US20080200524A1

US20080200524A1 - Amide compounds as ion channel ligands and uses thereof

Info

Publication number: US20080200524A1
Application number: US11/982,351
Authority: US
Inventors: Michael G. Kelly; Satyanarayana Janagani; Guoxian Wu; John Kincaid
Original assignee: Individual
Current assignee: Individual
Priority date: 2003-10-07
Filing date: 2007-11-01
Publication date: 2008-08-21
Also published as: GB0422296D0; EP1685109A2; GB2406856B; US20050192293A1; WO2005034870A2; BRPI0415167A; WO2005034870A3; CA2541299A1; US7338950B2; MXPA06003949A; GB2406856A; WO2005032493A8; US20050197364A1; WO2005032493A2; JP2007525482A; WO2005032493A3

Abstract

Compounds are disclosed that have a formula represented by the following:

The compounds may be prepared as pharmaceutical compositions, and may be used for the prevention and treatment of a variety of conditions in mammals including humans, including by way of non-limiting example, pain, inflammation, traumatic injury, and others.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a Continuation of Ser. No. 10/962,195, filed on Oct. 7, 2004, which in turn claims the priority of co-pending provisional applications U.S. Ser. No. 60/434,022, filed on Oct. 7, 2003, and U.S. Ser. No. 60/575,937, filed Jun. 1, 2004. The disclosures of said applications are incorporated by reference herein in their entireties. Applicants claim the benefits of both applications under 35 U.S.C. §119(e).

FIELD OF THE INVENTION

This invention relates to novel compounds and to pharmaceutical compositions containing such compounds. This invention also relates to methods for preventing and/or treating pain and inflammation-related conditions in mammals, such as (but not limited to) arthritis, Parkinson's disease, Alzheimer's disease, stroke, uveitis, asthma, myocardial infarction, the treatment and prophylaxis of pain syndromes (acute and chronic or neuropathic), traumatic brain injury, acute spinal cord injury, neurodegenerative disorders, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders, using the compounds and pharmaceutical compositions of the invention.

BACKGROUND OF THE INVENTION

Studies of signaling pathways in the body have revealed the existence of ion channels and sought to explain their role. Ion channels are integral membrane proteins with two distinctive characteristics: they are gated (open and closed) by specific signals such as membrane voltage or the direct binding of chemical ligands and, once open, they conduct ions across the cell membrane at very high rates.
There are many types of ion channels. Based on their selectivity to ions, they can be divided into calcium channel, potassium channel, sodium channel, etc. The calcium channel is more permeable to calcium ions than other types of ions, the potassium channel selects potassium ions over other ions, and so forth. Ion channels may also be classified according to their gating mechanisms. In a voltage-gated ion channel, the opening probability depends on the membrane voltage, whereas in a ligand-gated ion channel, the opening probability is regulated by the binding of small molecules (the ligands). Since ligand-gated ion channels receive signals from the ligand, they may also be considered as “receptors” for ligands.
Examples of ligand-gated ion channels include nAChR (nicotinic acetylcholine receptor) channel, GluR (glutamate receptor) channel, ATP-sensitive potassium channel, G-protein activated channel, cyclic-nucleotide-gated channel, etc.
Transient receptor potential (TRP) channel proteins constitute a large and diverse family of proteins that are expressed in many tissues and cell types. This family of channels mediates responses to nerve growth factors, pheromones, olfaction, tone of blood vessels and metabolic stress et al., and the channels are found in a variety of organisms, tissues and cell types including nonexcitable, smooth muscle and neuronal cells. Furthermore, TRP-related channel proteins are implicated in several diseases, such as several tumors and neurodegenerative disorders and the like. See, for example, Minke, et al., APStracts 9:0006P (2002).
Nociceptors are specialized primary afferent neurons and the first cells in a series of neurons that lead to the sensation of pain. The receptors in these cells can be activated by different noxious chemical or physical stimuli. The essential functions of nociceptors include the transduction of noxious stimuli into depolarizations that trigger action potentials, conduction of action potentials from primary sensory sites to synapses in the central nervous system, and conversion of action potentials into neurotransmitter release at presynaptic terminals, all of which depend on ion channels.
One TRP channel protein of particular interest is the vanilloid receptor. Also known as VR1, the vanilloid receptor is a non-selective cation channel which is activated or sensitized by a series of different stimuli including capsaicin, heat and acid stimulation and products of lipid bilayer metabolism (anandamide), and lipoxygenase metabolites. See, for example Smith, et al., Nature, 418:186-190 (2002). VR1 does not discriminate among monovalent cations, however, it exhibits a notable preference for divalent cations with a permeability sequence of Ca²⁺>Mg²⁺>Na⁺=K⁺=Cs⁺. Ca²⁺ is especially important to VR1 function, as extracellular Ca²⁺ mediates desensitization, a process which enables a neuron to adapt to specific stimuli by diminishing its overall response to a particular chemical or physical signal. VR1 is highly expressed in primary sensory neurons in rats, mice and humans, and innervates many visceral organs including the dermis, bones, bladder, gastrointestinal tract and lungs. It is also expressed in other neuronal and non-neuronal tissues including the CNS, nuclei, kidney, stomach and T-cells. The VR1 channel is a member of the superfamily of ion channels with six membrane-spanning domains, with highest homology to the TRP family of ion channels.
VR1 gene knockout mice have been shown to have reduced sensory sensitivity to thermal and acid stimuli. See, for example, Caterina, et al. Science, 14:306-313 (2000). This supports the concept that VR1 contributes not only to generation of pain responses but also to the maintenance of basal activity of sensory nerves. VR1 agonists and antagonists have use as analgesics for the treatment of pain of various genesis or etiology, for example acute, inflammatory and neuropathic pain, dental pain and headache (such as migraine, cluster headache and tension headache). They are also useful as anti-inflammatory agents for the treatment of arthritis, Parkinson's Disease, Alzheimer's Disease, stroke, uveitis, asthma, myocardial infarction, the treatment and prophylaxis of pain syndromes (acute and chronic [neuropathic]), traumatic brain injury, spinal cord injury, neurodegenerative disorders, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders, renal disorders, obesity, eating disorders, cancer, schizophrenia, epilepsy, sleeping disorders, cognition, depression, anxiety, blood pressure, lipid disorders, and atherosclerosis.
Compounds, such as those of the present invention, which interact with the vanilloid receptor can thus play a role in treating or preventing or ameliorating these conditions.
A wide variety of Vanilloid compounds of different structures are known in the art, for example those disclosed in European Patent Application Numbers, EP 0 347 000 and EP 0 401 903, UK Patent Application Number GB 2226313 and International Patent Application, Publication Number WO 92/09285. Particularly notable examples of vanilloid compounds or vanilloid receptor modulators are capsaicin or trans 8-methyl-N-vanillyl-6-nonenamide which is isolated from the pepper plant, capsazepine (Tetrahedron, 53, 1997, 4791) and olvanil or —N-(4-hydroxy-3-methoxybenzyl)oleamide (J. Med. Chem., 36, 1993, 2595).
International Patent Application, Publication Number WO 02/08221 discloses diaryl piperazine and related compounds which bind with high selectivity and high affinity to vanilloid receptors, especially Type I Vanilloid receptors, also known as capsaicin or VR1 receptors. The compounds are said to be useful in the treatment of chronic and acute pain conditions, itch and urinary incontinence.
International Patent Application, Publication Numbers WO 02/16317, WO 02/16318 and WO 02/16319 suggest that compounds having a high affinity for the vanilloid receptor are useful for treating stomach-duodenal ulcers.
WO04/56774 describe certain substituted biphenyl-4-carboxylic acid arylamide analogues having possible application as receptor modulators.
U.S. Pat. No. 3,424,760 and U.S. Pat. No. 3,424,761 both describe a series of 3-Ureidopyrrolidines that are said to exhibit analgesic, central nervous system, and pyschopharmacologic activities. These patents specifically disclose the compounds 1-(1-phenyl-3-pyrrolidinyl)-3-phenyl urea and 1-(1-phenyl-3-pyrrolidinyl)-3-(4-methoxyphenyl)urea respectively. International Patent Applications, Publication Numbers WO 01/62737 and WO 00/69849 disclose a series of pyrazole derivatives which are stated to be useful in the treatment of disorders and diseases associated with the NPY receptor subtype Y5, such as obesity. WO 01/62737 specifically discloses the compound 5-amino-N-isoquinolin-5-yl-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide. WO 00/69849 specifically discloses the compounds 5-methyl-N-quinolin-8-yl-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide, 5-methyl-N-quinolin-7-yl-1-[3-trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide, 5-methyl-N-quinolin-3-yl-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide, N-isoquinolin-5-yl-5-methyl-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide, 5-methyl-N-quinolin-5-yl-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide, 1-(3-chlorophenyl)-N-isoquinolin-5-yl-5-methyl-1H-pyrazole-3-carboxamide, N-isoquinolin-5-yl-1-(3-methoxyphenyl)-5-methyl-1H-pyrazole-3-carboxamide, 1-(3-fluorophenyl)-N-isoquinolin-5-yl-5-methyl-1H-pyrazole-3-carboxamide, 1-(2-chloro-5-trifluoromethylphenyl)-N-isoquinolin-5-yl-5-methyl-1N-pyrazole-3-carboxamide, 5-methyl-N-(3-methylisoquinolin-5-yl)-1-[3-(trifluoromethyl)phenyl]-1N-pyrazole-3-carboxamide, 5-methyl-N-(1,2,3,4-tetrahydroisoquinolin-5-yl)-1-[3-(trifluoromethyl)phenyl]-1H-pyrazole-3-carboxamide.
German Patent Application Number 2502588 describes a series of piperazine derivatives. This application specifically discloses the compound N-[3-[2-(diethylamino) ethyl]-1,2-dihydro-4-methyl-2-oxo-7-quinolinyl]-4-phenyl-1-piperazinecarboxamide.
We have now discovered that certain compounds have surprising potency and selectivity as VR-1 antagonists. The compounds of the present invention are considered to be particularly beneficial as VR-1 antagonists as certain compounds exhibit improved aqueous solubility and metabolic stability.

