulfonate Trifluoromethanesulfonic anhydride (5.0 mL, 30 mmol) was dissolved in CH 2
CI
2 (100 mL), and cooled to o0C 5-Chloro-3-pyridinol (3.10 g, 23.9 mmol), and triethylamine (6.5 mL, 47 mmol) were dissolved in CH 2 C2 (50 mL), and the resulting solution was added to the cold trifluoromethanesulfonic anhydride solution dropwise via cannula. The resulting dark brownish-red solution was stirred at 0°C for 5 minutes, and then the ice bath was removed and the reaction mixture was allowed to warm to ambient temperature. After stirring for 16 h at ambient temperature the reaction was quenched by pouring into water and basified by addition of saturated aqueous sodium carbonate. The basic aqueous phase was extracted with CH 2
CI
2 (2 x 50 mL), the combined organics were then dried over Na 2
SO
4 filtered and concentrated in vacuo. The resulting black viscous oil was filtered through a plug of silica gel and fractions were collected while eluting with 1:1 hexane:ethyl acetate. Fractions containing the desired product were combined, concentrated in vacuo, and further purified by column chromatography eluting with 15:1 then 10:1 hexane:ethyl acetate to afford 3-pyridinyl trifuoromethanesulfonate (3.68 g, 59% yield) as a golden liquid. 'H NMR (CDCI 3 300 MHz) 8 8.65 J=2 Hz, 1H), 8.52 J=2 Hz, 1H), 7.70 J=3 Hz, 111). MS (ESI) 261 263 (MI, "CI).
WO 01/16121 PCT/US00/23923 28 Example 4 Synthesis of 3-Chloro-5-l(trimethylsilvl)ethvnll pvridine 5-Chloro-3-pyridinyl trifluoromethanesulfonate (4.0 g, 15 mmol) and Cul (580 mg, 3.0 mmol) were combined in DME (100 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (10.6 mL, 76.5 mmol), and PdCI 2 (PPh3) 2 (1.lg, mmol) were added, then trimethylsilyl-acetylene (3.3 ml, 23 mmol) was added dropwise. The reaction mixture was stirred at ambient temperature for 1 h at which time GC/MS analysis indicated that the reaction was complete. The mixture was filtered through Celite
T
m, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo and the residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane then 99:1 hexane:ethyl acetate to afford 3-chloro-5-[(trimethylsilyl)ethynyl]pyridine (2.8 g, 87% yield) as a brown solid. 'H NMR (CDCI3. 300 MHz) 5 8.51 1H), 8.44 1H), 7.70(s, 1H), 0.22 9H). MS (El ionization) 209 Example Synthesis of (1.4g, 6.7mmol) was dissolved in methanol ml) and cooled to 0°C, to the resulting solution was added potassium carbonate (93 mg, 0.67 mmol).
The ice bath was removed and the reaction mixture was stirred at ambient temperature for 0.5 h at which time thin layer chromatography (TLC) and GC/MS analysis indicated that the reaction was complete. The solvent was removed in vacuo and the residue was dissolved in diethyl ether (50 mL), washed with water (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo to afford 3-chloro-5-ethynylpyridine (822mg, 90% yield) which was pure by GC/MS analysis. MS (El ionization) 137 ("CI M 4 139 37 CI This material was carried on to the next step without further purification.
Example 6 Synthesis of 3-Chloro-5-(1l3-thiazol-2-vlethvnvl)pyridine 2-Bromo-l,3-thiazole (980 mg, 6.0 mmol) and Cul (230 mg, 1.2 mmol) were combined in DME (15 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Tricthylamine (4.2 mL, 30 mmol) and PdC12(PPh 3 2 (420 mg, 0.60 mmol) were added, then 3-chloro-5-ethynylpyridine (820 mg, 19 mmol) was added dropwise. After stirring at ambient temperature for 16 h, GC/MS analysis showed starting material remaining. The reaction mixture was heated at reflux for 2 h. The mixture was filtered through CeliteTM, the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was WO 01/16121 PCT/USOO/23923 29 dissolved in ethyl acetate (100 mL), and washed with water (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane then 9:1 hexane:ethyl acetate to afford 3-chloro-5-(1,3-thiazol-2-ylethynyl)pyridine which contained some dimer. This material was crystallized from hot ethyl acetate to afford 3-chloro- 5-(1,3-thiazol-2-ylethynyl)pyridine (300 mg 23% yield) as light orange crystals M.p. 124-125°C. 'H NMR (CDC3. 300 MHz) 8 8.70 J=1.5 Hz, 1H), 8.59 J=3.0 Hz, 1H), 7.93 J=3.0 Hz, 1H), 7.88 J=2.0 Hz, 1H), 7.48 J=3.0 Hz, 2H). MS (ESI) 221.1 Example 7 Synthesis of 2-(Cvclohexylethvnvl)-1 3-thiazole 2-Bromo-l,3-thiazole (3.1 g, 19 mmol) and CuI (290 mg, 1.5 mmol) were combined in DME mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (13 mL, 95 mmol) and PdCl 2 (PPh 3 2 (530 mg, 0.76 mmol) were added and cyclohexylethyne (2.0 g, 19 mmol) was added dropwisc. The reaction mixture was stirred at ambient temperature for 16 h at which time GC/MS analysis indicated that the reaction was complete. The mixture was filtered through Celite T M and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo and the residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo.
The residue was purified by column chromatography eluting with hexane then 99:1 hexane:ethyl acetate to afford 2-(cyclohexylethynyl)-l,3-thiazole (1.6 g, 44% yield) as a yellow oil. 'H NMR
(CDCI
3 300 MHz) 7.76 J=9.0 Hz, 1H), 7.28 J=3.0 Hz, 1H), 2.68-2.59 1H), 1.91-1.28 (m, MS (ESI) 191.7 Example 8 Synthesis of 2-1-Pentynyl)-1 -thiazole 2-Bromo-l,3-thiazole (2.0 g, 12 mmol) and Cul (183 mg, 0.96 mmol) were combined in DME (30 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (8 mL, 60 mmol) and PdCl2(PPh 3 2 (337 mg, 0.48 mmol) were added and 1pentyne (979 mg, 14.4 mmol) was added dropwise. The reaction mixture was stirred at ambient temperature for 6 h at which time GC/MS analysis indicated that the reaction was not complete.
Additional 1-pentyne (3.0 mL, 29 mmol) was added and the reaction was heated to 35 0 C under a condenser. After heating for 16 h, GC/MS analysis indicated that the reaction was complete. The mixture was filtered through CeliteT, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo and the residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo.
The residue was purified by column chromatography eluting with hexane, 99:1, then 97:3 hexane:ethyl WO 01/16121 PCT/USO0/23923 acetate to 2-(l-pentynyl)-1,3-thiazole (820 mg, 44% yield) as a yellow oil. 'H NMR (CDCI 3 300 MHz) 8 7.76 J=3.0 Hz, I1H), 7.28 J=3.0 Hz, IHf), 2.47-2.42 (in, 214), 1.68-1.60 (in, 2H), 1.08 -0.99 (in, 3H). MS (ESI) 15 1.6 Example 9 Synthesis of 2-(3-Cvclohexvl-l-propvnvh -1 3-thiazole 2-Bromo-l,3-thiazole (2.0 g, 12 mmol) and Cul (185 mng, 0.97 inmol) were combined in DME mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamnine (8.5 mL, 61 mmol) and PdCI 2 (PPh 3 2 (340 mg, 0.49 mmol) were added and 3cyclohexyl-1 propyne (2.9 g, 24 mmol) was added dropwise. The reaction was stirred at ambient temperature for 16 h at which time GUIMS analysis indicated that the reaction was complete. The mixture was filtered through Celitem, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo. Thc residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 dried over Na 2
SO
4 filtered, and concentrated in vacuo.
The residue was purified by column chromatography eluting with hexane, then 98:2 hexane:ethyl acetate to afford 2-(3-cyclohexyl-l-propynyl)-1,3-thiazole (1.14 g, 46% yield) as ayellow oil. 'H NMR
(CDCI
3 300 MHz) 6 7.76 J=3.0 Hz, 7.27 J3.0 Hz, 1K), 2.35 J=6 Hz, 2M), 1.89-1.61 (in, 1.3 1.03 (in, 6H). MS (ESI) 205.9 Example Synthesis of 241I-Cyclohexen-l-vlethvnfl)-5-nitro-13-thizole 2-Bromo-5 -nitro-l1,3-th iazole (2.5 g, 12 minol) and Cul (460 mng, 2.5 mmol) were combined in DME (30 inL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (8.4 mL, 60 minol) and PdCI 2 (PPh 3 2 (840 mg, 1.2 inmol) were added and 1-ethynycyclohexerie (1.5 g, 14.4 inmol) was added dropwise. The reaction was heated under reflux for 16 h at which time GCUMS analysis indicated that the reaction was complete. The mixture was filtered through CeliterM%, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo and the residue was dissolved in ethyl acetate (300 miL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane, 99:1 then 98.5:1.5 hexane:ethyl acetate to afford 2-(l -cyclobexen- I-yiethynyl)-5 -nitro- 1,3-thiazole (1.4 g, 51.8% yield) as a yellow powder. M.p.
85-86 0 C. H NMR (CDC1 3 300 MHz) 8 8.5 114), 6.52 (br s, 114), 2.24 (br s, 414), 1.63 (br s, 4H). MS WO 01/16121 PCT/USO0/23923 31 Example 11 Synthesis of 2-(3,3-Dimethyl-l-butynvl)- 13-thiazole Triphenylphosphine (380 mg, 1.5 mmol) was dissolved in THF (20 mL), then argon was bubbled through the solution for several minutes to deoxygenate it. Palladium(II) acetate (82 mg, 0.37 mmol) was added, and the reaction mixture was heated to 60 0 C for 0.5 h, and then cooled to ambient temperature. Cul (210 mg, 1.1 mmol), 2-bromo-1,3-thiazole (1.6 g, 9.8 mmol), potassium carbonate (4.2 g, 31 mmol) and water (0.70 mL, 39 mmol) were dissolved in DME (30 mL) and argon was bubbled through the mixture for several minutes to deoxygenate the mixture. 3,3-dimethyl-l-butyne g, 12.2 mmol) was then added to mixture. The catalyst solution of triphenylphosphine and palladium (II) acetate in THF was added to the reaction flask which was heated to 30 0 C for 2h. After this time heating was discontinued and the mixture was allowed to stir at ambient temperature. After stirring for 16 h, GC/MS analysis showed the reaction to be complete. The mixture was filtered through Celite T M the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL), washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane, then 99:1 hexane:ethyl acetate to afford 2- (3,3-dimethyl-l-butynyl)-l,3-thiazole (0.45 g, 28% yield) as a yellow oil. 'H NMR (CDCI 3 300 MHz) 7.74 J=3.0 Hz, 1H), 7.28 J=3.0 Hz, 1H), 1.33 9H). MS (ESI) 166.1 Example 12 Synthesis of l-(13-Thiazol-2-vlethynyl)cvclopentanol 2-Bromo-l,3-thiazole (3.1 g, 19 mmol) and Cul (360 mg, 1.9 mmol) were combined in DME mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (13 mL, 94 mmol) and PdCi 2 (PPh 3 (660 mg, 0.94 mmol) were added and 1ethynycyclopentanol (2.5 g, 23 mmol) was added dropwise. The reaction was heated at 50 0 C for 16 h at which time GC/MS analysis indicated that the reaction was complete. The mixture was filtered through CeliteTM, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo and the residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane, 6:1 then 3:1 hexane:ethyl acetate to afford 1-(1,3-thiazol-2-ylethynyl)cyclopentanol (2.3 g, 52% yield) as a yellow powder. 'H NMR (CDCI 3 300 MHz) 8 7.80 J=3 Hz, 1H), 7.65 J=3 Hz, 1H), 2.04-1.73 10.8H). MS (El ionization) 193 WO 01/16121 PCT/US00/23923 32 Example 13 Synthesis of 2-(l-Cyclopenten-l-vlethynvl)-1 3-thiazole 1-(1,3-Thiazol-2-ylethynyl)cyclopentanol was dissolved in pyridine (20 ml) and phosphorus oxychloride (1.2 g, 6.2 mmol) was added dropwise under argon. The reaction was stirred at ambient temperature for Ih at which time a precipitate had appeared. At this time GC/MS analysis indicated that the reaction was complete and the pyridine was removed in vacua. The residue was dissolved in ethyl acetate (200 mL) and washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacua. The residue was purified by column chromatography eluting with hexane then 99:1 hexane:ethyl acetate to 2-(1-cyclopenten-l-ylethynyl)-1,3-thiazole (0.25 g, 24% yield) as a light brown solid. M.p. 70.5-72 0 C, 'H NMR (CDCI 3 300 MHz) 6 7.80 J=3.0 Hz, IH), 7.34 (d, IH), 6.31-6.30 IH), 2.60-2.45 4H), 2.00-1.90 2H). MS (ESI) 176.1 Example 14 Synthesis of Methyl 3-(1 3-thiazol-2-yl)-2-propynvl ether 2-Bromo-1,3-thiazole (2.0 g, 12 mmol) and Cul (456 mg, 2.4 mmol) were combined in DME (30 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (8.6 mL, 60 mmol) and PdCI 2 (PPh 3 2 (842 mg, 1.2 mmol) were added and methyl propargyl ether (1.00 g, 14.4 mmol) was added dropwise. The reaction was stirred at under a condenser. After stirring at 55°C for 16 h, GC/MS analysis indicated that the reaction was complete. The mixture was filtered through Celite"T, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacua and the residue was dissolved in ethyl acetate (300 mL), washed with water (300 mL), brine (300 mL), dried over Na 2
SO
4 filtered, and concentrated in vacua. The residue was purified by column chromatography eluting with hexane, 99:1, 97:3, then 96:4 hexane:ethyl acetate to afford methyl 3-(1,3-thiazol-2-yl)-2-propynyl ether (250 mg, 13% yield) as a yellow oil. 'H NMR (CDCI3. 300 MHz) 8 7.78 J=3.0 Hz, 1H), 7.37 J=3.0 Hz, 1H), 4.37 2H), 3.47 3H). MS (ESI) 154.1 Example Synthesis of 2-Methyl-4-(3-pyridinvl)-3-butvn-2-ol 3-Bromopyridine (3.0 mL, 31 mmol), triethylamine (22 mL, 160 mmol), Cul (1.2 g, 6.2 mmol), and PdCl2(PPh 3 2 (1.1 g, 1.5 mmol) were combined in DME (92 mL) and cooled to 0°C. 2-Methyl-3butyne-2-ol (9.0 mL, 93 mmol) was then added and the reaction was allowed to slowly warm to ambient temperature. The mixture was then heated to 55-600C for 16 h. The mixture was filtered through CeliteTM, and the pad was washed thoroughly with ethyl acetate. The combined filtrates were washed with brine (3 x 100 mL), dried over MgSO 4 and filtered. The solution was concentrated in vacuo, and the residue was purified by column chromatography eluting with 90:10 hexane:ethyl acetate WO 01/16121 PCT/US00/23923 33 then ethyl acetate to afford 2-methyl-4-(3-pyridinyl)-3-butyn-2-ol (2.0 g, 40% yield) as a brown oil 'H NMR (CDCI 3 300 MHz) 5 8.76 (br s, 1H), 8.52 (br s, 1H), 7.74-7.70 1H), 4.08 (br s, 1H), 1.63 (s, 3H). MS (El ionization) 161 Example 16 Synthesis of 3-Ethynylpyridine 2-Methyl-4-(3-pyridinyl)-3-butyn-2-ol (611 mg, 3.79 mmol) was dissolved in toluene (12 mL) at ambient temperature. A small amount (spatula tip) of NaH (60% dispersion in mineral oil) was added, and the reaction was heated to reflux. After 15 minutes the reaction was cooled to ambient temperature, and quenched by the addition of IM aqueous HCI (30 mL). Crude product from a previous preparation (-200 mg) was added to the workup mixture. The acidic aqueous was extracted with ethyl acetate (2 x 20 mL), basified by the addition of saturated aqueous NaHC0 3 and extracted with CH2C 2 The CH 2
C
2 extracts were dried over MgSO 4 filtered, and concentrated in vacuo to afford crude 3-ethynylpyridine (1.5 g, >100%) as a brown liquid., 'H NMR (CDC[ 3 300 MHz) 8 8.73 (br s, 1H), 8.58 (br s, 1H), 7.80-7.76 1H), 7.29-7.16 1H), 3.28 1H). A portion of this material was carried on to the next step without further purification.
Example 17 Synthesis of 3-(13-Thiazol-2-vlethvnyl)pvridine 2-Bromo-1,3-thiazole (0.15 mL, 1.6 mmol), Cul (98 mg, o.51 mmol), PdCl2(PPh 3 2 (120 mg, 0.17 mmol) and triethylamine (2.8 mL, 20 mmol) were combined in DMF (6.8 mL) and cooled in an ice bath. 3-Ethynylpyridine (520 mg, 5.04 mmol) was then added to the mixture as a solution in DMF mL). The ice bath was removed and the reaction was allowed to stir at ambient temperature for 16 h. The reaction mixture was filtered through a pad of Celite T M and the pad was washed thoroughly with ethyl acetate. The filtrate was washed with brine (3 x 20 mL). A partial emulsion was observed.
The mixture was concentrated in vacuo and the residue was taken up in CH 2
CI
2 washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with 80:20 followed by 30:20 hexane:ethyl acetate to afford 3- (1,3-thiazol-2-ylethynyl)pyridine (160 mg) as a mixture with another product exhibiting a mass of 204 in the GC/MS, assigned as pyridylalkyne dimer. A portion of the mixture (100 mg) was further purified by preparative reverse-phase HPLC eluting with a gradient of 80:20 to 0:100 water:acetonitrile over twenty minutes. The fractions containing the desired product were collected (detection by uv at 210 nm) to afford 3-(1,3-thiazol-2-ylethynyl)pyridine as a white waxy solid (15 mg). 'H NMR (CDCI 3 300 MHz) 8 9.3-8.5 (br s, 2 7.92-7.90 2H), 7.50-7.30 2H). MS (ESI) 187.0 WO 01/16121 PCT/US00/23923 34 Example 1 Synthesis of 3.3.5.5-Tetramthl--(2-pridinlethnyl)cyIohCxanlO.
To a solution of 2-ethynylpyridine (1.0 g, 10 minol) in THE at -78*C was added a 1.0 M solution of ethyl magnesium bromide in TI-IF (10 mL, 10 mmol). After stirring at reduced temperature for 30 minutes a solution of 3,3,5,5-tetramethylcyclohexanone (1.5 g, 10 inmol) in THF was addcd rapidly. The mixture was allowed to warm to ambient temperature over 16 hours, then partitioned between water and ethyl acetate. The organic layer was dried over anhydrous Na 2
SO
4 and concentrated in vacuo. The resultant product was purified by flash column chromatography on silica gel eluting with 1:1 hexane:ethyl acetate to afford 3,3,5,5-tctramethyl-lI- (2-pyridinylethynyl)cyclohexanol (250 mg, 10% yield) as a white solid. M.p. 126-127 0 C. 'H NMAR (DMSO-d 6 300 MHz) 6 8.57 (in, I 7.64 (in, I 7.39 J=5 Hz, 1 7.22 (in, IlH), 1.91 J=9 Hz, 2H), 1.71 J=9 Hz, 2H), 1.26 2H), 1. 14 6H), 1.09 6H1).
Example 19 Synthesis of 2-(.,.-ermty--yloee--lehnlprdn 3,3,5,5-Tetramethyl-1-(2-pyridinylethynyl)cyclohexanol (200 mg, 0.78 innol) was dissolved in pyridine. P0C13 (153 mng, 1.0 inmol) was added, and the mixture was heated to reflux for 6 h. After cooling, the P0C13 and pyridine were removed in vacuo. The residue was purified by flash column chromatography on silica gel eluting with 2:1 hexane:ethyl acetate to afford 2-[(3,3,5,5-tetramethyl-lcyclohexen-l-yl)ethynyljpyridine (148 mng, 80% yield) as a light tan solid. M.p. 55-56 0 C. 'H NMR
(CDCI
3 3 00 MHz) 5 8.5 6 (in, I1H), 7.62 (in, I1-H), 7.40 J=7 Hiz, I 7.18 (mn, IlH), 6.09 111), 2.00 2H), 1.35 2H), 1.05 611), 0.99 6H).
Exapl 2 Synthesis of 2-(-ehllgcoeen1yehntprdn and 2-i(3.methyl-l-cyclohexen-1-yl)ethyflyllpyridifle (1:1) Using the procedures for Examples 18 and 19 but with the appropriate starting materials, methyl-I -cyclohexen-1I-yl)ethynylJpyridine and 2-[(3-inethyl- i-cyclohexen- 1-yl)ethynyl]pyridine were obtained as a mixture of racemie regioisomers. 'H NMR (CDCI 3 300 MHZ) 8 8.56 (nm, IH), 7.62 (in, 1H), 7.40 (in, IH), 7.19 (in, IH), 6.32 0.5H), 6.20 0.5 2.25 (in, 3H), 1.73 (in, 1.22 (mn, I1H), 1.0 1 (in, 3H). MS (EI ionization) Two peaks: 197 (MW).
WO 01/16121 PCT/US00/23923 Example 21 General procedure for 2-pyridvlenvnes To a cooled a solution of 2-ethynylpyridine in THF to -78 0 C was added n-BuLi (1.6 M in hexane, 1 equiv). After 20 minutes stirring at reduced temperature this material was mixed with a solution of the appropriate ketone (1 equiv) in THF. The solution was allowed to warm slowly to ambient temperature. The reaction mixture was then quenched and partitioned between water and ethyl acetate. The organic layer was dried over Na 2 SO4, and concentrated in vacuo. The resultant product was purified by flash column chromatography on silica gel eluting with 1:1 hexane:ethyl acetate. The resulting product was dissolved in pyridine or a mixture of pyridine and methylene chloride
POCI
3 (1.2 equiv) was added and the solution refluxed for 4 to 8 hours. The resultant mixture was partitioned between 1M K 2
CO
3 and ethyl acetate. The organic layer was dried over Na 2
SO
4 and concentrated in vacuo. The resultant product was purified by flash column chromatography on silica gel eluting with 2:1 hexane:ethyl acetate.
Using this general procedure the following invention compounds (see Examples 22-33) were obtained.
Example 22 Synthesis of 2- [(4-Methyl- -cyclopenten-1-yl)ethynyl pyridine and 2-(3-Methyl-1-cyclopenten-1yl)ethynyllpyridine (1:1) Reactants: 2-ethynylpyridine (620 mg, 6.0 mmol), 3-methylcyclopentanone (0.64 mL, mmol); yields 2-[(4-methyl-l-cyclopenten-l-yl)ethynyl]pyridine and 2-[(3-methyl-l-cyclopenten-1yl)ethynyl]pyridine as a transparent oil (200 mg, 18% overall yield), as mixture of regio- and stereoisomers. 'H NMR (CDCI 3 300 MHz) 8 8.56 1H), 7.64 1H), 7.44 1H), 7.20 1H), 6.19 0.5H), 6.18 0.5H), 2.90 0.5H), 2.70 2.5H), 2.21 2H), 1.48 0.5H), 1.08 (app d, J= 7.5Hz, 3H). Two peaks: 182 167 Example 23 Synthesis of 2-(Bicyclol2.2.11hept-2-en-2-lethynyl)pyridine Reactants: 2-ethynylpyridine (1.0 g, 10.0 mmol), norcamphor (1.1 g, 10.0 mmol); yields 2- (bicyclo[2.2.1]hept-2-en-2-ylethynyl)pyridine as a black oil (215 mg, 11% over two steps). This material was mixed with fumaric acid (128 mg, 1.11 mmol), dissolved in Methanol and the resulting solution was concentrated in vacuo to afford a dark brown solid. This was triturated with a mixture of ethyl acetate:ethanol and the resultant solids were partitioned between aqueous K 2
CO
3 and ethyl acetate. The organics were dried over Na 2
SO
4 and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel eluting with 2:1 hexane:cthyl acetate to afford 2- (bicyclo[2.2.1 ]hept-2-en-2-ylethynyl)pyridine (30 mg, 1.5 overall yield) as a translucent brown oil.
WO 01/16121 PCT/USOO/23923 36 'H NMR (DMSO-d,, 300 M~z) 8 8.58 J= 5Hz, IH), 7.64 (in, LH), 7.40 (in, IH), 7.19 (in, 111), 6.48 3=4Hz, 111), 3.07 111), 2.97 Iif), 1.76 (mn, 211), 1.51 (in, 1H), 1.23 (in, IH), 1.11 (in, 1H). MS (El ionization) 195 (NC).
Example24 Synthesis of 2-I(2,6-Dimethydl-cyclohexen-1-yl)ethynyllpyridine Reactants: 2-ethynylpyridine (5.0 minol, 515 mg), 2,6-dimethylcyclopentanone inmol, 0.82 mL); yields 2-[(2,6-dimethyl-l-cyclohexen-1-yl)ethynyljpyridine as a transparent oil (200 mg, 19% overall yield). 'H NMR (CDCI 3 3 00 MHz) 8 8.5 6 (in, 11H), 7.60 (mn, IHM, 7.42 (mn, I1H), 7.19 (in, I 2.40 (in, I 2. 10 (mn, 2H), 2.01 3H), 1.76 2H), 1.56 (in, I 1.34 (in, IlH), 1.22 (app d, J= 7Hz, 3H). MS (El ionization) 211(M-).
Exam 2 Synthesis of 2-(1-Cyclohepten-1-ylethynyl)pyridine Reactants: 2-ethynylpyridine (5.0 innol, 515 mg), cycloheptanone (6.0 inmol, 0.71 inL); yields 2-(l-cyclohepten-1-ylethynyl)pyridine as a transparent oil (200 mg, 18% overall yield). 'H NMR
(GDCI
3 300 MWz) 8 8.54 (mn, 111), 7.59 (in, 111), 7.40 (mn, IH), 7.16 (in, IH), 6.52 J= 7 Hz, 111), 2.47 (mn, 2H), 2.26 (in, 211), 1.77 2H), 1.61 (in, 2H), 1.56 (in, 2H). MS (El ionization) 197 Synthesis of 2-(1-Cycloocten-1-ylethynyl)pyridine Reactants: 2-ethynylpyridine (5 15 mg, 5.0 minol), cyclooctanone (756 ing, 6.0 inmol); yields 2- (l-cycloocten-l-ylethynyl)pyridine as a transparent oil (250 mg, 24% ov'erall yield).'H NMR (GDCl 3 300 MHz) 8 8.57 (in, IH), 7.62 (in, 111), 7.40 (in, 111), 7.18 (in, 111), 6.33 J= 7 Hz, IH), 2.41 (in, 2H), 2.23 (in, 2H), 1.66 2H), 1.52 (br m, 6H). MS (El ionization) 211 (NO).
Exampl2 Synthesis of 2-I(4-Methyl-l-cyclohexen-1 -yo)ethynyllpyridine Reactants: 2-ethynylpyridine (6.0 inmol, 618 ing), 4-mctliylcyclohexanone (6.0 mmnol, 672 mng); yields 2-[(4-inethy1-1-cyclohexen-1-y1)ethynylIpyridine as a transparent oil (250 ing, 21% overall yield). 'H NMR (CDC1 3 300 Mffz) 5 8.57 (in, I 7.59 (in, I 7.39 (mn, I 7.20 (in, IlH), 6.30 (in, 1H), 2.22 (mn, 3H1), 1.72 (mn, 3H), 1.25 (mn, lH), 0.99 (in, 3H1). MS (El ionization) 197 WO 01/16121 PCT/USOO/23923 37 Example 28 Synthesis of 2-(3,6-Dihydro-2E-thiopyran-4-ylethynvl~pyridine Reactants: 2-ethynylpyridine (6.0 mmol, 618 mg), tetrahydrothiopyran-4-one (6.0 nimol, 696 mg); yields 2-(3,6-dihydro-2H-thiopyran-4-ylethynyl)pyridine as a transparent oil (150 mg, 12% overall yield). 'H NMR (CDCI 3 300 MIHz) 6 8.57 (in, 1H), 7.61 (in, IH), 7.40 (in, 1K), 7.21 (in, 1K), 6.46 (mn, 1KM, 3.27 (in, 211), 2.78 (mn, 2H4), 2.57 (in, 2H1). MIS (El ionization) 201 (Mr).
Synthesis of 24(3.6-Dihydro-2H-pyran-4-ylethynyflpyrfdine Reactants: 2-ethynylpyridine (6.0 mmol, 618 mg), tetrahydro-4H-pyran-4-one (6.0 mmol, 600 mng); yields 2-(3,6-dibydro-2H1-pyran-4-ylethynyl)pyridine as a transparent oil (200 mg, 18% overall yield). 'H NMR (CDCI 3 300 MHz) 5 8.57 (in, I1H), 7.63 (in, IH), 7.44 (in, 111), 7.21 (in, 1KM, 6.29 (in, IH), 4.25 (in, 2H), 3.81 (in, 2H), 2.36 (in, 2H). MS (El ionization) 185 (Md).
Exam 3 Synthesis of I(1R)-1 .7.7-Trimethylbicyclol2.2.l1 hept-2-en-2-yll ethynvlpyridine Reactants: 2-ethynylpyridine (6.0 inmol, 618 mg), (IR)-(+)-canxphor (6.0 inmol, 912 mg); yields 2- I ,7,7-trimethylbicyclo[2.2. I ]hept.2-en-2-yljethynyl) pyri dine as a transparent yellow oil (125 mg, 9% overall yield). 'H NMR (CDCI 3 300 MHz) 8 8.57 (in, IlH), 7.64 (in, 1KH), 7.43 (in, 1KH), 7.17 (in, 1H), 6.49 31Iz, 1K), 2.41 J= 3Hz, 111), 1.92 (br mn, IH), 1.65 (in, IH), 1.18 (mn, IH), 1.17 3H), 1.09 (br n, 1H), 0.84 3H), 0.82 3H). MIS (El ionization) 237 Examl 1 Syntheses of 2-[(3.5-Dimethyl-1 -cyclohexen-1 -yl)ethynyljpyridine Reactants: 2-ethynylpyridine (6.0 mxnol, 618 mg), 3,5-diniethylcyclohexanone (6.0 minol, 0.85 inL); yields 2-[(3,5-dimethyl-1I-cyclohexen- I-yi)ethynyl]pyri dine as a transparent yellow oil (500 mg, 39% overall yield) as a mixture of diastereomers. 1H NMR (CDCI 3 300 MHz) 8 8.57 (in, 1K), 7.62 (in, I1U), 7.40 (mn, 1H), 7.19 (in, 1K), 6.15 (br s, I 2.29 (in, 2H), 1.80 (br m, 2H), 1.00 (in, 6H), 0.88 (hr mn, 2H). MS (El ionization) 211 (Mr).
WO 01/16121 PCT/US00/23923 38 Example 32 Synthesis of 2-U[(5R)-5-Methyl-1-cyclohexen- 1-vilethynyll pyridine compound with 2-1 r(3R)-3mnethyl- I -cyclohexen-I-yll ethynvl Pyridine (1:1) Reactants: 2-ethynylpyridine (6.0 mmol, 618 ing), (3R)-(+)-3-methylcyclohexanone (6.0 mmol, 0.73 mL); yields 2- {[(5R)-5-methyl-l1-cyclohexen-1I-yllethynyl }pyridine and [(3R)-3-metbyl- 1cyclohexen-1-yllethynyl~pyridine as a transparent yellow oil (440 mng, 37% overall yield) as a mixture of regioisomers. 11H NMR (CDCI,, 300 Nfl-lz) 88.56 (in, IH), 7.62 (mn, 11-1), 7.40 (mn, IH), 7.18 (in, I 6.31 (in, 6.19 (in, 2.30 (in, 3H), 1.85 (mn, 2.5H), 1.22 (in, 1H), 0.98 (mn, MS (El ionization) 197 (M 4 two peaks resolved.
Example 33 Synthesis of 2-k(3E)-3-Methyl-3-penten-l-vnyll pyridine. 2-(3-ethyl-3-buten-l-yvl~pvridine and 2r(3Z-3-nethyl-3-venten-l-ynyll pyridine Reactants: 2-ethynylpyridine (6.0 inmol, 618 ing), 2-butanone (6.0 rmol, 0.54 mnL); yields 2- [(3E)-3-methyl-3 -penten- 1-ynyl]pyridine, 2-(3-ethyl-3-buten-1I-ynyl)pyridine and -ietbyl-3penten-l-ynyljpyridine as a transparent oil (135 mg, 14% overall yield) as a mixture of E, Z and exomethylene isomers. 'H NMR (CDCI 3 300 Mi~z) 58.59 (mn, IHM, 7.65 (in, IH), 7.44 (mn, 1H), 7.20 (in, 1IH), 5.88 (mn, 0.75H), 5.53 0.33H) 5.40 0.33 2.29 J= 7Hz, 0.65H), 1.93 (mn, 4.5H), 1.17 (t, J= 7Hz, I MS (El ionization) 15 7 two peaks resolved.
Example 34 Synthesis of 5-Ethvl-2-(p~henylethvnvl~tpyrimidine hydrochloride 2-Chloro-S-ethylpyrimidine (500 mng, 3.5 inmol), PdC] 2 (PPh 3 2 (250 mg, 0.35 minol), Gui (203 mng, 1.06 minol), tniethylamine (6.0 mL, 43 inmol), and n-Bu 4 NI (3.85 g, 10.4 inmol) were combined in dimethylforinamide (DMI) (30 mnL). The mixture was cooled in an ice bath and then phenylacetylene inL, 14 inmol) was added. The reaction mixture was then heated to 45-50*C and after 1.5 hi, additional phenylacetylene (1.5 mnL, 14 inmol) was added. After an additional 17 h the reaction was diluted with ethyl acetate, washed with brine (4 x 15 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The resulting black oil was purified by column chromatography eluting with hexane then 90:10 hexane:ethyl acetate to afford 5-ethyl-2-(phenylethynyl)pyri mid ine (770 mng, >100%) as a black oil. MS (El ionization) 208 This material was carried on to the salt formation without further purification.
5-Ethyl-2-(phenylethynyl)pyrimidine (730 ing, 3.7 iniol) was dissolved in CH 2
CI
2 (3.0 mL) and treated with HCI in diethyl ether (4.1 mL of a IN solution, 4.1 inmol). Upon addition of the HC! solution a solid precipitated from the solution. The mixture was diluted with diethyl ether (2 mL) and WO 01/16121 PCT/USOO/23923 39 the supernatant decanted. The resultant solid was dried under high vacuum at 50 0 C to afford 5-ethyl-2- (pheny lethynyl)pyri mid ine hydrochloride (450 mg, 49 yield) as an orange solid. M.p. 101-104 0 C. 'H NMR (CD 3 OD, 300 MHz) 6 8.75 2H), 7.58-7.55 (in, 2H), 7.4 1-7.32 (in, 3H), 2.67 J=7.6 Hz, 2R-), 1.21 J=7.6 Hz, 31).
E~xample Synthesis of 4.6-Dimethoxv-2-(phenvlethvnvl)nvrim idine hydrochloride 2-Chloro-4,6-dimethoxypyrimidine (500 mng, 2.9 mmol), PdCl 2 (PPh 3 2 (200 mg, 0.28 inmol), Gui (160 mng, 0.84 minol), triethylamine (4.8 mL, 34 minol), and Rz-Bu 4 N1 (3.2 g, 8.7 mmol) were combined in DMF (24 mL). The mixture was cooled in an ice bath and then phenylacetylene (1.25 m.L, 11.4 inmol) was added. The reaction mixture was allowed to warm to ambient temperature. After 2.5 h at ambient temperature the reaction mixture was heated to 4S-50*C. After 2 h, additional phenylacetylene (1.0 mL, 9.1 inmol) was added. After an additional 17 h stirring at 45-50*C, the reaction mixture was filtered through a pad of Celite'm, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were washed with brine (4 x 20 mL), dried over MgSO 4 filtered and concentrated in vacuo. The resulting black oil was purificd by column chromatography eluting with hexane, 90: 10, then 85:15 hexane:ethyl acetate to afford product contaminated with an impurity.
Careful column chromatography of this impure material eluting with hexane then 90:10 hexane:ethyl acetate afforded 4,6-dimethoxy-2-(phenylethynyl)pyrimidine (320 mg, 46% yield) as a yellow solid.
This material was carried on to the salt formation without fuirther purification.
4,6-Dimethoxy-2-(phenylethynyl)pyrimidine (320 mg, 1.3 minol) was dissolved in CH 2
CI
2 mL), and treated with 1-ICI in diethyl ether (1.6 mL of a I OM solution, 1.6 mmol). A yellow solid precipitated immediately. The mixture was diluted with ethyl acetate and allowed to stand in the freezer for 16 h. The cold supernatant was decanted and the remaining solids were triturated with ethyl acetate (1.5 mL), and then hexane (3 x 2 mL). The remaining solid was dried in vacuo to afford 4,6diinethoxy-2-(phenylethynyl)pyrimidine hydrochloride (174 mg, 47 yield) as a yellow solid. M.p.