SUMMARY OF THE INVENTION

It has now been found that compounds such as those set forth herein, are capable of modifying mammalian ion channels such as the VR1 cation channel. This finding leads to novel compounds having therapeutic value. It also leads to pharmaceutical compositions having the compounds of the present invention as active ingredients and to their use to treat, prevent or ameliorate a range of conditions in mammals such as but not limited to pain of various genesis or etiology, for example acute, chronic, inflammatory and neuropathic pain, dental pain and headache (such as migraine, cluster headache and tension headache).
Accordingly, in a first aspect of the invention, compounds are disclosed that are capable of modifying ion channels, in vivo, having a formula:
wherein:
A is N, CR⁴, a carbon atom bound to L, or is not an atom;
one of W, Z, B, Y and X is a carbon atom bound to L if A is not an atom, another of W, Z, B, Y and X is a carbon atom bound to G, and each of the remaining W, Z, B, Y and X is independently N or CR⁴;
L is a bond or —(CH₂)_n—, wherein n is an integer of 1-3;
G is C═O, C═S or SO₂;
R¹is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;
R²is hydrogen or substituted or unsubstituted alkyl;
R³is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl; and
each R⁴is independently hydrogen, alkyl, substituted or unsubstituted alkyl, acyl, acylamino, alkylamino, alkylthio, alkoxy, alkoxycarbonyl, alkylarylamino, arylalkyloxy, amino, aryl, arylalkyl, sulfoxide, sulfone, sulfanyl, aminosulfonyl, arylsulfonyl, sulfuric acid, sulfuric acid ester, dihydroxyphosphoryl, aminohydroxyphosphoryl, azido, carboxy, carbamoyl, carboxyl, cyano, cycloheteroalkyl, dialkylamino, halo, heteroaryloxy, heteroaryl, heteroalkyl, hydroxyl, nitro or thio,
or a pharmaceutically acceptable salt, solvate or prodrug thereof;
and isomers and stereoisomers thereof.
In a further embodiment of the invention, compounds are capable of modifying ion channels, in vivo, having a formula IA
In compounds of formula IA, L will be a bond and G will be carbonyl. R¹is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, heteroaryl, aralkyl, or heteroaralkyl; R²is hydrogen; and R³is substituted heteroaryl.
In a further embodiment of the compounds of formula IA, R³is of the formula
wherein R⁴is as described above; n is an integer of from 1-3; and A is independently selected from alkyl, substituted alkyl, acyl, substituted acyl, acylamino, substituted acylamino, alkylamino, substituted alkylamino, alkylthio, substituted alkylthio, alkoxy, substituted alkoxy, alkoxycarbonyl, substituted alkoxycarbonyl, alkylarylamino, substituted alkylarylamino, arylalkyloxy, substituted arylalkyloxy, amino, aryl, substituted aryl, arylalkyl, substituted arylalkyl, sulfoxide, substituted sulfoxide, sulfone, substituted sulfone, sulfanyl, substituted sulfanyl, aminosulfonyl, substituted aminosulfonyl, arylsulfonyl, substituted arylsulfonyl, sulfuric acid, sulfuric acid ester, dihydroxyphosphoryl, substituted dihydroxyphosphoryl, aminohydroxyphosphoryl, substituted aminohydroxyphosphoryl, azido, carboxy, carbamoyl, substituted carbamoyl, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, dialkylamino, substituted dialkylamino, halo, heteroaryloxy, substituted heteroaryloxy, heteroaryl, substituted heteroaryl, heteroalkyl, substituted heteroalkyl, hydroxyl, nitro or thio.
In a more particular embodiment of the invention, compounds capable of modifying ion channels, in vivo, have a formula II
wherein: R¹is substituted alkyl. In some compounds, R¹is —(CR² ₂)x-R^4′; R²is hydrogen or alkyl; R^4′ is R⁴; R⁴is as described for formula I; and x is an integer from 1-3.
In preferred embodiments of compounds of formula II, R^4′ is selected from t-butyl, aryl, cycloalkyl, cycloheteroalkyl and heteroaryl. In some of these preferred embodiments, R^4′ is substituted or unsubstituted phenyl, or naphthalene. In yet other of these preferred embodiments, R^4′ is substituted or unsubstituted cyclopropyl, cyclopentyl or cyclohexyl. In still others, R^4′ is substituted or unsubstituted pyrrolidinyl, piperidinyl, or morpholinyl. In yet others, R^4′ is substituted or unsubstituted pyridinyl, pyrimidinyl quinoline, benzodioxane, tetrahydroquinoline, indole, indazole or carbazole. Further preferred embodiments have R^4′ that is substituted or unsubstituted furanyl, imidazolyl, thiophenyl, pyrazolyl, or thiazolyl. In especially preferred embodiments, R^4′ is t-Bu.
In some specific compounds of Formula II, x is 1 or 2.
In some other specific compounds of Formula II, R¹is substituted or unsubstituted cycloalkyl, cycloheteroalkyl or heteroaryl. In some preferred of these embodiments, R¹is substituted or unsubstituted cyclopropyl, cyclopentyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, pyrimidinyl quinoline, benzodioxane, tetrahydroquinoline, indole, indazole, carbazole, furanyl, imidazolyl, thiophenyl, pyrazolyl or thiazolyl.
In additional specific compounds of formula II, A is halo, NR² ₂, trihaloalkyl, alkoxy, SO₂R², or SO₂NR². In some preferred embodiments, A is selected from the group consisting of Cl, CF₃, OMe, NMe₂, SO₂CF₃, and SO₂NMe₂.
In certain specific compounds where A is selected from the group consisting of Cl, CF₃, OMe, NMe₂, SO₂CF₃, and SO₂NMe₂, each of W, X, Y and Z is CR⁴. In other specific compounds, W is N and each of X, Y and Z is CR⁴. In still other specific compounds, Y is N and each of W, X and Z is CR⁴. In still further specific compounds, two of W, X, Y and Z are Ns.
In yet further particular embodiments, the compounds of the invention are set forth and may be selected from a comprehensive listing of such compounds, set forth later on herein in Table 1. The Table contains in excess of 1420 compounds that have been synthesized and have as a group, demonstrated activity in their capacity of modifying ion channels, in vivo, and thereby functioning in the therapeutic applications set forth herein in relation to capsaicin and the vanilloid receptor.
The compounds of the present invention are useful for the treatment of inflammatory pain and associated hyperalgesia and allodynia. They are also useful for the treatment of neuropathic pain and associated hyperalgesis and allodynia (e.g. trigeminal or herpetic neuralgia, diabetic neuropathy, causalgia, sympathetically maintained pain and deafferentation syndromes such as brachial plexus avulsion). The compounds of the present invention are also useful as anti-inflammatory agents for the treatment of arthritis, and as agents to treat Parkinson's Disease, Alzheimer's Disease, stroke, uveitis, asthma, myocardial infarction, traumatic brain injury, spinal cord injury, neurodegenerative disorders, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders, renal disorders, obesity, eating disorders, cancer, schizophrenia, epilepsy, sleeping disorders, cognition, depression, anxiety, blood pressure, lipid disorders, and atherosclerosis.
In one aspect, this invention provides compounds which are capable of modifying ion channels, in vivo. Representative ion channels so modified include voltage-gated channels and ligand-gated channels, including cation channels such as vanilloid channels.
In a further aspect, the present invention provides pharmaceutical compositions comprising a compound of the invention, and a pharmaceutical carrier, excipient or diluent. In this aspect of the invention, the pharmaceutical composition can comprise one or more of the compounds described herein.
In a further aspect of the invention, a method is disclosed for treating mammals, including humans, as well as lower mammalian species, susceptible to or afflicted with a condition from among those listed herein, and particularly, such condition as may be associated with e.g. arthritis, uveitis, asthma, myocardial infarction, traumatic brain injury, acute spinal cord injury, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders, which method comprises administering an effective amount of one or more of the pharmaceutical compositions just described. Correspondingly, the invention extends and includes the use of the compounds of the invention for the treatment of the mentioned maladies, as well as for the preparation of pharmaceutical compositions and like medicaments, which include among their applications and uses, the treatment of the stated maladies. Likewise the invention extends to compounds of the invention for use as pharmaceuticals and medicaments.
In yet another method of treatment aspect, this invention provides a method of treating a mammal susceptible to or afflicted with a condition that gives rise to pain responses or that relates to imbalances in the maintenance of basal activity of sensory nerves. Compounds have use as analgesics for the treatment of pain of various geneses or etiology, for example acute, inflammatory pain (such as pain associated with osteoarthritis and rheumatoid arthritis); various neuropathic pain syndromes (such as post-herpetic neuralgia, trigeminal neuralgia, reflex sympathetic dystrophy, diabetic neuropathy, Guillian Barre syndrome, fibromyalgia, phantom limb pain, post-masectomy pain, peripheral neuropathy, HIV neuropathy, and chemotherapy-induced and other iatrogenic neuropathies); visceral pain, (such as that associated with gastroesophageal reflex disease, irritable bowel syndrome, inflammatory bowel disease, pancreatitis, and various gynecological and urological disorders), dental pain and headache (such as migraine, cluster headache and tension headache).
In additional method of treatment aspects, this invention provides methods of treating a mammal susceptible to or afflicted with neurodegenerative diseases and disorders such as, for example Parkinson's disease, Alzheimer's disease and multiple sclerosis; diseases and disorders which are mediated by or result in neuroinflammation such as, for example traumatic brain injury, stroke, and encephalitis; centrally-mediated neuropsychiatric diseases and disorders such as, for example depression mania, bipolar disease, anxiety, schizophrenia, eating disorders, sleep disorders and cognition disorders; epilepsy and seizure disorders; prostate, bladder and bowel dysfunction such as, for example urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway disease and disorders such as, for example, allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders which are mediated by or result in inflammation such as, for example rheumatoid arthritis and osteoarthritis, myocardial infarction, various autoimmune diseases and disorders, uveitis and atherosclerosis; itch/pruritus such as, for example psoriasis; alopecia (hair loss); obesity; lipid disorders; cancer; blood pressure; spinal cord injury; and renal disorders method comprises administering an effective condition-treating or condition-preventing amount of one or more of the pharmaceutical compositions just described.
In additional aspects, this invention provides methods for synthesizing the compounds of the invention, with representative synthetic protocols and pathways disclosed later on herein.
Other objects and advantages will become apparent to those skilled in the art from a consideration of the ensuing detailed description, in conjunction with the following illustrative drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: A graph demonstrating the activity of