13 7-13 8. 'H NMR (CD 3 OD, 300 NMz) 5 7.65-7.62 (in, 2H), 7.46-7.42 (im, 3H), 6.16 1H), 3.97 (s, 6H-).
Example 36 Synthesis of 2-l(E)-2-(3-Fluorophenyl'lethenyll-6-nethylpyrazine 2,6-Dimethylpyrazine (5.0 g, 46 mmol) was dissolved in THE (200 mL) and cooled to 0 0
C.
Potassium t-butoxide (46 mL of a L OM solution in THF, 46 inmol) was added to afford a dark red solution. Thc solution was allowed to warm to ambient temperature and stir for I hr. The solution was thcn cooled to 0 0 C, and 3-fluorobcnzaldehyde (4.9 mL, 46 inmol) was added via syringe pump over 2 WO 01/16121 PCT/US00/23923 h. The reaction was then allowed to slowly warm to ambient temperature. After stirring at ambient temperature for 18 h, the reaction mixture was cooled to 0°C and quenched by the addition of concentrated aqueous HCI (10 mL). The resulting suspension was allowed to warm to ambient temperature for 15 minutes, then cooled to 0°C and brought to pH=8 by addition of solid NaHCO 3 The layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine (200 mL), dried over MgSO 4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography eluting with 90:10, 85:15, then 80:20 hexane:ethyl acetate to afford 2-[(E)-2-(3-fluorophenyl)ethenyl]-6-methylpyrazine (4.14 g, 42% yield) as a light yellow solid. M.p. 43-44 0 C. 'H NMR (CDCI 3 300 MHz) 8 8.44 1H), 8.31 1H), 7.29 J=16 Hz, IH), 7.37-7.26 3H), 7.12 J=16 Hz, IH), 7.05-6.98 IH), 2.59 3H). MS (ESI) 214.5 This material was carried on to the next step without further purification.
Example 37 Synthesis of 2- 1.2-Dibromo-2-(3-fluorophenvl)ethyll-6-methylpyrazine 2-[(E)-2-(3-Fluorophenyl)ethenyl]-6-methylpyrazine from Example 36 (4.14 g, 19.3 mmol) was dissolved in CCI 4 (40 mL). To this solution was added a solution of bromine (1.2 mL, 23 mmol) in CC14 (20 mL). The brown mixture was then heated to 60 0 C. After 6h the suspension was treated with saturated aqueous NaHCO 3 (200 mL) and diluted with ethyl acetate (700 mL). The organic layer was washed with 5% aqueous Na 2
S
2 03 (100 mL), brine (100 mL), dried over MgSO 4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography eluting with 80:20 hexane:ethyl acetate then 95:5, 94:6, and 90:10 CH 2 Cl 2 :ethyl acetate to afford 2-[1,2-dibromo-2-(3fluorophenyl)ethyl]-6-methylpyrazinc (2.97 g, 17% over two steps) as a white solid. This material was carried on to the next step without further purification.
Example 38 Synthesis of 2-1(3-Fluorophenyl)ethynyll-6-methylpyrazine hydrochloride 2-[1,2-Dibromo-2-(3-fluorophenyl)ethyl]-6-methylpyrazine (2.97 g, 7.94 mmol) was dissolved in THF (40 mL), treated with DBU (8.7 mL, 63 mmol), and heated to reflux. After 16 h the reaction mixture was cooled, filtered, concentrated in vacuo, and purified by column chromatography eluting with 80:20 then 75:25 hexane:ethyl acetate to afford 2-[(3-fluorophenyl)ethynyl]-6-methylpyrazine (427 mg, 25% yield). This material was carried on to the salt formation without further purification.
2-[(3-Fluorophenyl)ethynyl]-6-methylpyrazine (520 mg, 2.45 mmol) was dissolved in CH 2
CI
2 (3 mL), and the resulting solution was treated with HCI in diethyl ether (2.7 mL of a 1.OM solution, 2.7 mmol). The mixture was sonicated, and the solvent decanted. The remaining solid was dried under high vacuum to afford 2-[(3-fluorophenyl)ethynyl]-6-methylpyrazine hydrochloride (338 mg, WO 01/16121 PCT/USOO/23923 41 yield) as a light yellow solid. M.p. 62-63 0 C. 'H NMR (CDCI 3 300 MHz) 5 8.73 I 8.57 IK) 7.54-7.3 5 (in, 3H), 7.28-7.20 (in, 3H), 2.84 3H).
Example 39 Synthesis of I-Chloro-4-(l-cyclohexen-1-yl')-3-butvn-2-one Anhydrous ZnCI 2 (5.0 g, 37 minol) was dissolved in TI-F (25 m.L) and the solution cooled to 0 0 C in an ice bath. In another flask 1-ethynylcyclohexene (4.3 mL, 36.3 mmol) was dissolved in TI-IF mL), cooled to 0 0 G in an ice bath, and treated with n-butyllithiumn (15.7 mL of a 2.2M solution in hexane, 34.5 mmol). After 20 minutes the cyclohexenylethynyllithium solution was added via cannula to the ZnCI 2 solution. After an additional 20 minutes Pd(PPh 3 4 (620 mg, 0.54 mmol) was added to the alkynylzinc solution. The resulting yellow solution was treated with chioroacetyl chloride (4.2 mL, mmol) dropwise over 10 minutes. Aft er 2 h at 0 0 C the reaction mixture was quenched by the addition of saturated aqueous NH44CI (500 mL), and diluted with ethyl acetate. The aqueous phase was extracted with ethyl acetate (3 x 200 ml) and the combined organic layers were washed with water (200 ml), brine (200 ml), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo to afford a dark brown oil that was purified by column chromatography eluting with hexane, then 99:1 hexane:ethyl acetate to afford I-chloro-4-(l-cyclohexen--y)-3-butyn-2-one (4.4 g, 67% yield) as an orange oil. 1H NM (CDC1 3 300 M4Hz) 8 6.56 (in, 4.23 2H), 2.19 (in, 4H1), 1.68-1.62 (in, 4F-I. MS (El ioniz~ation) 1 82 (3"CI NC), 184 (31CI Mf). The material was carried on to the next step without fuirther purification.
Examnle Synthesis of 4-(l-Cyclohexen-l-ylethynvl)-2-methyl-1.3-thiazole.
P-toluenesulfonic acid salt I-Chloro-4-(l-cyclohexen-l-yl)-3-butyn-2-one (2.0 g, 11.0 inmol) was dissolved in DMF (10.0 mL), thioacetaniide (950 mng, 12.6 inmol) was added, and the resulting pale brown solution was stirred at ambient temperature for 64 h. The reaction mixture was diluted with ethyl acetate (300 mL), washed with saturated NaHCO 3 solution (300 mL), water (300 mL), brine (300 dried over Na2S0 4 filtered, and concentrated in vacuo. The residue was dissolved in ethyl acetate, adsorbed onto silica gel and purified by column chromatography eluting with hexane, 99:1 then 98:2 hexane:ethyl acetate to afford 4-(l-cyclohexen-1-ylethynyl)-2-inethyl-1,3-thiazole (620 mng, 28% yield) as a yellow powder.
'H NMR 300 MI-z) 8 7.22 1K), 6.27-6.24 (in, 1K), .2.7 3H) 2.22-2.12 (in, 4K4), 1.68- 1.58 (mn, 4H1).
1-Cyclohexen- 1-ylethynyl)-2-methyl- 1,3-thiazole (620 ing, 3.1 minol) was dissolved in ethanol (30 mL) at ambient temperature. p-Toluenesulfonic acid monohydrate (580 mg, 3.1 minol) was WO 01/16121 PCT/US00/23923 42 added in one portion to afford a brown solution. After all of the acid had dissolved the reaction mixture was stirred for several minutes and then concentrated in vacuo to afford a dark brown oil which solidified under high vacuum. The crude material was dissolved in hot ethyl acetate. After cooling to ambient temperature the material was stored in the freezer for few hours. The supematant solution was decanted and the crystalline solids were dried under high vacuum to afford crystalline 4-(l-cyclohexenl-ylethynyl)-2-methyl-1,3-thiazole p-toluenesulfonate salt (882 mg 74% yield) as yellow crystals.
M.p. 128-129 0 C. 'H NMR (CDOD, 300 MHz) 5 7.87 1H), 7.71-7.68 J=9 Hz, 2H), 7.24-7.21 (d, J=9Hz, 3H), 6.38 2.88, 3H), 2.36 3H), 2.21-2.17 4H), 1.68-1.64 4H).
Example 41 Synthesis of 4-(1-Cyclohexen-l-vlethynyl)-13-thiazol-2-ylamine, p-toluenesulfonic acid salt l-Chloro-4-(l-cyclohexen-l-yl)-3-butyn-2-one (2.0 g, 11 mmol) was dissolved in DMF (10.0 mL), thiourea (996 mg, 13.1 mmol) was added, and the resulting pale brown solution was stirred at ambient temperature for 16 h. The reaction mixture was diluted with ethyl acetate (200 mL), washed with saturated NaHCO 3 solution (100 mL), water (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The dark oil was dissolved in ethyl acetate, adsorbed onto silica gel and purified by column chromatography eluting with 9:1 then 3:1 hexane:ethyl acetate to afford 4-(1cyclohexen-l-ylethynyl)-l,3-thiazol-2-ylamine (1.1 g, 49% yield) as an off-white solid. MS (El ionization) 204 4-(1-Cyclohexen-l-ylethynyl)-1,3-thiazol-2-ylamine (1.lg, 5.4 mmol) was dissolved in ethanol mL) at ambient temperature. p-Toluenesulfonic acid monohydrate (1.0 g, 5.4 mmol) was added in one portion to afford a brown solution. After all of the acid had dissolved the reaction mixture was stirred for several minutes and then concentrated in vacuo to afford a dark brown oil which solidified under high vacuum. The crude material was dissolved in hot ethyl acetate. After cooling to ambient temperature the material was stored in the freezer. After several hours in the freezer, the supernatant solution was decanted and the crystalline solids were dried under high vacuum to afford 4-(1cyclohexen-l-ylethynyl)-l,3-thiazol-2-ylamine p-toluenesulfonate salt (1.84 g, 87% yield) as off-white powder. M.p. 188-189 0 C. 'H NMR (CD 3 OD, 300 MHz) 5 7.72 -7.69 J=9 Hz, 2H), 7.24-7.22 (d, J=6 Hz, 2H), 6.94 1H), 6.34-6.32 1H), 2.36 3H), 2.19-2.15 4H) 1.70-1.61 4H).
Example 42 Synthesis of 2-(l-Cyclohexen-l-ylethynyl)-6-methvlpyridine 2-Bromo-6-methyl pyridine (2.0 g, 12 mmol) and Cul (440 mg, 2.3 mmol) were combined in DME (30 mL), and argon gas was bubbled through the suspension for several minutes to deoxygenate WO 01/16121 PCT/US00/23923 43 the mixture. Triethylamine (8.0 mL, 58 mmol) and PdCI 2 (PPh 3 2 (814 mg, 1.16 mmol) were added, followed by the dropwise addition of 1-ethynylcyclohexene (1.7 g, 15 mmol). The reaction was stirred at ambient temperature overnight. GC/MS showed no starting 2-bromo-6-methylpyridine remaining.
The mixture was diluted with ethyl acetate (100 mL), and filtered through Celite T M The pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography eluting with hexane then 99:1, 98:2 hexane:ethyl acetate to afford 2-(l-cyclohexen-l-ylethynyl)-6-methylpyridine (1.8 g, 79% yield) as a red oil. 'H NMR (CDC1 3 300 MHz) 5 7.51-7.46 1H), 7.21 J-9 Hz, 1H), 7.03 J--9 IH), 6.32-6.29 (m, 1H), 2.53 3H), 2.24-2.21 2H), 2.14-2.12 2H), 1.67-1.57 4H). MS (ESI) 198.1 Example 43 Synthesis of 2-(Cyclohexylethynyl)-6-methylpyridine 2-Bromo-6-methyl pyridine (2.0 g, 11.6 mmol) and Cul (440 mg, 2.3 mmol) were combined in DME (30 mL), and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (8.0 mL, 58 mmol) and PdCI 2 (PPh 3 2 (814 mg, 1.16 mmol) were added, followed by the dropwise addition of cyclohexylethyne (1.25 g, 11.6 mmol). The reaction was stirred at ambient temperature overnight. GC/MS showed no starting 2-bromo-6-methylpyridine remaining.
The mixture was diluted with ethyl acetate (100 mL), and filtered through Celite T M The pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography eluting with hexane then 98:2, 96:4 hexane:ethyl acetate to afford 2-(cyclohexylethynyl)-6-methylpyridine (1.78 g, 77% yield) as a pale brown liquid that partially solidified on standing in the freezer. 'H NMR (CDCI 3 300 MHz) 8 7.52-7.46 1H), 7.20 J-9 Hz, 1H), 7.03 J-9 Hz, 1H), 2.6 (m,l 2.54 3H), 2.93-2.89 2H), 1.78-1.73 (m, 2H), 1.57-1.54 3H), 1.36-1.32 3H). MS (ESI) 200.1 Example 44 Preparation of 4-Methyl-2-l(E)-2-phenylethenyll-1 3-oxazole Cinnamamide (2.0 g, 14 mmol), chloroacetone (0.93 mL, 16 mmol), and K 2 CO3 (940 mg, 6.8 mmol) were combined under argon and the mixture was heated in a 1200 C oil bath. The reaction mixture solidified and stirring stopped upon heating. After 16 h, the cooled reaction mixture was quenched by the addition of water (20 mL) and then diluted with ethyl acetate (100 mL). The organic phase was dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography eluting with 90:10 hexane:ethyl acetate. The product that was obtained (280 mg) showed some close running impurities by thin layer chromatography (TLC). The material was WO 01/16121 PCT/US00/23923 44 further purified by preparative TLC via multiple elutions with 95:5 hexane:ethyl acetate. Isolation of the middle of the main band afforded 4-methyl-2-[(E)-2-phenylethenyl]-1,3-oxazole (47.0 mg, 2% yield) as a yellow oil. 'H NMR (CDCI 1 300 MHz) 5 7.53-7.32 7H), 6.91 J=16.4 Hz, IH), 2.21 3H). MS (ESI) 185.7 Example 4-Methvl-2-[2-phenvlethenvll-1.3-thiazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of Ethyl 4-methyl-1,3-thiazole-2-carboxylate: Ethyl thiooxamate (3.0 g, 22 mmol) was dissolved in ethanol (30 mL) under argon. Chloroacetone (1.8 mL, 22 mmol) was added and the resultant solution was heated at 800 C for 16 h. The reaction mixture was concentrated in vacuo, adsorbed onto silica gel, and purified by column chromatography on silica gel eluting with 97:3, then 95:5 hexane:ethyl acetate to afford ethyl 4-methyl-1,3-thiazole-2-carboxylate (850 mg, 22% yield) as an oil. 'H NMR (CDCI 3 300 MHz) 7.20 1H), 4.48 J=7.1 Hz), 2.56 3H), 1.47 3H). MS (El ionization) 171 Preparation of (4-Methyl-l,3-thiazol-2-yl)methanol: Ethyl 4-methyl-l,3-thiazole-2carboxylate (450 mg, 2.6 mmol) was dissolved under argon in tetrahydrofuran (THF) (5 mL). The solution was cooled to 00 C in an ice bath and treated with diisobutylaluminum hydride (5.3 mL of a solution in toluene, 7.9 mmol). The reaction mixture was allowed to slowly warm to ambient temperature over 16 h and then cooled to -780 C and cautiously quenched by the dropwise addition of methanol. After gas evolution had ceased, saturated aqueous sodium-potassium tartrate (10 mL) was added, and the reaction mixture was allowed to warm to ambient temperature. The aqueous phase was extracted with ethyl acetate (3 x 30 mL), the combined organics dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with 90:10 then 1:1 hexane:ethyl acetate to afford (4-methyl-l,3-thiazol-2-yl)methanol (180 mg, 53% yield) as an oil. H NMR (CDC 3 300 MHz) 6 6.81 IH), 4.84 3H), 2.36 3H). MS (El ionization) 129 Preparation of 4-Methyl-,3-thiazole-2-carbaldehyde: (4-Methyl-1,3-thiazol-2-yl)methanol (180 mg, 1.4 mmol) was dissolved in CH 2 Cl 2 (9 mL), and treated with Magtrieve T M (2.5 The resultant suspension was heated to reflux for 16 h. The reaction mixture was cooled and filtered through a pad of Celite T M The filter pad was washed thoroughly with CH 2
CI
2 and the combined filtrates were concentrated in vacuo to afford 4-methyl-1,3-thiazole-2-carbaldehyde (120 mg, 67% yield) as an oil. The material was carried on to the next step without further purification. MS (El ionization) 127 WO 01/16121 PCT/US00/23923 Preparation of 4-Methyl-2-[2-phenylethenyll-1,3-thiazole: Sodium hydride (102 mg of a suspension in mineral oil, 4.3 mmol) was slurried in dry I,2-dimethoxyethane (DME) under argon, cooled to 00 C in an ice bath, and diethylbenzyl phosphonate (0.89 mL, 4.3 mmol) was added dropwise to the suspension. Thirty minutes after the completion of the phosphonate addition, 4-methylthiazole-2carboxaldehyde (120 mg, 0.94 mmol) was added as a solution in DME (5 mL). After stirring for 4 h, further NaH (40 mg of a 60% suspension in mineral oil, 1.0 mmol) was added and the reaction mixture was allowed to warm to ambient temperature and stir for 16 h. The reaction mixture was quenched by the addition of water (20 mL) and ethyl acetate (50 mL). The aqueous phase was extracted with ethyl acetate (2 x 30 mL), the combined organics were dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with 90:10 hexane:ethyl acetate to afford 4-methyl-2-[2-phenylethenyl]-1,3-thiazole (30 mg, 16% yield) as an oil.
'H NMR analysis showed the material to be a 10:1 mixture of E and Z isomers (spectral data are reported for the major isomer). 'H NMR (CDCI 3 300 MHz) 8: 7.54-7.51 2H) 7.42-7.23 6.80 1H) 2.47 3H). MS (ESI) (minor isomer) 202.2 (major isomer) 201.6 (MN).
Example 46 2-Methvl-4-|(E)-2-phenylethenvll-1.3-thiazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of Ethyl 2-methyl-l,3-thiazole-4-carboxylate: Thioacetamide (7.6 g, 100 mmol) was added to ethanol (60 mL), and the resultant suspension was cooled to 0° C and treated with ethyl bromopyruvate (12.5 mL, 100 mmol). The resultant solution was stirred for five minutes at 0° C at which time the ice bath was removed and the solution was allowed to warm to ambient temperature.
After 0.5 h at ambient temperature, the solution was heated to reflux. After 12 h, the solvents were removed in vacuo, and the resultant crude product was taken up in ethyl acetate (300 mL). The organic phase was washed with saturated aqueous NaHCO 3 (50 mL), and the basic aqueous solution was then extracted with additionalethyl acetate (2 x 50 mL). The combined organic solutions were washed with brine (50 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo to afford a yellow waxy solid. The crude product was dissolved in a small amount of hot ethyl acetate, diluted with hot hexane, and the resultant solution was allowed to cool to ambient temperature, seeded with a small amount of crude product, and transferred to the freezer. After 16 h the crystalline product was collected, washed with cold 8:1 hexane:ethyl acetate, and allowed to dry under high vacuum to afford ethyl 2-methyl-1,3-thiazole-4-carboxylate (11.76 g, 69% yield) as large brownish crystals. M.p. 56- 58.5°C. IH NMR (CDC13, 300 MHz) 5 8.06 1H), 4.43 2H), 2.78 3H), 1.42 3H). MS (El ionization) 171 WO 01/16121 PCT/USOO/23923 46 Preparation of (2-Methyl-1,3-thiazol-4-yl),nethanol: Ethyl 2-methyl-I ,3-thiazole-4carboxylate (15 g, 60 mmol) was slurried in THF (40 mL) and cooled to 0 0 C. Lithium aluminum hydride (60 mL of a I M solution in TI-F) was added slowly and the reaction mixture was allowed to warm to 250 C. After 16 h the reaction was quenched by the dropwise addition of water (2.28 mL), 15% NaOI- solution (2.28 mL) and then water (6.84 mL). Ethyl acetate (100 mL) was added, the reaction mixture filtered, and the filtrate concentrated in vacuo. The crude residue was chromatographed on silica gel with ethyl acetate:hexane 1) as eluant to afford (2-methyl-I1,3-thiazol- 4-yl)methanol as an oil (4.41 g, This material was carried on to the next step without further purification.
Preparation of 2-Methyl-1,3-thiazole-4-carbaldehyde: (2-Methyl- 1,3 -thiazol-4-yl)methanol (4.4 g, 34 mmol) was dissolved in CH 2
CI
2 (400 mL). Magtrieve m (44 g) was added and the reaction was heated under reflux for 24 h. The mixture was filtered through CeliteTm, and the filter pad was washed thoroughly with CH 2
CI
2 The filtrate was concentrated in vacuo to afford 2-methyl-l,3thiazole-4-carbaldehyde (3.7g, 86% yield) as a yellow oil which was carried on to the next step without further purification. IH NM4R (CDCI3, 300 Mi-z) 5 9.98 8.06 2.79 MIS (El ionization) 127 Preparation of 2-Methyl-4-I(E)-2-phenylethenyll-1,3-thiazole: Sodium hydride (120 mg of a 60% suspension in mineral oil, 3.0 mmol) was slurried in DME (6 mL) and cooled to 00 C. Diethyl benzylphosphonate (1.1I g, 5.0 minol) was added dropwise and after 15 minutes, 2-mctbyl- 1,3 -thiazole- 4-carbaldehyde (320 mg, 2.5 mmol) in DME (5 mL) was added dropwise to the reaction mixture. After 3 h at 00 C the reaction mixture was diluted with ethyl acetate (10 mL) and washed with saturated aqueous NH 4 CI (10 mL), saturated aqueous NaIICO 3 (10 mL), brine (10 mL), dried over Na2S0 4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with 90:10 hexane:ethyl acetate to afford 2-methyl-4-[(E)-2-pheriylethenyl]-l,3thiazole (330 mg, 65% yield) as an oil. IH NMR (CDCI3, 300 MHz) 8 7-7.5 (in, 8H), 2.7 MS (ESI) 202.1 Example 47 2-Methvl-44nPhenvlethnl-1 .3-thiazole.
n-toluenesulfonic acid salt The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of I-Chloro-4-phenyl-3-butyn-2-one: Anhydrous ZnCI 2 (10 g, 73 minol) was dissolved in TI-F (50 mL) and the solution cooled to 00 C in an ice bath. In another flask phenylacetylene (8.0 mL, 73 mmol) was dissolved in THIF (50 mL), cooled to 00 C in an ice bath, and WO 01/16121 PCT/US0O/23923 47 treated with n-butyllithium (32 mL of a 2.2M solution in hexane, 70 mmol). After 20 minutes the phenylethynyllithium solution was added via cannula to the ZnCI 2 solution. After an additional minutes Pd(PPh 3 4 (1.23 g, 1.06 mmol) was added to the alkynylzinc solution. The resulting yellow solution was treated with chloroacetyl chloride (8.8 mL, 110 mmol) dropwise over 10 minutes. After 2 h at 0° C the reaction mixture was quenched by the addition of cold aqueous IM HCI (50 mL), and diethyl ether (500 mL). The acidic aqueous was extracted with diethyl ether (2 x 50 mL) and the combined organic extracts were washed with water (50 mL), saturated NaHCO 3 (50 mL), and brine mL). The dark solution was dried and decolorized over Na 2
SO
4 and charcoal and filtered through a pad of CeliteTM. The pad was washed thoroughly with ethyl acetate and the combined filtrates were concentrated in vacuo to afford a dark brown oil. The crude product was purified by column chromatography eluting with hexane, 99:1, 98:2, 96:4, then 94:6 hexane:ethyl acetate to afford 1chloro-4-phenyl-3-butyn-2-one (7.83 g, 60% yield) as an orange oil which darkened and partially solidified upon standing in the freezer. 1H NMR (CDCI3, 300 MHz) 5 7.63-7.60 2H), 7.53-7.39 3H), 4.33 3H). MS (El ionization) 178 (35C1 180 (37CI Preparation of 2-Methyl-4-(phenylethynyl)-1,3-thiazole, p-toluenesulfonic acid salt: 1- Chloro-4-phenyl-3-butyn-2-one (1.6 g, 9.1 mmol) was dissolved in dry acetonitrile (15 mL), treated with thioacctamide (680 mg, 9.1 mmol), and heated to reflux for 4 h. After cooling, the acetonitrile was removed in vacuo, and the residue was partitioned between water (50 mL) and ethyl acetate (150 mL).
The organic phase was dried over Na 2
SO
4 filtered, concentrated in vacuo, adsorbed onto silica gel, and purified by column chromatography on silica gel eluting with 99:1, 98:2, then 95:5 hexane:ethyl acetate to afford 2-methyl-4-(phenylethynyl)-1,3-thiazole (240 mg, 13% yield) as a red oil. 1H NMR (CDC13, 300 MHz) 5 7.58-7.54 2H), 7.40-7.34 4H), 2.74 3H). MS (El ionization) 199 2-Methyl-4-(phenylethynyl)-l,3-thiazole (270 mg, 1.35 mmol) was dissolved in ethanol (6 mL) at ambient temperature. p-Toluenesulfonic acid monohydrate (252 mg, 1.32 mmol) was added in one portion to afford a brown solution. After all of the acid had dissolved the reaction mixture was stirred for several minutes and then concentrated in vacuo to afford a dark brown oil which partially solidified under high vacuum. The crude material was triturated with diethyl ether, and the resulting solids were taken up in hot ethyl acetate. The hot solution was treated with decolorizing carbon and filtered hot to afford a pale brown solution, which was treated with hexane until cloudy, then heated to afford a clear solution. The solution was seeded with authentic product and allowed to crystallize. After cooling to ambient temperature the material was stored in the freezer for 16 h. The supernatant solution was decanted and the crystalline solids were pumped down under high vacuum to afford crystalline 2methyl-4-(phenylethynyl)-1,3-thiazole, p-toluenesulfonic acid salt (260 mg, 52% yield) as brown crystals. M.p. 131-132.5 0 C. 1H NMR (CD30D, 300 MHz) 8 7.99 1H), 7.70 J=8.1Hz), 7.60-7.57 2H), 7.46-7.43 3H), 7.23 J=8 Hz, 2H), 2.89, 3H), 2.36 3H). MS (ESI) 199.7 WO 01/16121 PCT/US00/23923 48 Example 48 Preparation of 4-(Phenylethvnvl)-1,3-thiazol-2-amine.
p-toluenesulfonic acid salt 1-Chloro-4-phenyl-3-butyn-2-one (245 mg, 1.37 mmol) was dissolved in DMF (1.0 mL), thiourea (126 mg, 1.66 mmol) was added, and the resulting pale brown solution was stirred at ambient temperature for 5 days. The reaction mixture was diluted with Ethyl acetate (50 mL), washed with saturated NaHC0 3 solution (10 mL), water (10 mL), brine (10 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The dark oil was dissolved in methanol, adsorbed onto silica gel and purified by column chromatography eluting with 4:1, 3:1, then 2:1 hexane:ethyl acetate to afford 4- (phenylethynyl)-l,3-thiazol-2-amine (56 mg, 20% yield) as an oil. 1H NMR (CDC13, 300 MHz) 8 7.53-7.50 2H), 7.37-7.32 3H), 6.77 1H), 5.41 (br s, 2H). MS (El ionization) 200 4-(Phenylethynyl)-l,3-thiazol-2-amine (700 mg, 3.5 mmol) was dissolved in ethanol (20 mL) at ambient temperature. p-Toluenesulfonic acid monohydrate (660 mg, 3.5 mmol) was added in one portion to afford a brown solution. After all of the acid had dissolved the reaction mixture was stirred for several minutes and then concentrated in vacuo to afford a dark brown oil which solidified under high vacuum. The crude material was dissolved in hot ethyl acetate containing a few drops of methanol. After cooling to ambient temperature the material was stored in the freezer for few hours.
The supernatant solution was decanted and the crystalline solids were dried under high vacuum to afford crystalline 4-(phenylethynyl)-l,3-thiazol-2-amine, p-toluenesulfonic acid salt (1.0 g 73% yield) as brown crystals. M.p. 131-132.5 0 C. 1H NMR (CD30D, 300 MHz) 8 7.73 -7.70 J=9 Hz, 2H), 7.56-7.52 (dd, J-9, 2 Hz, 2H), 7.48-7.37 3H), 7.23-7.20 J=9Hz, 2H), 7.12, 1H), 2.34 3H).
Example 49 4-Methyl-5-(phenylethynyl)-1,3-thiazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of 5-Bromo-4-methyl-1,3-thiazole: N-Bromosuccinimide (30 g, 170 mmol) was suspended in CC1 4 (150 mL) and 4-methylthiazole (15 g, 150 mmol) was added in one portion. The mixture was heated under reflux for 4 h then allowed to cool to ambient temperature. The reaction mixture was diluted with ethyl acetate (500 mL) and washed with water, then brine. The organics were dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 9:1 hexane:methylene chloride to afford 5-bromo-4-methyl-1,3-thiazole (6.8g, 24% yield) as a dark reddish oil. 1H NMR (CDC13, 300 MHz) 8 8.69 1H), 2.44 3H). MS (El ionization) 177 (35CI 179 (37C1 WO 01/16121 PCT/USOO/23923 49 Preparation of 4-Methyl-5-(phenylethynyl)-1,3-thiazole: 5-Bromo-4-methyl- 1,3-thiazole (570mg, 3.2 mmol) and Gui(120 mg ,0.63 mmol were combined in DME (5 mL )and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine mL, 14 mmol) and PdCI 2 (PPh 3 2 (220 ing, 0.32 mmcl) were added, followed by the dropwise addition of phenylacetylene (1.0 mL, 9.5 mmol After 16 h stirring at ambient temperature additional phenylacetylene (0.35 mL, 3.2 mmol), CuT, (61 mg, 0.32 mmol), and PdCI 2 (PPh 3 )2 (I110 mg, 0. 16 mmol) were added and the reaction mixture was stirred for a further 16 h. The reaction was then heated to reflux, and after 3 h the reaction mixture was cooled, diluted with ethyl acetate (20 and filtered through CeliteTMA. The filter pad was washed thoroughly with ethyl acetate and the combined filtrates were washed with water (20 mL), brine (20 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo.
The residue was purified by column chromatography on silica gel eluting with 95:5 hexane:ethyl acetate to afford 4-methyl-5-(phenylethynyl)-1,3-thiazole (166 mg, 26% yield) as a red oil. IH NMR (CDCI3, 300 MHz) 8 8.62 IH), 7.54-7.50 (in, 7.37-7.26 (in, 3H), 2.6 3H). MS (ESI) 200.1 Example 5-!(2-Fluorophenvl)ethynvll-4-methXl-1 .3-thiazole 5-Bromo-4-methyl-1,3-thiazole (500 mg, 2.8 mmol) and Cul (120 mg, 0.63 mmol) were combined in DM-E (5 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamnine (2.0 mL, 14 mmol) and PdC1 2 (PPh 3 )2 (220 mng, 0.32 mmol) were added, followed by the dropwise addition of (2-fluorophenyl)acetylene (1.0 g, 8.3 mmcl) The reaction mixture was heated and after 3 hi at reflux, allowed to cool, diluted with ethyl acetate (20 mL), and filtered through CeliteTh 4 The filter pad was washed thoroughly with ethyl acetate and the combined filtrates were washed with water (20 mL), brine (20 mL), dried over Na 2
SO
4 and filtered.
The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with 95:5 hexane:ethyl acetate to afford 5-[(2-fluorophenyl)ethynyl]4-methyl-1,3thiazole (690 mg, 56% yield) as a red oil. 1H NMR (CDCI3, 300 MIk) 68.65 IH), 7.53-7.47(in, I 7.3 6-7.26 (in, I 7.17-7.09 (mn, 2H), 2.62 3 MS (ESI) 217.3 Example 51 2-(Phenylethvnvl)-I.3-thiazole. o-toluenesulfonic acid salt 2-Bromo-1,3-thiazole (2.0 g, 12 mmcl) and Cull (460 mg, 2.4 minol) were combined in DME mL), and argon gas wvas bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (8.6 inL, 62 mmcl) and PdC12(PPh3)2 (856 mg, 1.22 mmol) were added and then phenylacetylene (3.7g, 36.5 minol) was added dropwise. The reaction was heated to reflux at which point the reaction mixture solidified. Additional DME (20 inL) was added to dissolve the solids and the reaction mixture was allowed to stir for 16 h at reflux, at which time GUIMS showed no WO 01/16121 PCT/USOO/23923 remaining 2-bromothiazole. After cooling, the mixture was diluted with 200 mL ethyl acetate, and filtered through Celite'T. The pad was then washed thoroughly with ethyl acetate and the combined filtrates were washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography eluting with hexane, then 97:3 hexane:ethyl acetate) to afford 2-(phenylethynyl)-l,3-thiazole (1.3 g, 60% yield) as a yellowish oil. 1H NMR (CDC13, 300 MHz) 8 7.86 J=3.2 Hz, 1H), 7.61-7.57 2H), 7.42-7.26 (m, 4H). MS (ESI) 185.2 2-(Phenylethynyl)-l,3-thiazole (3.00 g, 16.2 mmol) was dissolved in ethanol (75 mL) at ambient temperature. p-Toluenesulfonic acid monohydrate (3.08 g, 16.2 mmol) was added in one portion to afford a yellow solution. After all of the acid had dissolved the reaction mixture was stirred for several minutes and then concentrated in vacuo to afford a bright yellow solid. An attempt was made to recrystallize the material from hot ethyl acetate:hexane, but the recovery of solid after cooling was poor. Analysis of the crude material from the reaction showed it to be of sufficient purity. 2- (Phenylethynyl)-l,3-thiazole, p-toluenesulfonic acid salt (4.77 g, 82% yield) was obtained as a bright yellow powder that turns beige upon standing at ambient temperature. M.p. 130-132 0 C. 1H1 NMR 300 MHz) 8 8.17 J=3.7 Hz, 1H), 8.05 J=3.7 Hz, 1H), 7.72-7.67 4 7.56-7.46 3 7.22 J=8 Hz, 2 2.35 3 MS (ESI) 186.1 Example 52 4-Bromo-2-(phenylethynyll-13-thlazole 2,4-Dibromo-l,3-thiazole (2.0 g, 8.2 mmol) and CuI (312 mg, 1.64 mmol) were combined in DME (25 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (5.7 mL, 41 mmol) and PdCi 2 (PPh 3 2 (580 mg, 0.82 mmol) were added, and then phenylacetylene (0.90 mL, 8.6 mmol) was added dropwise. The reaction was stirred for 4 h at ambient temperature at which point GC/MS analysis showed the reaction to be complete The mixture was filtered through CeliteTM, the filter pad was washed thoroughly with Ethyl acetate and the combined filtrates were then concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL), washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane, then 97:3 hexane:ethyl acetate to afford pure 4-bromo-2-(phenylethynyl)-l,3-thiazole (1.0 g, 46% yield) as a colorless solid. M.p. 79-82 0 C; 1H NMR (CDC13, 300 MHz) 8 7.60-7.56 2H) 7.45-7.26 3H), 7.26 1H). MS (El ionization) 263 79Br), 265 81Br).
WO 01/16121 PCT/US00/23923 51 Example 53 5-Methyl-2-(phenylethvnyl)-1.3-thiazole 4-Bromo-2-(phenylethynyl)-l,3-thiazole (500 mg, 1.89 mmol) was dissolved in dry THF mL) under argon, cooled to -78 0 C, then t-butyllithium (1.7 mL of a 1.7M solution in pentane, 2.8 mmol) was added. After 30 min 0.4 mL of the reaction mixture was quenched with saturated NH 4
CI
and extracted with ethyl acetate. GC/MS of the crude product from this workup and comparison with a GC/MS of authentic 2-(2-phenylethynyl)-thiazole showed the two to be identical. This confirmed that the initial coupling with phenylacetylene (in Example 9) took place at the bromine at the 2 position rather than at the 4 position. The GC/MS also showed that the reaction was not complete. Additional tbutyllithium (1.7 mL of a 1.7M solution in pentane, 2.8 mmol) was added. After 30 minutes iodomethane (0.36 mL, 5.7 mmol) was added to the reaction mixture, and the reaction was allowed to warm to ambient temperature for 16 h. The solvents were then removed in vacuo, and residue dissolved in ethyl acetate (50 mL), washed with water (50 mL), brine (50 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography eluting with hexane, 99:1, 98:2, 97.5:2.5 hexane:ethyl acetate to afford 2-(2-phenylethynyl)-5-methyl-1,3thiazole (90 mg). 1H NMR analysis of the material showed it to be impure, despite its apparent homogeneity by TLC analysis. The crude compound was dissolved in DMSO and purified by preparative HPLC with a 30 min gradient from 70:30 water:acetonitrile to 100% acetonitrile to afford pure 5-methyl-2-(phenylethynyl)-1,3-thiazole as a white powder (44 mg, 24% yield). M.p. 82-83 0 C. 1 H NMR (CDC13, 300 MHz) 8 7.85-7.55 2H), 7.49 J=0.6 Hz, 1H), 7.37-7.32 3H), 2.49 3H).
MS (ESI) 200.1 Example 54 2-(3-Methylphenyl)ethvnyll-1,3-thiazole 2-Bromo-1,3-thiazole (2.0 g, 13 mmol) and Cul (330 mg, 1.7 mmol) were combined in DME (25 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (5.7 mL, 43 mmol) and PdCI 2 (PPh 3 2 (604 mg, 0.86 mmol) were added and mtolylacetylene (1.0 g, 8.6 mmol) was added dropwise. The reaction mixture was heated under reflux for 4 h at which time GC/MS showed the reaction was complete. The mixture was filtered through Celite
T
M, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL), washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane then 98:2 hexane:ethyl acetate to afford methylphenyl)ethynyl]-thiazole (280 mg, 16% yield) as a yellow oil which was still impure by 1H NMR analysis. The compound (280 mg) was dissolved in DMSO and purified by preparative HPLC with a 30 min gradient from 70:30 water:acetonitrile to 100% acetonitrile to afford pure WO 01/16121 PCT/US00/23923 52 methylphenyl)ethynyl]-1,3-thiazole (122 mg, 6% yield) as a yellow oil. lH NMR (CDCI3, 300 MHz) 6 7.84 J=3.0 Hz, 1H), 7.40-7.34 (in, 3H), 7.28-7.18 (in, 2H4), 2.34 31-1). MS (EST) 200.1 Example 2-1(4-Fluoronbhenvl)ethvnvll-1 .3-thiazole 2-Bromo- 1,3 -thiazole (1.0 g, 6.2 inmol) and Cul (152.4 mg, 0.8 minol) were combined in DME (17 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (3 mL, 21 mmol) and PdCI 2 (PPh 3 2 (280 mng, 0.40 mmol) were added and I1ethynyl-4-fluorobenzene (500 ing, 4.2 minol) was added dropwise. The reaction was heated under reflux for 4 h at which time GU/MS showed the reaction was complete. The mixture was filtered through CeliteTm, and the filter pad was washed thoroughly with Ethyl acetate. The combined filtrates were then concentrated in vacua. The residue was dissolved in ethyl acetate (100 mL), washed with water (100 brine (100 mL), dried over Na 2
SQ
4 filtered and concentrated in vacu. The residue was purified by column chromatography eluting with hexane then 98:2 hexane:ethyl acetate to 24[(4fluorophenyl)ethynyl]-1,3-thiazole as a white solid which was impure by NMR analysis. The compound (280 mg) was dissolved in DMS0 and purified by preparative HPLC with a 30 min gradient from 60:40 water:acetonitrile to 100% acetonitrile to afford 2-[(4-fluorophenyl)ethynyl]-1,3-thiazole (33 mng, 3% yield) as a white solid. M.p. 78-79 0 C. 'H NMR (CDC 3 300 MHz) 8 7.83 J=3.0 Hz, IM, 7.60-7.55(m, 2H), 7.39 J=3.0 Hz, 7.14-7.00 (in, 211). MS (ESI) 204 Example 56 2-1(2-Fluoroiphenvl)ethvnvll-1.3-thiazole 2-Broino-1,3-thiazole (2.0 g, 13 minol) and Cu! (316 mg, 1.6 iniol) were combined in DME m.L) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (5.8 mL,, 42 iniol) and PdCI 2 (PIh 3 2 (580 mg, 0.83 minol) were added and 1ethynyl-2-fluorobenzene (1.0 g, 8.3 inmol) was added dropwise. The reaction was heated under reflux for 4 h at which time GUIMS showed the reaction was complete. The mixture was filtered through CeliteTm, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacua. The residue was dissolved in ethyl acetate (200 mL) and washed with water (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane then 98:2 hexane:ethyl acetate to afford fluorophenyl)ethynyl]-1,3-thiazole (700 ing, 41% yield) as a yellow oil that was impure by 'H NMR analysis. The compound (140 mg) was dissolved in DMS0 and purified by preparative 1-PLC with a min gradient from 50:50 water:acetonitrile to 100% acetonitrile to afford 24[(2fluorophenyl)ethynyl]-1,3-thiazole (53.2 mg, 75% yield) as a yellow oil. NMR (CDC1 3 300 MIHz) 8 7.88 J=3.0 Hz, IH), 7.59-7.53 (in, IH), 7.42-7.35 (in, 7.18-7.09 (in, 2H). MIS (ESI) 204
(NC+H).
WO 01/16121 PCT/US00/23923 53 Example 57 4.5-Dimethyl-2-(phenvlethnyl)-1.3-thlazole.
The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of 2-Bromo-4,5-dimethyl-l,3-thiazole: 4,5-Dimethyl-l,3-thiazole (5.0 g, 44 mmol) was dissolved in carbon tetrachloride (60 mL), and N-bromosuccinimide (8.19 g, 46 mmol) was added. The resulting mixture was heated to reflux while protected from light with aluminum foil. After '2 h at reflux the dark suspension was cooled and allowed to stand at ambient temperature for 16 h. The reaction mixture was then heated to reflux for an additional 4 h and cooled to ambient temperature.
After standing at ambient temperature for 16 h, GC/MS analysis showed mostly monobromide, with a small amount of starting 4,5-dimethyl-1,3-thiazole present. The yellow supematant solution was decanted from the black solids that had deposited on the walls of the flask. The crude product was concentrated in vacuo, and then further purified by column chromatography eluting with hexane, 99:1, 98:2, 96:4, 94:6, then 90:10 hexane:ethyl acetate to afford 2-bromo-4,5-dimethyl-1,3-thiazole (1.34 g, 16 yield) as a white semi-solid. M.p. 55-60 0 C. 'H NMR (CDCI3. 300 MHz) 8 2.31 3H), 2.30 (s, 3H). MS (El ionization) 191 7 9 Br), 193 (M "Br).
Preparation 4,5-Dimethyl-2-(phenylethynyl)-l,3-thiazole: 2-Bromo-4,5-dimethyl- 1,3thiazole (1.3 g, 6.8 mmol) and CuI (190 mg, 1.36 mmol) were combined in DME (30 mL) and argon gas was bubbled through the suspension for several minutes to deoxygenate the mixture. Triethylamine (5.0 mL, 34 mmol) and PdCI2(PPh3)2 (477 mg, 0.68 mmol) were added and phenylacetylene (1.86 g, 18.9 mmol) was added dropwise. After stirring at ambient temperature for 16 h, GC/MS showed the reaction was complete. The mixture was filtered through Celite T M the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (100 mL), washed with water (100 mL), brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with hexane then 95:5 hexane:ethyl acetate to afford 4,5-dimethyl-2-(phenylethynyl)-l,3-thiazole (338 mg 24% yield) as a white solid. M.p. 55-60 0 C. 1H NMR (CDC13, 300 MHz) 5 7.56-7.53 2H), 7.35- 7.33 3H), 2.38 6H). MS (ESI) 214.0 Example 58 5-Methyl-3-j(E)-2-phenylethenyll-1.24-oxadiazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of (2E)-N-Hydroxy-3-phenyl-2-propenimidamide: To a solution of hydroxylamine hydrochloride (690 mg, 10 mmol) in ethanol was added NaOH (400 mg, 10 mmol), WO 01/16121 PCT/US00/23923 54 followed by cinnamonitrile (1.3 g, 10 mmol). This mixture was heated to reflux for 16 h. The ethanol was removed in vacuo, the residue was acidified with 3M HCI (5 mL) and the solution was boiled for minutes. The cooled solution was made basic (pH 8) with NH0OH, and partitioned between water and ethyl acetate. The organics were concentrated to give a sticky white mass which was used without further purification.
Preparation of 5-Methyl-3-[(E)-2-phenylethenyl]-l,2,4-oxadiazole: (2E)-N-Hydroxy-3phenyl-2-propenimidamide (1.6 g, 10 mmol) was mixed with excess acetyl chloride (30 mL), and heated to reflux. After 1 h the solution was cooled and the excess acetyl chloride was removed in vacuo. The resultant solids were triturated in hot ethyl acetate-hexane-acetone solution, and the supematant was removed. The remaining solids were dried under high vacuum to afford 5-methyl-3- [(E)-2-phenylethenyl]-1,2,4-oxadiazole (0.50 g, 27% yield from cinnamonitrile) as a pale yellow powder. M.p. 166-168 0 C. 'H NMR (DMSO-d, 300 MHz) 5 7.59 3H), 7.42 3H), 6.58 J=16 Hz, IH), 2.17 3H).
Example 59 5-l(E)-2-phenylethenvll-1.2.4-oxadiazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of N-[(Dimethylamino)methylenej-cinnamamide: Cinnamamide (4.4 g, mmol) and N,N-dimethylformamide dimethylacetal (9.9 mL, 75 mmol) were heated at reflux for 2 h.
Excess formamide and methanol formed in the reaction were removed in vacuo. The solids which formed were collected and washed with hexane, followed by two portions of ethyl acetate, then dried under vacuum at 400 C to afford N-[(dimethylamino)methylene]-cinnamamide (3.5 g, 58% yield) as a white crystalline solid. M.p. 91-93 0 C. 'H NMR (DMSO-d 6 300 MHz) 8 8.58 1H), 7.83 J=16 Hz, IH), 7.57 2H), 7.36 3H), 6.74 J=16 Hz, 1H), 3.18 6H).
Preparation of N-I(Hydroxyamino)methylenel-cinnamamide: N-[(Dimethylamino)methylene]-cinnamamide (1.0 g, 5.0 mmol) was added to a solution of hydroxylamine hydrochloride (415 mg, 6.0 mmol) in acetic acid (5 mL of 70% aqueous) and aqueous NaOH (1.2 mL, 5 M, 6.0 mmol). After stirring for 1.5 h water (5 mL) was added, and the reaction mixture was cooled to 50 C in an ice bath. The solution was partitioned between ethyl acetate and water, and the organics were dried over Na 2
SO
4 then concentrated in vacuo to afford N- [(hydroxyamino)methylene]-cinnamamide (850 mg, This material was carried on to the next step without further purification.
WO 01/16121 PCT/USOO/23923 Preparation of 5-L(E)-2-pbenylethenylJ-1,2,4-oxadiazole: N-[(Hydroxyamino)methyleneJcinnamamide (850 mg, 4.5 mnmol) was dissolved in acetic acid/dioxane (20 mL 1:1 and heated to reflux for 2 h. The cooled solution was made basic (pH 8) with K 2 C0 3 and partitioned between ethyl acetate and water. The organics were dried over Na 2 SOI, and concentrated in vacuo. The resultant solids were triturated with hot ethyl acetate, and the soluble portion was purified by flash column chromatography Qfn silica eluting with 2:1 hexane:ethyl acetate to afford 5-[(E)-2-phenylethenyl]- 1,2,4oxadiazole (60 mg, 8% yield) as a white powder. M.p. 53-55*C. 11H NMR (CDC13, 300 MHz) 8 8.43 1H1), 7.85 J=16 Hz, IH), 7.60 (in, 2H), 7.45 (in, 3H), 7.05 J=16 Hz, lH). MS (El ionization) 171 (Mi-H).
Eape6 Preparation of I -Methyl-5-j(E)-Z-phenylethenyU]- H-I .2.4-triazole N-[(Dimethylamino)methyleneJ-cinnamamide (2.0 g, 10 minol) was added to a mixture of acetic acid (20 mL) and methylhydrazine (0.6 m.L, 11 I mol). The mixture was heated to 90' C for 2 h.
The cooled mixture was concentrated in vacuo, and made basic (pH 9) with solid K 2 C0 3 The aqueous residue was partitioned between ethyl acetate and water, the organics were dried over Na 2
SO
4 and concentrated in vacuo. The resultant powder was recrystallized from a minimum of boiling Ethyl acetate. The collected solids were triturated and washed with ethyl acetate to afford l-methyl-5-[(E)-2phenylethenyl]-IH-1,2,4-triazole (0.9 g, 49 yield) as a pale yellow powder. M.p. 67-70'C from Ethyl acetate. 'H NMR (CDCl 3 300 MHz) 8 7.88 1H), 7.73 J=16 Hz, lH), 7.57 (in, 211), 7.39 (mn, 3H), 6.92 J=I6Hz, I 3.96 3H).
Example 61 H-I ,2.4-triazole The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of N-I(Dimethylamino)ethylideuel-cdnnamamide: Cinnainaiide (1.8 g, 12 mxnol) and NN-diinethylacetamide diinethylacetal (2.8 inL, 19 nimol) were heated to 1200 C for 2 h.
Excess amide and methanol formed in the reaction were removed in vacua. The resultant material was purified by flash colurm chromatography on silica eluting with ethyl acetate to afford N- [(dimethylamino)ethylidene]-cinnamamide (2.1 g, 78 yield) as a pale yellow solid containing one equivalent of ethyl acetate. 'H NMR (DMSO-d 6 300 Mvffz) 8 7.61 J=16 Hz, lH), 7.53 (in, 211), 7.36 (im, 3H), 6.68 J=16 Hz, 111), 3.13 (app d, J=15 Hz, 611), 2.30 3H).
Preparation of 3-Metbyl-5-[(E)-2-phenyletaenyl-1 H-1,2,4-triazole: N-[(Diinethylamino)ethylidene]-cinnamamide (2.0 g, 9.0 minol) was added to a solution of hydrazine hydrate (0.59 mL, 10 minol) and acetic acid (25 mQL. The mixture was heated to 90*C for 3 h, cooled, WO 01/16121 PCT/USOO/23923 56 and concentrated in vacua. The resulting material was basified (pH 8) with solid K 2 C0 3 and partitioned between ethyl acetate and water. The organics were dried over Na 2
SO
4 and concentrated in vacuo. The resultant material was purified by flash column chromatography on silica eluting with 1:2 hexane-ethyl acetate to afford 3-methyl-5-[(E)-2-phenylethenyl]-IH-l,2,4-triazole (1.6 g, 96% yield) as a white powder. M.p. 131-134 0 C. 'H NlvMR (CDCI,, 300 MHz) 6 7.68 J=16 Hz, 0.3H), 7.59 (d, J=16 Hz, 0.7H), 7.49 (in, 2H1), 7.32 (mn, 3H), 7.05 J=16 Hz, 111), 6.49 J=16 Hz, 0.4H), 6.21 (hr s, 0.3H), 5.88 (hr s, 0.3H), 2.53 3H). MIS (El ionization) 184 Example 62 Ethyl 3-I(E)-2-phenylethenyll-1.2.4-thiadlazole-5-carboxylae The title compound was prepared in several steps, with the preparation of key intermediates set forth herein as follows: Preparation of 5-[(E)-2-Phenylethenyl]-1,3,4-oxathiazol-2-one: Cmnnamamide (1.9 g, 13 mmol) was mixed with chlorocarbonylsuiphenylchioride (1.0 mL, 13 inmol) in chloroform and heated to reflux for 18 h. The reaction mixture was concentrated in vacua and the resultant material was taken up in boiling hexane (100 mL). The solvent was decanted from the remaining solid, the volume of this solution was reduced by half, and the solution was allowed to cool. The crystalline solids which formed were collected and washed with hexane to afford 5-[(E)-2-phenylethenylJ- 1,3,4-oxathiazol-2one (1.93 g, 72% yield) as pale yellow needles. M.p. 105-106 0 C 'H NMR (DMSO-d 6 300 M~z) 8 7.52 (in, 3H), 7.46 0.5H1), 7.41 (in, 2.5H), 6.63 3=1 6 Hz, I H).
Preparation of Ethyl 3-[(E)-2-phenylethenyl]-1,2,4-thiadiazole-5-carboxylate: Phenylethenyl]-1,3,4-oxathiazol-2-one (1.4 g, 7.0 inmol) was mixed with ethyl cyanoformate (2.4 g, inmol) in xylenes, and heated to reflux. for 3 h. The reaction mixture was concentrated in vacuo, and the resulting material was recrystallized from ethyl acetate-ethanol to give ethyl phenylethenyl]- 1,2,4-thiadiazole-5-carboxylate (1.2 g, 66% yield) as a light yellow solid. M.p. 79- 80 0 C. 'H NMR (CDCl 3 300 MHz) 8 7.98 J=16 Hz, 111), 7.61 (in, 2H1), 7.40 (in, 3M1, 7.29 J=1 I Hz, 11H), 4.56 J=7 Hz, 2H), 1.50 J=7 Hz, 3H). MS (El ionization) 259 Exm 6 13.i(E't-2-phenylethenyll-1.2.4-thiadiazol-5-yllmethan-oI Ethyl 3-[(E)-2-phenylethenyl]-i,2,4-thiadiazole-5-carboxylate (1.0 g, 3.8 mmol) was suspended in methanol (50 mL), and NaBH.
4 (220 mng, 5.8 minol) was added in portions. The mixture was stirred for 16 h at ambient temperature. The reaction was concentrated in vacuo, acidified with aqueous HCI (4 M) to pH 2, and partitioned between Ethyl acetate and water. The organics were dried over Na 2
SO
4 concentrated in vacua, and the resultant material was rerystallized from ethyl acetate to afford 2-phenylethenyll- 1,2,4-thiadiazol-5-yl) methanol (0.6 g, 71% yield) as a pale yellow solid. M.p. 119- WO 01/16121 PCT/USO01/23923 57 120 0 C. 'H NMR (CDC1 3 300 MHz) 5 7.79 J=16 Hz, 1H), 7.57 2H), 7.38 3H), 7.23 J=16 Hz, IH), 5.14 J=5 Hz, 2H), 3.29 J=5 Hz, 1H). MS (El ionization) 217 Example 64 2-(l-Cvclohexen-1-vlethynyl)-5-methythiophene 2-Iodo-5-methylthiophene (1.0 g, 4.5 mmol) was mixed with PdCI 2 (44 mg, 0.25 mmol), PPh 3 (200 mg, 0.75 mmol), Cul (140 mg, 0.75 mmol) and K 2
CO
3 (1.66 g, 12 mmol) in a solution of DME and water Argon gas was bubbled through the suspension for twenty minutes to deoxygenate the mixture. 1-Ethynyl-l-cyclohexene (1.06 g, 10.0 mmol) was added and the mixture was heated to reflux. After 16 hours the reaction was cooled and filtered through Celite T M The resultant solution was partitioned between water and ethyl acetate. The organic layer was dried over anhydrous Na 2
SO
4 and concentrated in vacuo. The resulting product was purified by flash column chromatography on silica gel eluting with hexane to afford 2-(l-cyclohexen-l-ylethynyl)-5-methylthiophene (0.96 g, 86% yield) as a light yellow oil. 'H NMR (CDCI 3 300 MHz) 5 6.94 J= 3.5 Hz, IH), 6.60 1H), 6.18 (m, IH), 2.45 3H), 2.13 4H), 1.68-1.57 4H). MS (El ionization) 202 Example Synthesis of 2-(1H-inden-2-ylethvnyl)pyridine 2-Indanone (1.0g, 7.6 mmol) was dissolved in THF (60 mL) and cooled to -78 0 C in an argon blanketed flask. Potassium hexamethyl disilazide (9.1 mmol, 18.2 mL of a 0.5M solution in toluene) was added dropwise to this stirred solution. After 30 min, N-phenyl trifluoromethanesulfonimide (4.05 g, 11.4 mmol) was added as a solution in THF (15 mL). The reaction was stirred for 15 min at -78 0
C,
then brought to ambient temperature and stirred for an additional 1 h, after which time it was quenched with H20 (15 mL) and diluted with ethyl acetate (500 mL). The ethyl acetate solution was washed with (3 x 100 mL) and brine (100 mL), then dried (MgSO4), filtered, and concentrated in vacuo. The residue was chromatographed on silica gel, eluting with 30:1 hexanes:ethyl acetate to afford 2- [(trifluoromethyl)sulfonyl]-lH-indene (1.28 g, 64% yield) as a brown oil. The enol triflate (500 mg, 1.89 mmol) and 2-ethynyl pyridine (579 mg, 5.62 mmol) were dissolved in DME (10 mL) and deoxygenated via argon bubbling for 20 min and then added via syringe to a deoxygenated DME mL) solution of triphenylphosphine (100 mg, 0.38 mmol), bis-triphenylphosphine palladium dichloride (133 mg, 0.19 mmol), Cul (72 mg, 0.38 mmol), and triethylamine (954 mg, 1.32 mL, 9.45 mmol). The reaction was capped with a reflux condenser and stirred at 80°C for 1.5 h, after which time it was cooled to ambient temperature and poured in to a separatory funnel containing ethyl acetate (300 mL), where it was washed with H 2 0 (2 x 100 mL) and brine (100 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 6:1 hexanes:ethyl acetate to afford 2-(1H-inden-2-ylethynyl)pyridine (309 mg, 75% yield) as the free base, which was then solubilized in diethyl ether (25 mL) and precipitated as the WO 01/16121 PCT/USOO/23923 58 hydrochloride salt 170-172*C) upon treatment with I M HCI in diethyl ether (5 mL). '1H NMR
(CD
3 OD, 300 MHz) 5 8.98 J='5.7 Hz, 1H), 8.76 J=8.0 Hz, 1H), 8.33 J=8.0 Hz, 1K), 8.17 (t, J=6.3 Hz, I1H), 7.82 1KH), 7.72 (in, 2H), 7.53 (in, 2H), 3.96 2H). MS (ESI) 218.1 Example 66 Synthesis of 243.4-dihydro-2-nanhthalenvlethvnyl)Dvnidine 2,6-Lutidine (547 mg, 594 p±L, 5.1 mmol) was added neat, via syringe to a stirred, argon blanketed solution of P-tetralone (500 mg, 3.4 mmol) in methylene chloride (30 mL) at ambient temperature. After 5 mini, trifluoroinethanesulfonic anhydride (1.439 g, 858 tiL, 5.1 inmol) was added slowly via syringe. After 1.5 h, the reaction was quenched with saturated aqueous NaHCO 3 (5 mL) and partitioned between ethyl acetate (200 mL) and H 2 0 (50 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacu. The residue was then chromatographed on silica gel, eluting with 20:1 hexanes:ethyl acetate to afford 3-[(trifluoromethyl)sulfonyl]-1,2-dihydronaphthalene (361 mg, 38% yield) as a brown oil. Following the procedure described for Example 65, the 13-tetralone enol triflate (275 mg, 1.0 mmcl) was cross-coupled with 2-ethynylpyridine (337 mg, 3.27 mmol) in a reaction over 16 h at 70*C. Upon completion, the reaction was concentrated in vacuo, and the residue was chromatographed on silica gel, eluting with 7:1 hexanes:ethyl acetate to afford 2-(3,4-dihydro-2naphthalenylethynyl)pyridine (193 mg, 84% yield) as an off white semi-solid. 'H NMR (CDCI 3 300 MIHz) 8 8.62 J=4.5 Hz, 110), 7.67 (ddd, J=7.7 Hz, 7.7 Hz, 1.8 Hz, 7.48 J=7.9 Hz, IK), 7.07- 7.28 (in, 5H), 7.00 I 2.91 J=7.8 Hz, 214), 2.59 J=8.4 Hz, 2H). MS (ESI) 232.1 (M 4 ExarnDle 67 Synthesis of 1-(2-Dyridinylethynyl)l-indanoI n-Butyllithium (18.93 inmol, 11.8 ml of 1.6M solution in hexanes) was added dropwise to a stirred solution of 2-ethynylpyridine (1.95 g, 18.93 mmol) in THF (80 mL) at -78'C. After 15 mini, solid anhydrous CeC1 3 (4.66 g, 18.93 mmol) was added and the reaction was stirred for an additional 1 h, at which time a solution of 1-indanone (1.00 g, 7.57 minol) in THIF (10 mL) was added via syringe.
The reaction was then warmed to ambient temperature, stirred for I h, and quenched with 5 mL H 2 0.
The reaction was then diluted with ethyl acetate (250 mL) and extracted with H 2 0 (3 x 75 mL). The combined aqueous portions were back-extracted with ethyl acetate (100 mL). The ethyl acetate layers were combined, dried over MgSO 4 filtered, and concentrated in vacua. The crude residue was chromatographed on silica gel, eluting with 1:1 hexanes:ethyl acetate to afford 1-(2-pyridinylethynyl)- I-undanol (1.1 g, 62% yield) as a thick amber liquid. 114NMR (CDCI 3 300 MIHz) 8 8.42 J=4.9 Hz, 1KH), 7.63 1=6.6 Hz, I1K), 7.54 (ddd, J=7.7 Hz, 7.7 Hz, 1.7 Hz, I 7.33 J=7.8 Hz, I1H), 7. 7.25 (in, 4H), 5.50 1K), 3.12 (in, 1K), 2.91 (in, 1K), 2.68 (in, IM), 2.53 (mn, 1H). MS (ESI) 236.1 WO 01/16121 PCT/US00/23923 59 Example 68 Synthesis of I-fluoro-2-{2-pyridinylethynyl)-2-indanol Potassium hexamethyldisilazide (19.9 mmol, 39.8 mL of 0.5M solution in toluene) was added slowly to a stirred solution of2-indanone (2.20 g, 16.6 mmol) in THF (60 mL) at -78 0 C. After 15 min, trimethylsilyl chloride (2.64 mL, 20.8 mmol) was added to generate the IH-inden-2-yl trimethylsilyl ether. The reaction was stirred for 10 min at -78 0 C, then brought to ambient temperature and stirred for an additional 30 min, after which time the solvents were removed in vacuo and replaced with anhydrous acetonitrile (80 mL). Solid Selcctfluor" (8.82 g, 24.9 mmol) reagent was then added and the resulting slurry was stirred for 16 h at ambient temperature. The reaction was then diluted with ethyl acetate (400 mL) and extracted with H 2 0 (3 x 100 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacuo. The residue was chromatographed on silica gel, eluting with 4:1 hexanes:ethyl acetate to afford the 1-fluoro-2-indanone (1.05 g, 42% yield) product as a sticky brown solid. Following the procedure outlined for Example 67, the intermediate 1-fluoro-2-indanone (1.05 g, mmol) was cross-coupled with 2-ethynyl pyridine (1.44 g, 14.0 mmol) to afford 1-fluoro-2-(2pyridinylethynyl)-2-indanol (320 mg, 18% yield, 8% overall yield) as a tan semi-solid. 'H NMR
(CDCI
3 300 MHz) 8 8.57 J=4.8 Hz, 1H), 7.64 (ddd, J=7.8 Hz, 7.8 Hz, 1.8 Hz, 1H), 7.49 J=7.3 Hz, 1H), 7.22-7.43 5H), 5.98 9 F] 56.0 Hz, 1H), 4.29 1H), 3.51 2H). MS (ESI) 254.1 Example 69 Synthesis of 2-[(3-fluoro-l H-inden-2-vl)ethynyllpyridine n-Butyllithium (2.39 mmol, 1.5 mL of 1.6M solution in hexanes) was added slowly to a stirred solution of 1-fluoro-2-(2-pyridinylethynyl)-2-indanol (550 mg, 2.17 mmol) and LiBr (188 mg, 2.17 mmol) in THF (15 mL) at -78 0 C. After 30 min, methanesulfonyl chloride (274 mg, 185 pL, 2.39 mmol) was added and the reaction was stirred for an additional 1.5 h at ambient temperature before quenching with H 2 0 (5 mL). The reaction flask contents were then transferred to a separatory funnel containing ethyl acetate (100 mL) that was subsequently washed with H 2 0 (2 x 50 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was then dissolved in methylene chloride (25 mL), treated with DBU (1.982 g, 1.947 mL, 13.02 mmol), and warmed to reflux. After 16 h, the reaction mixture was cooled to ambient temperature, diluted with methylene chloride (100 ml) and washed with H 2 0 (2 x 50 mL). The methylene chloride layer was dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 5:1 hexanes:ethyl acetate to afford 2-[(3-fluoro-lH-inden-2-yl)ethynyl]pyridine mg, 15% yield) as a yellow oil which was then solubilized in ether (15 mL) and precipitated as the hydrochloride salt 150-152C) upon treatment with IM HCI in diethyl ether (2 mL). 'H NMR
(CD
3 OD, 300 MHz) 8 8.81 J=5.3 Hz, IH), 8.59 (ddd, J=8.0 Hz, 8.0 Hz, 1.4 Hz, 1H), 8.16 J=8.
WO 01/16121 PCT/USO0/23923 Hz, 8.00 (dd, J=6.4 Hz, 6.4 Hz, lH), 7.45-7.58 (in, 4H), 3.76 9 F] =6.6 Hz). MS (ESI) 236.1 (MW+H).
Example Synthesis of 2-1(tert-butoxvcarhonvl)aminol benzoic acid Anthranilic acid (1.37 g, 10.0 mmol) and di-tert-butyl dicarbonate (3.12 g, 14.3 mmol) were added to a stirred mixture of 0.5M NaOH (20.0 dioxane (10.0 mL), and CH 3 CN (2.0 m-L) at 0 0
C.
The cold bath was removed, and the reaction mixture was stirred at ambient temperature for 16 h before quenching with 10% aqueous citric acid (30 mL). The mixture was diluted with H 2 0 (100 mL) in a separatory funnel and extracted with methylene chloride (3 x 100 mL). The combined methylene chloride extracts were dried (MgSO 4 filtered, and concentrated in vacuo to obtain 2-[(Ier!butoxycarbonyl)aminojbenzoic acid (2.33 g, 98% yield) as a white solid that was suitably pure to carry on to the next synthetic step.
Example 71 Synthesis of tert-butyl 2-iethoxv(metbyl~aminolcarbonyl1 phenylcarbamate To a stirred solution of the N-BOC-anthranilic acid from Example 70 (2.33 g, 9.8 mmol) in methylene chloride (100 mL) was added EDC (2.82 g, 14.7 mmol), hydroxybenzotriazole dihydrate (1.459 g, 10.8 minol), diisopropyl ethylamine (6.0 mnL, 34.3 mmcl), and NO-dimethylhydroxylamine hydrochloride (1.43 g, 14.7 mmol). The reaction was stirred for 16 h, after which time it was concentrated in vacuo and diluted with ethyl acetate (500 mL). The ethyl acetate solution was washed with IM HCI (100 mL), saturated NaHCO 3 (100 mL), brine (100 mL), and 1120 (2 x 100 mL), then dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 4:1 hexanes:ethyl acetate to afford tert-butyl 2-{[methoxy(methyl)amino]carbonyl)phenylcarbamate (2.00 g, 73% yield) as a pale yellow solid.
Example 72 Synthesis of tert-butyl 2-formylphenvicarbamate LiAIH4 (6.67 mmol, 6.67 mL of a I OM solution in THF) was added to an Ar-purged, stirred solution of the tert-butyl 2-{[methoxy(methyl)arnino]carbonyl~phenylcarbamate from Example 71 (1.87 g, 6.67 mmcl) in THF (50 mL) at -78'C. Stirring was continued at -78*C for 15 min, after which time the reaction was brought to ambient temperature and stirred for an additional I h. The reaction was then cooled to 0 0 C and treated with IN aqueous HCI (25 mL) to quench. The mixture was poured in to a separatory funnel containing H 2 0 (250 mL) and extracted with methylene chloride (3 x 100 mL).
The combined methylene chloride layers were dried (MgSOA) filtered, and concentrated in vacuo to afford tert-butyl 2-formyiphenylcarbamnate (1.48 g, 100% yield) as a tan solid.
W001/16121 PCT/USOO/23923 61 Example 73 Synthesis of tert-butvl 24r 1-hyd roxv-3-(2-nvridinvl)-2-v~rovynyll phenvlcarbamate n-Butyllithium (14.0 mmol, 8.75 mL of 1.6M solution in hexanes) was added dropwise to a stirred solution of 2-ethynylpyridine (1.44 g, 14.0 mmol) in TI-f (80 mL) at -781C. After 15 min, solid anhydrous CeC1 3 (3.45 g, 14.0 mmol) was added and the reaction was stirred for an additional I h, after which time a solution of the terl-butyl 2-formylphenylcarbamate from Example 72 (1.48 g, 6.7 mmol) in THIF (20 mL) was added via syringe. The reaction was warmed to ambient temperature, stirred for I h, quenched with H 2 0 (5 mL), then diluted with ethyl acetate (250 mL) and extracted with H 2 0 (3 x mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacua, resulting in a crude residue that was chromatographed on silica gel, eluting with 2:1 hexanes:ethyl acetate to afford the racemic 2' alkanol (1.71 g, 79% yield) as a light peach colored amorphous solid.
Example 74 Synthesis of 4-(2-pyrid invlethynyl)-1 .4-dihvdro-2H-3.l-benzoxazin-2-one The tert-butyl 2-formylphenylcarbamate from Example 73 (250 mg, 0.77 mmol) was deprotected without incident by dissolving the compound in dioxane (10 mL), treating it with 4M HCI in dioxane (30 mL, 120 mmol HCI), and stirring for 2.5 h, resulting in precipitation from solution of the l-(2-aminophenyl)-3-(2-pyridinyl)-2-propyn-1-ol as the hydrochloride salt. The dioxane solvent was decanted, the precipitate was triturated with diethyl ether (3 x 20 mL) and dried in vacua to obtain the crude deprotected material as a pale pink solid. The deprotection was assumed to be quantitative, and the crude material was therefore carried on to the next step, dissolving it in a solution of diisopropylethylamine (500 mg, 674pjL, 3.87 mmol) in methylene chloride (20 mL). The reaction flask was cooled to 0 0 C, and phosgene (1.935 mmol, 1.02 mL of a 1.89M solution in toluene) was added dropwise to the stirred solution. Stirring at 0 0 C was continued for 2.5 h, followed by an H-20 (5 mL) quench. The biphasic mixture was diluted with ethyl acetate (100 mL) and washed with H 2 0 (2 x mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacua. The crude residue was chromatographed on silica gel, eluting with 2:1 hexanes:ethyl acetate to obtain 4-(2pyridinylethynyl)-l,4-dihydro-2H-3,1-benzoxazin-2-one (140 mg, 73% yield) as a tan, semi-solid glass.
'H NMR (CDCI 3 300 MHz) 8 9.13 I1-ED, 8.61 J=4.4 Hz, I1H), 7.69 (ddd, J=7.8 Hz, 7.8 Hz, 1.8 Hz, IH), 7.51 J=7.8 Hz, 1H), 7.41 J=7.6 Hz, IH), 7.27-7.34 (in, 7.11 (ddd, J=7.6 Hz, 7.6 HL, 0.8 Hz, 1H), 6.93 J=7.8 Hz, 1H), 6.34 1H). MS (ESI) 251.1 (M 4 Example Synthesis of ethyl 3-42-pyvridinylethnyl)--azabicclo3.2.1 loct-2-ene-8-carboxvlate N-Carbethoxy-4-tropinone (500 mg, 2.54 mmol) was dissolved in TI-F (25 mL) and cooled to 78'C in an argon blanketed flask. Potassium hexamethyldisilazide (3.05 inmol, 6.1 mL of a solution in toluene) was added dropwise to this stirred solution. After 15 min, N-phenyl WO 01/16121 PCT/US00/23923 62 trifluoromethanesulfonimide (1.36 g, 3.81 mmol) was added as a solution in THF (10 mL). The reaction was stirred for 15 min at -78 0 C, then brought to ambient temperature and stirred for an additional 1 h, after which time it was quenched with H 2 0 (10 mL) and diluted with ethyl acetate (200 mL). The ethyl acetate solution was washed with H20 (3 x 50 mL), dried (MgSO 4 filtered, and concentrated in vacuo. The residue was chromatographed on silica gel, eluting with a gradient from to 50% ethyl acetate in hexanes to afford the ethyl 3-[(trifluoromethyl)sulfonyl]-8-azabicyclo[3.2. ]oct- 2-ene-8-carboxylate (820 mg, 98% yield) as a tan liquid. The ethyl 3-[(trifluoromethyl)sulfonyl]-8azabicyclo[3.2.l]oct-2-ene-8-carboxylate (820 mg, 2.50 mmol) and 2-ethynylpyridine (516 mg, 5.00 mmol) were dissolved in DME (15 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated DME (25 mL) solution of triphenylphosphine (131 mg, 0.50 mmol), bis-triphenylphosphine palladium dichloride (176 mg, 0.25 mmol), Cul (95 mg, 0.50 mmol), and triethylamine (1.265 g, 1.74 mL, 12.50 mmol). The reaction was warmed to 50°C and stirred for 1.5 h, after which time it was cooled to ambient temperature and poured into a separatory funnel containing ethyl acetate (350 mL), where it was washed with H 2 0 (2 x 100 mL) and brine (100 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 1:1 hexanes:ethyl acetate to afford ethyl 3-(2pyridinylethynyl)-8-azabicyclo[3.2. 1]oct-2-ene-8-carboxylate (400 mg, 57% yield) as a free base. The toluenesulfonate salt was then prepared by adding solid toluenesulfonic acid to a solution of ethyl 3-(2pyridinylethynyl)-8-azabicyclo[3.2.1]oct-2-ene-8-carboxylate (200 mg, 0.69 mmol) in ethanol (10 mL).