Compounds

18 and 24 in inhibiting a capsaicin induced intracellular current. The functional activity of compounds at a VR1 receptor may be determined by measuring changes in intracellular calcium levels in neurons such as those in the dorsal root ganglion (DRG neurons).

FIG. 2: A graph showing that Compound 18 at a dose of 30 mg/kg significantly blocks capsaicin induced plasma extravasation in rats. The results are expressed in μg EB/mg tissue. The results are represented for the delivery vehicle HPBCD, control compound BCTC, compound 18 at two concentrations and capsaicin alone in a delivery vehicle.

FIG. 3: A dose response curve demonstrating the increasing effectiveness of Compound 18 in inhibiting a capsaicin induced calcium ion influx at higher concentrations tested, namely 50 nM, 100 nM and 250 nM.

FIG. 4: A dose response curve demonstrating the increasing effectiveness of Compound 24 in inhibiting a capsaicin induced calcium ion influx at higher concentrations tested, namely 20 nM, 40 nM and 100 nM and 200 nM.

FIG. 5: A graph showing the times per second test subjects lick their paw when a delivery vehicle is administered alone, capsaicin is administered, capsaicin is administered with a control compound, and capsaicin is administered with Compound 18 to the affected area.

FIG. 6: A graph demonstrating that a dose of Compound 18 at 30 mg/kg significantly increases latency of paw withdrawal demonstrating reversal of thermal hyperalgesia. The figure depicts the time in seconds until animals withdraw from thermal stimulation at baseline and two hours after administration of a delivery vehicle, a control compound, and three concentrations of Compound 18.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