The mixture was stirred for 5 min until all solids were dissolved, after which time the solution was concentrated in vacuo. The resulting red oil was triturated with ether (3 x 10 mL) and placed under high vacuum, where it foamed to a dark red semi-solid. 'H NMR (CD30D, 300 MHz) 8 8.86 J=5.3 Hz, 1H), 8.64 (ddd, J=8.0 Hz, 8.0 Hz, 1.5 Hz, 1H), 8.16 J=8.1 Hz, 1H), 8.07 (dd, J=6.3 Hz, 1H), 7.80 J=8.2 Hz, 2H), 7.31 J=8.0 Hz, 2H), 6.96 J=5.3 Hz, 1H), 4.64 br., 1H), 4.52 br., IH), 4.24 J=7.0 Hz, 2H), 2.92-3.08 br., 1H), 2.45 3H), 2.07-2.41 4H), 1.86 br., 1H), 1.36 J=7.0, 3H). MS (ESI) 283.2 Example 76 Synthesis of 1-chloro-4-(trimethylsilyl)-3-butyn-2-one Aluminum trichloride (21.9 g, 164 mmol) was suspended in CH 2 Cl2 (250 mL) and cooled in an ice bath. Bis(trimethylsilyl)acetylene (20.0 g, 117 mmol) and chloroacetyl chloride (10.3 mL, 129 mmol) were combined in CH 2 Cl 2 (150 mL) and the solution was added to the AICI 3 suspension dropwise from an addition funnel over I h. The dark brownish-red solution was stirred at 0°C for 1 h, then the ice bath was removed. After I h at ambient temperature, the reaction was cooled to 0°C and quenched by slow addition of IM HCI.(250 mL). The acidic solution was extracted with CH 2 Cl 2 (2 x 500 mL), the combined organic layers were washed with H 2 0 (500 mL), NaHCO 3 (500 mL), brine (500 mL) and dried over Na 2
SO
4 The organic layer was treated with silica gel, and filtered to afford a clear WO 01/16121 PCT/US00/23923 63 solution which was concentrated in vacuo. The residue was distilled under high vacuum through a vigreaux column. The main part of the distillate was collected at a head temperature of 58*C (the lower thermometer was 68'C) to afford 1 -chloro-4-(trimethylsilyl)-3-butyn-2-one (11.25 g, 54% yield) as a light yellow oil. 'H NMR (CDCI 3 300 MHz) 6 4.24 214), 0.28 914).
Examyle 77 Synthesis of 2-methyl-4-1(trimethylsilyl)ethvnyll-1 .3-thiazole The 1-chloro-4-(trimethylsilyl)-3-butyn-2-one from Example 76 (10 g, 57.2 mmol) was dissolved in DMF (100 mL), then thioacetamide (5.6 g, 74 mmol) was added in one portion. The mixture was allowed to stir for 16 h at ambient temperature, at which time TLC showed no remaining 1-chloro-4-trimethylsilyl)-3-butyn-2-one. The mixture was diluted with ethyl acetate (400 mL), and washed with 1-20 (3 x 300 mL), brine (300 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography eluting with hexane, 98:2, then 96.5:3.5 hexane:ethyl acetate to afford 2-methyl-4-[(trimethylsilyl)ethynyl]-1,3-thiazole g, 72% yield) as reddish-brown oil. H NMR (CDCI 3 300 MI-lz) 8 7.32 IH), 2.70 31-1), 0.26 (s, 9H). MS (El ionization)195 Example 78 Synthesis of 4-F(trimethylsilyl~ethynyll- 1,3-thiazol-2-ylamine The 1-chloro-4-(trimethylsilyl)-3-butyn-2-one from Example 76 (4.25 g, 24.3 mmol) was dissolved in DMF (20 mL), then thiourea (2.45 g, 32.2 mmol) was added in one portion. The mixture was allowed to stir for 16 h at ambient temperature, at which time TLC showed no remaining I1-chloro- 4-(trimethylsilyl)-3-butyn-2-one. The mixture was diluted with ethyl acetate (200 mL), and washed with H 2 0 (3 x 200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography eluting with hexane, 9:1, then 4:1 hexane:ethyl acetate to afford 4-[(trimethylsilyl)ethynyl]-1,3-thiazol-2-ylamine (4.1 g, 86% yield) as a yellow solid. H NMR (CDCI 3 300 M.Hz) 8 6.70 1H), 5.47 (br s, 2H), 0.20 9H). MS (El ionization) 196 Example 79 Synthesis of 4-f(2-methyl-i .3-thiazol-4-yl)ethynyllisopuinoline 4-Bromoisoquinoline (276 mg, 1.33 mmol) and 2-methyl-4-[(trimethylsilyl)ethynyl]- 1,3thiazole from Example 77 (200 mg, 1.02 mmol) were dissolved in DMF (5 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated, 40 0 C DMF mL) solution of triphenylphosphine (71 mg, 0.27 mmol), bis-triphenylphosphine palladium dichloride (93 mg, 0.13 mmol), Cul (51 mg, 0.27 mmol), tetrabutylamnmonium iodide (377 mg, 1.02 mmol), and triethylamine (515 mg, 710 piL, 5.1 mmol). The reaction mixture was warmed to 60*C, and WO 01/16121 PCT/US00/23923 64 tetrabutylammonium fluoride (1.33 mmol, 1.33 mL of a 1.0 M solution in THF) was added slowly over h. The reaction was then cooled to ambient temperature and poured into a separatory funnel containing 1:1 hexanes:ethyl acetate (200 mL) where it was washed with 50% dilute brine (3 x 75 mL), dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 1.5:1 hexanes:ethyl acetate to afford 4-[(2-methyl-1,3-thiazol-4yl)ethynyl]isoquinoline (195 mg, 76% yield) as an off-white solid that was then solubilized in ether mL) and precipitated as the white hydrochloride salt 209-210 0 C) upon treatment with IM HCI in diethyl ether (5 mL). 'H NMR (CD 3 0D, 300 MHz) 8 9.85 1H), 8.93 1H), 8.73 J=8.4 Hz, 1H), 8.64 J=8.4 Hz, 1H), 8.42 (ddd, J=7.2 Hz, 7.2 Hz, 1.1 Hz, 1H), 8.17 (dd, J=7.1 Hz, 7.1 Hz, 1H), 8.16 1H), 2.85 3H). MS (ESI) 251.1 Example Synthesis of 4-{4-isoquinolinvlethynyl)-l-3-thiazol-2-amine Following the procedure and mole equivalents indicated for Example 79, 4-bromoisoquinoline (691 mg, 3.32 mmol) and 4-[(trimethylsilyl)ethynyl]-l,3-thiazol-2-amine from Example 78 (500 mg, 2.55 mmol) were cross-coupled to obtain 4-(4-isoquinolinylethynyl)-1,3-thiazol-2-amine (262 mg, 41% yield) as a tannish-orange solid after eluting with 2:1 hexanes:ethyl acetate from a silica gel column.
This material was then solubilized in ether (15 mL) and precipitated as the yellow hydrochloride salt >150 0 C, dec.) upon treatment with IM HCI in diethyl ether (5 mL). 'H NMR (CD 3 OD, 300 MHz) 8 9.85 1H), 8.96 1H), 8.69 J=8.4 Hz, 1H), 8.62 J=8.3 Hz, 1H), 8.40 (ddd, J=7.1 Hz, 7.1 Hz, 1.2 Hz, 1H), 8.16(dd, J=8.1 Hz, 8.1 Hz, IH), 7.58(s, 1H). MS (ESI) 252.0 Example 81 Synthesis of2-l(trimethvlsilyl)ethynyllpyrimidine PdCI 2 (166 mg, 0.93 mmol), Cul (481 mg, 2.52 mmol), and triethylamine (18 mL, 129 mmol) were combined in DME (50 mL) under argon. Argon gas was bubbled through the resulting dark suspension while it was warmed to 70 0 C in an oil bath. Triphenylphosphine (978 mg, 3.73 mmol) was added and the argon flow was continued for 10 min. The argon flow was discontinued, the heating bath was removed, and 2-bromopyrimidine (5.14 g, 32.3 mmol), and trimethylsilylacetylene (9.1 mL, 64 mmol), were added as a solution in DME (30 mL) followed by a rinse of the flask and syringe with DME (10 mL). Solids appeared in the flask after the addition was completed. The reaction mixture was then heated to 45 0 C. After 1 h the heating was discontinued and the reaction mixture was allowed to cool to ambient temperature. After 16 h at ambient temperature TLC analysis showed no starting 2bromopyrimidine present. The reaction mixture was concentrated in vacuo, diluted with diethylether (300 mL), and filtered. The filtrate was washed with saturated aqueous NaHCO 3 (100 mL), H 2 0 (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo to afford a dark oil which partially solidified when pumped down under high vacuum. The crude product was purified by column WO 01/16121 PCT/USOO/23923 chromatography eluting with hexane, 9:1, 8:1, 6:1, 4: 1, then 3:1 hexane:ethyl acetate to afford 2- [(trimethylsilyl)ethynyl]pyrimidine (5.2 g, 91 yield) as a yellow solid. 'H NMR (CDCI 3 300 M1-z) 6 8.72 J=~4.9 Hz, 2H), 7.25 (in, I 0.30 9H). MS (El ionization) 176 Example 82 Synthesis of 4-(2-pyrimidinvlethvnyl)isoa uinoli ne 4-Bromoisoquinoline (499 mg, 2.40 mmol) and 2-[(trimethylsilyl)ethynyljpyrimidine from Example 81 (352 mg, 2.00 mmol) were dissolved in DMF (10 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (84 mg, 0.32 mmol), bis-triphenylphosphine palladium dichloride (112 mg, 0. 16 minol), Cu! (61 mg, 0.32 inmol), tetrabutylammonium iodide (369 mg, 1.00 inmol), and triethylamine (1.01 g, 1.39 mL, 10.00 inmol) in DMF (20 mL) at 40'C. The reaction was warmed to 50 0 G, and tetrabutylammonium fluoride 10 mmol, 2. 10 mL of a 1.OM solution in THF) was added slowly over h. The reaction was then cooled to ambient temperature and poured in to a separatory funnel containing 1: 1 hcxanes:cthyl acetate (200 inL) where it was washed with 50% dilute brine (3 x 75 m1L), dried (MgSO4), filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 1.5:1 hexanes:ethyl acetate to afford 4-(2-pyrimidinylethynyl)isoquinoline (210 mg, yield) as an off-white solid that was then solubilized in diethyl ether (15 mL) and precipitated as the tan hydrochloride salt 158-159*C) upon treatment with of IM HCI in diethyl ether (3 mL).
'14 NMR (CD 3 OD, 300 MHz) 8 9.95 I M, 9.06 I1H), 8.99 J=5.0 Hz, 2M), 8.75 J=8.4 Hz, 1lH), 8.67 J=8.3 Hz, 114f), 8.42 (ddd, J=7.2 Hz, 7.2 Hz, 1. 1 H-z, I1-H), 8.17 (dd, J=7.3 1-z, 7.3 Hz, I1H), 7.67 (dd, J=5.0 Hz, 5.0 Hz, I MS (ESI) 232.0 Example 83 Synthesis of 2-1(6.7-dimethoxy-3.4-dihydro-2-naphthalenyl)ethynyllpyvridine Using the procedure described in Example 66 but using 6,7-dimethoxy-l-tetralone in place of P-tetralone, 6,7-dimethoxy-3,4-dihydro-1-naphthaleny trifluoromethanesulfonate was prepared in yield. 6,7-Dimethoxy-3,4-dihydro-l-naphthalenyl trifluoromethanesulfonate (1.48 g) was crosscoupled with 2-ethynylpyridine without incident following the procedure described for Example 66.
The crude reaction material was chromatographed on silica gel, eluting the product with 2:1 hexanes:ethyl acetate to obtain 2-(6,7-dimethoxy-3,4-dihydro-1-naphthalenylethynyl)pyridine (265 mng, 40% overall yield, 2 steps) as a pale yellow solid that was then solubilized in diethyl ether (15 mL) and precipitated as the yellow-orange hydrochloride salt upon treatment with of IM HCI in diethyl ether mL). M.p. 148-150*C; 'H NMR (CDOD, 300 MHz) 8 8.76 J=5.7 Hz, I 8.52 (ddd, J=7.9 Hz, 7.9 Hz, 1.5 Hz, 8.14 J=8.0 Hz, 1H), 7.94 (dd, J=6.3 Hz, 6.3 Hz, 1H), 7.11 IH), 6.80 Hz, 11H), 6.79 I 3.83 3 3.80 3H4), 2.74 J=8.2 Hz, 2H), 2.43 (in, 2H). MS (ESI) 292.1
(MW+H).
WO 01/16121 PCT/US00/23923 66 Example 84 Synthesis of methyl 5-1(2-methyl- 3-thiazol-4-yl)ethynyl nicotinate Concentrated sulfuric acid (18 mL) was added to a stirred solution of 5-bromonicotinic acid (10.1 g, 50.0 mmol) in methanol (300 mL). The reaction was warmed to reflux and stirred for 18 h, then cooled to ambient temperature and quenched with saturated NaHCO 3 adjusting the pH to The methanol was removed in vacuo, and the remaining aqueous mixture was further diluted with H 2 0 (250 mL) and extracted with ethyl acetate (3 x 100 mL). The ethyl acetate layers were combined, dried (MgSO4), filtered, and concentrated in vacuo to obtain methyl 5-bromonicotinate (10.09 g, 93% yield) as a white crystalline solid 98-99 0 C) without further purification. This material (3.02 g, 14.0 mmol) and 2-methyl-4-[(trimethylsilyl)cthynyl]-1,3-thiazole from Example 77 (2.48 g, 12.7 mmol) were dissolved together in DMF (20 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated, 40 0 C DMF (60 mL) solution of triphenylphosphine (534 mg, 2.04 mmol), bis-triphenylphosphine palladium dichloride (713 mg, 1.02 mmol), Cul (388 mg, 2.04 mmol), tetrabutylammonium iodide (1.88 g, 5.08 mmol), and triethylamine (6.414 g, 8.8 mL, 63.5 mmol). The reaction was warmed to 50*C, and tetrabutylammonium fluoride (14.0 mmol, 14.0 mL of a l.OM solution in THF) was added via syringe pump over 1.5 h. The reaction was then cooled to ambient temperature and poured into a separatory funnel containing 1:1 hexanes:ethyl acetate (400 mL) where it was washed with 50% dilute brine (3 x 100 mL). The aqueous portion was back-extracted with 1:1 hexanes:ethyl acetate (100 mL). The organic layers were then combined, dried (MgSO 4 filtered, and concentrated in vacuo, and the crude residue was chromatographed on silica gel, eluting with 3:1 hexanes:ethyl acetate to afford methyl 5-[(2-methyl-1,3-thiazol- 4-yl)ethynyl]nicotinate (2.56 g, 78% yield) as an off-white solid. M.p. 124-125 0 C. 'H NMR (CDCI 3 300 MHz) 5 9.15 1H), 8.93 1H), 8.40 (dd, J= 2.0 Hz, 2.0 Hz, 1H), 7.49 IH), 3.98 3H), 2.76 3H). MS (ESI) 259.0 Example Synthesis of N-hydroxyethanimidamide hydrochloride Hydroxylamine hydrochloride (13.8 g, 200 mmol) was dissolved in a IM solution of NaOH in ethanol (200 mL). To this was added acetonitrile (8.2 g, 10.43 mL, 200 mmol) neat, via syringe. The reaction was stirred at reflux for 17 h, then cooled to ambient temperature at which time 12M HCI was added (35.4 mL, 425 mmol). The mixture was concentrated in vacuo to afford a white solid to which was added boiling ethanol (200 mi). The insoluble material was then filtered, and the filtrate was collected and concentrated in vacuo to obtain N-hydroxyethanimidamide hydrochloride (19.45 g, 88% yield) as a white crystalline solid.
WO 01/16121 PCT/USOO/23923 67 Example 86 Synthesis of 3-(3-methyl-1 2.4-oxad iazol-5-yl)-5-1(2-niethyl-I .3-thiazol-4-vl)ethynyll nyridine NaH (23.4 mmol, 936 mg of a 60% suspension in mia oil) was added to a suspension of Nhydroxyethanimidamide hydrochloride from Example 85 (1.27 g, 11.5 mmol) in THF (50 mL). The mixture was warmed to 50*C and stirred for 30 min, after which time methyl 5-[(2-methyl-1,3-thiazol- 4-yl)ethynyl]nicotinate from Example 84 (1.00g, 3.9 mmol) was added as a solution in TI-F (20 mL).
After 45 min, the reaction was quenched with H 2 0 (15 mL), and partitioned between ethyl acetate (250 mL) and 1H20 (100 mL). The ethyl acetate layer was dried (MgSOA) filtered, and concentrated in vacuo. The crude residue was filtered through a short plug of silica gel, eluting with 2.5% methanol in methylene chloride, then recrystallized from 1:1 hexanes:ethyl acetate to obtain 3-(3-methyl- 1,2,4oxadiazol-S-yl)-S-[(2-methyl- 1,3 -thiazol-4-yl)ethynyljpyridine (596 mg, 54% yield) as white crystalline leaves, M.p. 153-154 0 C. 'H NMR (CDCI 3 300 MI-L) 6 9.27 J=2.0 Hzl, I1H), 8.95 J=2.0 Hz, I1-1), 8.52 (dd, J=2.0 Hz, 2.0 Hz, I 7.50 I 2.77 3H), 2.51 3H). MS 283.0 (NC+H).
Exam ple 87 Synthesis of I-(methylsufonyl')-3-U(2-methyl-1,3-thiazol-4-vl~ethvnyll-1 H-indole To a vigorously stirred solution of indole (1.0 g, 8.6 mmol) in DMF (40 mL) at ambient temperature was added KOH pellets (1.8 g, 32.1 mmol) followed by iodine (4.34 g, 17.1 mnmol). After min, the reaction was poured in to a separatory funnel containing saturated aqueous sodium thiosulfate (200 mL), which was then extracted with 1:1 hexanes:ethyl acetate (3 x 100 mL). The combined organic layers were first back-extracted with 1: 1 H 2 0:brine (3 x. 75 mL), then dried (MgS0 4 filtered, and concentrated in vacuo. The crude 3-iodo-IH-indole product was dissolved in benzene (IS mL) and to this solution was added tetrabutylammonium iodide (332 mg, 0.9 mmol), H 2 0 (10 mL), and v/v aqueous NaQI- (10 mL). This biphasic mixture was stirred vigorously at ambient temperature while a solution of methanesulfonyl chloride (1.47 g, 993 gL, 12.8 mmol) in benzene (15 m.L) was added dropwise via syringe. After this addition, the reaction was stirred for 1 h, then partitioned with ethyl acetate (200 mL) and H 2 0 (100 mL). The ethyl acetate layer was dried (MgSO 4 filtered, and concentrated in vacuo leaving a crude residue that was eluted through a short plug of silica with 4:1 hexanes:ethyl acetate. The resulting brown solid (2.26 g) was recrystallized from methanol to provide 3-iodo-1-(methylsulfonyl)-IH-indole (1.44 g, 43% yield, 2 steps) as tan-colored needles. Following the procedure and mole equivalents indicated above for Example 79 the 3-iodo-1-(methylsulfonyI)-IHindole (350 mg, 1.09 mmol) and 2-mnethyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 mg, 0.77 mmol) were cross-coupled to obtain 1-(methylsulfonyl)-3-[(2-methyl-1,3-thiazolyl)ethynyl]-IH-indole (55 mg, 23% yield) as a tan solid after eluting with 4:1 hexanes:ethyl acetate from a silica gel column. This material was then solubilized in ether (10 m.L) and precipitated as the off-white hydrochloride salt 152-154'C) upon treatment with of I M HCI in diethyl ether (2 mL).
'H NMR (CD 3 QD, 300 Mliz) 5 8.02 I1H), 7.96 1KH), 7.95 (dd, J=7.5 Hz, 1L1 Hz, 114), 7.81 (dd, WO 01/16121 PCT/US00/23923 68 J=~7.1 Hz, 1.2 Hz, 1H), 7.49 (ddd, J=7.8 Hz, 7.8 Hz, 1.2 Hz, 111), 7.43 (ddd, J=7.5 Hz, 7.5 H4, 1.1 Hz, 3.36 314i), 2.91 3H). MS 317.0 (NC+H).
Example 88 Synthesis of 2-chloro-5-1(2-methyl-1.3-th iazol-4-vl)ethvnvll Dyrdine 2-Chloro-5-iodo-pyridine (3.0g, 12.53 mmol) and 2-methyl-4-[(trimethylsilyl)ethynyl]-1,3thiazole from Example 77 (2.57 g, 13.16 mmol) were dissolved in DMF (15 m.L) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (394 mg, 1.50 mmol), bis-triphenylphosphine palladium dichloride (528 mg, 0.75 mmol), Cul (286 mg, 1.50 mmol), tetrabutylammonium iodide (927 mg, 2.51 mmol), and triethylamine (6.33 g, 8.7 mL, 62.7 mmol) in DMF (60 mL) at 40'C. The reaction was warmed to 50 0 C, and tetrabutylammonium fluoride (13.8 mmol, 13.8 mL of a L.OM solution in THIF) was added via syringe pump over 1.5 h. The reaction was then cooled to ambient temperature and poured into a separatory funnel containing 1: 1 hexanes:ethyl acetate (200 mL) where it was washed with 50% dilute brine (3 x mL), dried (MgSO4, filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 0.5% to 1.0% methanol in methylene chloride to afford 2-chloro-5-[(2methyl-1,3-thiazol-4-yl)ethynyllpyridine (2.29 g, 78% yield) as an off-white solid, M.p. 138-139-C. 'H NMR (CDC1 3 300 MI-z) 8 8.56 J=2.1 Hz, IlH), 7.78 (dd, J=8.3 Hz, 2.3 Hz, 1WH, 7.45 Il-H), 7.3 3 J=8.2 Hz, 1WH, 2.75 3H). MIS (ESI) 234.9 Example 89 Synthesis of 5-1(2-methyl-I ,3-thiazol-4-Yl)ethynyll-2-phenylpyridine Phenylboronic acid (569 mg, 4.26 mmol) and 2-chloro-5-[(2-methyl-1I,3-thiazol-4yi)ethynyllpyridine from Example 88 (1.00 g, 4.26 mmol) were dissolved in DNME (10 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (112 mg, 0.43 mmol), bis-triphenylphosphine palladium dichloride (150 mg, 0.21 mmol), and potassium carbonate (1.18 g, 8.52 mmol) in DME (15 mE) and
H
2 0 (25 mL) at 40'C. The reaction was stirred at reflux for 3 h, then cooled to ambient temperature and poured into a separatory funnel containing ethyl acetate (250 mL). The ethyl acetate layer was washed with saturated NaHCO 3 (50 mL), and H 2 0 (2 x 50 mL), then dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 4:1 hexanes:ethyl acetate to afford 5-[(2-methyl-l,3-thiazol-4-yl)ethynylJ-2-phenylpyridine (1.00 g, 85% yield) as a pale yellow solid, M.p. 102-103 0 C. 'H NMvR (CDCI 3 3 00 MHz) 8 8.86 J=2.l1, 1IH), 8.00 (in, 2 H, 7.89 (dd, J=8.3 Hz, 2.2 Hz, 11H), 7.73 J=8.3 H7, I 7.45 I1H, 7.43-7.52 (in, 3H), 2.76 3H). MIS (ESI) 277.0 WO 01/16121 PCT/USO0/23923 69 Example Synthesis of 2-(4-chlorophenyl)-5-I(2-methyl- ,.3-thiazol-4-YI)ethynyll pyridine Following the procedure and mole equivalents indicated above for Example 89, 4-chloro phenylboronic acid (74 mg, 0.47 mmol) and 2 -chloro-5-[(2-methyl-1,3-thiazol..4yl)ethynyl]pyridine from Example 88 (100 mg, 0.43 mmol) were cross-coupled to obtain 2-(4-chlorophenyl)-5-[(2-methyl- 1,3-thiazol-4-yl)ethynyl~pyridine (105 mg, 79% yield) as a white solid after eluting with 3:1 hexanes:ethyl acetate from a silica gel column. This material was solubilized in ethanol (10 mL) and acidified with I M HCI in diethyl ether (4 mL). The pale yellow hydrochloride salt 193-194 0
C)
was then obtaincd upon concentration of this solution in vacuo and trituration with diethyl ether (3 x mL). 1 H NMR (CD 3 OD, 300 MHz) 8 8.86 I1H), 7.98 J=8.5 Hz, 2M), 7.95 J=8.6 Hz, IlH), 7.73 J=8.2 Hz, 11K), 7.47 1K), 7.47 J=8.4 Hz, 2H), 2.76 3H). MS (ESI) 311.0 Example 91 Synthesis of 2-(4-methowvphenyl)-5-[(2-metbyl-1.3-thiazol-4.yl)ethynyll Dyrdine Following the procedure and mole equivalents indicated above for Example 89, 4-methoxy phenylboronic acid (106 mg, 0.70 mmol) and 2 -chloro-5-[(2-methyl-1,3-thiazol-4-yl)ethynyllpyridine from Example 88 (150 mg, 0.64 mmol) were cross-coupled to obtain 2-(4-methoxyphenyl)-5-[(2methyl-1,3-thiazol-4-yl)ethynyllpyridine (156 mg, 80% yield) as a white solid, M.p. 125-126*C, after eluting with 2:1 hexanes:ethyl acetate from a silica gel column. NNM (CDC1 3 300 MHz) 8 8.81 (d, J= 1.6 Hz, I1H), 7.96 J=8.8 Hz, 2H), 7.82 (dd, J=8.3 Hz, 2.2 Hz, I 7.63 J=8.2 H4z, 1KH), 7.42 (s, 1IH), 6.98 J=8.8 Hz), 3.84 3H), 2.73 3 MIS (ESI) 3 07.0 Example 92 Synthesis of 243-nvridl)-5-1(2-methyl-1 .3-thiazol-4-yl)ethynyll pynidine Following the procedure and mole equivalents indicated above for Example 89, pyridine-3boronic acid (86 mg, 0.70 mmol) and 2-chloro-5-[(2-methyl-1,3-thiazol-4-yl)ethynyl~pyridine from Example 88 (150 mg, 0.64 mmol) were cross-coupled to obtain 2-(3-pyridyl)-5-[(2-methyl-l,3-thiazol.
4-yI)ethynyl]pyridine (80 mg, 45% yield) as a pale yellow solid after eluting with 1:1 hexanes:ethyl acetate from a silica gel column. This material was then solubilized in diethyl ether (10 m.L) and precipitated as the pale pink hydrochloride salt upon treatment with of I M HCI in diethyl ether (2 mL) and trituration with fresh diethyl ether (3 x 5 mL). 'H NMR (CD 3 OD, 300 MfHz) 8 9.63 I1-H), 9.3 7 J=8.2 Hz, 8.97 1K), 8.95 J=5.8 Hz, 1K), 8.21-8.3 1 (in, 31H), 8.07 1H), 2.88 314I).
MIS (ESI) 278.0 WO 01/16121 PCT/US00/23923 Exarnyle 93 Synthesis of 2-(4-pyridvl)-5-I(2-methvI-1 3-thiazol-4-Yl?)ethynyll pyrdine Following the procedure and mole equivalents indicated above for Example 89, pyridine-4boronic acid (86 mg, 0.70 mmol) and 2-chloro-5-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine from Example 88 (150 mg, 0.64 mmol) were cross-coupled to obtain 2-(4-pyridyl)-5-[(2.methyl-l,3-thiazol- 4-yl)ethynyl]pyridine (30 mg, 17% yield) as an off-white solid after eluting with 2:1 ethyl acetate:hexanes from a silica gel column. This material was then solubilized in ether (10 mL) and precipitated as the pale yellow hydrochloride salt (M.p.>185*C, dec.) upon treatment with of I M HCI in diethyl ether (2 mL) and trituration with fresh diethyl ether (3 x 5 mL). 1H- NMR (CD 3 OD, 300 MI-z) 5 9.01 1H), 8.98 J=7.0 H-lz, 2H), 8.85 J=6.7 Hz, 2H), 8.40 J=8.2 H4z), 8.22 (dd, J=8.2 Hz, 2.0 1-Iz, I 7.94 I1-H), 2.88 3H). MS (ESI) 278.0 Example 94 Synthesis of 3-ch loro-6-l (2-methyl-I 3-thiazol-4-vl)ethvnvll Dyndazine 3,6-Dichloropyridazine (299 mg, 2.00 mmol) and 2-methyl-4-[(trimethylsilyl)ethynyl]-1,3thiazole from Example 77 (391 mg, 2.00 mxnol) were dissolved in DM F (10 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (105 mg, 0.40 mmol), bis-triphenyiphosphine palladium dichloride (140 mg, 0.20 mmol), Gui (76 mg, 0.40 mmol), tetrabutylammonium iodide (369 mg, 1.00 mmol), and triethylatnine (1.01 g, 1.39 mL, 10.0 mmol) in DMF (15 mL) at 40'C. The reaction was warmed to 501C, and teti-abutylammon ium fluoride 10 mxnol, 2. 10 mL of a 1.OM solution in THF) was added via syringe pump over 1.5 h. The reaction was then cooled to ambient temperature and poured into a separatory funnel containing 1: 1 hexanes:ethyl acetate (200 mnL) where it was washed with 50% dilute brine (3 x mL), dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 1.5:1 hexanes:ethyl acetate, then recrystallized from 2:1 ethyl acetate:hexanes to afford 3-chloro-6-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridazine (182 mg, 39% yield) as pale pink needle crystals, M.p. 197-198 0 C. 'H NMR (CDCI 3 300 MHz) 5 7.64 J=8.8 Hz, I 7.63 Il-H), 7.52 J=8.8 H-z, IH), 2.77 3H). MIS (ESI) 235.9 Example Synthesis of 3-1(2-methyl-i .3-thiazol-4-VI)ethynvll-6-phenylpyridazine Phenylboronic acid (47 mg, 0.38 mmol) and 3-chloro-6-[(2-methyl-1,3-thiazol-4yI)ethynyl]pyridazine from Example 94 (75 mg, 0.32 mniol) were dissolved in DME (2 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (8.4 mg, 0.032 mmol), bis-triphenylphosphine palladium dichloride (11.2 mg, 0.0 16 mmol), and potassium carbonate (89 mg, 0.64 mmol) in DME (2 mL) and
H
2 0 (4 mL) at 40'C. The reaction was stirred at reflux for 16 h. then cooled to ambient temperature WO 01/16121 PCT/USOO/23923 71 and poured into a separatory funnel containing ethyl acetate (100 mL). The ethyl acetate layer was washed with saturated NaHCO 3 (50 mL), and H 2 0 (2 x 50 mL), then dried (MgSOA) filtered, and concentrated in vacuo. The crude residue was chromnatographed on silica gel, eluting with 1:1 hexanes:ethyl acetate to afford 3-[(2-methyl-l,3-thiazol-4-yl)ethynyl]-6-phenylpyridazine (43 mg, 49% yield) as a white solid. 1 H NMR (CDCI 3 300 MHz) 858.11 (in, 2H), 7.86 J=8.8 H4z, I 7.72 (d, J=8.8 Hz, 1I1M, 7.62 I 7.51-7.56 (mn, 311), 2.78 314). MS 278.0 (MW+l).
Example 96 Synthesis of 4-1(6-nhenyl-3-Dyvridinvl)ethvyll-l thiazol-2-amine Phenylboronic acid (566 mg, 4.4.64 inmol) and 2,5-dibromopyridine (1.00 g, 4.22 mmol) were dissolved in DME (10 mL) and deoxygenated via argon bubbling for 20 min. This solution was then added via syringe to a deoxygenated solution of triphenylphosphine (I111 mg, 0.42 minol), bistriphenyiphosphine palladium dichloride (148 mg, 0.21 inmol), and potassium carbonate (1.17 g, 8.44 mmol) in DME (15 mL) and H120 (25 mL) at 401C. The reaction was stirred at reflux for I h, then cooled to ambient temperature and poured into a separatory funnel containing ethyl acetate (250 mnL).
The ethyl acetate layer was washed with saturated NaHCO 3 (50 mL), and H20 (2 x 50 miL), then dried (MgSO 4 filtered, and concentrated in. vacuo. The crude residue was chromatographed on silica gel, eluting with 14:1 hexanes:ethyl acetate to afford 5-bromo-2-phenylpyridine (783 mg, 78% yield) as a white crystalline solid.
5-Bromo-2-phenylpyridine (300 mg, 1.28 mmol) and 4-[(trimethylsilyl)ethynyl]-1,3-thiazol-2ylamine from Example 78 (210 mg, 1.07 inmol) were added as solids to a deoxygenated solution of triphenylphosphine (56 mg, 0.21 mmol), bis-triphenylphosphine palladium dichloride (75 mg, 0. 11 inmol), CuT (41 mg, 0.21 inmol), tetrabutylammonium iodide (198 mng, 0.54 iniol), and triethylamine (540 mg, 740 p.L, 5.4 mmol) in DMF (15 mL) at 40*C. The reaction was warmed to 60*C, and tetrabutylanimoniuin fluoride 17 minol, 1. 17 mL of a 1.OM solution in THF) was added slowly over 1 h. The reaction was then cooled to ambient temperature and poured into a separatory funnel containing 1: 1 hexanes:ethyl acetate (250 mL) where it was washed with 50% dilute brine (3 x 100 mL), dried (MgSO 4 filtered, and concentrated in vacuo. The crude residue was chromatographed on silica gel, eluting with 1: 1 hexanes:ethyl acetate to afford 4-[(6-phenyl-3-pyridinyl)ethynyl]-l1,3-thiazol- 2-amine (186 mg, 63% yield) as a pale yellow solid, M.p. 194-195 0 C. 'H NMR (CD 3 0D, 300 MHz) 8.74 111), 7.91-7.96 (in, 3H1), 7.78 J=8.3 Hz, 1H), 7.45-7.54 (in, 3H), 6.84 111). MS (ES!) 278.0 WO 01/16121 PCT/USO0/23923 72 Example 97 Synthesis of 3, 4 dihydro-l-naphthalenyll trifluoromethanesulfonate To a stirred solution of ax-tetralone (1.00g, 6.84 mmol) in CH 2 C1 2 (7OmL) under argon at ambient temperature was added lutidine (1.2OmL, 6.84 mmol) followed by dropwise addition of trifluoromethanesulfonic anhydride (1.73mL, 10.3 mmol). The reaction mixture was stirred for lb and then concentrated in vacuo and purified by flash chromatography on silica gel eluting with 10:1 hexane:ethyl acetate to afford 3, 4 dihydro-l-naphthalenyl trifluoromethanesulfonate (1 .64g, 87% yield) as a pale yellow oil. 'H NMR (CDCI 3 300M1-Iz) 8 7.36-7.32 (in, I 7.29-7.24 (in, 214), 7.20- 7.16 (in, I1H), 6.03-6.00 (in, I 2.90-2.85 (in, 2H), 2.5 5-2.48 (in, 2H).
Example 98 Synthesis of 2-(3.4-dihvd ro-l-nanhthalenyl)ethynylpyridine hydrochloride A stirred suspension of Cul (159 mng, 0.834 mmnol), triphenylphosphine (219 mg, 0.834 minol), PdCI 2 (PPh 3 2 (293 mg, 0.417 inmol), and triethylamine (2.9OinL, 20.8 minol) in DME (3OmL), was degassed with a stream of argon for several min. A solution of 2-ethynyl pyridine (1.30g, 12.5 rmol) and 3, 4 dihydro-lI-naphthalenyl trifluoromethanesulfonate from Example 97 (1.15g, 4.17 mmol) in DME (IOmL) was added to the mixture. The mixture was heated to 80'C for 1.5 h. The reaction mixture was cooled to ambient temperature and filtered through a pad of CeliteThm. The filtrate was concentrated in vacuo and the crude material was purified by flash chromatography on silica gel eluting with 8:1 then 5: 1 hexane:ethyl acetate to afford 2-(3,4-dihydro-1I-naphthalenyl)ethynylpyridine (798mg, 72% yield) as a pale yellow oil.
2-(3,4-Dihydro-l-naphthalenyl)cthynylpyridine (300mg, 1.29 mmol) was dissolved in diethyl ether and treated with HCI in diethyl ether (2.00 m.L of a 1 M solution, 2.00 inmol). Upon addition of the [-CI solution a white solid precipitated from the solution. The mixture was concentrated in vacua the pale yellow solid was recrystallized from Methanol/diethyl ether. The mother liquor was decanted and the resulting pale yellow solid was dried under highi vacuum to afford 2-(3,4-dihydro-1naphthalenyl)ethynylpyridine hydrochloride (205mg, 18% yield) as a pale yellow solid, M.p. 153- 155 0 C. 'H NMR (CD 3 OD, 300MHz) 8 8.76-8.74 (mn, I1W, 8.43-8.38 (mn, lH), 8.08-8.05 J=8 Hz, IH), 7.88-7.83 (in, 1H), 7.66-7.63 J=8.3 Hz, IH), 7.31-7.18 (in, 3H), 6.91-6.88 (in, IH), 2.88-2.82 (in, 214, 2.54-2.47 (in, 2H). MS 232 (M14).
Example 99 Synthesis of To a stirred solution of 5-bromonicotinic acid (2.5g, 12.4 minol) in CH 2
CI
2 (lOOinL) was added N, 0-dimethyihydroxylainine hydrochloride (1 .44g, I 4.8mmol), HOBT (1 .84g, 13 .6minol), and EDC (2.61g, 13.6 mmcl) sequentially as solids followed by diisopropylethylamine (6.46mL, 37.lmmol).
WO 01/16121 PCT/USOO/23923 73 The mixture was stirred at ambient temperature under argon for 1 h. The reaction mixture was concentrated in vacuo and redissolved in ethyl acetate. The organic material was washed with I M HCI and brine (3 x 25mL), dried over MgSO 4 filtered and concentrated in vacua. The crude material was purified by flash chromatography on silica gel eluting with 1: 1 hexane:ethyl acetate to afford N-methoxy-N-methyinicotinamide (2.99g, 51% yield) as a colorless oil. '-INMR (CDCI 3 300MHz) 8 8.21-8.20 (in,IH), 7.85-7.82 IH), 7.73-7.70 (in,IH), 3.58 3.40 3H-).
Example 100 Synthesis of N-methoxy-N-methyl-5-[(2-methvl-1. 3-thiaiol-4-yl)ethynyll nicotinamide A stirred suspension of Cul (257 mng, 1.35 minol), triphenylphosphine (354 mg, 1.35 minol), PdCI 2 (PPh,) 2 (467 mg, 0.665 mmol), tetrabutylammonium iodide (1.89g, 5.12 mmol) and triethylamine (3.57 mL, 25.6 minol) in DMF (50mL), was degassed with a stream of argon for several minutes and warmed to 40'C. 2-Methyl-4[(trimethylsilyl)ethynyl]-1,3-thiazole from Example 77 (1.00g, 5.12 inmol) and 5-broino-N-methoxy-N-methylnicotinanide from Example 99 (1.63 g, 6.65 minol) was added to the reaction mixture. The mixture was heated to 70'C and TBAF in TI-F (6.65 mL, 1.0 M solution, 6.65 minol) was added via syringe pump over 2 h. The reaction mixture was cooled to ambient temperature and diluted with 1.:1 hexane:ethyl acetate. The organic material was washed with dilute brine (3 x 25 inL), dried over MgSO 4 filtered and concentrated in vacuo. The crude material was purified by flash chromatography on silica gel (2:1 then 1:1 hexane:ethyl acetate) to afford Nmethoxy-N-methyl-5-[(2-methyl-l, 3-thiazol-4-yl)ethynyl] nicotinamide (708 mng, 48% yield) as an orange oil. 'H NMR (CDCI 3 300Mz) 8 8.89-8.84 (mn, 2H), 8.19-8.17 (in, 114), 7.55-7.53 (in, 11H), 3.57 3H), 3.40 3H), 2.74 3H). MS 288 Example 101 Synthesis of (4-fluorophenvl'N5-i(2-methvl-1 .3-thiazol-4-YI~ethynyll-3-DvridinvI1 uetbanone To a stirred solution of 'N-methoxy-N-methyl-5-[(2-methyl-1, 3-thiazol-4-yl)ethynyl] nicotinamide from Example 100 (50 mng, 0. 174 mniol) at ambient temperature under argon was added a solution 4-fl uorophenyl magnesium bromide in THIF (0.696 m.L, I OM solution, 0.696 minol). The reaction mixture was stirred for 24 h and then diluted with 1-20. The aqueous solution was extracted with ethyl acetate (3 x 25mL). The combined organic layers were washed with brine (2 x 25inL), dried over MgS0 4 filtered and concentrated in vacua. The crude material was purified by flash chromatography on silica gel eluting with 3:1 then 2.1 hexane:ethyl acetate to afford (4fluorophenyl){ 5-[(2-methyl-1I,3-thiazol-4-yl)ethynyl]-3-pyridinyl }methanone (15 mg, 27% yield) as a pale yellow solid, M.p. 132-134 0 C. 'HNMR (CDCI 3 300MHz) 8 9.02-8.95 (in, 8.20-8.19 (in, I 7.89-7.85 (in, 2H), 7.48 I 7.25-7.19 (in, 2H), 2.76 3H). MS 323 WO 01/16121 PCT/USOO/23923 74 Example 102 Synthesis of (4-.methoxvphenylM(5-I(2-methvl-1,3-thiazol-4-vl)ethynvll-3-Dvridinvii methanone To a stirred solution of N-mcthoxy-N-methyl-5-[(2-rrethyl-l, 3-thiazol-4-yl)ethynyl] n icotinamide from Example 100 (50 mg, 0. 174 mmol) at ambient temperature under argon was added a solution 4-anisylmagnesium bromide in THE (1 .39mL, 0.5M solution, 0.696 mmol). The reaction mixture was stirred for 24 h and then diluted with H 2 0. The aqueous solution was extracted with ethyl acetate (3 x 25mL). The combined organic layers were washed with brine (2 x 25mL), dried over MgSO 4 filtered and concentrated in vacua. The crude material was purified by flash chromatography on silica gel eluting with 3:1 then 2:1 hexane:ethyl acetate to afford (4-methoxyphenyl){5-[(2-methyl- 1,3-thiazol-4-yl)ethynyl]-3-pyridinyl~methanone (13 mg, 22% yield) as a pale yellow solid, M.p. 97- 99"C. 'H NMR (CDCI1 3 300OMHz) 5 8.94-8.90 (in, 2H), 8.18-8.17 (in, I 7.85 -7.82 (in, 2 7.47 (s, IH), 7.02-6.99 (in, 2HM, 3.92 3HM, 2.76 3M). MIS 335 Example 103 Synthesis of 2-(2-cvclorpropylethvnvl)thiazole 2-Bromo-l,3-thiazole (1.31 g, 8.0 minol), Cul (143 mg, 0.75 minol), PdCI 2 (45 mg, 0.75 inmol), PPh 3 (197 mng, 0.75 inmol), and K 2 C0 3 (4.5 g, 33 inmol) were combined in DME (50 mL) and
H
2 0 (25 mL), and argon gas was bubbled through the suspension for several min to deoxygenate the mixture. Cyclopropyl trimethylsilylacetylene (1.8 g, 13.3 minol) was added and the reaction was heated at reflux for 12 h. The mixture was filtered through Celiterm, the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL), washed with H 2 0 (200 mL), brine (100 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. This material was purified by column chromatography eluting with 4:1 hexane:ethyl acetate to afford 2-(2-cyclopropylcthynyl)thiazole (800 mg, 67% as a light brown oil.
p-Toluenesulfonic acid (1.02 g, 5.4 mmol) and 2-(2-cyclopropylethynyl)thiazole (800 nmg, 5.3 mmol) wvere dissolved in Methanol (50 mL), and the solution was concentrated in vacu. The resulting black viscous oil was triturated with diethylether with sonication. The diethylether layer was decanted and the remaining brown gum was dried in vacua. Material was free based with aqueous K 2 C0 3 and extracted with ethyl acetate (2 x 75 mL), dried over Na 2
SO
4 filtered and concentrated in vacu. The residue was purified by column chromatography on silica gel eluting with 4:1 hexane:ethyl acetate to afford 2-(2-cyclopropylethynyl)thiazole (150 mng, 19%) as an oil. 'H NMR (CDCI 3 300 MHz) -5 7.75 J= 3 Hz, IH), 7.25 J= 3 Hz, 1H) 1.50 (mn, 1H), 0.93 (mn, 4H). MS (API-ES Positive) 150 WO 01/16121 PCT/US00/23923 Example 104 Synthesis of 3-(2-pyridinvlethynyl)-2-cvclohexen-l-one A solution of 2-ethynylpyridine (10.3 g, 100 mmol) in THF (200 mL) was cooled to -50 0 C and a solution of n-BuLi (100 mmol, 2.0 M in hexane) was added slowly keeping the solution temperature below -400 C. After 30 min at reduced temperature the solution became opaque, brown and slightly viscous. 3-Ethoxy-2-cyclohexen-l-one (15.4 g, 110 mmol) was added all at once and the solution was allowed to warm slowly to ambient temperature with stirring. After 12 h the dark solution was acidified with HCI (400 mmol, 2.0 followed after 30 min by basification with solid K 2
CO
3 The mixture was partitioned between ethyl acetate and H 2 0, the H20 layer was washed with a further portion of ethyl acetate, the combined organic layers were washed with H 2 0 (200 mL), brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 2:1 to 1:1 hexane:ethyl acetate to afford a yellow oil (10.5 g, This material was crystallized from ethyl acetate to afford two crops of light yellow flakes (5.9 g, 30%) M.P, 74-75° C. 'H NMR (CDC3. 300 MHz) 5 8.64 1H), 7.72 1H), 7.51 1H), 7.30 1H), 6.37 IH), 2.59 2H), 2.46 2H), 2.09 2H). MS (ESI) 197 Example 105 Synthesis of3-(2-pyridinylethynvl)-2-cyclohexen-l-one oxime Hydroxylamine hydrochloride (350 mg, 5.0 mmol) and KOH (560 mg, 10 mmol) were dissolved in wet ethanol (15 mL). 3-(2-Pyridinylethynyl)-2-cyclohexen-l-one from Example 104 g, 2.5 mmol) was added and the mixture heated at reflux for 2 h. The solution was allowed to cool and the solids filtered. The ethanol soluble portion was concentrated in vacuo and the residue was purified by flash column chromatography on silica gel eluting with ethyl acetate to afford 3-(2pyridinylethynyl)-2-cyclohexen-l-one oxime (375 mg, 70 as a tan solid. M.p. 125-127 0 C. 'H NMR
(CDCI
3 300 MHz) 5 8.58 1H), 7.68 1H), 7.47 1H), 7.24 1H), 6.75 1H), 2.65 (m, 2H), 2.41 2H), 1.84 2H). MS (API-ES positive) 213 Example 106 Synthesis of Z)-3-(2-pyridinylethynyl)-2-cyclohexene-l-one O-methyloxime To a solution of 3-(2-pyridinylethynyl)-2-cyclohexene-1-one from Example 104 (10 mg, 0.05 mmol) in ethanol (0.3 mL) was added methoxylamine hydrochloride (30-35 wt. in H 2 0; 7jI, 0.05 mmol) in a 10 ml Teflon reaction tube. Piperidinomethyl polystyrene (28 mg of 3.63 mmol/g beads, 0.1 mmol) was added and the resulting suspension was heated at 80 0 C in an orbital shaker. After shaking at 0 C for 16 h, the reaction was cooled to ambient temperature and chloroformate polystyrene (47.5 mg of 1.0 mmol/g beads, 0.05 mmol) was added. The reaction was allowed to shake at 40°C for 1 hour, then cooled to ambient temperature, and tris (2-aminoethyl)amine polystyrene 1 mg of 4.5 mmol/g beads, 0.05 mmol) was added. The reaction was then allowed to shake at 40 0 C for 1 hour, then cooled, WO 01/16121 PCT/US00/23923 76 and diethyl ether (0.3 ml) was added. The reaction was vortexed and filtered using a fritted syringe attached to a vacuum block. The collected sample was then concentrated in vacuo using a Savant rotary evaporator to afford 3-(2-pyridinylethynyl)-2-cyclohexene-1-one O-methyloxime as a mixture of E, Z, isomers (12 mg, 90% yield) as a light brown oil. 1 H NMR (CDC13, 300 MHz) 8 8.61-8.59 1H), 7.68-7.67 1H), 7.47-7.44 1H), 7.28-7.23 1H), 7.17-6.57 1H), 3.90 (d J=10.9 Hz, 1H), 2.57-2.52 2H), 2.47-2.39 2H), 1.91-1.88 2H), 1.83-1.79 2H); MS (ESI) 226.28 (MV+H).
Example 107 Synthesis of Z)-3-(2-pyridinylethynyl)-2-cyclohexen-1-one O-ethyloxime To a solution of 3-(2-pyridinylethynyl)-2-cyclohexene- -one from Example 104 (20 mg, 0.10 mmol) in ethanol (0.6 mL) was added O-ethylhydroxylamine hydrochloride (10 mg, 0.10 mmol) in a ml Teflon reaction tube. Piperidinomethyl polystyrene (56mg of 3.63 mmol/g beads, 0.2 mmol) was added and the resulting suspension was heated at 80 0 C in an orbital shaker. After shaking at 80 0 C for 16 h, the reaction was cooled to ambient temperature and chloroformate polystyrene (95 mg of mmol/g beads, 0.10 mmol) was added. The reaction was allowed to shake at 40 0 C for 1 hour, cooled to ambient temperature, and treated with tris (2-aminoethyl)amine polystyrene (22 mg of 4.5 mmol/g beads, 0.10 mmol). The reaction was then allowed to shake at 40 0 C for 1 hour, then cooled, and diethyl ether (0.6 ml) was added. The reaction was vortexed and filtered using a fritted syringe attached to a vacuum block. The collected sample was then concentrated in vacuo using a Savant rotary evaporator to afford 3-(2-pyridinylethynyl)-2-cyclohexen-1-one O-ethyloxime as a mixture of E, Z isomers (22 mg, 91% yield) as a light brown oil. 'H NMR (CDC13, 300 MHz) 8 8.61-8.59 1H), 7.70-7.63 (m, 1H), 7.47-7.43 1H), 7.28-7.23 1H), 7.22-6.58 1H), 4.20-4.10 2H), 2.56 (t J=6.6 Hz, 2H), 2.47-2.38 2H), 2.47-2.39 2H), 1.94-1.87 2H), 1.85-1.76 2H), 1.29 (t J=6.0 Hz, 3H); MS (ESI) 240.30 Example 108 Synthesis of E/Z-3-(2-pyridinvlethvnyl)-2-cyclohexen-l-one 0-allyloxime To a solution of 3-(2-pyridinylethynyl)-2-cyclohexen-l-one from Example 104 (513 mg, 2.60 mmol) in ethanol (15 mL) was added O-allylhydroxylamine hydrochloride (393 mg, 3.59 mmol).
Piperidinomethyl polystyrene (1.54 g of 3.63 mmol/g beads, 5.6 mmol) was added and the resulting suspension was heated in an oil bath at 80 0 C. After stirring at 80 0 C for 16 h, TLC analysis showed no starting 3-(2-pyridinylethynyl)-2-cyclohexen- -one remaining. The reaction mixture was cooled, diluted with ether, and filtered through a glass frit to remove the resin. The filtrate was concentrated to afford a yellow oil which was purified by column chromatography eluting with hexane, 20:1, 9:1, then WO 01/16121 PCT/US00/23923 77 8.5:1.5 hexane:ethyl acetate to afford E,Z-3-(2-pyridinylethynyl)-2-cyclohexen-1I-one O-allyloxime (550 mg, 83% yield) as a light brown oil. 'H NMR (CDCI 3 300 MHz) 8 8.63-8.60 (in, I 7.70-7.64 (in, IM), 7.48-7.44 (in, 11-1), 7.28-7.22 (in, 6.60-6.59 (in, 11H), 6.07-5.96 (in, IH), 5.36-5.22 (in, 2H-1), 4.64-4.59 (in, 2H), 2.62-2.58 (in, IM), 2.49-2.40 (in, 2H), 1.95-1.78 (in, 2H); MS (ESI) 253.1 Example 109 Synthesis of (Z-methyl-1342-Dyridinvlethynyl)-2-cyclohexen-l-vlidenelethanoate Trimethylphosphonoacetate (462 mg, 2.5 mmol) was dissolved in THIF (20 mL) and the solution was cooled to 00 C at which time LiHDMS (2.6 mL, 1.0 M in THF) was added slowly. After 30 min 3-(2-pyridinylethynyl)-2-cyclohexen-1 -one from Example 104 (0.5 g, 2.5 mmol) was added and the solution was allowed to warm to ambient temperature. After a further 3 h the mixture was partitioned between ethyl acetate and H 2 0, the aqueous layer was washed with a further portion of ethyl acetate, the combined organic layers were washed with H 2 0 (50 mL), brine (30 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography eluting on silica gel with 1.5:1 then 1: 1 hexane:ethyl acetate to yield methyl-[3-(2-pyridinylethynyl)-2cyclohexen-1-ylidene]ethanoate as a mixture of E and Z double bond isomers. This mixture was further purified by reverse phase 1-IPLC to obtain a major isomer and a minor isomer. The major isomer was identified as the Z-isomer based on the lack of NOESY correlation between the two vinyl protons. M P 65-68 0 C; 'H NMR (CDC 3 ,300 MI-z) 8 8.60 J=5 Hz, 7.98 (in, IH), 7.67 (in, IH), 7.47 (in, LH), 7.23 (mn, IM), 5.57 11M), 3.73 3M), 2.44 (in, 3H), 1.84 (in, 3H); MS (API-ES positive) 254 Example 110 Synthesis of 2-W13S)-3-methyl-l-cyclopenten-l-yllethynylhpyridine 2-fl(4S)-4-methYl-1cyclopenten-l-yllethynvlI pyridine To a solution of 2-ethynylpyridine (0.6 mL, 6.0 inmol) in TI-F (2 mL) at -40' C was added nbutyllithium (3.75 mL, 6.0 inmol). After stirring at reduced temperature for 30 min the solution was added rapidly to a Solution of (R)-(+)-3-methylcyclopentanone (0.65 mL, 6.0 minol) in TI-F (10 mL).
The mixture was allowed to warm to ambient temperature over 16 h, then partitioned between H 2 0 and ethyl acetate. The organic layer was dried over anhydrous Na1 2
SO
4 and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel eluting with 1: 1 hexane:ethyl acetate to afford (R)-(+)-3-methyl-1-(2-pyridinylethynyl)cyclopentano as a dark oil. This material was dissolved in pyridine/CH 2
CI
2 (10 mIL, P01 3 (0.55 mL, 6.0 minol) was added, and the mixture was heated to reflux for 4 h. After cooling, the POC1 3 and pyridine were removed in vacuo, and the residue was partitioned between H 2 0 and ethyl acetate. The organic layer was dried over anhydrous Na 2
SO
4 and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel WO 01/16121 PCT/US00/23923 78 eluting with 2:1 hexane:ethyl acetate to afford 2- {[(3S)-3-methyl-lI-cyclopenten-1I-yllethynyl) pyridine and 2-{[(4S)-4-mcthyl-1-cyclopenten-I-yljethynyl~pyridine (0.425 g, 38% as a dark brown oil. 1H NMR (CDCI 3 300 MHz) 6 8.57 J--9 Hz, I 7.63 (in, I1H), 7.42 (in, I1-H), 7.20 (in, 1KH), 6.17 (mn, 1K), 2.90 (in, 0.5K), 2.61 (mn, 2H), 2.16 (mn, 1.47 (mn, 1.06 J=12 Hz, 311). GUIMS two peaks 11.88 and 11.91 min. (ESI) 182 Example 111 Synthesis of 2-1(3,5--dimethvl-l-cyclohexen-1-vl)ethynyll yridine The procedure was carried out as for Example 110 using 3,5-cis-dimethylcyclohexanone (756 mg, 6.0 mmol), to give 2-[(3,5-diinethyl-1-cyclohcxcn-1-yl)ethynyljpyridine (0.325 g, 25% as a yellow oil. 'K NMR (CDCI 3 ,300 MII-z) 6 8.57 (mn, IK), 7.63 (in, 1K), 7.41 (in, IH), 7.18 (mn, 1H), 6.15 (in, IH), 2.27 214), 1.79 3H), 1.01 (mn, 7H). GCIMS 12.96 min. (ESI) 211 Example 112 Synthesis of 2-i(3,4-dimethyl-1-cyclonenten-1-Yl)ethynyl nyridine The procedure was carried out as for Example 110 using 3,4-dimethylcyclopentanone (672 mg, 6.0 mmol), to yield a mixture of cis and trans 2-[(3,4-diinethyl-1-cyclopenten-1-yl)ethynyljpyridine (0.51 g, 43%) as a yellow oil. 'H NMR (CDCI 3 ,300 MI-z) 5 8.57 (mn, 1K), 7.63 (mn, 1K), 7.43 (mn, IH), 7.19 (in, 1K), 6.14 (in, 1K), 2.64-2.80 (in, 2K), 2.43 (mn, 1K), 2.22 (in, 111), 1.89 (in, 1H), 1.07 (in, 3H-), 0.94 (in, 3K). GUIMS two peaks 12.15 and 112.44 min. (ESI) 196 Example 113 Synthesis of -115-(trilluoroinethyl)-l-cyclohexen-l-yllethvnyvll pyndine 2-113-(trifluoromethvi)- 1-cyclohexen-l-yllethynylhoyrjdine The procedure was carried out as for Example 110 using 3-trifluoroinethylcyclohexanone (960 mg, 6.0 iniol), to yield a 3:1 mixture of 2-{[5-(trifluoromethyl)-1-cyclohexen-1-yl]ethynyl~pyridine and 2-{[3-(trifluoromethyl)-1-cyclohexen-1-yI]ethynyl~pyridine (0.51 g, 43% as a brown oil. 'K NMR (CDCI 3 .300 MHz) 8 8.57 (in, 1H), 7.65 (mn, 1K), 7.43 (in, 1K), 7.21 (mn, 114), 6.32 (in, 0.75K), 6.21 (mn, 0.25K), 2.98 (mn, 0.25K), 2.25-2.57 (in, 3.5H), 2.01 (mn, 2H), 1.58 (mn, 1KR), 1.29 (mn, 0.25K).
GCUMS three peaks 12.22, 12.37 and 12.49 min. (ESI) 251 252 Example 114 Synthesis of 2-(1.4.4a.5.6.78.8a-octahvdro-2-naphthalenylethvnvl)Dynidine and 2-(3.4.4a.5,6,8,8a-octahydro-2-naphthalenvlethynvl)pyvridine The procedure was carried out as for Example 110 using cis/frans 2-decalone (1.83 g, 12 inmol) to yield a mixture of four stereoisoiners each of 2-(1,4,4a,5,6,7,8,8a-octahydro-2naphthalenylethynyl)pyridine 2-(3,4,4a,5 ,6,7,8,8a-octahydro-2-naphthalenylethynyl)pyridine (0.50 g, as a brown oil. 'K NMR (CDC 3 300 MHz) 5 8.57 J=5 Hz, 11-1), 7.63 (in, 1K), 7.37 (in, I1K), WO 0 1/16121 PCT/USOO/23923 79 7.18 (in, IH), 6.27 (mn, 6.20 (in, 0.5H1), 2.05-2.34 (in, 1.79 (in, 1.48 (in, 7H), 1.00 (in, IlH). MIS (API-ES positive) 238 Exam~le 115 Synthesis of 2-i(3-methvl-l-cvclohexen- 1-YI)ethvnyllpvridine A solution of 3-methyl-2-cyclohexen-lI-one (990 mg, 9.0 mmol) in THF was chilled to -780 C and L-selectride (9.5 minol, 1.0 M in TI-F) was added slowly via syringe. After I h at reduced temperature N-phenyltriflimide (3.2 g, 9.0 mmol) was added all at once. The reaction was allowed to warm to ambient temperature while stirring overnight. The reaction was diluted with two volumes of hexane, and the organic phase was washed with H20, then 10% aqueous NaOH and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel eluting with hexane to afford 3-methyl- I-cyclohexen-lI-yl trifluoromethanesulfonate (400 mg, 18% 3-methyl- I -cyclohexen- I -yl trifluoromethanesulfonate (400 mg, 1.6 inmol), Gui (30 mg, 0. mmol), PdCI 2 (9 Ing, 0.05 ITmol), PPh 3 (40 mg, 0.15 inmol), and K 2 C0 3 (552 mng, 4.0 inmol) were combined in DME (15 mL) and H-20 (15 mL), and argon gas was bubbled through the suspension for several mini to deoxygenate the mixture. 2-Ethynylpyridine (412 mg, 4.0 mmol) was added and the reaction mixture was stirred at ambient temperature for 16 h and then heated at reflux for I h. The mixture was filtered through Celite T m the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 mL), washed with H 2 0 (1 00 brine (50 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 3:1 hexane:ethyl acetate to afford 2-[(3-inethyl-1-cyclohexen-1-yl)ethynyl]pyridine (250 mng, 80%) as a clear oil containing 5% of a regioisomer. 'H NMIR (CDC1 3 300 MHz) 8 8.56 (in, 111), 7.62 (in, I 7.40 (in, 1I-1), 7.18 (mn, I1H), 6.18 (in, 111), 2.31 (in, I11-0, 2.21 (in, 2H), 1.79 (in, 1.55 (in, I 1. 18 (mn, I1H), 1.02 J=7 I-z, 3H). MS (API-ES positive) 198 Example 116 Synthesis of 5-methyl-l-cyclohexen-1-vl tnifluoromethanesulfonate To a solution of 5-methyl-1,3-cyclohexanedione (1.0 g, 7.9 minol) in ethanol (25 mL) was added p-toluenesulfonic acid (85 mng, 0.5 minol), and the reaction was heated to 60' C for 16 h. The reaction was cooled and concentrated in vacuo. The residue was purified by flash column chromatography on silica eluting with hexane to give 3-ethoxy-S-methyl-2-cyclohexen-1 -one (1.2 g, quantitative yield) as a clear oil.
3-Ethoxy-5-methyl-2-cyclohexen- I-one (1.2 g, 7.8 inmol) was dissolved in TI-IF (25 mL).
LiAIH 4 (3.5 minol, I OM in TI-F) was added, and the reaction was allowed to stir at ambient temperature for 3 h. 112S04 (10% aqueous, 25 mL) was added slowly and the mixture was partitioied WO 01/16121 PCT/US00/23923 between H 2 0 and ethyl acetate. The organics were concentrated in vacuo to give 5-methyl-2cyclohexen-I -one (715 mg, 83%) as a clear oil.
A solution of 5-methyl-2-cyclohexen-1-one (715 mg, 6.5 mmol) in THF was chilled to -78' C and L-selectride (6.0 mmol, 1.0 M in TI-f) was added slowly via syringe. After I h at reduced temperature N-phenyltriflimide (1.8 g, 5.0 mmol) was added all at once. The reaction was allowed to warm to ambient temperature while stirring overnight. The reaction was diluted with two volumes of hexane, and the organ ics were washed with H 2 0, then 10% aqueous NaOH and concentrated in vacuo.
The residue was purified by flash column chromatography on silica gel eluting with hexane to afford methyl-1-cyclohexen-l-yl trifluoromethanesulfonate (450 mg, 28% 1H NMR (CDCI 3 300 MHz) 8 5.74 (in, I 2.34 (in, 2.20 (in, 2H), 2.02 (in, IlH), 1.90 (mn, I 1.70 (mn, I 1.23 (in, lI-H), 1.03 J=6.5 Hz, 3H4). MS (El ionization) 244 Example 117 Synthesis of trifluoromethanesulforiate (450 mng, 1.8 mmol), Gui (57 mng, 0.3 minol), PdC12 (18 Mng, 0. 1 inmol), PPh 3 (79 mng, 0.3 mmol), and K 2 C0 3 (640 ing, 4.7 mmol) were combined in DME (20 mL) and H 2 0 (20 mL), and argon gas was bubbled through the suspension for several min to deoxygenate the mixture. 2-Ethynylpyridine (484 mg, 4.7 mmol) was added and the reaction stirred at ambient temperature for 16 h, then heated at reflux for I h. The mixture was filtered through Celite" h1 the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were concentrated in vacuo. The residue was dissolved in ethyl acetate (200 washed with H 2 0 (100 mL), brine (50 dried over Na 2
SO
4 filtered and concentrated in vacu. The residue was purified by column chromatography on silica gel eluting with 3:1 hexane:ethyl acetate to afford methyl-l-cyclohexen-l-yl)ethynyl]pyridine (290 ing, 82%) as a light yellow oil. 'H NMR (CDC 3 300 MHz) 8 8.56 (in, I1H), 7.62 (in, I 7.40 (in, I 7.17 (in, I 6.31 (in, IRH), 2.33 (in, I 2.20 (mn, 2H), 1.89 I 1.71 (m,211), 1.22 Il-H), 0.99 J= 6.5 Hz, 3H).
Example 118 Synthesis of 3-(2-pyridinyletbynyl)-2-cyclohexen-1-oI 3-(2-Pyridinylethynyl)-2-cyclohexen-1 -one from Example 104 (294 mg, 1.5 iniol) and CeCI 3 heptahydrate (381 mg, 1.0 inmol) were dissolved in CH 3 0H (16 mL). NaB-L (127 mg, 3.4 inmol) was added portionwise over 5 min. After 15 min. the reaction was quenched with H 2 0 and partitioned between H 2 0 and ethyl acetate. The organics were washed with aq. NH 4 CI, dried over Na 2
SO
4 filItered and concentrated in vacua. The residue was purified by column chromatography eluting with 1: 1 hexane:ethyl acetate to afford a clear oil (275 mng, 92% WO 01/16121 PCT/USOO/23923 81 p-Toluenesulfonic acid (263 mg, 1.4 mmol) and 3-(2-pyridinylethynyl).2-cyclohexen-lI-ol (275 mg, 1.4 mmol) were dissolved in ethanol:Methanol 1, 40 mL), and the solution was concentrated in vacua. The resulting viscous oil was triturated with diethylether and sonicated. The diethylether layer was decanted and remaining oil was dried in vacua. This material was free based with aqueous K 2 C0 3 and extracted with ethyl acetate (2 x 35 mL), dried over Na 2
SO
4 filtered and concentrated in vacua.
The residue was purified by column chromatography silica gel eluting with 3:2 hexane:ethyl acetate to afford 3-(2-pyridinylethynyl)-2-cyclohexen-lI-ol (160 mg, 57% as a clear oil. 'H NMR (CDC1 3 300 MI-z) 8 8.57 (in, 7.65 (mn, 7.42 (in, IH), 7.22 (in, IH), 6.33 (mn, IH), 4.32 (in, IH), 2.25 (in, 2H), 2.13 (in, I 1.87 (hr mn, 2H), 1.63 (in, 214). MS (API-ES Positive) 199 Example 119 Synthesis of 4. 6-d imethyl-2-pyrimidinyl trifluoromethanesulfonate To a stirred solution of 4,6-dimethyl-2-hydroxypyrimidine (5.0 g' 40 minol) in anhydrous
CH
2
CI
2 (100 mL) was added triethylaniine (11.2 mL, 80 inmol), followed by slow addition of trifluoromethanesulfonic anhydride (6.8 mL, 40 mmol) at 0 0 C under argon. The reaction mixture was allowed to warm to 22'C and stirred overnight. The reaction mixture was then diluted with CH 2
CI
2 (100 mL). The organic phase was washed with sat. NaCI (3 x 20 mL), dried (MgSOA) filtered and concentrated under reduced pressure to give a dark oil. Purification by flash chromatography on silica gel eluting with 3:1 hexane:ethyl acetate afforded 4, 6-dimcthyl-2-pyrimidinyl trifluoromethanesulfonate as a brown oil (6.0 g, 'H NMR (CDCI 3 8: 7. 10 I 2.45 6 Hf).
Example 120 Synthesis of 4,6-dimethyl-2-(Dhenylethynyil~primidine hydrochloride A stirred solution of 4, 6-dimethyl-2-pyrimidinyl trifluoromethanesulfonate from Example 119 g, 19.5 minol) in 2:1 DME:H 2 0 (100 mL) was degassed with argon for 10 min. Then K 2 C0 3 (6.7 g, 48.8 minol), GuI (0.37 g, 1.95 mmol), PdCI 2 (Ph 3
P)
2 (0.68 g, 0.98 inmol) and phenylacetylene (5.4 mL, 48.8 mmol) were added at 220C2. The resulting mixture was then heated at 900(2 for 2 h under argon. The reaction mixture was then cooled to 220(C and filtered through a pad of Celitem. The filtrate was concentrated under reduced pressure to give, after purification by flash chromatography on silica gel eluting with 3:1 hcxane:ethyl acetate, the desired compound as a brown crystal which was subsequently treated with a solution of IM HCI in diethyl ether (20 mL) to yield 4,6-dimethyl-2- (phenylcthynyl)pyrimidine hydrochloride as a yellow solid (3.0 g, M.P 149-1500C. 'H NMR
(CD
3 OD) 5: 7.76-7.73 (in, 2 7.67 I 7.60-7.49 (in, 3 1H), 2.69 3 H) ppm; MS(ES): 209 WO 01/16121 PCT/US04)/23923 82 Example 121 Synthesis of 3-(6-methyl-2-yyridinvl)2-nropyn-1 -ol A solution of 2-bromo-6-methylpyridine (2.5 g, 14.5 mmol) in 2:1 DME:H 2 0 (30 mL) was degassed with argon for 10 min. Then PdCl 2 (Ph 3
P)
2 (1.0 g, 1.4 mmol), CuT (0.8 g, 4.3 mmol), K 2 C0 3 (5.0 g, 36.3 mmol) were added followed by propargyl alcohol (2.1 mnL, 36.3 mmol). The resulting mixture was heated at 90'C under argon for 2 h, allowed to cool to 22*C, then filtered through a pad of CeliteTIA. The filtrate was concentrated under reduced pressure and the residue purified by flash chromatography on silica gel eluting with 2:1 ethyl acetate:hexane to give the cross-coupled product 3- (6-methyl-2.pyridinyl)2-propyn- t-ol as a yellow solid (1.0 g, 'H NMIR (CDCI 3 8: 7.58-7.53 (in, 1 7.27-7.25 J 7.5 Hz, I 7.13-7. 10 J 7.8 Hz, I 4.54 2 2.55 3 H4).
Example 122 Synthesis of 3-(6-methyl-2-oyridinyl)-2-propvnvl methanesulfonate To a stirred solution of 3-{6-methyl-2-pyridinyl)2-propyn-l-ol from Example 121 (1.0 g, 6.8 mmol) in anhydrous CH2C1 2 (20 m.L) was added triethylamine (1.0 mL, 7.5 mmol) followed by methanesulfonyl chloride (0.6 mL, 7.5 mmol) at 0 0 C under argon. After 2 h the reaction mixture was diluted with CH 2
CI
2 (50 mL), the organic phase was washed with sat. NaHCO 3 (3 x 10 mL) and sat.
NaCI (3 x 10 mL), dried (MgSO 4 filtered and concentrated under reduced pressure to afford 3-(6methyl-2-pyridinyl)-2-propynyl methanesulfonate as a brown oil (1.4 g, 89%) that was used in the next step without further purification. 'H NM4R (CDCI 3 6: 7.68-7.62 (in, I 7.28 J 7.5 Hz, I H), 7.15 J 7.8 Hz, I 4.55 2 3.09 3 2.57 3 H).
Example 123 Synthesis of 2-methyI-6-(3-nhenvl-1-nroo~vnvl~ovridine To a stirred solution of 3-(6-methyl-2-pyridinyl)-2-propynyl methanesulfonate from Example 122 (1.2 g, 5.3 mmcl) in anhydrous TI-F (10 mL) under argon at O 0 C was added phenylmagnesium bromide (2.1 mL, 6.4 mmol). The reaction mixture was then warmed to 22'C, stirred for I h then diluted with ethyl acetate (50 mL). The organic phase was washed with sat. NaHCO 3 (3 x lOmL), H 2 0 (3 x 10 mL) and sat. NaCI (3 x 10 inL), dried (MgSQ 4 filtered and concentrated under reduced pressure to give a dark oil. Purification by flash chromatography on silica gel eluting with 4:1 hexane:ethyl acetate to afford 2-methyl-6-(3-phenyl-1-propynyl)pyridine as a brown oil (360 mng, 33%).
NMR (CDCI 3 8: 7.54-7.50 (mn, I 7.37-7.19 (in, 5 M, 7.09-7.06 J 7.8 Hiz, I HI), 6.93-6.90 J 7.5 Hz, 1 3.79 2H), 2.5 5 3 H).
WO 01116121 PCT/US00/23923 83 Example 124 Synthesis of 2-methyl-6-(3-phenyl-1.2-propadienyl)pyridine To a stirred solution of 2-methyl-6-(3-phenyl-lI-propynyl)pyridine from Example 123 (100 mg, 0.48 mmol) in anhydrous THE (10 m.L) at -78'C under argon was slowly added n-BuLi (2.5 M in hexane, 0.23 mL, 0.58 mmol). The resulting reddish reaction mixture was stirred for 30 min, then quenched with Methanol (1 mL). The reaction mixture was then warmed to 22"C and taken up in ethyl acetate (50 The organic phase was washed with H 2 0 (3 x 15 mL) and sat. NaCI (3 x 15 mL), dried (MgSOA) filtered and concentrated under reduced pressure. Purification by flash chromatography on silica gel eluting with 6:1 hexane:ethyl acetate afforded 2-methyl-6-(3-phenyl-1,2-propadienyl)pyridine as a yellow oil (40 mg, MS(ES): 208 'H NMR (CDCI 3 8: 7.5 1-7.45 (in, 1 H4), 7.39- 7.21 (mn, 6 6.99-6.96 J 7.5 Hz, I 6.77-6.7 5 J 6.6 Hz,lI 6.65-6.62 J 6.6 Hz, I 2.55 3 1-1).