When describing the compounds, pharmaceutical compositions containing such compounds and methods of using such compounds and compositions, the following terms have the following meanings unless otherwise indicated. It should also be understood that any of the moieties defined below may be substituted with a variety of substituents, and that the respective definitions are intended to include such substituted moieties within their scope. By way of non-limiting example, such substituents may include e.g. halo (such as fluoro, chloro, bromo), —CN, —CF₃, —OH, —OCF₃, C_2-6alkenyl, C_3-6alkynyl, C_1-6alkoxy, aryl and di-C_1-6alkylamino.
“Acyl” refers to a radical —C(O)R, where R is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl as defined herein. Representative examples include, but are not limited to, formyl, acetyl, cylcohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl and the like.
“Acylamino” refers to a radical —NR′C(O)R, where R′ is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl and R is hydrogen, alkyl, alkoxy, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl or heteroarylalkyl, as defined herein. Representative examples include, but are not limited to, formylamino, acetylamino, cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino, benzylcarbonylamino and the like.
“Acyloxy” refers to the group —OC(O)R where R is hydrogen, alkyl, aryl or cycloalkyl.
“Substituted alkenyl” includes those groups recited in the definition of “substituted” herein, and particularly refers to an alkenyl group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Alkoxy” refers to the group —OR where R is alkyl. Particular alkoxy groups include, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.
“Substituted alkoxy” includes those groups recited in the definition of “substituted” herein, and particularly refers to an alkoxy group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, heteroaryl, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Alkoxycarbonylamino” refers to the group —NRC(O)OR′ where R is hydrogen, alkyl, aryl or cycloalkyl, and R′ is alkyl or cycloalkyl.
“Aliphatic” refers to hydrocarbyl organic compounds or groups characterized by a straight, branched or cyclic arrangement of the constituent carbon atoms and an absence of aromatic unsaturation. Aliphatics include, without limitation, alkyl, alkylene, alkenyl, alkenylene, alkynyl and alkynylene. Aliphatic groups typically have from 1 or 2 to about 12 carbon atoms.
“Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groups particularly having up to about 11 carbon atoms, more particularly as a lower alkyl, from 1 to 8 carbon atoms and still more particularly, from 1 to 6 carbon atoms. The hydrocarbon chain may be either straight-chained or branched. This term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, tert-butyl, n-hexyl, n-octyl, tert-octyl and the like. The term “lower alkyl” refers to alkyl groups having 1 to 6 carbon atoms. The term “alkyl” also includes “cycloalkyls” as defined below.
“Substituted alkyl” includes those groups recited in the definition of “substituted” herein, and particularly refers to an alkyl group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, heteroaryl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂—, and aryl-S(O)₂—.
“Alkylene” refers to divalent saturated aliphatic hydrocarbyl groups particularly having up to about 11 carbon atoms and more particularly 1 to 6 carbon atoms which can be straight-chained or branched. This term is exemplified by groups such as methylene (—CH₂—), ethylene (—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like.
“Substituted alkylene” includes those groups recited in the definition of “substituted” herein, and particularly refers to an alkylene group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Alkenyl” refers to monovalent olefinically unsaturated hydrocarbyl groups preferably having up to about 11 carbon atoms, particularly, from 2 to 8 carbon atoms, and more particularly, from 2 to 6 carbon atoms, which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 sites of olefinic unsaturation. Particular alkenyl groups include ethenyl (—CH═CH₂), n-propenyl (—CH₂CH═CH₂), isopropenyl (—C(CH₃)═CH₂), vinyl and substituted vinyl, and the like.
“Alkenylene” refers to divalent olefinically unsaturated hydrocarbyl groups particularly having up to about 11 carbon atoms and more particularly 2 to 6 carbon atoms which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 sites of olefinic unsaturation. This term is exemplified by groups such as ethenylene (—CH═CH—), the propenylene isomers (e.g., —CH═CHCH₂— and —C(CH₃)═CH— and —CH═C(CH₃)—) and the like.
“Alkynyl” refers to acetylenically unsaturated hydrocarbyl groups particularly having up to about 11 carbon atoms and more particularly 2 to 6 carbon atoms which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 sites of alkynyl unsaturation. Particular non-limiting examples of alkynyl groups include acetylenic, ethynyl (—C≡CH), propargyl (—CH₂C≡CH), and the like.
“Substituted alkynyl” includes those groups recited in the definition of “substituted” herein, and particularly refers to an alkynyl group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Alkanoyl” or “acyl” as used herein refers to the group R—C(O)—, where R is hydrogen or alkyl as defined above.
“Aryl” refers to a monovalent aromatic hydrocarbon group derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexylene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene and the like. Particularly, an aryl group comprises from 6 to 14 carbon atoms.
“Substituted Aryl” includes those groups recited in the definition of “substituted” herein, and particularly refers to an aryl group that may optionally be substituted with 1 or more substituents, for instance from 1 to 5 substituents, particularly 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkenyl, substituted alkenyl, alkoxy, substituted alkoxy, alkoxycarbonyl, alkyl, substituted alkyl, alkynyl, substituted alkynyl, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Fused Aryl” refers to an aryl having two of its ring carbon in common with a second aryl ring or with an aliphatic ring.
“Alkaryl” refers to an aryl group, as defined above, substituted with one or more alkyl groups, as defined above.
“Aralkyl” or “arylalkyl” refers to an alkyl group, as defined above, substituted with one or more aryl groups, as defined above.
“Aryloxy” refers to —O-aryl groups wherein “aryl” is as defined above.
“Alkylamino” refers to the group alkyl-NR′R″, wherein each of R′ and R″ are independently selected from hydrogen and alkyl.
“Arylamino” refers to the group aryl-NR′R″, wherein each of R′ and R″ are independently selected from hydrogen, aryl and heteroaryl.
“Alkoxyamino” refers to a radical —N(H)OR where R represents an alkyl or cycloalkyl group as defined herein.
“Alkoxycarbonyl” refers to a radical —C(O)-alkoxy where alkoxy is as defined herein.
“Alkylarylamino” refers to a radical —NRR′ where R represents an alkyl or cycloalkyl group and R′ is an aryl as defined herein.
“Alkylsulfonyl” refers to a radical —S(O)₂R where R is an alkyl or cycloalkyl group as defined herein. Representative examples include, but are not limited to, methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl and the like.
“Alkylsulfinyl” refers to a radical —S(O)R where R is an alkyl or cycloalkyl group as defined herein. Representative examples include, but are not limited to, methylsulfinyl, ethylsulfinyl, propylsulfinyl, butylsulfinyl and the like.
“Alkylthio” refers to a radical —SR where R is an alkyl or cycloalkyl group as defined herein that may be optionally substituted as defined herein. Representative examples include, but are not limited to, methylthio, ethylthio, propylthio, butylthio, and the like.
“Amino” refers to the radical —NH₂.
“Substituted amino” includes those groups recited in the definition of “substituted” herein, and particularly refers to the group —N(R)₂where each R is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, cycloalkyl, substituted cycloalkyl, and where both R groups are joined to form an alkylene group. When both R groups are hydrogen, —N(R)₂is an amino group.
“Aminocarbonyl” refers to the group —C(O)NRR where each R is independently hydrogen, alkyl, aryl and cycloalkyl, or where the R groups are joined to form an alkylene group.
“Aminocarbonylamino” refers to the group —NRC(O)NRR where each R is independently hydrogen, alkyl, aryl or cycloalkyl, or where two R groups are joined to form an alkylene group.
“Aminocarbonyloxy” refers to the group —OC(O)NRR where each R is independently hydrogen, alkyl, aryl or cycloalkyl, or where the R groups are joined to form an alkylene group.
“Arylalkyloxy” refers to an —O-arylalkyl radical where arylalkyl is as defined herein.
“Arylamino” means a radical —NHR where R represents an aryl group as defined herein.
“Aryloxycarbonyl” refers to a radical —C(O)—O-aryl where aryl is as defined herein.
“Arylsulfonyl” refers to a radical —S(O)₂R where R is an aryl or heteroaryl group as defined herein.
“Azido” refers to the radical —N₃.
“Carbamoyl” refers to the radical —C(O)N(R)₂where each R group is independently hydrogen, alkyl, cycloalkyl or aryl, as defined herein, which may be optionally substituted as defined herein.
“Carboxy” refers to the radical —C(O)OH.
“Carboxyamino” refers to the radical —N(H)C(O)OH.
“Cycloalkyl” refers to cyclic hydrocarbyl groups having from 3 to about 10 carbon atoms and having a single cyclic ring or multiple condensed rings, including fused and bridged ring systems, which optionally can be substituted with from 1 to 3 alkyl groups. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, and the like, and multiple ring structures such as adamantanyl, and the like.
“Substituted cycloalkyl” includes those groups recited in the definition of “substituted” herein, and particularly refers to a cycloalkyl group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Cycloalkoxy” refers to the group —OR where R is cycloalkyl. Such cycloalkoxy groups include, by way of example, cyclopentoxy, cyclohexoxy and the like.
“Cycloalkenyl” refers to cyclic hydrocarbyl groups having from 3 to 10 carbon atoms and having a single cyclic ring or multiple condensed rings, including fused and bridged ring systems and having at least one and particularly from 1 to 2 sites of olefinic unsaturation. Such cycloalkenyl groups include, by way of example, single ring structures such as cyclohexenyl, cyclopentenyl, cyclopropenyl, and the like.
“Substituted cycloalkenyl” includes those groups recited in the definition of “substituted” herein, and particularly refers to a cycloalkenyl group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Fused Cycloalkenyl” refers to a cycloalkenyl having two of its ring carbon atoms in common with a second aliphatic or aromatic ring and having its olefinic unsaturation located to impart aromaticity to the cycloalkenyl ring.
“Cyanato” refers to the radical —OCN.
“Cyano” refers to the radical —CN.
“Dialkylamino” means a radical —NRR′ where R and R′ independently represent an alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, heteroaryl, or substituted heteroaryl group as defined herein.
“Ethenyl” refers to substituted or unsubstituted —(C═C)—.
“Ethylene” refers to substituted or unsubstituted —(C—C)—.
“Ethynyl” refers to —(C≡C)—.
“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo. Preferred halo groups are either fluoro or chloro.
“Hydroxy” refers to the radical —OH.
“Nitro” refers to the radical —NO₂.
“Substituted” refers to a group in which one or more hydrogen atoms are each independently replaced with the same or different substituent(s). Typical substituents include, but are not limited to, —X, —R¹⁴, —O⁻, ═O, —OR¹⁴, —SR¹⁴, —S⁻, ═S, —NR¹⁴R¹⁵, ═NR¹⁴, —CX₃, —CF₃, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —S(O)₂O—, —S(O)₂OH, —S(O)₂R¹⁴, —OS(O₂)O⁻, —OS(O)₂R¹⁴, —P(O)(O⁻)₂, —P(O)(OR¹⁴)(O⁻), —OP(O)(OR¹⁴)(OR¹⁵), —C(O)R¹⁴, —C(S)R¹⁴, —C(O)OR¹⁴, —C(O)NR¹⁴R¹⁵, —C(O)O⁻, —C(S)OR¹⁴, —NR¹⁶C(O)NR¹⁴R¹⁵, —NR¹⁶C(S)NR¹⁴R¹⁵, —NR¹⁷C(NR¹⁶)NR¹⁴R¹⁵and —C(NR¹⁶)NR¹⁴R¹⁵, where each X is independently a halogen; each R¹⁴, R¹⁵, R¹⁶and R¹⁷are independently hydrogen, alkyl, substituted alkyl, aryl, substituted alkyl, arylalkyl, substituted alkyl, cycloalkyl, substituted alkyl, cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, substituted heteroarylalkyl, —NR¹⁸R¹⁹, C(O)R¹⁸or —S(O)₂R¹⁸or optionally R¹⁸and R¹⁹together with the atom to which they are both attached form a cycloheteroalkyl or substituted cycloheteroalkyl ring; and R¹⁸and R¹⁹are independently hydrogen, alkyl, substituted alkyl, aryl, substituted alkyl, arylalkyl, substituted alkyl, cycloalkyl, substituted alkyl, cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl.
Examples of representative substituted aryls include the following
In these formulae one of R^6′ and R^7′ may be hydrogen and at least one of R^6′ and R^7′ is each independently selected from alkyl, alkenyl, alkynyl, cycloheteroalkyl, alkanoyl, alkoxy, aryloxy, heteroaryloxy, alkylamino, arylamino, heteroarylamino, NR¹⁰COR¹¹, NR¹⁰SOR¹¹, NR¹⁰SO₂R¹⁴, COOalkyl, COOaryl, CONR¹⁰R¹¹, CONR¹⁰R¹¹, NR¹⁰R¹¹, SO₂NR¹⁰R¹¹, S-alkyl, S-alkyl, SOalkyl, SO₂alkyl, Saryl, SOaryl, SO₂aryl; or R^6′ and R^7′ may be joined to form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms, optionally containing one or more heteroatoms selected from the group N, O or S. R¹⁰, R¹¹, and R¹²are independently hydrogen, alkyl, alkenyl, alkynyl, perfluoroalkyl, cycloalkyl, cycloheteroalkyl, aryl, substituted aryl, heteroaryl, substituted or hetero alkyl or the like.
“Hetero” when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group have been replaced by a nitrogen, oxygen, or sulfur heteroatom. Hetero may be applied to any of the hydrocarbyl groups described above such as alkyl, e.g. heteroalkyl, cycloalkyl, e.g. cycloheteroalkyl, aryl, e.g. heteroaryl, cycloalkenyl, cycloheteroalkenyl, and the like having from 1 to 5, and especially from 1 to 3 heteroatoms.
“Heteroaryl” refers to a monovalent heteroaromatic group derived by the removal of one hydrogen atom from a single atom of a parent heteroaromatic ring system. Typical heteroaryl groups include, but are not limited to, groups derived from acridine, arsindole, carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and the like. Preferably, the heteroaryl group is between 5-20 membered heteroaryl, with 5-10 membered heteroaryl being particularly preferred. Particular heteroaryl groups are those derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole, oxazole, and pyrazine.
Examples of representative heteroaryls include the following:
wherein each Y is selected from carbonyl, N, NR⁴, O, and S.
Examples of representative cycloheteroalkyls include the following
wherein each X is selected from CR⁴, NR⁴, O and S; and each Y is selected from N, NR⁴, O and S, and where R^6′ is R.
Examples of representative cycloheteroalkenyls include the following:
wherein each X is selected from CR⁴, NR⁴, O and S; and each Y is selected from carbonyl, NH, NR⁴, O and S.
Examples of representative aryl having hetero atoms containing substitution include the following:
wherein each X is selected from C(R⁴)₂, NR⁴, O and S; and each Y is selected from carbonyl, NR⁴, O and S.
“Hetero substituent” refers to a halo, O, S or N atom-containing functionality that may be present as an R⁴in a R⁴C group present as substituents directly on A, B, W, X, Y or Z of the compounds of this invention or may be present as a substituent in the “substituted” aryl and aliphatic groups present in the compounds.
Examples of hetero substituents include:
-halo,
—NO₂, —NH₂, —NHR, —N(R)₂,
—NRCOR, —NRSOR, —NRSO₂R, OH, CN, CO₂R,
—CO₂H,
—R—OH, —O—R, —COOR,
—CON(R)₂, —CONROR,
—SO₂H, —R—S, —SO₂N(R)₂,
—S(O)R, —S(O)₂R, wherein each R is independently an aryl or aliphatic, optionally with substitution. Among hetero substituents containing R groups, preference is given to those materials having aryl and alkyl R groups as defined herein. Preferred hetero substituents are those listed above.
As used herein, the term “cycloheteroalkyl” refers to a stable heterocyclic non-aromatic ring and fused rings containing one or more heteroatoms independently selected from N, O and S. A fused heterocyclic ring system may include carbocyclic rings and need only include one heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, piperazinyl, homopiperazinyl, piperidinyl and morpholinyl, and are shown in the following illustrative examples:
optionally substituted with one or more groups selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—. Substituting groups include carbonyl or thiocarbonyl which provide, for example, lactam and urea derivatives. In the examples, M is CR⁷, NR₂, O, or S; Q is O, NR₂or S. R⁷and R⁸are independently selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Dihydroxyphosphoryl” refers to the radical —PO(OH)₂.
“Substituted dihydroxyphosphoryl” includes those groups recited in the definition of “substituted” herein, and particularly refers to a dihydroxyphosphoryl radical wherein one or both of the hydroxyl groups are substituted. Suitable substituents are described in detail below.
“Aminohydroxyphosphoryl” refers to the radical —PO(OH)NH₂.
“Substituted aminohydroxyphosphoryl” includes those groups recited in the definition of “substituted” herein, and particularly refers to an aminohydroxyphosphoryl wherein the amino group is substituted with one or two substituents. Suitable substituents are described in detail below. In certain embodiments, the hydroxyl group can also be substituted.
“Thioalkoxy” refers to the group —SR where R is alkyl.
“Substituted thioalkoxy” includes those groups recited in the definition of “substituted” herein, and particularly refers to a thioalkoxy group having 1 or more substituents, for instance from 1 to 5 substituents, and particularly from 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.
“Sulfanyl” refers to the radical HS—. “Substituted sulfanyl” refers to a radical such as RS— wherein R is any substituent described herein.
“Sulfonyl” refers to the divalent radical —S(O₂)—. “Substituted sulfonyl” refers to a radical such as R—(O₂)S— wherein R is any substituent described herein. “Aminosulfonyl” or “Sulfonamide” refers to the radical H₂N(O₂)S—, and “substituted aminosulfonyl” “substituted sulfonamide” refers to a radical such as R₂N(O₂)S— wherein each R is independently any substituent described herein.
“Sulfone” refers to the group —SO₂R. In particular embodiments, R is selected from H, lower alkyl, alkyl, aryl and heteroaryl.
“Thioaryloxy” refers to the group —SR where R is aryl.
“Thioketo” refers to the group ═S.
“Thiol” refers to the group —SH.
One having ordinary skill in the art of organic synthesis will recognize that the maximum number of heteroatoms in a stable, chemically feasible heterocyclic ring, whether it is aromatic or non aromatic, is determined by the size of the ring, the degree of unsaturation and the valence of the heteroatoms. In general, a heterocyclic ring may have one to four heteroatoms so long as the heteroaromatic ring is chemically feasible and stable.
“Pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly in humans.
“Pharmaceutically acceptable salt” refers to a salt of a compound of the invention that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. Such salts include: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like. The term “pharmaceutically acceptable cation” refers to a non toxic, acceptable cationic counter-ion of an acidic functional group. Such cations are exemplified by sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, and the like.
The term “solvate” refers to forms of the compound that are associated with a solvent, usually by a solvolysis reaction. Conventional solvents include water, ethanol, acetic acid and the like. The compounds of the invention may be prepared e.g. in crystalline form and may be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric solvates and non-stoichiometric solvates.
“Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant, excipient or carrier with which a compound of the invention is administered.
“Preventing” or “prevention” refers to a reduction in risk of acquiring a disease or disorder (i.e., causing at least one of the clinical symptoms of the disease not to develop in a subject that may be exposed to or predisposed to the disease but does not yet experience or display symptoms of the disease).
“Subject” includes humans. The terms “human,” “patient” and “subject” are used interchangeably herein.
“Therapeutically effective amount” means the amount of a compound that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The “therapeutically effective amount” can vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject to be treated.
“Treating” or “treatment” of any disease or disorder refers, in one embodiment, to ameliorating the disease or disorder (i.e., arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment “treating” or “treatment” refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, “treating” or “treatment” refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In yet another embodiment, “treating” or “treatment” refers to delaying the onset of the disease or disorder.
“Prodrugs” refers to compounds, including derivatives of the compounds of the invention, which have cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention which are pharmaceutically active in vivo. Such examples include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like.
Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but in the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well know to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendant on the compounds of this invention are preferred prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters. Preferred are the C₁to C₈alkyl, C₂-C₈alkenyl, aryl, C₇-C₁₂substituted aryl, and C₇-C₁₂arylalkyl esters of the compounds of the invention.
It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed “isomers”. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers”.
Stereoisomers that are not mirror images of one another are termed “diastereomers” and those that are non-superimposable mirror images of each other are termed “enantiomers”. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (−)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture”.
The compounds of this invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art.

The Compounds

As set forth earlier herein, the compounds of the present invention are useful for preventing and/or treating a broad range of conditions, among them, arthritis, Parkinson's disease, Alzheimer's disease, stroke, uveitis, asthma, myocardial infarction, the treatment and prophylaxis of pain syndromes (acute and chronic or neuropathic), traumatic brain injury, acute spinal cord injury, neurodegenerative disorders, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders or conditions in mammals.
In order that the invention described herein may be more fully understood, the following structures representing compounds typical of the invention are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.
Accordingly, additional groups of particular compounds are provided. Thus, and as discussed earlier herein, suitable compounds capable of modifying ion channels in vivo, may be selected from those listed in Table 1, below, and may be prepared either as shown or in the form of a pharmaceutically acceptable salt, solvate or prodrug thereof; and isomers and stereoisomers thereof. All such variants are contemplated herein and are within the scope of the present invention.
In certain aspects, the present invention provides prodrugs and derivatives of the compounds according to the formulae above. Prodrugs are derivatives of the compounds of the invention, which have cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention, which are pharmaceutically active, in vivo. Such examples include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like.
Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well know to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendant on the compounds of this invention are preferred prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters. Preferred are the C₁to C₈alkyl, C₂-C₈alkenyl, aryl, C₇-C₁₂substituted aryl, and C₇-C₁₂arylalkyl esters of the compounds of the invention.

Pharmaceutical Compositions

When employed as pharmaceuticals, the amide compounds of this invention are typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.
Generally, the compounds of this invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.
The pharmaceutical compositions of this invention can be administered by a variety of routes including by way of non limiting example, oral, rectal, transdermal, subcutaneous, intravenous, intramuscular and intranasal. Depending upon the intended route of delivery, the compounds of this invention are preferably formulated as either injectable or oral compositions or as salves, as lotions or as patches all for transdermal administration.
The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the furansulfonic acid compound is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing form.
Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As before, the active compound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like.
Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s), generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as a ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope of this invention.
The compounds of this invention can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.
The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pa., which is incorporated herein by reference.
The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.
The following formulation examples illustrate representative pharmaceutical compositions of this invention. The present invention, however, is not limited to the following pharmaceutical compositions.

Formulation 1

Tablets

A compound of formula I is admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 240-270 mg tablets (80-90 mg of active compound per tablet) in a tablet press.

Formulation 2

Capsules

A compound of formula I is admixed as a dry powder with a starch diluent in an approximate 1:1 weight ratio. The mixture is filled into 250 mg capsules (125 mg of active compound per capsule).

Formulation 3

Liquid

A compound of formula I (125 mg), sucrose (1.75 g) and xanthan gum (4 mg) are blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color are diluted with water and added with stirring. Sufficient water is then added to produce a total volume of 5 mL.

Formulation 4

Tablets

The compound of formula I is admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 450-900 mg tablets (150-300 mg of active compound) in a tablet press.

Formulation 5

Injection

The compound of formula I is dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg/ml.

Formulation 6

Topical

Stearyl alcohol (250 g) and a white petrolatum (250 g) are melted at about 75° C. and then a mixture of a compound of formula I (50 g) methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate (10 g), and propylene glycol (120 g) dissolved in water (about 370 g) is added and the resulting mixture is stirred until it congeals.