Example 125 Synthesis of methyl 2-Wltrifluoromethyl)sulfonvlloxyl-1-cvclooentene-I -carboxylate To a stirred solution of methyl 2-oxocyclopentane carboxylate (5.0 g, 35.2 minol) in anhydrous
CH
2
CI
2 under argon at 0 0 C (40 mL) was added NaH (60 in oil, 1.4 g, 35.2 iniol). After stirring for min the resulting yellow cloudy suspension was treated with trifluoromethanesulfonic anhydride (7.lmL, 42.2 minol). The reaction mixture was then warmed to 22'C and after 2 h the reaction was treated with 10% HCI (100 The aqueous phase was extracted with CH 2
CI
2 (3 x 40 mL), the combined organic extracts were washed with sat. NaCI (3 x 50 dried (MgSO 4 filtered and concentrated. The resulting residue was purified by flash chromatography on silica gel eluting with 20:1 hexane:ethyl acetate to afford methyl 2- {[(trifluoromethyl)sulfonyl]oxy}-l1-cyclopentene- 1-carboxylate as a colorless oil (6.5 g, 'H NMR (CDCI 3 5: 3.80 3 2.78-2.69 (in, 4 2.00-2.00 (mn, 2 Example 126 Synthesis of methyl 2-42-nvridinvlethynyfl--cydlopentene-1-carhoxylate A solution of methyl 2- ([(trifluoromethyl)sulfonyl]oxy}-l1-cyclopentene- 1-carboxylate from.
Example 125 (2.0 g, 7.2 iniol) in DMF at 22"C was degassed with argon for 10 min Triethylaniine mL, 1S mmol), Cul (0.41 g, 2.2 inmol), PdCI 2 (Ph 3
P)
2 (0.5 g, 0.72 minol), n-BU 4 N1 (8.0 g, 21.6 inmol) and 2-ethynylpyridine (1.9 g, 18.0 inmol) were added at 22'C and the resulting mixture was heated at 90'C for 2 h under argon. The reaction mixture was allowed to cool to 22*C and filtered through a pad of CeliteT"4. The filtrate was concentrated under reduced pressure to give, after purification by flash chromatography on silica gel eluting with 4:1 hexane:ethyl acetate, methyl 2-(2pyridinylethynyl)-l-cyclopentene-1-carboxylate as a brown solid (1.2 g, 72 M.P. 45-46*C.
WO 01/16121 PCT/USOO/23923 84 MS(ES): 228 1 H NivLR (CDCI 3 8: 8.63-8.6 1 (in, I 7.68-7.65 (in, I 7.5 5-7.53 (in, IH) 7.26-7.23 (in, I 3.81 3 2.81-2.75 (in, 4 2.01-1.90 (mn, 2 H).
Example 127 Synthesis of 2-(2-pynidinylethynyl)-l-cyclopentene-l-carboxylic acid To a solution of methyl 2-(2-pyridinylethynyl)-l-cyclopentene-l-carboxylate from Example 126 (1.0 g, 4.4 mol) in 3:1 Methanol:H 2 0 (20 ml) was added LiOH.H 2 0 (0.55 g, 13.2 mmol). After stirring at 22'C for 5 h the reaction mixture was treated with 10% HC1 (100 mL). The aqueous phase was extracted with CH 2
CI
2 (3 x 40 mL) and the combined organic extracts were washed with sat. NaCI (3 x 50 mL), dried (MgSO 4 filtered and concentrated under reduced pressure to give a crude solid.
Purification by flash chromatography on silica gel eluting with 10:1 CH 2
CI
2
:CH
3 OH afforded 2-(2pyridinylethylnyl)-1-cyclopentene-1-carboxylic acid as a gray solid (330 mng, M.P. 151-152*C.
MS(ES): 214 'H NMR (CD 3 OD) 5: 8.53-8.50 (in, 1 7.88-7.82 (mn I M, 7.62-7.59 (in, I 7.43-7.38 (in, 1 H4), 2.81-2.72 (in, 4 2.06-1.96 (in, 2 H).
Example 128 Synthesis of 1 ff2-(2-qyridinylethvlnyl)-l-cyclopentene-l-yllcarbonvyl iperid e hydrochloride To a stirred solution of 2-(2-pyridinylethylnyl)-1-cyclopentene-l-carboxylic acid from Example 127 (100 mg, 0.47 minol) in CH 2
CI
2 (2 mL) was added HOBT (95 mng, 0.70 mmol), EDCI (135 mng, 0.70 mmol), triethylamine (0.2 mL, 1.4 minol) and piperidine (0.07 mL, 0.70 inmol) at 22TC.
The reaction mixture was stirred for 18 h, then diluted with C1-1 2 C1 2 (50 mL). The organic phase was washed with sat. NaHCO 3 (2 x 25 mL) and sat. NaCl (2 x 25), dried (MgSO 4 filtered and concentrated in vacuo to give a yellow oil. Purification by flash chromatography on silica gel eluting with 1:1 hexane:ethyl acetate afforded I {[2-(2-pyridinylethylnyl)-1-cyclopentene-l-yljcarbonyl~piperidine as a yel low oil which was treated with I M I ICI in diethyl ether (5 mL) to yield 1 {[2-(2-pyridinylethyinyl)l-cyclopentene-1-yl]carbonyl~piperidine hydrochloride as a yellow foam (40 mng, MS(ES): 281 H NMR (CDC1 3 5: 8.75 (br, I 8.21 (br, 1 1H), 7.74-7.73 (in, 2 3.71 (in, 2 3.53 (in, 2 2.84-2.79 (in, 4 2.11-2.05 (in, 2 1.69-1.61 (in, 6 H).
Example 129 Synthesis of 1-methyl-4-f 2-(2-nyvridinylethvlnvl)-l-cvclooente e-1-yilcarbonylviyjnerazine hydrochloride To a stirred solution of 2-(2-pyridinylethylnyl)-l-cyclopentene-l-carboxylic acid from Example 127 (100 mg, 0.47 minol) in CH 2
CI
2 (2 inL) was added HOBT (95 ing, 0.70 inmol), EDGI (135 ing, 0.70 iniol), triethylamine (0.2 inL, 1.4 inmol) and 1-inethylpiperazine (123 mg, 0.70 iniol) at 22*C. The reaction mixture was stirred for 18 h and then diluted with an additional amount of CH 2
CI
2 mL). The organic phase was washed with sat. NaHCO 3 (2 x 25 mL) and sat NaCI (2 x 25), dried WO 01/16121 PCT/USOO/23923 (MgSO 4 filtered and concentrated in vacuo to give a yellow oil. Purification by flash chromatography on silica gel eluting with 10:1 CH 2
CI
2
:CH
3 OH afforded 1-methyl-4-{[2-(2-pyridinylethylnyl)-1cyclopentene-1-yl]carbonyl~piperazine as a yellow oil which was treated with I M HCI in diethyl ether mL) to yield 1 -methyl-4- ([2-(2-pyridinylethylriyl)- 1-cyclopentene-1I-yl]carbonyl} piperazine hydrochloride as a yellow foam (130 mg, MS(ES): 296 'H NMR (CDCI 3 5: 8.66 (br, 1 8.31 (br, I 7.82 (br, 2 1H), 4.75 (in, I 4. 10 (mn, 2 3.80 (mn, 3 3.36 (mn, 2 3.07 3 2.79 (in, 4 2.15 (mn, 2 H).
Example 130 Synthesis of 4-(4-pvyrimidinyl)nhenyl trifluoromethanesulfonate To a stirred solution of 4-(4-pyrimidinyl)phenol (250 mng, 1.45 mmol) in anhydrous CH 2
CI
2 (100 mL) was added triethylainine (0.4 mL, 2.9 minol) followed by slow addition of trifluoromethanesulfonic anhydride (0.25 mL, 1.45 minol) at 00(2 under argon. The reaction mixture was allowed to warm to 220(C and stirred overnight and then diluted with CH 2
CI
2 (100 mL). The organic phase was washed with sat. NaCI (3 x 20 inL), dried (MgSO 4 filtered and concentrated under reduced pressure to give a dark oil. Purification by flash chromatography on silica gel eluting with 4:1 hexane:ethyl acetate afforded 4-(4-pyrimidinyl)phenyl trifluoromethanesulfonate a brown oil (210 ing, MS(ES): 305 'H NMR (CDCI3) 5: 9.31-9.30 J 1.2 Hz, I 8.85-8.83 (mn, I H), 8.22-8.19 (mn, 2 7.73 (in, 1 7.46-7.43 (in, 2 H).
Examp~le 131 Synthesis of 4-14-[(2-methvl-1 ,3-thiazol-4-vl)ethvnvylnhenylIpyuimidine A stirred solution of 4-(4-pyrimidinyl)phenyl trifluoromethanesulfonate from Example 130 (200 mg, 0.66 mmol) in DMF (8 mL) was degassed with argon for 10 min. PdCI 2 (Ph 3
P)
2 (46 mng, 0.07 inmol), Cul (38 mg, 0.2 mmol), triethylamine (0.23 mL, 1.64 mmol) and n-Bu 4 NBr 243 mg, 0.66 mmol) were added, followed by 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 (169 mg, 0.86 inmol). The reaction mixture was heated at 700(2 under argon and then T7BAF (1.0 M in THF, 0.86 mL, 0.86 mmol) was added slowly over 20 min. The reaction mixture was allowed to cool to 220(C, then filtered through a pad of Celite'm. The filtrate was concentrated under reduced pressure and the residue purified by flash chromatography on silica gel eluting with 1: 1 hexane:ethyl acetate to give the cross-coupled product 4 -{4-[(2-methyl-1,3-thiazol-4-yl)ethynyljphenyl~pyrijmidine as a yellow solid. This material was treated with 1 M HCI in diethyl ether to afford a yellow solid (130 mng, 5 M.P. 190-192 0 C. MS(ES): 278 'H NMR (CD 3 OD) 8: 9.53 (br, I 9.12 (br, 1 8.57 (mn, I 8.47 1 8.45 1 7.92 I 7.84 I 7.81 1 2.81 3 H).
WO 01/16121 PCT/US00/23923 86 Exam~le 132 Synthesis of 3-bromo-5-1U2-methyl-l thiazol-4-yl)ethynyll Dyrdine 3,5-bisl(2-m ethyl- 1.3thiazol-41-yI)thynvll Dyridine A solution of 3,5-dibromopyridine (5.1 g, 21.5 mmol) in DMIF (100 m.L) was degassed with argon for 10 min. PdCI 2 (Ph 3
P)
2 (0.75 g, 1. 1 mmol), Cul (0.61 g, 3.2 mmol) and triethylamine (3.7 mL, 26.9 mmol) were added followed by 2-methyl-4-[Qtrimethylsilyl)ethynyl]-1,3-thiazole from Example 77 (2.1 g, 10.7 mmol). The reaction mixture was heated at 73*C under argon. Then TBAF (1.0 M in TI-f, 11.8 mL, 11.8 mmol) was added slowly over 20 min. The reaction mixture was allowed to cool to 22'C, filtered through a pad of Celitem and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with 4:1 hexane:ethyl acetate to give two cross-coupled products: 3-bromo-5-[(2-methyl-l,3-thiazol.4-yl)ethynyllpyridine as a white solid (1.7 g, M.P. 123-124-C, MS(ES): 280 'H NMvR (CDCI 3 5: 8.69-8.68 J 1.7 Hz, I H), 8.63-8.23 J =2.2 Hz, I 7.99-7.98 (in, I 7.46 1 H4), 2.76 3 H) ppm; and 3,5-bis[(2methyl-1,3-thiazol-4-yl)ethynyl]pyridine as a yellow solid (160 mg 5 M.P. 171-172*C, MS(ES): 322 'H NMR (CDCI 3 8: 8.72-8.71 (in, 2 7.95 (in, I 7.46 2 2.76 6 14).
Example 133 Synthesis of 3-(3-nyridinv)-5-I(2-methvl-1 .3-thiazol-4-vflethvnvllpyridine hydrochloride A solution of 3-bromo-5-[(2-methyl-l,3-thiaol-4-yl)etbynyllpyridine from Example 132 rng, 0.27 mmol) in TI-F (15 mL) was degassed with argon for 10 min. Pd(Ph 3
P)
4 (15 mg, 0.013 minol), KOH (45 mg, 0.81 mmol) and n-Bu 4 NBr (43 mng, 0.13 inmol) were added, followed by diethyl(3pyridyl)borane (51 mng, 0.35 minol). The reaction mixture was heated at reflux for 5 h under argon, then allowed to cool to 22'C and filtered through a pad of Celite'~'. The filtrate was concentrated under reduced pressure and the residue purified by flash chromatography on silica gel eluting with 2:1 ethyl acetate:hexane to give 3-(3.pyridinyl)-5-[(2-methyl- 1,3 -thiazol-4-yl)ethynyllpyridine as a brown oil.
This material was treated with I M HCI in diethyl ether to yield 3-(3-pyridinyl)-5-[(2.methyl-l,3thiazol-4-yl)ethynyljpyridine hydrochloride as a yellow solid (60 mng, M.P. 164-166'C. MS(ES): 278 'H NMR (CDCI 3 6: 8.97 (in, 1 8.90 1.5 Hz, I 8.83-8.82 2.1 Hz, 1 H), 8.80-8.77 (in, 2 8.30 (in, I 8.11 (in, 1 7.82 (in, I ED, 7.52 I 7.27 I 2.78 3
H).
Example 134 Synthesis of 3-(4-pyridinVl')-5-i(2-methyl-1 .3-thiazol-4-yI)ethynyll Dyrdine hydrochloride A solution of 3-bromo-5-[(2-methyl-l,3-thiazo-4-yl)ethynyl]pyridine from Example 132 (48 mg, 0.17 mmol) in 2:1 DME:H 2 0 (30 mL) was degassed with argon for 10 mini. Pd(Ph 3
P)
4 (10 ing, 0.009 minol), K 2 C0 3 (59 mg, 0.43 mmol) and n-Bu 4 NBr (48 mg, 0. 15 inmol) were added, followed by pyridine-4-boronic acid (32 mg, 0.26 inmol). The reaction mixture was heated at 90*C under argon for WO 01/16121 PCT/USOO/23923 87 1 h, then allowed to cool to 22 0 C, filtered through a pad of Celite'm and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with 2:1 ethyl acetate:hexane to give 3-(4-pyridinyl)-5-[(2-methyl- 1,3-thiazol-4.yl)ethynyl]pyridine as a colorless oil. This material was treated with I M HCI in diethyl ether to afford 3-(4-pyridinyl)-5-[(2methyl-l,3-thiazol-4-yl)ethynyl]pyridine hydrochloride as a yellow solid (20 mg, M.P. 115- 11 7 0 C. MS(ES): 278 'H NMR (CD 3 OD) 8: 9.48 (br, I 9.28 (br, 1 9.18 I 9.09 2 8.63 2 8.08 1 2.83 3 H).
Exampl~e 135 Synthesis of 5-f5-r(2-methyl-1,3-thiazoI-4-vl)ethvnvll-3-oviidinvl1 pvrm idine hydrochloride A solution of 3-bromo-5-[(2-methyl-1,3-thiazol-4-yl)ethynyljpyridine from Example 132 (100 mg, 0.36 mmol) in 2:1 DMIF:H 2 0 (5 mL) was degassed with argon for 10 min. Pd(Ph 3
P)
4 (21 mg, 0.0 18 mmol), K 2 C0 3 (124 mg, 0.9 mmol) and n-Bu 4 Nllr (115S mg, 0.36 mmcl) wvere added, followed by pyrimidinylboronic acid (67 mg, 0.54 mmcl). The reaction mixture was heated at 90*C under argon for I h, then allowed to cool to 22'C and filtered through a pad of CeliteTm. The filtrate was concentrated under reduced pressure and the residue purified by flash chromatography on silica gel eluting with 2:1 ethyl acetatc:hexane to give 5-[(2-methyl-1 ,3-th iazol-4-yl)ethynylj-3pyridinyl~pyrimidine as a colorless oil. This material was treated with I M HCI in diethyl ether to afford 5- {5-[(2-methyl- 1,3-thiazol-4-yl)ethynyl]-3-pyridinyl }pyrimidine hydrochloride as a yellow solid (20 mg, M.P. 135-137"C. MS(ES): 279 'H NMR (CD 3 OD) 8: 9.37-8.77 (in, 7.97 (mn, 1 7.73 (mn, I 2.78-2.77 J 1.6 Hz, 3 H).
Example 136 Synthesis of 3-(3,5-dimethyl-4-isoxazoly)-5-i(2-methyl-1 ,3-thiazo-4-yl)ethynyllpyridine hydrochloride A solution of 3-bromo-5-[(2-methyl-l,3-thiazol4yl)ethynyllpyridine from Example 132 (100 mng, 0.36 mmol) in 2:1 DMF:H- 2 0 (5 mL) was degassed with argon for 10 min. Pd(Ph 3
P)
4 (21 mg, 0.018 inmol), K 2 C0 3 (124 mng, 0.9 inmol) and n-Bu 4 NBr (11 Ing, 0.3 6 mnmol) were added followed by diinethyl-4-isoxazolylboronic acid (78 mng, 0.54 inmol). The reaction mixture was heated at 90*C under argon overnight, allowed to cool to 22*C and filtered through a pad of Celite-. The filtrate was concentrated under reduced pressure and the residue purified by flash chromatography on silica gel eluting with 2:1 hexane:ethyl acetate to give 3-(3,5-dimethyl.4-isoxazolyl)-5-[(2-inethyl-1,3-thiazo-4yl)ethynyl]pyridine as a yellow solid. This material was treated with 1 M HCI in diethyl ether to afford 3-(3,5-dimethyl-4-isoxazolyl)-5-[(2-methyl- 1,3 -thiazo-4-yl)ethynyl]pyridine hydrochloride as a yellow solid (19 mg, MS(ES): 296 'H NMR (CD 3 OD) 8: 9.20-8.81 (in, 3 8.01 (br, 1 H), 2.80 3 2.58 3 2.39 3 H).
WO 01/16121 PCT/USOO/23923 88 Examnple137 Syntheses of -4-methoxyphenyl)-5-f(2-methyl-l .3-thiazol-4-yl)ethynyllpyridine hydrochloride A solution of 3-bromo-5-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine from Example 132 (100 mg, 0.36 mmol) in 2:1 DMfF:H 2 0 (5 m.L) was degassed with argon for 10 min. Pd(Ph 3
P)
4 (21 mg, 0.018 mmol), K 2 G0 3 (124 mg, 0.9 mmol) and n-Bu 4 NBr (115 mg, 0.36 mmol) were added followed by 4methoxyphenylboronic acid (82mg, 0.54 mmol). The reaction mixture was heated at 90'C under argon for 1 h, then allowved to cool to 22 0 C and filtered through a pad of Celitem. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel eluting with 1.1 hexane:ethyl acetate to give 3-(4-methoxyphenyl)-5-[(2-methyl-l,3-thiazol-4yl)ethynyllpyridine as a yellow solid. This material was trcated with I M HC1 diethyl ether to afford 3- (4-methoxyphenyl)-5-[(2-methyl-1,3-thizol-4-yl)ethynyjpyridine hydrochloride as a yellow foam mg, MS(ES): 307 'H NMR (CDOD) 5: 9.12 1 9.00-8.98 (in, 2 7.96 1 7.84 I 7.81 1 7.17 I 7.14 I 3.89 3 2.76 3 H).
Using the synthetic and purification procedures described in Examples 132 to 137, and using the appropriate reagents, compounds described in Examples 138 to 145 were prepared.
Exampl 3 3-1(2-Methyl-i .3-thiazol-4-yIoethynyll-5-(2-thienyl)pyridinc Isolated as an off white solid. M.P 175-176TC. MS(ES): 283 'H -NMvR (CDOD) 9.16 (br, I 8.92-8.91 (in, 2 7.96 I 7.86-7.84 (in, I 7.77-7.75 (mn, I 7.28-7.26 (in, 1 2.78 3 H).
E~xamle13 3-(2-Furyl)-1-(2-methyl-1.3-thiazol-4-ylethynyllpyridine Isolated as a colorless glass. MS(ES): 267 'H NMR (CDCI 3 5: 8.87 (mn, I Hf), 8.64 (in, I 8.10-8.08 (in, 1 7.54-7.54 (in, I 7.46 I11H), 8.78-6.77 (in, 1 6.53 (in, I 2.76 3 H).
Example 140 3-1(2-Methyl-I .3-thlazol-4-yUothynll-5-(-trlfluoromethyl)phenvllpyridine Isolated as an off-white foam. MS(ES): 267 'H NMR (CD 3 OD) 5: 9.13 (br, I 9.02 (br, I 8.85 (br, 1 8.03 I 8.00 I 7.91 2 7.88 1 2.75 3 H).
Exampfle 41 3-(1-Benzothlen-2-yI)-5-1(2-methyl-1-3-thiazol-4-yl)ethynyl pvrldlne Isolated as an off-white solid. MS(ES): 333 NMR (CD 3 OD) 8: 9.12 (br, I 8.84 (br, 1 H1), 8.70 I 1I), 8.03 I 7.92-7.90 (in, 3 7.45-7.42 (mn, 2 2.76 3 H).
WO 01/16121 PCT/USOO/23923 89 Example 142 3-(2-Methvl- 1 3-thiazol-4-yl)-5-(1H-pyrazol-3-yl)nyridine Isolated as an off-white solid. M.P. 184-186 0 C, MS(ES): 267 'H NMR (CD 3 OD) 8: 9.28 I 9.13 1 9.01 I 8.03 1 7.88-7.87 J 2.25 Hz, I 7.07-7.06 J 2.25 Hz, I 2.80 3 H).
Example 143 3-42.4-Difluoroohenyl)-5-(2-methvl-1,3-thiazol-4-vl)pvridine Isolated as a white solid. MS(ES): 313 'H NMR (CD 3 OD) 8: 8.87 (br, 2 8.45 (br, I 7.86 I1-H), 7.72-7.68 (in, 1 7.24-7.16 (in, 2 2.74 3 H Example 144 3-(4-Fluorophenv)-5-(2-methvl-1.3-thiazol4-l)Dyridine Isolated as a low melting/hygroscopic solid. MS(ES): 295 'H NMR (CD 3 OD) B: 9.16 (in, I 9.06 (in, I 9.10 (in, 1 7.96 I 7.93-7.89 (in, 2 7.39-7.33 (in, 2 2.76 3
H).
Example 145 3-(2-methyl-0,-thiazol-4-yl) Isolated as a hygroscopic solid. MS(ES): 277 'H NMR (CD 3 OD) 8: 9.10 (br, 2 H), 8.76 I 7.90 I 7.82-7.79 (mn, 2 7.60-7.54 (in, 3 2.75 3 H).
Example 146 Synthesis of 2-fluorocvclohexanone To a stirred solution of (1-cyclohexen-1-yloxyXtrimethyl)silane (3.5 g, 21 inmol) in dry MeCN (200 mL) under argon was added Selecffluor reagent (8.0 g, 23 iniol). The mixture was stirred at ambient temperature for 3 h, then stored at -20'C overnight, at which time GUIMS showed the reaction to be complete. The mixture was diluted with ethyl acetate (600 mL), washed with dilute brine (300 mL) and then saturated brine (100 inL), dried over Na 2
SO
4 and concentrated in vacuo. The resulting liquid was purified by Kugelrohr distillation (130'C air bath at 20 Torr) to afford 2fluorocyclohexanane (1.6 g, 67%) as a clear, colorless oil. 11-I NMR (CDC1 3 300 N4IHz) B 4.90 (ddd, Jji.=49.1 Hz, J 2 =1 1.3 Hz,4 J 3 =6.4 Hz, IH), 2.61-1.67 (in, 8R). MS (El ionization) 116 (M)J.
Example 147 Synthesis of 2-fluoro-1-l(trimethvlsilvl)ethvnyllcyclohexanoI A solution of n-BuLi (7.6 inL, 2.2M in hexanes, 17 inmol) was added slowly at -40'C to a stirred solution of triinethylsilylacetylene (2.3 mL, 16 inmol) in dry THE (30 inL) under argon. The mixture was stirred at -40*C for 20 min, cooled to -78*C, then a solution of 2-fluorocyclohexanone WO 01/16121 PCT/US00/23923 from Example 146 (1.6 g, 14 mmol) in dry THF (20 mL) was added via syringe. The mixture was warmed to ambient temperature gradually over I h and then stirred at ambient temperature for 4 h.
Analysis of the reaction mixture by GCIMS at this time showed the reaction to be complete. The reaction was quenched by the addition of saturated aqueous NH 4 CI (20 mL), stirred for 5 min, then poured into H 2 0 (50 mL), and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with H 2 0 (30 mL), brine (30 mL), dried over Na 2
SO
4 filtered and concentrated in vacua to afford 2-fluoro-l-[(trimethylsilyl)ethynyljcyclohexanol (3.4 g, 97%) as a light brown oil as a mixture of diastereomers. 1H NMR (CDCI 3 300 MHz) 5 1.89-1.25 (in, I1OH), 0. 15 9H). MS (El ionization) 214 Example 148 Synthesis of 2-fluoro-1-(1.3-thiazol-2-vlethvnyl)cvclohexanol PdCI 2 (45 mng, 0.25 mmol) was suspended in DME (20 mL), then argon was bubbled through the solution for several min to deoxygenate it. H-20 (6.6 K 2 C0 3 (4.5 g, 32 inmol), CH 3 OH mL), 2-bromo-l,3-thiazole (2.63 g, 16.1 mmol), PPh 3 (280 mg, 1.07 minol) and Cul (204 mg, l.O7mmol) were then added and the reaction mixture was heated to 50 0 C. After a few min 2-fluoro-l- ((trimethylsilyl)ethynyl]cyclohexanol from Example 147 (2.3 g, 11I minol) was added to the dark brown suspension. The reaction was then heated to 601C and after 16 h at 601C, GUIMS showed no remaining 2-bromo-1,3-thiazole. The reaction mixture was concentrated in vacuo, diluted with ethyl acetate (200 mL), and filtered through Celitel. The filter pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with H 2 0 (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel on silica gel eluting with hexane, 9:1, then 7.5:2.5 hexane:ethyl acetate to afford 2-fluoro-1-(1,3-thiazol-2ylethynyl)cyclohexanol (1.1 g, 45%) as a yellowish solid, and a 5:1 mixture of diastereomers. 'H NMR (CDC1 3 300 M[Hz) 5 7.83 J=3 Hz, lH), 7.37 J=3 Hz, IH), 4.80-4.33 (mn, IH), 3.06 2.21- 1.23 (in, 6H). MS (El ionization) 225 Example 149 Synthesis of 3-ethoxy-5-methvl-2-cyclohexen-l-one A solution of 5-methyl-1,3-cyclohexanedione (9.83 g, 77.9 minol) and TsOH.H 2 0 (636 mng, 6 in ethanol (80 mL) was heated to reflux. After 15 min, triethyl orthoformate (5 mnL, 30 inmol) was added. After I 0mmn TLC showed the reaction was incomplete, and additional triethyl orthoformate mL, 30 minol) was added. After a further 10 min GCIMS showed the reaction to be complete. The reaction mixture was concentrated in vacua, then diluted with diethylether (300 mL) and washed with NaHCO 3
H
2 0 (300 mL), brine (2 x 300 dried over Na 2 S0 4 and filtered. The filtrate was concentrated in vacua, to afford 3-ethoxy-5-methyl-2-cyclohexen-1I-one as an yellowish-orange oil WO 01/16121 PCT/USOO/23923 91 (11.04 g, 'H NMR (CDCI 3 300M1-Iz) 8 5.34 I 3.94-3.86 (in, 2H), 2.44-1.95 (in, 6H), 1.36 J=7 Hz, 3H), 1.08 J=6 Hz, 3H). MS (El ionization) 154 Example 150 Synthesis of 5-mnethvl-3-(2-pyid inylethynyl)-2-cvclohexen- I-one A solution of n-BuLi (8 m-L, 2.2M in hexanes, 18 nmol) was added slowly at 0 0 C to a stirred solution of dilsopropylamine (2.8 mL, 19 mmol) in dry THF (20 mL) under argon. The mixture was stirred at 0 0 C for 30 min then a solution of 2-ethynylpyridine (2.27 g, 22.0 mmol) in dry THF (20 mL) was added via syringe. The mixture was stirred at 0 0 C for I h, then 5-methyl-3-ethoxy-2-cyclohexene- I-one from Example 149 (2.0 g, 13 mmol) was added. After stirring for 16 h at ambient temperature GC/MS showed the reaction was complete. The reaction mixture was quenched by the addition of saturated aqueous NH 4 CI (100 mL), stirred for 5 min and poured into H 2 0 (50 mL), and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with H20 (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude material was purified by column chromatography on silica gel eluting with hexane, 9:1, 4:1, then 7:3 hexane:ethyl acetate to afford 5-methyl-3-(2-pyridinylethynyl)-2-cyclohexen-I -one (1.0 g, 36%) as a light green solid. M.p.= 62.5-65 0 C. H NMR (CDCl 3 300 MHz) 5 8.64 J=6 Hz, I1H), 7.74-7.68 (mn, I1-I), 7.51 J-9 Hz, I 7.32-7.26 (in, I1H), 6.37 I1H), 2.67-2.50 (in, 2H), 2.33-2.09 (in, 3 1. 11 3H). MS (ESI) 212.0 Example 151 Synthesis of 3-ethoxy-5,5-dimethvl-2-cyclohexen- I-one To a solution of 5,5-dimethyl-l,3-cyclohexanedione (25.2 g, 180 minol) in ethanol (200 mL), was added TsOH- H 2 0 (1.06 g, 4.2 then triethyl orthoformate (30 m.L, 180 minol). The resulting pale yellow solution was then heated to reflux. After heating for 15 min the solution had turned bright orange. GCUMS analysis of the reaction after 1 h showed no starting material present. The orangebrown solution was concentrated in vacuo, diluted with ethyl acetate (300 mL), washed with saturated aqueous NaHCO 3 (2 x 75 mL), brine (150 dried over Na 2
SO
4 filtered and concentrated in vacuo to afford an orange semi-solid. The crude product was purified by column chromatography on silica gel eluting with hexane, then 3:1 hexane:ethyl acetate to afford 3-ethoxy-5,5-dimethyl-2-cyclohexen-1I-one (13.5 g, 44%) as an orange oil that slowly solidified. 1'H NMR (CDC1 3 3 00 MHz) 8 5.34 I1H), 3.90 J 7.0 Hz, 2H), 2.28 IH), 2.21 114), 1.37 J=7.0 Hz, 3H), 1.08 MIS (El ionization) 168 (M) WO 01/16121 PCT/USOO/23923 92 Example 152 Synthesis of 5.5-d imethyl-3-(2-Dyvridinylethynyl)-2-cyclohexen-l-one A solution of n-BuLi (7.0 mL, 2.2M in hexanes, 15 mmol) was added slowly at 0 0 C to a stirred solution of diisopropylamine (2.5 mL, 18 mmol) in dry TI-F (20 mL) under argon. The mixture was stirred at 0 0 )C for 30 min then a solution of 2-ethynylpyridine (2.1 g, 2Ommol) in dry TI-F (20 miL) was added via syringe. The mixture was stirred at 0 0 C for I h then 3-ethoxy-5,5-dimethyl-2-cyclohexen-lone from Example 151 (2.0 g, 12 mmol) was added and the reaction mixture was allowed to warm to ambient temperature. After 16 h at ambient temperature GCUMS showed the reaction to be complete.
The reaction was quenched by the addition of saturated aqueous NH 4 CI (100 mL). After 5 min the reaction was poured into H-20 (50 mL), and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with H 2 0 (100 mL), brine (100 ML), dried over Na 2
SO
4 filtered and concentrated in vacuo. The crude material was purified by column chromatography on silica gel eluting with hexane, 9:1, then 4:1 hexane:ethyl acetate to afford 5,5-dimethyl-3-(2-pyridinylethynyl)-2cyclohexen-l-one (0.563 g, 12%) as a red oil.' H NMR (CDCI 3 300 MIHz) 8 8.64 J=6 H-z, IHf), 7.72-7.71 (in, 7.51 J=6 Hz, IH), 7.28-7.27 (in, 1H), 6.38 114), 2.47 2H), 2.31 2M), 1.09 MS (ESI) 226.1 Example 153 Synthesis of 2-(trimethylsilvl)ethynyll bicyclo[2 .2.11 heotan-2-oI A solution of n-BuLi (46 mL of a 2.2M solution in hexanes, 100 mmol) was added slowly at -78'C to a stirred solution of trimethylsilylacetylene (10 g, 100 mmol) in dry TI-F (100 mL) under argon. The mixture was stirred at -78'C for 30 min, then a solution of bicyclo[2.2.1]heptan-2-one (norcamphor) (7.7 g, 70 inmol) in dry TI-F (70 mL) was added via syringe. The mixture was allowed to reach ambient temperature gradually and stirred for 1.5 hr, at which time GC/MS showed the reaction to be complete. The reaction was quenched by the addition of saturated aqueous NH 4 CI (300 mL), the mixture was stirred for 5 min and poured into H 2 0 (100 mL), and extracted with ethyl acetate (3 x 300 mL). The combined organic layers were washed with H 2 0 (100 mL), brine (100 dried over Na.
2
SO
4 and filtered. The crude material was purified by column chromatography on silica gel eluting with hexane, then 20:1 hexane:ethyl acetate to afford 2-[(trimethylsilyl)ethynyl]bicyclo[2.2.1]heptan-2ol (10 g, 47%) as a red oil. H NMR (CDC1 3 300 MHz) 8 2.06-1.31 (in, I IH), 0. 16 9H). MS (El ionization) 207.
Example 154 Synthesis of 2-41 .3-thiazol-2-ylethvnyflhicvlo2.2.11 henDtan-2-oI PdCI 2 (27 mg, 0.l1inmol) was suspended in DME (20 inL), then argon was bubbled through the solution for several min to deoxygenate it. H 2 0 (10 mL), K 2 C0 3 (2.5 g, 18 minol), CH- 3 0H (20 mL), 2bromo-1,3-thiazole (1.0 g, 6.1 minol), PPh 3 (160 mg, 0.61 mmol) and CuT (118 mg, 0.61 minol) were WO 01/16121 PCT/USOO/23923 93 added and the reaction was heated to 50*C. After a few min 2-[(trimethylsilyl)ethynyl]bicyclo[2.2.1lheptan-2-ol from Example 153 (1.3 g, 6.1 mmol) was added to the dark brown suspension. The reaction was heated to 60'C and allowed to stir for 16 h, at which time GU/MS showed no remaining 2-bromo-l,3-thiazole. The reaction mixture was diluted with ethyl acetate (200 mL), and filtered through CeliteTM. The filter pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with H 2 0 (200 mL), brine (200 mnL), dried over Na 2
SO
4 and filtered.
The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, 9:1, then 4:1 hexane:ethyl acetate to afford 2-(1,3-thiazol-2yiethynyl)bicyclo[2.2.1I]heptan-2-ol (550 mg, 42%) as a red oil. 'H NMR (CDCI 3 3 00 MHz) 5 7.79 (d, J=3 H7, I 7.34 J=3 Hz, I1H), 2.55-1.26 (in, I11K). MIS (El ionization) 219.
Example 155 Synthesis of 2-(bicyclo!2.2.llhept-2-en-2-vlethynyl)1,3-thiazole To a solution of 2-(1,3-thiazol-2-ylethynyl)bicyclo[2.2.1]heptan-2-ol from Example 154 (300 mng, 1.37 inmol) in CH 2
CI
2 (20 mL) under argon, was added a catalytic amount of 4-(dimethylamino)pyridine, and triethylaniine (0.57 mL, 4.1 inmol). After 2 min methanesulfonyl chloride 16 mL, mmol) was added. After I h GU/MS showed the reaction to be complete. The mixture was diluted with ethyl acetate (200 mL), and washed with NaHCO 3 (200 mL), 1-420 (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, then 98:2 hexane:ethyl acetate to afford 2- (bicyclo[2.2.1]hept-2-en-2-ylethynyl)-1,3-thiazole (203 mg, 74%) as a red oil. 'H NMAR (CDC1 3 300 MHz) 5 7.81 J=3 Hz, 11-1), 7.33 J=3 Hz, IH), 6.55 J=3 Hz, 111), 3.09 1ff), 2.99 J=1.2 H-z, I1K) 1.81-1.67 (mn, 2 1.53 J=8.5 Hz, I1H), 1.25-1.09 (mn, 3M1. MS (EST) 202.1 Example 156 Synthesis of 1-[(6-methyl-2-Dyridinvl~ethynvicyclopentanoI PdCI 2 (PPh 3 2 (320 mg, 0.45 inmol) was suspended in DME (20 mL) then argon was bubbled through the solution for several min to deoxygenate it. 2-Bromo-6-methylpyridine (1.9 g, I11 mmol), triethylamine (6.3 inL, 45 inmol) and GuI (170 mg, 0.91 inmol) were added and the reaction was heated to 50'C. After a few min 1-ethynyl cyclopentanol (1.0 g, 9.1 minol) was added to the dark brown suspension. The reaction was heated to 60*C, and after 16 h at 60*C GCIMS and TLC analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (200 mL), and filtered through CeliteM". The filter pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with 1120 (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, 9:1, 8:2, then 7:3 hexane:ethyl acetate to afford I -[(6-methyl-2-pyridinyl)ethynyl]cyclopentanol (1.2 g, 57%) as a yellow solid. MS (El ionization) 201 (Wy).
WO 01/16121 PCT/USOO/23923 94 Example 157 Synthesis of 2.41-cyclopenten-l-yiethynyl)-6-methylnynidine To a solution of 1-[(6-methyl-2-pyridinyl)ethynylJcyclopentanol from Example 156 (190 mg, mmol) in CH 2
CI
2 (10 mL) under argon was added phosphorus pentoxide (355 mg, 2.5 mmol). The mixture was allowed to stir at ambient temperature for 16 h at which time TLC showed the reaction to be complete. The reaction was quenched by the addition of saturated aqueous NaICO 3 solution (to basic pH), stirred for 5 min, and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with H 2 0 (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacua.
The crude material was purified by column chromatography on silica gel eluting with hexane, then 9:1 hexane:ethyl acetate to afford 2-(l-cyclopenten-l-yethynyl)-6-methylpyridine (110 mg, 63%) as a yellow solid. 1 H NMR (CDCI 3 300 M]Hz) 8 7.54-7.48 (in, 114), 7.25-7.23 (mn, IH) 7.07-7.03 (in, 1H), 6.25-6.23 (in, IiH), 2.60-2.43 (in, 71-1), 1.99-1.90 (mn, 2HM.MS (ESI) 184.1 (NrM1.
Example 158 Synthesis of Racemic 2-f l(cis)-3,4-dimethyl-I -c'vclonenten-l-yllethvnvll pyrdine A solution of n-BuLi (16.8 mL of a 2.5M solution in hexanes, 42.0 minol) was added gradually at 0 0 C to a stirred solution of diisopropylamine (5.9 mL, 42 mmol) in dry TH4F (40 mL) under argon.
The mixture was stirred at 0 0 C for 30 min then cooled to -78*C and a solution of cis-3,4dimethylcyclopentanone [Mori, Ueda, Tetrahedron, (1982), pp 1227-1233] (3.6 g, 32 mmol) in dry TI-f (20 inL) was added via syringe. After the mixture stirred at -78 0 C for I h a solution of Nphenyltrifluoromethanesulfonimide (13.8 g, 38.6 minol) in dry THF (40 mL) was added via syringe during 15 min. The reaction was allowed to warm gradually to ambient temperature, and after stirring for 16 h at ambient temperature GC/MS showed no remaining cis-3,4-dimethylcyclopentanone. The reaction was quenched with NaHCO 3 (2mb), DM-E (30 mL) was added, and argon was bubbled through the solution for several min to deoxygenate it. To the resulting solution of racemic (cis)-3,4-dimethyl-1cyclopenten-l-yl trifluoromethanesulfonate PdC] 2 (PPh 3 2 (1.1 g, 1.6 minol), triethylamine (22 mL, 160 minol), Cul (1.2 g, 6.4 inmol), Ph 3 P (0.50 g, 1.9 minol), and 2-ethynylpynidine (4.0 g, 38 mmol) were added, and the reaction was heated to 50*C. After stirring for 16 h at 50 0 C, TLC analysis showed no remaining racemic (cis)-3,4-dimethyl-l-cyclopenten-1-yl trifluoromethanesulfonate. The mixture was diluted with ethyl acetate (400 mL), and filtered through a Celite~' pad. The filter pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with 1120 (200 mL), brine (200 inL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, then 96:4 hexane:ethyl acetate to afford racemic 2-{[(cis)-3,4-dimethyl-l-cyclopenten-1-yl]ethynyl~pyridine (4.6 g, 73%) as a light brown oil. 1 H NMR (CDC1 3 300 M]Hz) 5 7.58-8.56 (in, 7.66-7.60 (in, 1K) 7.42 J=9 Hz, 11K), WO 01/16121 PCT/US00/23923 Example 159 Synthesis of 2-Hf(3S.4R)-3,4-dimethyl-l-cvclopenten-l-yllethynvlli~vridine A solution of n-BuLi (6.7 mL of a 2.5M solution in hexanes, 17 mmol) was added slowly to a stirred solution of diisopropylamine (2.4 mL, 17 mmol) in dry 11-F (20 mL) at 0 0 C under argon. The mixture was stirred at 0 0 G for 30 min then cooled to -78'C and a solution of (3S,4S)-3,4dimethylcyclopentanone [Kokke, W. C. M. Varkevisser, F. J1. Org. Chem. (1974), pp 1535-1539; Hcathcock, C. Davis, B. Hadley, C. J. Med. Chem. (1989), pp 197-202) (1.45 g, 13.0 mmolJ in dry THIF (10 mL) was added via syringe. After the mixture was stirred at -78*C for 1 h, a solution of N-phenyltrifluoromethanesulfonimide (5.5 g, l6mmol) in dry TI-F (20 mL) was added via syringe during 15 min. The reaction was stirred and allowed to gradually warm to ambient temperature. After stirring for 16 h, GC/MS showed no remaining (3S,4S)-3,4-dimethylcyclopentanone. The reaction was quenched with saturated aqueous NaHCO 3 (1 mL), DME (15 mL) was added, then argon was bubbled through the solution for several min to deoxygenate it. To the resulting solution of (3R,4S)-3,4dimethyl-1-cyclopenten-l-yl trifluoromethanesulfonate PdC1 2 (pPh 3 2 (460 mg, 0.65 mmol), triethylamine (9.0 mL, 65 mmol), Gui (0.50 g, 2.6 mmol), Ph 3 P (0.46 g, 0.65 mmol), and 2ethynylpyridine (1.3 g, 13 mmol) were added and the reaction was heated to 50*G. After stirring for 16 h at 50*C, TLC showed no remaining (3R,4S)-3,4-dimethyl-1-cyclopenten-1-yl trifluoromethanesulfonate. The mixture was diluted with ethyl acetate (200 mL), and filtered through CeliteTM. The filter pad was then thoroughily washed with ethyl acetate and the combined filtrates were washed with H 2 0 (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, then 98:2, then 96:4 hexanc:ethyl acetate to afford 2- ([(3S,4R)-3,4-dimethyl- I-cyclopenten-1I-y1lethynyl }pyridine g, 64%) as a yellow oil. 'H NN{R (GDGI 3 300 MHz) 5 8.57 (in, li1i), 7.66-7.60 (in, I1H) 7.41 J--9 Hz, 1K), 7.21-7.18 (in, 1H), 6.09 J=3 Hz, 1K), 2.74-1.85 (in, 4K), 1.09-1.04 (in, 6K). MS (ESI) 198.1 Example 160 Synthesis of Racemic 2- ii(trans)-3.4-dimethvl-l-cyclonten-l-vllethvnvllpvridine A cis-trans mixture of 2-[(3,4.-dimethyl-I-cyclopenten-1-yl)ethynyllpyridine was separated by preparative reverse phase HPLG (Zorbax. SB-G18 15cm x 21 mm x 5gm dp; Mobile phase A: 100:0.1
H
2 0:TFA, Mobile phase B: 100:0.1 acetonitrile:TFA; 30% B at 20 mL/min.) The resulting trifluoroacetic acid salt of the trans isomer was suspended in aqueous K 2 C0 3 and the resulting aqueous suspension was extracted with ethyl acetate. The aqueous layer was then saturated with solid NaCI and extracted with ethyl acetate (5 x 100 mL). The combined extracts were dried over Na 2
SO
4 filtered, and concentrated in vacuo to afford a brown oil. The dark product was further purified by column chromatography on silica gel eluting with 6:1 then 4:1 hexane:ethyl acetate to afford racemic 2- {[(trans)-3,4-dimethyl-l-cyclopenten-1-yl]ethynylpyridine (155.3 mg) as a pale yellow oil. 'H NMR WO 01/16121 PCT/USOO/23923 96
(CDCI
3 300 MI-z) 8 8.58-8.56 (in, 114), 7.66-7.59 (mn, 7.43-7.40 (mn, 11-1), 7.22-7.16 (in, 11-1), 6.12-6.10(in, 2.79-2.71 (mn, IH), 2.42-2.35(in, IH), 2.25-2.16(in, IH), 1.91-1.85 (in, 1.07 (t, Hz, MIS 198.1 Example 161 Synthesis of 2-methyl-4-(1.3-thiazol-2-vlethynyl)-l.3-thiazole GuI (50 mg, 0.26 minol), PdCl 2 (PPh 3 2 (93 mg, 0.13 mmol), PPb 3 (70 mng, 0.26 inmol), and tetrabutylaxnmonium iodide (377 mng, 1.02 mmol) were combined in dry DMF (20 mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Tricthylaminc (0.8 mL, 5 minol), was added, and the reaction mixture was warmed to 40*C, then a solution of 2-bromo-l1,3-thiazole (218 mg, 1.32 mmol) and 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 (200 mg, 1.02 inmol) in DMF (10 mi.) added. The reaction was warmed to 70"C and tetrabutylammonium fluoride (1.3 ml, of a I OM solution in THF, 1.3 inmol) was added by syringe pump over I h. After stirring for an additional 2 h, GUIMS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (30 mL), filtered through CeliteTM, and the filter pad was washed thoroughly with ethyl acetate. The combined filtrates were washed with H 2 0 (2 x. 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 9:1, 4:1, then 3:1 hexane:ethyl acetate to afford 2-methyl-4-(l,3-thiazol- 2-ylethynyl)-l,3-thiazole (165 mg, 78%) as a brown solid. 57-58*C. NMR (CDC1 3 .300 MHz) 8 7.88 J=3.0 Hz, IH), 7.55 7.41 J=3.0 H-l, 111), 2.76 3H). MIS (ESI) 207.0 Example 162 Synthesis of 4-(3-pyridinylethynyl)-1,3-thiazol-2-amine GuI (51 mg, 0.26 minol), PdC1 2 (PPh 3 2 (93 mng, 0.13 mmol), PPh 3 (70 mg, 0.26 minol), tetrabutylammonium iodide (377 mg, 1.02 inmol) were combined in dry DMF (20 mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylaniine (0.8 m.L, 5 mmcl), was added, the reaction mixture was warmed to 40*C, then a solution of 3-bromopyridine (210 mg, 1.32 inmol) and 4-[(trimethylsilyl)ethynyl]-1,3-thiazol-2-ylamine from Example 78 (200 mg, 1.02 inmol) in DMF (10 mL) added. The reaction was warmed to 70'C and tetrabutylammoniumn fluoride (1.3 ml, of a I OM solution in TI-F, 1.3 inmol) was added by syringe pump over I h. After stirring for an additional 2 h, GU/MS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (30 mL), filtered through CeliteTm, and the filter pad washed thoroughly with ethyl acetate.
The combined filtrates were washed with H 2 0 (2 x 200 niL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 4:1, 1:1, then 1:4 hexane:ethyl acetate. The product was further purified by recrystallization from hot CHC1 3 to afford 4-(3-pyridinylethynyl)-l,3-thiazol-2-amine (90 mg, 43%) as WO 01/16121 PCT/USOO/23923 97 a brown solid. 165-170 0 C. 'H NMR (CDCI 3 .300 MI-z) 8 8.65 I14), 8.50 J=6.0 Hz, IH), 7.95-7.92, (in, I 7.47-7.44 (in, I 6.93 IT-H). MS (El ionization) 20 1.0 (NI 4 Example 163 Synthesis of 4-l(2-methyl-1,3-thiazol-4-YI)ethynyllisothiazole GuI (60.0 mg, 0.31 mmol), PdCI 2 (PPh 3 2 (109 Mng, 0. 15 mmol), PPh 3 (82 mg, 0.31 minol), and tetrabutylanimoniumn iodide (440 mg, 1.2 mmol) were combined in dry DMF (20 miL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylaxnine (0.8 m.L, 5 mmol), was added, and the reaction mixture was warmed to 40'C, then a solution of 4-bromoisothiazole (200 mng, 1.2 inmol) and 2-methyl-4-[(trimethylsilyl)ethynylj-l,3-thiazoc from Example 77 (360 mg, 1.8 mmol) in DMF (10 mL) was added. The reaction was warmed to 70"C and tetrabutylammoniumn fluoride (1.6 mL of a L.0M solution in THF, 1.6 inmol) was added by syringe pump over I h. After stirring for an additional 2 h, GUIMS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (30 mL) and filtered through CeliteTM. The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 m.L), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 9:1, then 4:1 hexane:ethyl acetate to afford mcthyl-1,3.thiazol-4-yl)ethynyl]isothiazole (140 mng, 37%) as a yellow solid. 113.1 14*C. 'H NMR (CDCI 3 300 MIHz) 6 8.79 1H), 8.61 114, 7.41 IH), 2.75 3H). MS (ESI) 207.0 Example 164 Synthesis of 2-methvl-4-(1.3-thiazol-4-vlethvnvfl -13-thiazole GuI (60 mg, 0.31 inmol), PdCI 2 (PPh 3 2 (110 Mng, 0. 15 minol), PPh 3 (82 mng, 0.31 inmol), and tetrabutylainmonium iodide (440 mg, 1.2 mmol) were combined in dry DMF (20 mL), and argon gas was bubbled through the suspension for several min to deoxygenate it. Triethylamine (0.8 m.L, minol), was added, the reaction mixture was warmed to 40'C, then a solution of 4-bromo-1,3-thiazole (200 mng, 1.2 mmol) and 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 (360 mng, 1.8 inmol) in DMF (10 mL) was added. The rraction was warmed to 70*C and tetrabutylanimonium fluoride (1.6 m.L of a L.OM solution in THIF, 1.6 mmoi) was added via syringe pump over 1 h. After stirring for an additional 2 h, GUIMS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (30 mL) and filtered through CeliteTM, the filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 9:1, 4:1, then 3:1 hexane:ethyl acetate to afford 2methyl-4-(1,3-thiazol-4-ylethynyl)-1,3-thiazole (112 mng, 30%) as a yellow solid. 102-1031C. 'H WO 01/16121 PCT/USOO/23923 98 NMR (CDC1 3 3 00 MHz) 6 8.82 J=2 Hz, 1ffi), 7.64 J=2 Hz, I 7.46 I 2.75 3 MS (ESI) 207.0 (NC+H).
Example 165 Synthesis of 5-4(2-methvl-1 3 -thiazol-4.yl)eth yllisothistiole CuI (60 mg, 0.31 mmol), PdCI 2 (PPh 3 2 (110 Mg, 0. 15 mmol), PPh 3 (82 mg, 0.31 mmol), and tetrabutylammoniumn iodide (440 mg, 1.2 mmol) were combined in dry DMF (20 mL) and argon gas was bubbled through the mixturc for several min to deoxygenate it. Triethylamine (0.8 m.L, 5 mmol), was added, and the reaction mixture was warmed to 40*C, then a solution of S-bromoisothjazole (200 mg, 1.2 mmol) and 2 -methyl-4-[(trimethylsilyl)edhynyll13-thiazole from Example 77 (360 mg, 1.8 mmol) in DMF (10 mL) was added. The reaction was warmed to 70'C and tetrabutylammonium fluoride (1.6 mL of a I OM solution in THF, 1.6 mmol) was added by syringe pump over I h. After stirring for an additional 2 h, GU/MS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (30 mL) and filtered through Celite"~'. The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 9:1, then 4:1 hexane:ethyl acetate to afford mehl13tizl4y~tyyjstizl (160 mog, 42%) as a yellow solid. 79-80'C. 1H NNM
(CDCI
3 300 M Hz) 5 8.46 J=2 Hz, I 7.48 I 7.39, J=2 Hz, I 2.75 3H). MS (ESI) 207.0 Example 166 Synthesis of 2-(Fl1.1I '-iihenyll-4-vethynyl)pyrnmidin- Gui (84 mg, 0.44 mmol), PdCI 2 (PPh 3 2 (155 mng, 0.22 mmol), PPh 3 (110 Mg, 0.44 ramol), and tetrabutylammonium iodide (630 mg, 1.7 mmol) were combined in dry DMF (25 m.L) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylamine (I mL, 8 mmol), was added, and the reaction mixture was warmed to 40'C, then a solution of 4-bromobiphenyl (515 mg, 2.2 mmol) and 2 -[(trimethylsilyl)ethynyljpyrimidine from Example 81 (300 mg, 1.7 mmol) in DMF mL) was added. The reaction was warmed to 70 0 C and tetrabutylammoniumn fluoride (2.2 mL, of a 1.OM solution in TI-F, 2.2 mmol) was added by syringe pump over 2 h. After stirring for an additional 2 h, GCIMS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate O0 mL) and filtered through Celite Tm The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 9:1, then 8.5:1.5 hexane:ethyl acetate to afford 2-[,'bpey]-yehnlprmdn (200 mg, 46%) as a yellow solid. The product was dissolved in diethyl ether (20 mL) and HCI in diethyl ether (12 nil of a L.OM solution, 12 mmol) was added to afford a light yellow precipitate. The WO 01/16121 PCT/US00/23923 99 precipitate was filtered, and dried in vacuo to afford 2-([l,1'-biphenyl]-4-ylethynyl)pyrimidine hydrochloride (170 mg, 34%) as a yellow solid. 135-137 0 C. 'H NMR (CDC3. 300 MHz) 5 9.01 J=6 Hz, 2H), 7.76-7.66 7H), 7.49-7.39, 3H). MS (ESI) 257.0 Example 167 Synthesis of N'-{3-[2-(2-pyridinyl)ethynyll-2-cyclohexen-l-ylidene}-2-furohydrazide A solution of 2-furohydrazide (61 mg, 0.48 mmol) in ethanol (1 mL) was treated with acetic acid (1 drop), and 3-(2-pyridinylethynyl)-2-cyclohexen-l-one from Example 104 (86 mg, 0.44 mmol) was added as a solution in ethanol (1 mL) followed by a rinse of the flask and syringe (0.5 mL). The resulting solution was then heated in a 90 0 C oil bath for 45 min. The reaction solution was cooled to ambient temperature and concentrated in vacuo to afford an orange oil. The crude oil was adsorbed onto silica gel and purified by column chromatography on silica gel eluting with 40:1 CHCI 3
:CH
3 OH to afford N'-{3-[2-(2-pyridinyl)ethynyl]-2-cyclohexen-1-ylidene}-2-furohydrazide (40.2 mg, 30%) as a yellow oil. The material appeared to be one component by LC/MS analysis, but showed some extra peaks in the NMR spectra possibly caused by a mixture of double bond isomers being present. 'H NMR
(CDCI
3 300 MHz) 8 9.56 (br s, 0.1 9.33 (br s, 0.9H), 8.60 1H, J=4.7 Hz), 7.68 1H, J=8.6 Hz), 7.55-7.43 2H), 7.35-7.20 2H), 6.96 (br s, 0.1H), 6.86 (br s, 0.9H), 6.56 (dd, J=1.6, 3.5 Hz, 1H), 2.67-2.62 0.1H), 2.56-2.40 3.8H), 2.26 (br s, 0.2H), 2.03-1.93 1.8H). MS (ESI) 306.1 Example 168 Synthesis of N-{4-[2-(l-cyclohexen-l-yl)ethvnyll-13-thiazol-2-vl benzamide To a suspension of 4-(1-cyclohexen- -ylethynyl)-1,3-thiazol-2-ylamine tosylate from Example 41 (65 mg, 0.17 mmol) in THF (2.0 mL) was added triethylamine (0.10 mL, 0.72 mmol) to afford a yellow solution. Benzoyl chloride (40 uiL, 0.34 mmol) was then added to afford a suspension. After stirring for 16 h at ambient temperature, TLC and LC/MS analysis showed starting amine present. The reaction mixture was then heated in a 50 0 C oil bath. After 3 h at 50 0 C, TLC and LC/MS analysis showed little progress in the reaction. Additional benzoyl chloride (50 pL, 0.43 mmol) was added to the reaction. After stirring for 26 h after the addition of the second portion of benzoyl chloride the reaction was cooled to ambient temperature, quenched by the addition of saturated aqueous NaHC0 3 (10 mL), and diluted with ethyl acetate (30 mL). The aqueous layer was extracted with ethyl acetate (2 x 10 mL), the combined organics were washed with brine, dried over Na 2
SO
4 filtered, and concentrated in vacuo.
The crude product was purified by column chromatography on silica gel eluting with 12:1 then 10:1 hexane:ethyl acetate to afford N-{4-[2-(1-cyclohexen- -yl)ethynyl]-1,3-thiazol-2-yl}benzamide (37 mg, as an oil that partially solidified under high vacuum. The material was not completely pure by NMR analysis, and was then carefully purified by column chromatography on silica gel eluting with 15:1 then 12:1 hexane:ethyl acetate to afford N-(4-[2-(l-cyclohexen-l-yl)cthynyl]-l,3-thiazol-2- WO 01/16121 PCT/US00/23923 100 yl}benzamide (30 mg, 57%) as a white foam. 'H NMR (CDCI 3 300 MHz) 8 7.98 J=7.3 Hz, 2H), 7.58 J=7.3 Hz, 1H), 7.46 J=7.8 Hz, 2H), 7.09 1H), 5.92-5.80 1H), 2.05-2.00 2H), 1.95- 1.87 2H), 1.59-1.49 2H). MS (ESI) 309.1 331.0 Example 169 Synthesis of 3-|(2-methyl-1,3-thiazol-4-vl)ethynyllpyridine PdCI 2 (15 mg, 85 pmol), and Cul (23 mg, 120 pmol) were combined in DME (3 mL) under argon. H 2 0 (1 mL) was added and argon was bubbled through the resulting dark suspension while it was warmed to 40 0 C in an oil bath. Triphenylphosphine (84 mg, 320 pmol) was added and the argon flow was continued for 10 min. The argon flow was discontinued, solid potassium carbonate (277 mg, 2.0 mmol) was added, followed by 2-methyl-4-[(trimethylsilyl)ethynyl]-1,3-thiazole from Example 77 mg, 0.46 mmol) and 3-bromopyridine (56 gL, 0.58 mmol) as a solution in DME (2 mL) followed by a rinse of the flask and syringe with DME (1 mL). The temperature was raised to 75 0 C. After 16 h there was still 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole visible by GC/MS analysis. At this time Bu 4 NF (1.0 mL ofa 1.OM solution in THF, 1.0 mmol) was added to the reaction mixture. After min GC/MS analysis showed the reaction was complete. The reaction mixture was cooled to ambient temperature, diluted with ethyl acetate (50 mL), and H 2 0 (10 mL). The aqueous layer was extracted with ethyl acetate (2 x 10 mL), the combined organics were washed with brine (20 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with 2:1, 1:1, then 2:3 hexane:ethyl acetate to afford product contaminated with some close-running impurities. The material was then carefully purified by column chromatography on silica gel eluting with 2:1, then 1:1 hexane:ethyl acetate to afford 3-[(2-methyl-1,3thiazol-4-yl)ethynyl]pyridine (48.6 mg, 52 as a pale brown oil that partially solidified after pumping down under high vacuum. 75-76 0 C. 'H NMR (CDCI 3 300 MHz) 8 9.0-8.7 (br s, 1H), 8.7-8.45 (br s, 1H), 7.84 J=7.9 Hz, IH), 7.45 IH), 7.35-7.22 IH), 2.75 1H). MS (El ionization) 200 Example 170 Synthesis of 2.4-dimethyl-6-l(trimethylsilyl)ethynyllpyrimidine A solution of trifluoromethanesulfonic anhydride (6.5 mL, 39 mmol) was added slowly to a stirred solution of 2,4-dimethyl-6-hydroxypyrimidine (4.0 g, 32 mmol) and triethylamine (7.5 mL, 53 mmol) in dry CHzC2 (125 mL) at 0°C under argon. The mixture was stirred at ambient temperature for 16 h at which time TLC analysis showed the reaction to be complete. The mixture was diluted with
CH
2
C
2 (200 mL), washed with saturated aqueous NaHCO 3 (200 mL), brine (200 mL), dried over Na 2
SO
4 filtered and concentrated in vacuo. Half of the residue (4 g, 16 mmol) was dissolved in DME mL), and argon was bubbled through the solution for several min to deoxygenate it.
Trimethylsilylacetylene (2.9 g, 30 mmol) was added. The resulting solution was added to a WO 01/16121 PCT/US00/23923 101 deoxygenated mixture of PdCI 2 (83 mg, 0.47 mmol), triethylaniine (8.7 mL, 1.8 mmol), Cul (259 mg, 1.35 mmol), and triphenylphosphine (483 mg, 1.84 mmcl), in DME (20 mL) at 45*C. After stirring for 16 h at 451C, TLC showed no remaining 2,6-d imethyl-4-pyri mid inyl trifluoromethanesulfonate. The mixture was diluted with ethyl acetate (200 mL), and filtered through Celitem. The filter pad was then thoroughly washed with ethyl acetate and the combined filtrates were washed with H 2 0 (200 mL), brine (200 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacua, and the residue was purified by column chromatography on silica gel eluting with hexane, 20:1, then 9:1 hexane:ethyl acetate to afford 2,4-dimethyl-6-[(trimethylsilyl)ethynyl]pyrimidine (600 mg, 18%) as a yellow oil. 1'H NMvR (CDC1 3 300 MHz) 5 7. 10 I 2.69 I 2.48 I1H), 0.26 914).
Example 171 Synthesis of 4-411.1 '-biphenyll-4-ylethynyl)-2.6-dimethvll~vrimidine Cul (73 mg, 0.38 mmcl), PdC1 2 (PPh 3 2 (130 mng, 0. 19 mmcl), triphenylphosphine (100 mg, 0.3 8 mmcl), and tetr-abutylammonium iodide (543 mg, 1.47 mmol) were combined in dry DMF (25 mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylamine (1.0 mL, 7.4 mmol), was added, the reaction mixture was warmed to 40'C, and a solution of 4-bromo-1,1'biphenyl (446 mg, 1.9 mmcl) and 2,4-dimethyl-6-[(trimcthylsilyl)ethynyl]pyrimidine from Example 170 (300 mg, 1.47 mmcl) in DMF (10 mL) was added. The reaction mixture was warmed to 70'C and tetrabutylammonium fluoride (1.9 mL of a 1.0 M solution in THE, 1.3 mmol) was added by syringe pump over 2 h. After stirring for an additional 2 h, TLC and GC/MS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (50 mL) and filtered through Celite The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacua. The residue was purified by column chromatography on silica gel eluting with hexane, 18:1, then 9:1 hexane:ethyl acetate to afford 4-([l,1'-biphenyl]-4-yethynyl)-2,6-dimethylpyrimidine (12 ing, as a yellow solid.
145-148'C. 'H NMR (CDCI 3 300 M~lz) 8 7.70-7.60 (in, 5H), 7.49-7.36 (in, 4H), 7.19 I H), 2.73 3H), 2.52 3H). MS (ESI) 285.0 Example 172 Synthesis of 2-1(trimethylsilvl)ethynyll Dyrazine PdCI 2 (60 mng, 0.34 mmol), Cul (151 mng, 0.79 mmcl), and triethylainine (3.8 mL, 27 inmol) were combined in DME (10 mL) under argon. Argon was bubbled through the suspension, and after min triphenylphosphine (349 mng, 1.33 mmcl) was addcd and the reaction flask was immersed in a oil bath. The argon flow was discontinued after an additional 5 min, then trimethylsilylacetylene (2.4 mL, 17 mmcl), and 2-iodopyrazine (1.66 g, 8.06 mmcol) were added as a solution in DME (5 mL), followed by a rinse of the flask and syringe with DME (3 mL). After stirring for 16 h at 50'C, GC/MS analysis of the dark brown suspension showed no 2-iodopyrazine remaining. The reaction mixture was WO 01/16121 PCT/US00/23923 102 cooled to ambient temperature, filtered, and the solids were washed thoroughly with ethyl acetate. The combined filtrates were concentrated in vacuo to afford a dark oil, which was diluted with diethylether (100 mL), and washed with saturated aqueous NaHC0 3 (25 mL). The basic organic layer was extracted with diethylether (50 mL), and the combined organics were washed with brine (25 mL), dried over Na 2
SO
4 filtered, concentrated in vacuo, then diluted with benzene and concentrated in vacuo again to remove any remaining H 2 0 as its azeotrope. The crude material was purified by column chromatography on silica gel eluting with hexane, 30:1, then 20:1 hexane:ethyl acetate to afford 2- [(trimethylsilyl)ethynyl]pyrazine (1.2 g, 88%) as a yellow oil. H NMR (CDC1 3 300 MHz) 8 8.89 (s, 1H), 8.55 J=3 Hz, 1H), 8.49 J=3 Hz, 1H), 0.29 9H). MS (El ionization) 176 Example 173 Synthesis of 2-(I11 '-biphenyll-4-vlethvnvl)vrazine Cul (84 mg, 0.44 mmol), PdCI 2 (PPh 3 2 (155 mg, 0.22 mmol), triphenylphosphine (116 mg, 0.442 mmol), and tetrabutylammonium iodide (630 mg, 1.7 mmol) were combined in dry DMF mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylamine (0.8 mL, 8 mmol), was added, the reaction mixture was warmed to 40 0 C, and a solution of 4-bromo- 1,1'-biphenyl (515 mg, 2.2 mmol) and 2-[(trimethylsilyl)ethynyl]pyrazine from Example 172 (300 mg, 1.7 mmol) in DMF (10 mL) was added. The reaction was warmed to 70 0 C and tetrabutylammonium fluoride (2.2 mL of a 1.0 M solution in THF, 2.2 mmol) was added by syringe pump over 2 h. After stirring for an additional 2 h, TLC and GC/MS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (50 mL) and filtered through Celite T M The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 18:1, 9:1, then 8.5:1.5 hexane:ethyl acetate to afford 2-([l,l'-biphenyl].4-ylethynyl)pyrazine (100 mg, 23%) as a yellow solid. 112-113C.
'H NMR (CDC 3 300 MHz) 8 8.88 1H), 8.78 IH), 8.64 1H), 7.62-7.61 6H), 7.50-7.38 (m, 3H). MS (ESI) 257.0 Example 174 Synthesis of 46-dimethyl-2-i(trimethylsilyl)ethnyl Dyrimidine PdCI 2 (234 mg, 1.32 mmol), CuI (737 mg, 3.87 mmol), and triethylamine (23 mL, 165 mmol) were combined in DME (50 mL) under argon. Argon was bubbled through the suspension, and after min triphenylphosphine (1.37 g, 5.22 mmol) was added and the reaction flask was immersed in a oil bath. The argon flow was discontinued after an additional 5 min, then trimethylsilylacetylene (12 mL, 85 mmol), and 4,6-dimethyl-2-pyrimidinyl trifluoromethanesulfonate (11.3 g, 44.1 mmol), were added as a solution in DME (10 mL) followed by a rinse of the flask and syringe with DME (10 mL).
The reaction mixture rapidly turned from a dark orange color to black as the 4,6-dimethyl-2- WO 01/16121 PCT/US00/23923 103 pyrimidinyl trifluoromethanesulfonate solution was added. After stirring for 30 min at 50 0 C, GC/MS analysis of the black suspension showed no 4,6-dimethyl-2-pyrimidinyl trifluoromethanesulfonate remaining. The reaction mixture was allowed to cool to ambient temperature. The reaction mixture was concentrated in vacuo, diluted with diethylether (200 mL), filtered, and the solids were washed thoroughly with diethylether. The combined filtrates were washed with saturated aqueous NaHCO 3 mL), at which time a large quantity of solids appeared. The suspension was again filtered. The two layers of the filtrate were separated, and the aqueous layer was extracted with diethylether (2 x 50 mL).
The combined organics were washed with brine, dried over Na 2
SO
4 filtered, and concentrated in vacuo.
The crude material was purified by column chromatography on silica gel eluting with hexane, 20:1, 10:1, then 8:1 hexane:ethyl acetate to afford an orange oil with some white fluffy solids present. This was diluted with hexane and filtered through a plug of Celite M to remove the white solids. The filtrate was concentrated in vacuo to afford 4,6-dimethyl-2-[(trimethylsilyl)ethynyl]pyrimidine (4.63 g, 51%) as an orange oil. During the elution of the column, while the desired compound was eluting, the flow rate dropped and the column plugged so that elution was no longer possible. The silica gel from the column was slurried with ethyl acetate and CH2Cl, and filtered. The silica gel was then washed thoroughly with ethyl acetate. The combined filtrates were combined and concentrated in vacuo to afford a dark oil, which was quite clean by NMR analysis showing desired product contaminated with some material exhibiting extra phenyl signals (possibly triphenylphosphine). 'H NMR (CDCl 3 300 MHz) 8 6.98 1H), 2.49 3H), 0.28 9H). MS (El ionization) 204 Example 175 Synthesis of 2-(I1.'-biphenyll-4-ylethynyl)-4.6-dimethylpyrimidine Cul (84.0 mg, 0.442 mmol), PdCI 2 (PPh 3 2 (155 mg, 0.22 mmol), triphenylphosphine (116 mg, 0.44 mmol), and tetrabutylammonium iodide (630 mg, 1.7 mmol) were combined in dry DMF (25 mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylamine (0.8 mL, 8 mmol), was added, the reaction mixture was warmed to 40 0 C, and a solution of 4-bromo-l,l'biphenyl (515 mg, 2.2 mmol) and 4,6-dimethyl-2-[(trimethylsilyl)ethynyl]pyrimidine from Example 174 (340 mg, 1.7 mmol) in DMF (10 mL) was added. The reaction was warmed to 70 0 C and tetrabutylammonium fluoride (2.2 mL of a 1.0 M solution in THF, 2.2 mmol) was added by syringe pump over 2 h. After stirring for an additional 2 h, TLC and GC/MS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (50 mL) and filtered through Celite T m The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 18:1, 9:1, 8.5:1.5 hexane:ethyl acetate to afford 2-([l,l'-biphenyl]-4-ylethynyl)-4,6-dimethylpyrimidine (180 mg, 42%) as a yellow solid. The 2-([l,l'-biphenyl]-4-ylethynyl)-4,6-dimethylpyrimidine was dissolved in diethylether mL) and HCI (12 mL of a 1.0 M solution in diethylether, 12 mmol) was added. The resulting WO 01/16121 PCT/US00/23923 104 precipitate was filtered, and crystallized from boiling ethyl acetate to afford 2-([1,1'-biphenyl]-4ylethynyl)-4,6-dimethylpyrimidine hydrochloride (50 mg 11%) as yellow crystals. 163-165*C.
'H NMR 300 MHz) 8 7.82 J=3 Hz, 4H), 7.72-7.67 (in, 3H), 7.51-7.41 (in, 3H), 2.70(s, 6H).
MS (ESI) 285.1 Example 176 Synthesis of 1-chloro-64triniethylsilvl)-3.5-hexadivn-2-one Aluminum trichloride (6.85 g, 51.4 mmol) was suspended in CH 2
CI
2 (100 mL) and coaled in an ice bath. A solution of 4-bis(trimethylsilyl)1,3-butadiyne (10.0 g, 51.4 mmol) and chloroacetyl chloride (4.1 inL, 51 inmol) in CH 2
CI
2 (100 mL) was added to the AIC1 3 suspension dropwise from an addition funnel over I h. The dark brownish-red solution was stirred at O 0 C for I h, then the ice bath was removed. After I h at ambient temperature, the reaction was cooled to 0 0 C and quenched by slow addition of IM HCI (150 mL). The acidic aqueous solution was extracted with CH 2
CI
2 (2 x 300 mL), the combined organic layers were washed with H 2 0 (500 mL), saturated aqueous NaHCO 3 (500 mL), brine (500 mL) and dried over Na 2
SO
4 The organic layer was filtered to afford a clear solution which was concentrated in vacuo. The crude 1-chloro-6-(trimethylsilyl)-3,5-hexadiyn-2-one (10 g) was used in the next step without purification. MS (EI ionization) 183 Example 177 Synthesis of 2-methvl-4-[4-(trimethylsilvl)-1 .3-butadivnvll-1 .3-thiazole Crude l-chloro-6-(trimethylsilyl)-3,5-hexadiyn-2-one from Example 176 (5.0 g, 25 mmol) was dissolved in OMIF (70 mL), then thioacetamide (2.4 g, 33 mmol) was added in one portion. The mixture was allowed to stir for 16 h at ambient temperature, at which time TLC showed no remaining l-chlomo-6-(triinethylsily)-3,5-hexadiyn-2-one. The mixture was diluted with ethyl acetate (200 mL), washed with H 2 0 (3 x 300 mL), brine (300 mL), dried over Na 2
SO
4 and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography on silica gel eluting with hexane, then 20:1 hexane:ethyl acetate to afford 2-methyl-4-[4-(trimethylsilyl)- I,3-butadiynyl]- 1,3-thiazole (1.2 g, 24%) as reddish-brown oil. 1'H NMR (CDCI 3 300 MHz) 6 7.39 111), 2.68 (s, 3H), 0.26 9H). MS (El ionization)21.9 (M 4 Example 178 Synthesis of N-hydroxvbenzenecarboximidovl chloride To a solution of syn-benzaldehyde oxime (12 g, 99 mmol) in DMIF (70 mL) was added Nchlorosuccinimide (15.8 g, I118 mmol). After stirring at ambient temperature for 64 h, TLC analysis showed no starting oxime present. The reaction mixture was diluted with diethylether (600 m.L), washed with a 1:1 mixture of H2O:saturated aqueous NaCI (2 x 200 mL), brine (100 mL), dried over Na 2
SO
4 filtered and concentrated in vacua to afford an oil that was diluted with toluene and concentrated in vacuo to remove any remaining H 2 0. The resulting crude product was purified by WO 01/16121 PCT/USOO/23923 105 column chromatography on silica gel eluting with hexane, 30: 1, then 20:1 hexane:ethyl acetate to afford N-hydroxybenzenecarboximidoyl chloride (5.77 g, 37%) as white crystals. 52-53 0 C. 1 H NMIR
(CDCI
3 300 MHz) 8 9.00 IH), 7.84 J=6 Hz, 2H1), 7.49-7.3 7 (in, 314).