Methods of Treatment

The present compounds are used as therapeutic agents for the treatment of conditions in mammals. Accordingly, the compounds and pharmaceutical compositions of this invention find use as therapeutics for preventing and/or treating neurodegenerative, autoimmune and inflammatory conditions in mammals including humans.
In a method of treatment aspect, this invention provides a method of treating a mammal susceptible to or afflicted with a condition associated with arthritis, uveitis, asthma, myocardial infarction, traumatic brain injury, acute spinal cord injury, alopecia (hair loss), inflammatory bowel disease and autoimmune disorders, which method comprises administering an effective amount of one or more of the pharmaceutical compositions just described.
In yet another method of treatment aspect, this invention provides a method of treating a mammal susceptible to or afflicted with a condition that gives rise to pain responses or that relates to imbalances in the maintenance of basal activity of sensory nerves. Compounds have use as analgesics for the treatment of pain of various geneses or etiology, for example acute, inflammatory pain (such as pain associated with osteoarthritis and rheumatoid arthritis); various neuropathic pain syndromes (such as post-herpetic neuralgia, trigeminal neuralgia, reflex sympathetic dystrophy, diabetic neuropathy, Guillian Barre syndrome, fibromyalgia, phantom limb pain, post-masectomy pain, peripheral neuropathy, HIV neuropathy, and chemotherapy-induced and other iatrogenic neuropathies); visceral pain, (such as that associated with gastroesophageal reflex disease, irritable bowel syndrome, inflammatory bowel disease, pancreatitis, and various gynecological and urological disorders), dental pain and headache (such as migraine, cluster headache and tension headache).
In additional method of treatment aspects, this invention provides methods of treating a mammal susceptible to or afflicted with neurodegenerative diseases and disorders such as, for example Parkinson's disease, Alzheimer's disease and multiple sclerosis; diseases and disorders which are mediated by or result in neuroinflammation such as, for example traumatic brain injury, stroke, and encephalitis; centrally-mediated neuropsychiatric diseases and disorders such as, for example depression mania, bipolar disease, anxiety, schizophrenia, eating disorders, sleep disorders and cognition disorders; epilepsy and seizure disorders; prostate, bladder and bowel dysfunction such as, for example urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway disease and disorders such as, for example, allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders which are mediated by or result in inflammation such as, for example rheumatoid arthritis and osteoarthritis, myocardial infarction, various autoimmune diseases and disorders, uveitis and atherosclerosis; itch/pruritus such as, for example psoriasis; alopecia (hair loss); obesity; lipid disorders; cancer; blood pressure; spinal cord injury; and renal disorders method comprises administering an effective condition-treating or condition-preventing amount of one or more of the pharmaceutical compositions just described.
In further aspect of the invention there is provided the present compounds for use in the treatment of the above mentioned conditions; there is also provided use of the present compounds in the treatment of the above mentioned conditions; there is also provided use of the present compounds in the manufacture of a medicament for the treatment of the above mentioned conditions.
Injection dose levels range from about 0.1 mg/kg/hour to at least 10 mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to 96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 2 g/day for a 40 to 80 kg human patient.
For the prevention and/or treatment of long-term conditions, such as neurodegenerative and autoimmune conditions, the regimen for treatment usually stretches over many months or years so oral dosing is preferred for patient convenience and tolerance. With oral dosing, one to five and especially two to four and typically three oral doses per day are representative regimens. Using these dosing patterns, each dose provides from about 0.01 to about 20 mg/kg of the compound or its derivative, with preferred doses each providing from about 0.1 to about 10 mg/kg and especially about 1 to about 5 mg/kg.
Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses.
When used to prevent the onset of a neurodegenerative, autoimmune or inflammatory condition, the compounds or their derivatives of this invention will be administered to a patient at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Patients at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.
The compounds of this invention can be administered as the sole active agent or they can be administered in combination with other agents, including other active derivatives.

General Synthetic Procedures

The compounds of this invention can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures.
Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts, Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and references cited therein.
The target compounds are synthesized by known reactions outlined in the following schemes. The products are isolated and purified by known standard procedures. Such procedures include (but are not limited to) recrystallization, column chromatography or HPLC. The target compounds, for example, may be prepared by the reaction of an appropriately substituted halopyridine with an appropriately functionalized carboxy boronic acid to obtain the desired biaryl carboxylic acid. The carboxylic acid intermediate thus obtained can be conveniently converted to its corresponding amide by activation followed by reacting with an appropriately substituted amine. The products are isolated and purified by known standard procedures. Such procedures include (but are not limited to) recrystallization, column chromatography or HPLC.

Synthesis of intermediate pyridin-2-yl-benzoic acids

Intermediate

1

Synthesis of 3-chloro-[2,3′]bipyridinyl-6′-carboxylic acid

1a) Synthesis of 5-bis(hydroxyl)boron-2-methylpicoline

5-Hydroxy-2-methylpyridine (1.0 g, 9.2 mmol) was dissolved in 20 ml of dichloromethane and stirred at 0° C. To the reaction mixture was added 1.10 ml of anhydrous pyridine (13.8 mmol), followed by 2.32 ml of triflic anhydride (13.8 mmol). The reaction mixture was warmed to room temperature and allowed to stir until completion (monitored by LC-MS/TLC). The mixture was poured into a separatory funnel and washed with water three times. The organic layer was washed with brine, dried over MgSO₄, filtered and evaporated under vacuum. The material was used as crude material for the next step.
The triflate (4.6 mmol) was dissolved in acetonitrile (30 ml) and placed into a 5 ml microwave vessel. To the solution was added 1.5 eq of bis(pinacolato)diboron (6.9 mmol; 1.71 g). The mixture was stirred on a magnetic stir plate until dissolution. To the mixture was added KOAc (13.8 mmol; 1.35 g) and 98 mg of [1,1′-bis(diphenylphosphino)-ferrocene]dichloropalladium(II) (0.03 mol %). The reaction mixture was heated at 160° C. for 2×600 s. After completion (monitored by LC-MS), the acetonitrile was evaporated to give a black solid. The solid was dissolved in DMSO, ms and purified by HPLC to give the boronic acid (580 mg, 92%; 4.2 mmol).
MS: MH+=138

1b) Synthesis of 3-chloro-6′-methyl-[2,3′]bipyridinyl

The above boronic acid (4.2 mmol) was dissolved in acetonitrile (2 ml) and added to a 5 ml microwave vessel. To the solution was 6.9 mmol of 2,3-dichloropyridine (1.01 g), 53 mg of tetrakis(triphenylphosphine)palladium(0). After stirring until dissolution, 13.8 mmol of potassium carbonate (1.90 g) was added, followed by 1 ml of water. The mixture was then heated at 160° C. for 300 seconds. After reaction completion, the solvents were evaporated under vacuum. The target compound was purified by HPLC to give a yellow solid (800 mg; 85%).
MS: MH+=205

1c) Synthesis of 3-chloro-[2,3′]bipyridinyl-6′-carboxylic acid

The above 3-chloro-6′-methyl-[2,3′]bipyridinyl (2.5 mmol) was added to a 5 ml microwave vessel, followed by 3 ml of water. 3.75 mmol of potassium permanganate was added and the mixture was heated at 120° C. for 600 seconds. An additional 3.75 mmol of potassium permanganate was added and the mixture was resubmitted to microwave heating (same temperature) for 600 s. After completion (monitored by LC-MS), the mixture was filtered through celite and the manganese salts were washed with water. The water was evaporated to ˜10 ml and the product was purified by HPLC to give 293 mg as a white solid (51%).
MS: MH+=235

Intermediate 2

Synthesis of 4-(3-chloro-pyridin-2-yl)-3-methoxybenzoic acid

2a) Synthesis of 3-methoxy-4-boronic acid-benzoic acid methyl ester

1.0 g of 4-hydroxy-3-methoxybenzoic acid methyl ester (5.5 mmol) was dissolved in 20 ml of dichloromethane and stirred at 0° C. To the reaction mixture was added 0.66 ml of anhydrous pyridine (8.25 mmol), followed by 1.39 ml of triflic anhydride (8.25 mmol). The reaction mixture was warmed to room temperature and allowed to stir until completion (monitored by LC-MS/TLC). The mixture was poured into a separatory funnel and washed with water three times. The organic layer was washed with brine, dried over MgSO₄, filtered and evaporated under vacuum to give the triflate (used as crude for boronic acid formation).
The triflate (5.5 mmol) was dissolved in acetonitrile (30 ml) and placed into an 80 ml microwave vessel. To the solution was added 1.5 eq of Bis(pinacolato)diboron (8.25 mmol; 2.08 g). The mixture was stirred on a magnetic stir plate until dissolution. To the mixture was added KOAc (16.5 mmol; 1.62 g) and 134 mg of [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.03 mol %). The reaction mixture was heated at 160° C. for 2×600 s. After completion (monitored by LC-MS), the acetonitrile was evaporated to give a black solid. The solid was dissolved in EtOAc and washed with water, brine and dried over MgSO₄. After filtration, the solvent was evaporated under vacuum. The solid material was then dissolved in chloroform and filtered through silica. The chloroform was evaporated to give a dark green solid (used as crude for the next reaction).