Example 179 Synthesis of 5-1(2-methyl-I .3-thiazol-4-Yl)ethynyl 1-3-nhenylisoxazole To a solution of 2.methyl-4-[4-(trimethylsilyl)- 1,3-butadiynyl]-1I,3-thiazole from Example 177 (220 mg, 1.0 mmol) in dry diethylether (20 mL) under argon, tetrabutylammonium fluoride (1.1I mL of a 1.0 M solution in THF, 1. 1 mmol) was added by syringe during 15 min. After stirring for 0.5 h, TLC and GC/MS analysis showed the reaction to be complete. Then N-hydroxybenzenecarboximidoyl chloride from Example 178 (233 mng, 1.5 mmol) was added in one portion under argon. Triethylamnine (0.20 mL in 20 mL dry diethylether, 1.5 mmol) was added by addition funnel during 2 h. After stirring for an additional 2 h, TLC and GC/MS analysis showed the reaction was not complete, another inmol of N-hydroxybenzenecarboximidoyl chloride and triethylamine was added as described above.
After stirring for an additional 2h, the mixture was diluted with diethylether (50 mL) and washed with
H
2 0 (100 mL), brine (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, I8: 1, then 9:1 hexane:ethyl acetate to afford 5-[(2-methyl-1,3-thiazol-4-yl)ethynylj-3-phenylisoxazole (195 mg, 73%) as a yellow solid. The product was dissolved in diethylether (20 mL) and HCI (15 mL of a 1.0 M solution in diethylether, 15 mmol) was added, the resulting precipitate was filtered and washed with diethylether to givc 5-[(2-methyl-1,3-thiazol-4-yl)ethynyl]-3-phcnylisoxazole (90 mg, 34%) as yellow crystals. M.p.= I I 8-120"C. Elemental analysis of the material for C, H, N and CI showed the material to be the free base, not the hydrochloride salt. 'H NMR (CDC1 3 300 M]Hz) 8 7.94 1IH), 7.88-7.84 (mn, 2H), 7.50- 7.47 (in, 3 7.21 I1H), 2.73 3 MIS (ESI) 267.0 Example 180 Synthesis of Ethyl 5-[(2-methyl-1,3-thiazol-4-yI)ethynyll-3-isoxazolecarboxvlate To a solution of 2-methyl-4-[4-(triniethylsilyl)-1,3-butadiynyl]-1,3-thiazole from Example 177 (150 mg, 0.68 inmol) in dry diethylether (20 inL) under argon, tetrabutylamnmonium fluoride (0.75 mL of a 1.0 M solution in THF, 0.75 mmol) was added by syringe during 15 min. After stirring for 0.5 h, TLC and GCIMS analysis showed the reaction to be complete. Then commercially available ethyl (2- EZ)-chloro(hydroxyimino)ethanoate (155 mng, 1.0 inmol) was then added in one portion under argon.
Triethylamine (0.14 mL, 1.0 inmol) was added as a solution in dry diethylether (20 m.L) by addition funnel during 2 h. After stirring for an additional 2 h, TLC and GUIMS analysis showed the reaction was not complete. Another portion of ethyl (2-E.Z)-chloro(hydroxyimino)ethanoate (155 mg, inmol) and triethylamine (0.14 mL, 1.0 mmol) was added as described above. After stirring for an additional 2 h, the mixture was diluted with diethylether (50 mL) and washed with H 2 0 (100 mL), brine WO 01/16121 PCT/US00/23923 106 (100 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 18:1, then 9:1 hexane:ethyl acetate to afford ethyl [(2-methyl-1,3-thiazol-4-yl)ethynyl]-3-isoxazolecarboxylate (120 mg, 72%) as a yellow solid. The ethyl 5-[(2-methyl-1,3-thiazol-4-yl)ethynyl]-3-isoxazolecarboxylate was dissolved in diethylether mL) and HCI (16 mL of a 1.0 M solution in diethylether, 16 mmol) was added. The resulting precipitate was filtered and washed with diethylether to give ethyl 5-[(2-methyl-l,3-thiazol-4-yl)ethynyl]-3-isoxazolecarboxylate (75 mg, 42%) as yellow crystals. 125-127'C. Elemental analysis for C, H, N, and Cl showed the compound to be the free base, not the hydrochloride salt. 'H NMR (CDCI3, 300 MHz) 8 8.22 IN), 7.21 111), 4.43 J=9 Hz, 2H), 2.87 311), 1.40(t, J=6 Hz, 3H). NIS (ESI) 263.0 Eamnpl 81 Synthesis of 2-methoxy-5- [(2-methyl-i .3-thlazol-4-vhethynyll pyridine GuI (40 mg, 0.2 mmol), PdCI 2 (PPh 3 2 (72 mg, 0.10 mmol), triphenylphosphine (54 mg, 0.2 mmol), and tetrabutylammnonium iodide (376 mg, 1.0 mmol) were combined in dry DMF (20 mL) and argon gas was bubbled through the mixture for several min to deoxygenate it. Triethylamine (0.71 mL, 5.1 mmol) was added, the reaction mixture was warmed to 40'C, and a solution of 5-bromo-2methoxypyridine (231 mg, 1.23 mrnol) and 2-methyl-44[(trimethylsilyl)ethynyl]-1,3-thiazole from Example 77 (200 mg, 1.0 mmol) in DMF (10 mL) was added. The reaction was warmed to 70*C and tetrabutylammoniuni fluoride (1.3 mL of a 1.0 M solution in THE, 1.3 mmol) was added by syringe pump over 2 h. After stirring for an additional 2 h, TLC and GCIMS analysis showed the reaction to be complete. The mixture was diluted with ethyl acetate (100 mL) and filtered through Celite TM The filter pad was washed thoroughly with ethyl acetate, and the combined filtrates were washed with H 2 0 (2 x 200 mL), brine (200 mL), dried over Na 2
SO
4 filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with hexane, 18:1, then 9:1 hexane:ethyl acetate to afford 2-methoxy-5-[(2-methyl-1,3.thiazol-4-yl)ethynyllpyridine (150 mg, 20%) as a yellow solid. The product was further purified by crystallization from boiling hexane to give 2-methoxy-S-[(2methyl-1,3-thiazol-4-yl)ethynyljpyridine (120 mg, 5 as yellow crystals. 110-11 t 0 C. 'H NMR
(CDCI
3 300 MHz) 8 8.38 J=2 Hz, lEO), 7.71 (dd, J=2, 9 Hz, 11H), 7.37 111), 6.73 J=9 Hz, IHM.
MIS (ESI) 230.9 Example 182 Synthesis of 5-1(2-methyl-I .3-thlazol-4vI)ethy Lryll pvrlmidine PdCl 2 (27.8 mg, 160 A~mol), Cul (77 mg, 400 jimol), and triethylamine (3.0 mL, 22 mmol) were combined in DME (10 mL) under argon. Argon was bubbled through the suspension, and after 10 min triphenylphosphine (166 mg, 630 pmol) was added and the reaction flask was heated on a heating mantle. The argon flow was discontinued after an additional 5 min, then 5-bromopyrimidine (830 mg, WO 01/16121 PCT/US00/23923 107 5.2 mmol), and 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 (826 mg, 4.2 mmol), were added as a solution in DME (5 mL) followed by a rinse of the flask and syringe with DME (2 mL). Tetrabutylammonium fluoride (5.0 mL of a 1.0 M solution in THF, 5.0 mmol) was then added to the reaction mixture dropwise over 10 min. After stirring for an additional 2.5 h at 65 0 C, GC/MS analysis showed no remaining 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole, nor desilylated alkyne present. The reaction mixture was allowed to slowly cool to ambient temperature over 16 h, concentrated in vacuo, diluted with ethyl acetate (150 mL), H 2 0 (50 mL) and saturated aqueous NaHCO3 (25 mL). The basic aqueous layer was extracted with ethyl acetate (2 x 50 mL). The combined organics were washed with brine (50 mL), dried over Na 2
SO
4 filtered, and concentrated to afford a brown solid. The crude product was purified by column chromatography on silica gel eluting with hexane 3:1, 2:1, then 3:2 hexane:ethyl acetate to afford 5-[(2-methyl-l,3-thiazol-4yl)ethynyl]pyrimidine (687 mg, 81 as a brownish-yellow solid. M.p. 118.5-119.5°C. 'H NMR
(CDCI
3 300 MHz) 89.17 1H), 8.89 2H), 7.51 1H), 2.76 3H). MS (El ionization) 201 Example 183 Synthesis of 5-(2-methyl-1,3-thiazol-4-yl)ethynvllnlcotinonitrile PdCI 2 (25 mg, 140 mol), Cul (78 mg, 410 mol), and triethylamine (3.0 mL, 22 mmol) were combined in DME (10 mL) under argon. Argon was bubbled through the suspension, and after 10 min triphenylphosphine (150 mg, 570 mol) was added and the reaction flask was heated on a heating mantle. The argon flow was discontinued after an additional 5 min, then 5-bromonicotinonitrile (980 mg, 5.3 mmol), and 2-methyl-4-[(trimethylsilyl)ethynyl]-l,3-thiazole from Example 77 (867 mg, 4.4 mmol), were added as a solution in DME (5 mL) followed by a rinse of the flask and syringe with DME (2 mL). Tetrabutylammonium fluoride (5.0 mL of a 1.0 M solution in THF, 5.0 mmol) was then added to the reaction mixture dropwise over 10 min. After stirring for 16 h at 55 0 C, GC/MS analysis showed no remaining 2-methyl-4-[(trimethylsilyl)ethynyl]-1,3-thiazole, nor desilylated alkyne present. The reaction mixture was allowed to cool to ambient temperature overnight, concentrated in vacuo, diluted with ethyl acetate (150 mL), H 2 0 (50 mL) and saturated aqueous NaHCO 3 (25 mL). The basic aqueous layer was extracted with ethyl acetate (2 x 50 mL). The combined organics were washed with brine mL), dried over Na 2
SO
4 filtered, and concentrated to afford a brown solid. The crude product was purified on a Biotage Flash system eluting with 100:1 CHCI 3 :CHQH to afford 5-[(2-methyl-1,3thiazol-4-yl)ethynyl]nicotinonitrile (201 mg, 20 as a yellow solid. (there were a large number of impure fractions from the column) M.p. 173-175 0 C. 'H NMR (CDCI 3 300 MHz) 8 8.94 J=2.0 Hz, 1H), 8.82 J=2.0 Hz, 1H), 8.08 J=2.0 Hz, 1H), 7.52 1H), 2.77 3H). MS (El ionization) 225 WO 01/16121 PCT/USO0/23923 108 Example 184 Calcium Flux Assay The activity of compounds was examined against the hmGluR5a receptor stably expressed in mouse fibroblast Ltk" cells (the hmGluR5a/L38-20 cell line). See generally Daggett et al., Neuropharmacology 34:871-886 (1995). Receptor activity was detected by changes in intracellular calcium ([Ca 1 measured using the fluorescent calcium-sensitive dye, fura-2. hmGluR5a/L38-20 cells were plated onto 96-well plates, and loaded with 3 pM fura-2 for 1 h. Unincorporated dye was washed from the cells, and the cell plate was transferred to a custom-built 96-channel fluorimeter (SIBIA-SAIC, La Jolla, CA) which is integrated into a fully automated plate handling and liquid delivery system. Cells were excited at 350 and 385 nm with a xenon source combined with optical filters. Emitted light was collected from the sample through a dichroic mirror and a 510-nm interference filter and directed into a cooled CCD camera (Princeton Instruments). Image pairs were captured approximately every 1 s, and ratio images were generated after background subtraction. After a basal reading of 20 s, an ECso concentration of glutamate (10 iM) was added to the well, and the response evaluated for another 60 s. The glutamate-evoked increase in [Ca"]i in the presence of the screening compound was compared to the response of glutamate alone (the positive control). Results obtained from testing certain exemplary compounds from this assay are presented in Table 1.
Example 185 Phosphatidylinositol hydrolysis (TP1) assays Inositol phophate assays were performed as described by Berridge et al. (1982) (Berridge et al, (1982) Biochem. J. 206: 587-5950; and Nakajima et al.. JBiol. Chem. 267:2437-2442 (1992)) with slight modifications. Mouse fibroblast Ltk cells expressing hmGluR5 (hmGluR5/L38-20 cells) were seeded in 24-well plates at a density of 8xl0 5 cells/well. One gCi of 3 H]-inositol (Amersham PT6-271; Arlington Heights, Ill.; specific activity 17.7 Ci/mmol) was added to each well and incubated for 16 h at 37 0 C. Cells were washed twice and incubated for 45 min in 0.5 ml of standard Hepes buffered saline buffer (HBS; 125 mM NaCI, 5 mM KCI, 0.62 mM MgSO 4 1.8 mM CaCI 2 20 mM HEPES, 6 mM glucose, pH to The cells were washed with HBS containing 10 mM LiCI, and 400 pl buffer added to each well. Cells were incubated at 37 0 C for 20 min. For testing, 50 pl of 10X compounds used in the practice of the invention (made in HBS/LiCI (100 mM)) was added and incubated for 10 minutes.
Cells were activated by the addition of 10 pM glutamate, and the plates left for 1 hour at 37 0
C.
The incubations were terminated by the addition of 1 ml ice-cold methanol to each well. In order to isolate inositol phosphates (IPs), the cells were scraped from wells, and placed in numbered glass test tubes. One ml of chloroform was added to each tube, the tubes were mixed, and the phases separated by centrifugation. IPs were separated on Dowex anion exchange columns (AG I-X8 100-200 mesh formate form). The upper aqueous layer (750 ul) was added to the Dowex columns, and the WO 01/16121 PCT/US00/23923 109 columns eluted with 3 ml of distilled water. The eluents were discarded, and the columns were washed with 10 mis of 60 mM ammonium formate/5 mM Borax, which was also discarded as waste. Finally the columns were eluted with 4 ml of 800 mM ammonium formate/0.1 M formic acid, and the samples collected in scintillation vials. Scintillant was added to each vial, and the vials shaken, and counted in a scintillation counter after 2 hours. Phosphatidylinositol hydrolysis in cells treated with certain exemplary compounds was compared to phosphatidylinositol hydrolysis in cells treated with control and the results are shown in Table 1.
Example 186 Analgesic Animal Model (CFA Model) Compounds that modulate receptor-mediated analgesic responses were tested in an animal model. Robust inflammation was induced by the injection of complete Freund's adjuvant (CFA, mg/ml) subcutaneously into the hind paw of male Sprague-Dawley rats (150-175 The inflammatory response to CFA began approximately 12-hrs post-CFA injection and persisted for several days following inoculation. The inflamed hind paw became sensitive to noxious (paw pinch, plantar test) or innocuous (cold plate, Von Frey filaments) stimuli compared to the contralateral hind paw. Test compounds (240 pmol/kg) were administered via intraperitoneal injection. The analgesic activity of compounds was evaluated one to 24-hours post administration by assessing the duration of analgesic effect; threshold to elicit response; and time from painful stimulus to response. A general increase in duration of analgesic effect, threshold to painful stimuli or time to respond following compound administration suggested analgesic efficacy. For each group of animals, responses following administration of certain exemplary compounds were compared to responses prior to compound administration. Results are presented in Table 1.
TABLE 1* .2 N M Z E 44 ND
ND
46 47 48 ND 49
ND
51 52 ND 53 ND 54 55 ND 56 ND 57 ND 58 ND 59 ND 60 ND 61 ND WO 01/16121 WO 0116121PCT/USO0/23923 62 ND 63 ND 64 ND 66 ND 67 ND ND 69 4+ ND 74 ND ND
ND
79 s0 82 ND 83 ND 86 87 ND 88 ND 89
ND
91 ND ND 92 ND 93 ND 94 ND ND 96 ND 98 ND 100 ND ND 101 103 ND ND 104 ND 105 ND 106 ND 107 ND 108 109 ND 110 III 112 113 114 ND 115 ND 117 ND 118 ND 120 124 ND ND 126 ND 127 ND 131
ND
133 134 ND 135 136 ND 137 ND 138 ND 139 ND 140 ND ND 141 ND 142 ND 143 ND 144 ND 145 ND 148 ND ND 150 152 ND 155 ND 157 158 ND 159 ND 160 ND 161 ND 162 ND 163 ND 164 ND 165 ND 166 167 ND 168 N WO 01/16121 PCT/US0O/23923 169 173 ND 179 180 ND 181 ND ND 182 ND 183 *Table Legend: Ca2' Flux: 0.I nmol IC 5 o 10 mol IC5> 10 tmol PI Hydrolysis: 1.0 nmol <IC 50 10 pmol ICso 10 umol CFA Model: analgesic efficacy analgesic inefficacy ND not determined Data presented in Table 1 demonstrate that the activity of metabotropic glutamate receptor assessed by measuring receptor-activated changes in calcium flux, may be modulated by compounds contemplated for use in the practice of the invention. In assays using cells transfected with mGluR5a, a majority of invention compounds are effective at decreasing intracellular calcium flux, and decreasing phosphoinositol hydrolysis.
Example 187 Salt forms of 2-(Phenylethynyl)-l1--thiazole Various salt forms of 2-(phenylethynyl)-l,3-thiazole were prepared, and their solubility, hygroscopicity, and physical state were assessed. The compound 2-phenylethynl-1,3-thiazole was prepared (see Example 8) as well as hydrochloric acid, fumaric acid, methylsulfonic acid and toluene sulfonic acid salt forms of the compound. The hydrochloric acid salt form of 2-(phenylethynyl)-1,3thiazole is an oil. A fumaric acid salt form was not obtained by preparative steps including trituration with diethylether. Preparation of the methylsulfonate form results in a semi solid, hygroscopic material.
These salt forms are typically unsuitable for certain pharmaceutical formulations, such as for tablets, capsules, and the like. In contrast to the salt forms of 2-(phenylethynyl)-l,3-thiazole, described previously, the tosylate salt form the toluene sulfonic acid salt) is a solid having a melting point of about 129 up to 131 0 C and is non-hygroscopic. Therefore, this form is presently preferred for pharmaceutical formulations of tablets, capsules, and the like.
WO 01/16121 PCT/US00/23923 112 While the invention has been described in detail with reference to certain preferred embodiments thereof, it will be understood that modifications and variations are within the spirit and scope of that which is described and claimed.

Claims (33)

1. A compound having the structure: A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, wherein: A is a thiazolyl ring having the structure: Y-X wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, Scarbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and 15 B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl; provided that the following compounds are excluded: compounds wherein A is a thiazolyl ring, wherein each R is independently 20 hydrogen, nitro, halogen, Ci-C 4 -alkyl, CI-C 4 -haloalkyl, Ci-C 4 -alkoxy, Cl-C 4 -haloalkoxy, Ci-C 4 -alkylthio, Ci-C 4 -haloalkylthio, C 3 -C 6 -alkenyl, or C 3 -Cs-cycloalkyl; L is alkynylene; and B is substituted or unsubstituted aryl, wherein substituents are independently nitro, cyano, Ci-C 6 -alkyl, Ci-C 4 -haloalkyl, Ci-C 4 -alkoxy, Ci-C 4 haloalkoxy, Cl-C 4 -alkylthio, C -C 4 -haloalkylthio, C -C 4 -alkoxycarbonyl, C 3 -C 6 -alkenyl, phenyl, or phenoxy, wherein phenyl and phenoxy may bear further substituents; compounds wherein Y and Z are CR; X is S; R is phenyl; L is unsubstituted alkenylene, and B is unsubstituted phenyl; compounds wherein X and Z are CR, Y is S; R is phenyl; L is unsubstituted alkenylene, and B is unsubstituted phenyl; and compounds wherein A is unsubstituted thiazolyl; L is alkenylene or alkynylene; and B is unsubstituted phenyl.
2. A compound according to claim 1, wherein L is an azo group.
3. A compound according to claim 1, wherein L is alkenylene.
4. A compound according to claim 1, wherein L is alkynylene. fR:\LIBH]041 114 A compound according to claim 1, wherein B is substituted or unsubstituted hydrocarbyl.
6. A compound according to claim 5, wherein L is alkenylene.
7. A compound according to claim 5, wherein L is alkynylene.
8. A compound according to claim 1, wherein B is cyclohydrocarbyl.
9. A compound according to claim 8, wherein L is alkenylene. A compound according to claim 8, wherein L is alkynylene.
11. A compound according to claim 1, wherein B is substituted or unsubstituted aryl.
12. A compound according to claim 11, wherein L is alkenylene.
13. A compound according to claim 11, wherein L is alkynylene.
14. A compound according to claim 1, wherein B is substituted or unsubstituted heterocycle optionally containing one or more double bonds.
15. A compound according to claim 14, wherein L is alkenylene.
16. A compound according to claim 14, wherein L is alkynylene.
17. A compound according to claim 15, wherein B is substituted or unsubstituted heterocycle containing no double bonds or one double bond.
18. A compound according to claim 16, wherein B is substituted or unsubstituted heterocycle containing no double bonds or one double bond. 20 19. A compound having the structure: A-L-B, as defined in claim 1, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, of pharmaceutically acceptable salts thereof, substantially as hereinbefore described with reference to any one of the examples.
20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19, and a pharmaceutically acceptable carrier therefor.
21. A method of antagonizing the activity of metabotropic glutamate receptors, said method comprising contacting said receptors with at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, in an amount sufficient to antagonize the activity of said metabotropic glutamate receptor, wherein A is a thiazolyl ring having the structure: Y- wherein one of X, Y and Z is S; and [R:\LIBH]04155.doc:UG 115 the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryi, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
22. A method according to claim 21, wherein said metabotropic glutamate receptor is a Group I metabotropic glutamate receptor.
23. A method for treating a disease condition related to the activity of metabotropic glutamate receptors, said method comprising administering to a patient having said disease condition a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, or administering a pharmaceutical composition comprising said compound together with a pharmaceutically acceptable carrier therefore, wherein A is a thiazolyl ring having the structure: wherein one of X, Y and Z is S; and the remainder ofX, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
24. A method according to claim 23, wherein said disease condition is cerebral ischemia, chronic neurodegeneration, psychiatric disorders, schizophrenia, mood disorders, emotion disorders, disorders of extrapyramidal motor function, obesity, disorders of respiration, motor control and function, attention deficit disorders, [R:\LIBHj04155.doc:LG 116 concentration disorders, pain disorders, neurodegenerative disorders, epilepsy, convulsive disorders, eating disorders, sleep disorders, sexual disorders, circadian disorders, drug withdrawal, drug addiction, compulsive disorders, anxiety, panic disorders, depressive disorders, skin disorders, retinal ischemia, retinal degeneration, glaucoma, disorders associated with organ transplantation, asthma, ischemia or astrocytomas. A method according to claim 24, wherein said mood disorder is anxiety, depression, psychosis, drug withdrawal, tobacco withdrawal, memory loss, cognitive impairment, dementia, or Alzheimer's disease.
26. A method according to claim 24, wherein said extrapyramidal motor function is Parkinson's disease, progressive supramuscular palsy, Huntington's disease, Gilles de la Tourette syndrome, or tardive dyskinesia.
27. A method according to claim 24, wherein said pain disorder is neuropathic pain, chronic pain, acute pain, painful diabetic neuropathy, post-herpetic neuralgia, cancer-associated pain, pain associated with chemotherapy, pain associated with spinal cord injury, pain associated with multiple sclerosis, causalgia and reflex sympathetic dystrophy, phantom pain, post-stroke (central) pain, pain associated with HIV or AIDS, trigeminal neuralgia, lower back pain, myofacial disorders, migraine, osteoarthritic pain, postoperative pain, dental pain, post-bum pain, pain associated with systemic lupus, entrapment neuropathies, painful polyneuropathies, ocular pain, pain associated with 20 inflammation or pain due to tissue injury.
28. A method for preventing a disease condition related to the activity of metabotropic glutamate receptors in a subject at risk thereof, said method comprising administering to said subject a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, or administering a pharmaceutical composition comprising said compound together with a pharmaceutically acceptable carrier therefore, wherein A is a thiazolyl ring having the structure: Y-X wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, meicapto, nitro, carboxyl, [R:\LIBH]04155.doc:UG 117 carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
29. A method according to claim 28, wherein said disease is a disease of the pulmonary system, a disease of the nervous system, a disease of the cardiovascular system, a disease of the gastrointestinal system, a disease of the endocrine system, a disease of the exocrine system, a disease of the skin, cancer, or a disease of the ophthalmic system. Use of a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, for the manufacture of a S 15 medicament for antagonizing the activity of metabotropic glutamate receptors, wherein A is a thiazolyl ring having the structure: Y-x N wherein one ofX, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein 20 each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl.
31. The use according to claim 30, wherein said metabotropic glutamate receptor is a Group I metabotropic glutamate receptor.
32. Use of a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for treating a disease condition related to the activity of metabotropic glutamate receptors, wherein [R:\LIBH041 118 A is a thiazolyl ring having the structure: Y-x wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or Smore double bonds, or substituted aryl.
33. The use according to claim 32, wherein said disease condition is cerebral ischemia, chronic neurodegeneration, psychiatric disorders, schizophrenia, mood s15 disorders, emotion disorders, disorders of extrapyramidal motor function, obesity, disorders of respiration, motor control and function, attention deficit disorders, concentration disorders, pain disorders, neurodegenerative disorders, epilepsy, convulsive disorders, eating disorders, sleep disorders, sexual disorders, circadian disorders, drug withdrawal, drug addiction, compulsive disorders, anxiety, panic disorders, depressive 20 disorders, skin disorders, retinal ischemia, retinal degeneration, glaucoma, disorders associated with organ transplantation, asthma, ischemia or astrocytomas.
34. The use according to claim 33, wherein said mood disorder is anxiety, depression, psychosis, drug withdrawal, tobacco withdrawal, memory loss, cognitive impairment, dementia, or Alzheimer's disease.
35. The use according to claim 33, wherein said extrapyramidal motor function is Parkinson's disease, progressive supramuscular palsy, Huntington's disease, Gilles de la Tourette syndrome, or tardive dyskinesia.
36. The use according to claim 33, wherein said pain disorder is neuropathic pain, chronic pain, acute pain, painful diabetic neuropathy, post-herpetic neuralgia, cancer- associated pain, pain associated with chemotherapy, pain associated with spinal cord injury, pain associated with multiple sclerosis, causalgia and reflex sympathetic dystrophy, phantom pain, post-stroke (central) pain, pain associated with HIV or AIDS, trigeminal neuralgia, lower back pain, myofacial disorders, migraine, osteoarthritic pain, postoperative pain, dental pain, post-bur pain, pain associated with systemic lupus, [R:\LIBH]04155.doc:UG 119 entrapment neuropathies, painful polyneuropathies, ocular pain, pain associated with inflammation or pain due to tissue injury.
37. Use of a therapeutically effective amount of at least one compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for preventing a disease condition related to the activity of metabotropic glutamate receptors, wherein A is a thiazolyl ring having the structure: Y-X N wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, I. or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl. S. 20 38. The use according to claim 37, wherein said disease is a disease of the pulmonary system, a disease of the nervous system, a disease of the cardiovascular system, a disease of the gastrointestinal system, a disease of the endocrine system, a disease of the exocrine system, a disease of the skin, cancer or a disease of the ophthalmic system.
39. A pharmaceutically acceptable salt form of a compound having the structure A-L-B, or enantiomers, diastereomeric isomers or mixtures of any two or more thereof, wherein A is a thiazolyl ring having the structure: Y-x wherein one of X, Y and Z is S; and the remainder of X, Y and Z are each independently CR; wherein each R is independently hydrogen, halogen, substituted or unsubstituted hydrocarbyl, substituted or unsubstituted aryl, heterocycle, mercapto, nitro, carboxyl, [R:\LIBH]04155.doc:UG 120 carbamate, carboxamide, hydroxy, ester, cyano, amine, amide, amidine, amido, sulfonyl, or sulfonamide; L is substituted or unsubstituted alkenylene, alkynylene, or azo; and B is substituted or unsubstituted hydrocarbyl, substituted or unsubstituted cyclohydrocarbyl, substituted or unsubstituted heterocycle, optionally containing one or more double bonds, or substituted aryl; and the salt is acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, heptanoate, hexanoate, 2-hydroxyethanesulfonate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, tartrate, toluenesulfonate, undecanoate, sulfate, bisulfate, hemisulfate, hydrochloride, hydrobromide, hydroiodide, an ammonium salt, an alkali metal salt, an alkaline earth metal salt, a dicyclohexylamine salt, N-methyl-D-glucamine, phenylethylamine, or an amino acid salt. Is 40. The pharmaceutically acceptable salt form of the compound according to claim 39, wherein the salt is a toluene sulfonic acid salt. Dated 2 November, 2004 **Merck Co., Inc. *fee Se* ~Patent Attorneys for the Applicant/Nominated Person S 20 SPRUSON FERGUSON 999S4 [R:\LBH]04155.doc:UG
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Families Citing this family (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2383524C (en) * 1999-08-31 2010-09-28 Merck & Co., Inc. Thiazolyl alkynyl compounds and methods of use thereof
US7365074B2 (en) 1999-08-31 2008-04-29 Merck & Co., Inc. Pyridazine, pyrimidine and pyrazine ethyne compounds
US6774138B2 (en) * 1999-08-31 2004-08-10 Merck & Co., Inc. Thiazolyl(pyridyl)ethyne compounds
US7462619B2 (en) 1999-08-31 2008-12-09 Merck & Co., Inc. Pyridazine, pyrimidine and pyrazine ethyne compounds
CA2430696C (en) 2000-12-04 2009-01-27 F. Hoffmann-La Roche Ag Phenylethenyl or phenylethinyl derivatives as glutamate receptor antagonists
AU2003222738B2 (en) 2002-05-07 2009-09-17 Neurosearch A/S Novel azacyclic ethynyl derivatives
WO2004038374A2 (en) * 2002-10-24 2004-05-06 Merck & Co., Inc. Alkyne derivatives as tracers for metabotropic glutamate receptor binding
US8044206B2 (en) 2003-03-07 2011-10-25 Astellas Pharma Inc. Nitrogen—containing heterocyclic derivatives having 2,6-disubstituted styryl
TWI292318B (en) 2003-03-10 2008-01-11 Hoffmann La Roche Imidazol-4-yl-ethynyl-pyridine derivatives
US7531529B2 (en) 2003-06-05 2009-05-12 Roche Palo Alto Llc Imidazole derivatives
UA80888C2 (en) * 2003-06-05 2007-11-12 Hoffmann La Roche Imidazole derivatives as glutmate receptor antagonists
ATE495168T1 (en) * 2003-06-12 2011-01-15 Hoffmann La Roche HETEROARYL-SUBSTITUTED IMIDAZOLE DERIVATIVES AS GLUTAMA RECEPTOR ANTAGONISTS
US7452909B2 (en) * 2003-09-04 2008-11-18 Hoffman-La Roche Inc. Imidazole derivatives
GB0322612D0 (en) * 2003-09-26 2003-10-29 Novartis Ag Organic compounds
EP1677790A1 (en) * 2003-10-31 2006-07-12 AstraZeneca AB Alkynes ii
JP2007510645A (en) * 2003-10-31 2007-04-26 アストラゼネカ アクチボラグ Alkynes I
CN1871001A (en) * 2003-10-31 2006-11-29 阿斯利康(瑞典)有限公司 Alkynes III
RU2367661C2 (en) * 2004-03-05 2009-09-20 Тайсо Фармасьютикал Ко., Лтд. Thiazole derivatives
EP1729771B1 (en) 2004-03-22 2009-10-14 Eli Lilly &amp; Company Pyridyl derivatives and their use as mglu5 receptor antagonists
EP1756086B1 (en) * 2004-06-01 2008-06-04 F.Hoffmann-La Roche Ag Pyridin-4-yl-ethynyl-imidazoles and pyrazoles as mglu5 receptor antagonists
ES2307182T3 (en) * 2004-06-01 2008-11-16 F. Hoffmann-La Roche Ag PIRIDIN-4-IL-ETINIL-IMIDAZOLES AND PIRAZOLES AS RECEIVING ANTAGONISTS OF MGLUR5.
GB0413605D0 (en) 2004-06-17 2004-07-21 Addex Pharmaceuticals Sa Novel compounds
ATE520692T1 (en) 2004-10-07 2011-09-15 Merck Sharp & Dohme THIAZOLYL-MGLUR5 ANTAGONISTS AND METHODS OF USE THEREOF
JP5405311B2 (en) 2006-12-21 2014-02-05 エフ．ホフマン−ラ ロシュ アーゲー Polymorphs of mGluR5 receptor antagonists
US8420661B2 (en) 2010-04-13 2013-04-16 Hoffmann-La Roche Inc. Arylethynyl derivatives
AR084457A1 (en) * 2010-12-22 2013-05-15 Lundbeck & Co As H BICYCLE DERIVATIVES [3,2,1] OCTILAMIDE
CN107955052A (en) * 2011-12-23 2018-04-24 晶体制药独资有限公司 The method of alkynyl 16- substitution -17- ketone group steroids
WO2014121883A1 (en) * 2013-02-07 2014-08-14 Merck Patent Gmbh Substituted acetylene derivatives and their use as positive allosteric modulators of mglur4
CN106146391A (en) * 2015-04-15 2016-11-23 中国科学院上海药物研究所 Substituted benzamide compound of 5-fragrance alkynyl and preparation method thereof, pharmaceutical composition and purposes
AU2016273751B2 (en) 2015-06-03 2020-04-02 F. Hoffmann-La Roche Ag Ethynyl derivatives
JP2019081709A (en) * 2016-03-18 2019-05-30 石原産業株式会社 Pest control agent

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996003406A1 (en) * 1994-07-28 1996-02-08 Agouron Pharmaceuticals, Inc. Compounds useful as antiproliferative agents and garft inhibitors
WO1999002497A2 (en) * 1997-07-11 1999-01-21 Novartis Ag Pyridine derivatives

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3705153A (en) * 1969-05-14 1972-12-05 Sumitomo Chemical Co Novel thiazolyacetic acids and salts thereof
DE2426878A1 (en) * 1974-06-04 1976-01-08 Basf Ag 3,5-Substd-1,2,4-oxadiazoles prodn - from acylamidine or guanidine salts by N-halogenation then cyclising with base
JPS58219169A (en) * 1982-06-15 1983-12-20 Takeda Chem Ind Ltd Oxazoleacetic acid derivative
JPS62142168A (en) * 1985-10-16 1987-06-25 Mitsubishi Chem Ind Ltd Thiazole derivative and leukotriene antagonistic agent containing said derivative as active ingredient
JP2745073B2 (en) * 1989-10-27 1998-04-28 チッソ株式会社 Phenylacetylene compound
FR2663850B1 (en) * 1990-07-02 1994-01-14 Gird Galderma PHARMACEUTICAL OR COSMETIC COMPOSITION CONTAINING A RETINOUIDE AND A STEROL IN COMBINATION.
DE4023742A1 (en) * 1990-07-26 1992-01-30 Gruenenthal Gmbh SUBSTITUTED PHENYLACETYLENES, MEDICINAL PRODUCTS CONTAINING THEM AND METHOD FOR PRODUCING THESE COMPOUNDS AND MEDICINAL PRODUCTS
US5270312A (en) * 1990-11-05 1993-12-14 Warner-Lambert Company Substituted piperazines as central nervous system agents
ATE334120T1 (en) * 1995-01-06 2006-08-15 Targacept Inc PHARMACEUTICAL COMPOSITIONS FOR THE PREVENTION AND TREATMENT OF CENTRAL NERVOUS SYSTEM DISORDERS
HUP9900926A3 (en) * 1995-09-07 2002-12-28 Hoffmann La Roche New 4-(oxyalkoxyphenyl)-3-oxy-piperidines for treating heart and kidney insufficiency
JPH09235276A (en) * 1995-12-27 1997-09-09 Takeda Chem Ind Ltd Oxazole derivative, production of the same and use of the same
JP4056589B2 (en) * 1996-07-19 2008-03-05 武田薬品工業株式会社 Heterocyclic compounds, their production and use
ZA986594B (en) * 1997-07-25 1999-01-27 Abbott Lab Urokinase inhibitors
ATE404539T1 (en) * 1997-10-02 2008-08-15 Eisai R&D Man Co Ltd CONDENSED PYRIDINE DERIVATIVES
US6187777B1 (en) * 1998-02-06 2001-02-13 Amgen Inc. Compounds and methods which modulate feeding behavior and related diseases
JP2000119253A (en) * 1998-08-11 2000-04-25 Dai Ichi Seiyaku Co Ltd Novel sulfonyl derivative
CA2383524C (en) * 1999-08-31 2010-09-28 Merck & Co., Inc. Thiazolyl alkynyl compounds and methods of use thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996003406A1 (en) * 1994-07-28 1996-02-08 Agouron Pharmaceuticals, Inc. Compounds useful as antiproliferative agents and garft inhibitors
WO1999002497A2 (en) * 1997-07-11 1999-01-21 Novartis Ag Pyridine derivatives

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BRIT J PHARMACOL. 126, SUPPL PROC, MARCH 1999, P 249 *

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