2b) Synthesis of 4-(3-chloro-pyridin-2-yl)-3-methoxybenzoic acid

Boronic acid 3 (5.5 mmol) was dissolved in acetonitrile (10 ml) and added to an 80 ml microwave vessel. To the solution was 8.25 mmol of 2,3-dichloropyridine (1.2 g), 63 mg of tetrakis(triphenylphosphine)palladium(0). After stirring until dissolution, 5 ml of a 1 M potassium carbonate solution (aqueous) was added and the mixture was heated at 160° C. for 300 s. After reaction completion, the acetonitrile was evaporated under vacuum and 2 N KOH was added (20 ml), followed by 10 ml of THF. The reaction was heated until hydrolysis was complete (15 minutes). The solution was acidified (cHCl) and extracted 3× with EtOAc. After drying over MgSO₄, the organic layer was filtered and evaporated under vacuum. The compound was purified by HPLC to give a yellow solid (263 mg, 18%).
MS: MH+=264

Intermediate 3

Synthesis of 4-(3-Trifluoromethylpyridin-2-yl)benzoic acid

4-Bis(hydroxyl)boron-1-methylbenzoate (2.8 mmol) was dissolved in acetonitrile (2 ml) and added to a 5 ml microwave vessel. To the solution was 3.5 mmol of 2-chloro-3-trifluoromethylpyridine (633 mg), 34 mg of tetrakis(triphenylphosphine)-palladium(0). After stirring until dissolution, 8.4 mmol of potassium carbonate (1.16 g) was added, followed by 1 ml of water. The mixture was then heated at 160° C. for 300 seconds. After reaction completion, the solvents were evaporated under vacuum. The residue was dissolved in 2N KOH and THF and heated for 10 min. After hydrolysis, the THF was evaporated and the basic layer was washed with EtOAC. The aqueous layer was then acidified and extracted 3 times with EtOAc. The organic layers were combined and washed with water and brine. After drying, filtration and evaporation, the residue was purified by HPLC to give of the target compound as a white solid (602 mg; 81%).
MS: MH+=268

Intermediate 4

3-Fluoro-4-(3-trifluoromethylpyridin-2-yl)benzoic acid

The 4-Bromo-3-fluorobenzoic acid methyl ester (700 mg; 2.45 mmol) was dissolved in acetonitrile (3.0 ml) and placed into a 2 ml microwave vessel. To the solution was added 1.5 eq of Bis(pinacolato)diboron (3.67 mmol; 1.08 g). The mixture was stirred on a magnetic stir plate until dissolution. To the mixture was added KOAc (7.33 mmol; 7.16 mg) and 60 mg of [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.03 mol %). The reaction mixture was heated at 160° C. for 2×600 s. After completion (monitored by LC-MS), the acetonitrile was evaporated to give a black solid. The solid was dissolved in EtOAc and washed with water, brine and dried over MgSO₄. After filtration, the solvent was evaporated under vacuum. The solid material was then dissolved in chloroform and filtered through silica. The chloroform was evaporated to give a dark green solid (used as crude for the next reaction).
The boronic acid (2.45 mmol) was dissolved in acetonitrile (2.4 ml) and 3-(trifluoromethyl)-2-chloropyridine was added. After mixing, tetrakis palladium was added (25 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(912 mg; 3.0 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a white solid (530 mg; 76%).
MS: MH+=286

Intermediate 5

4-(3-Trifluoromethylpyridin-2-yl)-3-methoxybenzoic acid

The boronic acid was prepared as described in Intermediate 2 to give 410 mg (78%). The boronic acid (2.55 mmol) was dissolved in acetonitrile (2.4 ml) and 3-(trifluoromethyl)-2-chloropyridine was added. After mixing, tetrakis palladium was added (29 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(912 mg; 6.6 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a white solid (495 mg; 68%).
MS: MH+=298

Intermediate 6

4-(3-Methoxypyridin-2-yl)-3-fluorobenzoic acid

The boronic acid was prepared as described in Intermediate 4 to give 2.2 mmole. (quant. crude). The boronic acid (2.2 mmol) was dissolved in acetonitrile (2.4 ml) and 3-methoxy-2-chloropyridine (380 mg; 2.88 mmol) was added. After mixing, tetrakis palladium was added (25 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(912 mg; 6.6 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a beige solid (312 mg; 57%).
MS: MH+=248

Intermediate 7

4-(3-Methoxypyridin-2-yl)benzoic acid

4-bis(hydroxyl)boron-1-methylbenzoate (2.8 mmol; 582 mg) was dissolved in acetonitrile (2 ml) and added to a 5 ml microwave vessel. To the solution was 3.5 mmol of 2-chloro-3-methoxypyridine (482 mg), 34 mg of tetrakis(triphenylphosphine)palladium(0). After stirring until dissolution, 8.4 mmol of potassium carbonate (1.16 g) was added, followed by 1 ml of water. The mixture was then heated at 160° C. for 300 seconds. After reaction completion, the solvents were evaporated under vacuum. The residue was dissolved in 2N KOH and THF and heated for 10 min. After hydrolysis, the THF was evaporated and the basic layer was washed with EtOAC. The aqueous layer was then acidified and extracted 3 times with EtOAc. The organic layers were combined and washed with water and brine. After drying, filtration and evaporation, the residue was titrated with ether to give the desired product as a yellow solid (423 mg; 77%).
MS: MH+=230

Intermediate 8

4-(3-Trifluoromethylpyridin-2-yl)-3-chlorobenzoic acid

8a) 4-bromo-3-chloromethylbenzoate

The 4-bromo-3chlorotoluene (4.0 g; 19.5 mmol) was added to a 250 ml round bottom flask, followed by 50 ml of water. To the mixture was added 3.22 g of potassium permanganate (23.4 mmol) and the reaction was refluxed until completion. After cooling, the mixture was filtered through celite. The aqueous layer was acidified and extracted three times with ethylacetate. The organic layers were washed with brine, dried over MgSO₄. After filtration and evaporation, the resulting white solid was dissolved in 1.0M HCl in ether and stirred overnight. The methanol was removed under vacuum and the residue was dissolved in ethylacetate and washed with saturated sodium bicarbonate. The organic layer was dried over MgSO₄, filtered and evaporated to give the desired material as a clear oil (713 mg; 15%).

8b) 4-(3-Trifluoromethylpyridin-2-yl)-3-chlorobenzoic acid

The boronic acid was prepared as described in Intermediate 4 to give 2.8 mmole. (quant. crude). The boronic acid (2.8 mmol) was dissolved in acetonitrile (2.4 ml) and 2-chloro3-trifluoromethylpyridine (620 mg; 3.3 mmol) was added. After mixing, tetrakis palladium was added (33 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(1182 mg; 6.6 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a white solid (460 mg; 55%).
MS: MH+=303

Intermediate 9

4-(3-methoxypyridin-2-yl)-3-methoxybenzoic acid

The boronic acid was prepared as described in Intermediate 5 (2.5 mmol; crude) and was dissolved in acetonitrile (2 ml) and added to a 5 ml microwave vessel. To the solution was 3.0 mmol of 2-chloro-3-methoxypyridine (432 mg), 29 mg of tetrakis(triphenylphosphine)palladium(0). After stirring until dissolution, 7.5 mmol of potassium carbonate (1.06 g) was added, followed by 1 ml of water. The mixture was then heated at 160° C. for 300 seconds. After reaction completion, the solvents were evaporated under vacuum. The residue was dissolved in 2N KOH and THF and heated for 10 min. After hydrolysis, the THF was evaporated and the basic layer was washed with EtOAC. The aqueous layer was then acidified and extracted 3 times with EtOAc. The organic layers were combined and washed with water and brine. After drying, filtration and evaporation, the residue was titrated with ether to give the desired product as a yellow solid (310 mg; 47%).
MS: MH+=260

Intermediate 10

4-(3-Methoxypyridin-2-yl)-3-chlorobenzoic acid

The boronic acid was prepared as described in Intermediate 4 to give 2.8 mmole. (quant. crude). The boronic acid (2.14 mmol) was dissolved in acetonitrile (2.4 ml) and 3-methoxy-2-chloropyridine (368 mg; 2.6 mmol) was added. After mixing, tetrakis palladium was added (25 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(887 mg; 6.5 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a yellow solid (425 mg; 76%).
MS: MH+=264

Intermediate 11

4-(3-Chloropyridin-2-yl)-3-fluorobenzoic acid

The boronic acid was prepared as described in Intermediate 4 (2.45 mmol) was dissolved in acetonitrile (2.4 ml) and 2,3-dichloropyridine was added. After mixing, tetrakis palladium was added (25 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(912 mg; 3.0 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a white solid (403 mg; 56%).
MS: MH+=252

Intermediate 12

4-(3-Chloropyridin-2-yl)-2-aminobenzoic acid

12a) 4-Bromo-2-aminomethylbenzoate

4-Bromo-2-nitromethylbenzoate (300 mg; 1.15 mmol) was dissolved in 25 ml of methanol and shaken with 5% Pd(c) under hydrogen atmosphere (50 PSI) for 1 hour. The reaction was filtered through celite and evaporated to give the product as a white solid (255 mg; 96%).
MS: MH+=230

12b) 4-(3-Chloropyridin-2-yl)-2-aminobenzoic acid

The boronic acid was prepared as described in Intermediate 4 to give 3.4 mmole. (quant. crude). The boronic acid 656 mg (3.4 mmol) was dissolved in acetonitrile (2.4 ml) and 2,3-dichloropyridine (600 mg; 4.08 mmol) was added. After mixing, tetrakis palladium was added (40 mg; 0.01 mol %), followed by 0.8 ml of water and K₂CO₃(1.40 g; 10.8 mmol). The reaction mixture was heated at 160° C. in a Personal Chemistry Emrys Microwave for 300 s. After reaction completion, the solvents were evaporated and the residue was dissolved in EtOAc and washed with water and brine. The organic layer was dried over MgSO₄, filtered and evaporated. The residue was then dissolved in a 1:1 mixture of THF/2NKOH and heated until saponification was complete. The basic layer was then extracted with EtOAc and acidified with conc. HCl. The aqueous layer was then extracted three times with EtOAc. The combined organic layers were dried over MgSO₄, filtered and evaporated to give the desired material as a yellow solid (466 mg; 55%).
MS: MH+=249

Intermediate 13

4-(3-Chloropyridin-2-yl)-2-N-methylaminobenzoic acid

13a) 4-(3-Chloropyridin-2-yl)-2-(trifluoromethylacetamide)methylbenzoate

The 4-(3-Chloropyridin-2-yl)-2aminomethylbenzoate (1.43 mg; 0.55 mmol) was dissolved in 1,2-dichloroethane (5 ml). To the mixture was added trifluoroacetic anhydride (7.2 mmol; 11.0 ml) and pyridine (7.7 mmol; 0.63 ml) and the reaction was stirred for 2 hrs. The solution was diluted with dichloromethane and washed with water. The organic layer was dried over MgSO₄, filtered and evaporated to give the desired product as a white solid (1.5 g; 79%).
MS: MH+=359

13b) 4-(3-Chloropyridin-2-yl)-2-N-methylaminobenzoic acid

4-(3-Chloropyridin-2-yl)-2-(trifluoromethylacetamide)methylbenzoate (540 mg; 1.5 mmol) was dissolved in dry tetrahydrofuran. Triphenylphosphine (446 mg; 1.7 mmol) and DIAD (0.36 ml; 1.7 mmol), followed by anhydrous methanol (0.07 ml; 1.7 mmol). The THF was evaporated and the residue was dissolved in EtOAc. The organic layer was washed with water, brine, dried over MgSO₄, filtered and dried. The crude material was purified by flash chromatography using 2:1 hexanes:ethylacetate to give 465 mg of the desired product (83%).
The methylated product (464 mg; 1.24 mmol) was dissolved in methanol (8 ml) and 4 ml of aqueous lithium hydroxide (4 ml; 10%). The methanol was evaporated and the mixture was acidified and extracted with EtOAc. The organic layer was washed with water, dried over MgSO₄, filtered and evaporated to give the desired product as a yellow solid (338 mg; 91%).
MS: MH+=263

Intermediate 14

4-(3-Chloropyridin-2-yl)-2-N-methoxyethylaminobenzoic acid

The 4-(3-Chloropyridin-2-yl)-2-(trifluoromethylacetamide)methylbenzoate (1.2 g; 3.3 mmol) was dissolved in dry tetrahydrofuran. Triphenylphosphine (2.2 g; 8.3 mmol) and DIAD (1.7 ml; 8.3 mmol), followed by anhydrous methoxyethanol (0.66 ml; 8.3 mmol). The THF was evaporated and the residue was dissolved in EtOAc. The organic layer was washed with water, brine, dried over MgSO₄, filtered and dried. The crude material was purified by flash chromatography using 2:1 hexanes:ethylacetate to give 676 mg of the desired product (66%).
The alkylated product (525 mg; 1.24 mmol) was dissolved in methanol (8 ml) and 4 ml of aqueous lithium hydroxide (4 ml; 10%). The methanol was evaporated and the mixture was acidified and extracted with EtOAc. The organic layer was washed with water, dried over MgSO₄, filtered and evaporated to give the desired product as a yellow solid (260 mg; 66%).
MS: M+=307

Intermediate 15

4-(3-Chloropyridin-2-yl)-2-N-benzylaminobenzoic acid

The 4-(3-Chloropyridin-2-yl)-2-(trifluoromethylacetamide)methylbenzoate (436 m; 1.2 mmol) was dissolved in dry tetrahydrofuran. Triphenylphosphine (349 mg; 1.33 mmol) and DIAD (0.28 ml; 1.33 mmol), followed by anhydrous benzylalcohol (0.14 ml; 1.33 mmol). The THF was evaporated and the residue was dissolved in EtOAc. The organic layer was washed with water, brine, dried over MgSO₄, filtered and dried. The crude material was purified by flash chromatography using 2:1 hexanes:ethylacetate to give 435 mg of the desired product (80%).
The alkylated product (68 mg; 0.15 mmol) was dissolved in methanol (6 ml) and 4 ml of aqueous lithium hydroxide (2 ml; 10%). The methanol was evaporated and the mixture was acidified and extracted with EtOAc. The organic layer was washed with water, dried over MgSO₄, filtered and evaporated to give the desired product as a yellow solid (51 mg; 75%).
MS: M+=339

Intermediate 16

4-(3-Chloropyridin-2-yl)benzoic acid

Tetrakis palladium (0.12 g, 0.1 mMol) was added to a suspension of carboxybenzene boronic acid (0.33 g, 2.0 mMol) and 2,3-dichloropyridine (0.296 g, 2.0 mMol) in a mixture of 0.4M K₂CO₃(10 mL) and acetonitrile (10 mL) and the mixture was heated at 90° C. for 12 hrs under a blanket of nitrogen. The hot suspension was filtered, the filtrate concentrated to about half the original volume before being washed with methylene chloride. The aq. layer was carefully acidified with conc. HCl and the precipitate was collected, washed with water and vacuum dried to obtain the product as a white solid.
MS: m/z=232 (M−1)
Following the procedure described above for Intermediate 1-16 and the appropriate reagents, starting materials and purification methods known to those skilled in the art, the other benzoic acids, employed in synthesizing amide compounds of this invention, were prepared.

Amidation of Carboxylic Acid

EXAMPLE 1

A Representative Synthesis of Benzamide

4-(3-Chloropyridin-2-yl)-N-(4-trifluoromethyl-phenyl)benzamide (Compound 18)

To a suspension of 4-(3-chloro-pyridin-2-yl)-benzoic acid (5.0 g, 21.4 mmol) in methylene chloride (100 mL) at ambient temperature was added oxalyl chloride (5.43 g, 42.79 mmol) followed by two drops of DMF and the mixture was heated to reflux for 30 minutes. The clear solution was then concentrated to dryness, dissolved in methylene chloride (100 mL) and was treated with 4-trifluoromethyl aniline (4.14 g, 25.68 mmol) followed by triethylamine (2.6 g, 25.68 mmol) and the mixture was gently heated to reflux for 30 minutes and agitated overnight at ambient temperature. After treating the mixture with sat. Na₂CO₃, the organic layer was separated, washed with water, dried and concentrated to give the crude product which was chromatographed on silica gel to obtain 4.0 g (49.6%) of the title compound as a white solid.
MS: m/z=367 (M+1)
¹H NMR (DMSO-d6): δ 7.45-7.53 (m, 1H); 7.75 (d, 8.8 Hz, 2H); 7.86 (d, 5.2 Hz, 2H); 8.04-8.13 (m, 5H); 8.67 (dd, 4.8 Hz, 1.6 Hz, 1H); 10.72 (s, 1H).

EXAMPLE 2

A Representative Synthesis of Benzamides Using an Automated Parallel Synthesis Method

The appropriate benzoic acid (2 mmol) was dissolved or suspended in 15 ml of chloroform and treated with 20 mmol of thionyl chloride. The reaction mixture was refluxed for fifteen minutes and the solvents were removed under vacuum. The residue was dissolved in 4 ml of anhydrous chloroform and 60 μl (30 μmole) of this solution was added to each well of the 96 well glass plates. Appropriate amine was then added to the corresponding well (60 μmole), followed by N,N-diisopropylethylamine (120 μmole). The plate was then heated at 65° C. for 15 minutes. The solvents were removed using an HT-12 Genevac centrifugal evacuator and 100 μl of DMSO was added to each well and the compounds were transferred to a 96-well polypropylene reaction plate. The plates were then sealed using an ABgene plate sealer and submitted to LC-MS purification.

General Method for Automated Parallel LC-MS Purification of Libraries

The libraries were purified using a Perkin Elmer API100 mass spectrometer coupled to Shimadzu LC pumps. The chromatographic method employed was 10-100% gradient of acetonitrile to water over 8 minutes at a flow rate of 6 ml per minute. The column used was a 10×50 mm YMC C18 and the compounds were collected using a Gilson 204 fraction collector.
Following the procedure described above for Example 1 or 2 and the appropriate reagents, starting materials and purification methods known to those skilled in the art, the amide compounds of this invention were prepared.
The following synthetic and biological examples are offered to illustrate this invention and are not to be construed in any way as limiting the scope of this invention. In the examples below, all temperatures are in degrees Celsius (unless otherwise indicated).
The compounds that have been prepared in accordance with the invention, are presented in tabular form below. The syntheses of these representative compounds were carried out in accordance with the methods set forth above.

Exemplary Compounds of the Invention

The following compounds have been prepared according to the methods of the invention.

TABLE 1

AMIDE COMPOUNDS

					HPLC	HPLC
				HPLC	START	END	%
		MW	MS	RT	TIME	TIME	Inhibition @
ID	STRUCTURE	(Calc)	(Obs)	(Min)	(Min)	(Min)	1 μM

1	288.78	289.07	2.92	2.85	3.35

2	322.80	323.12	2.99	2.92	3.36

3	352.82	353.23	3.08	2.95	3.39	+

4	312.76	313.08	2.75	2.68	3.11	+

5	276.72	277.08	1.99	1.86	2.39

6	372.86	373.10	3.39	3.29	4.07	+

7	323.78	324.28	1.99	1.89	2.22

8	302.81	303.08	3.13	3.03	3.66	+

9	371.27	371.04	3.31	3.22	3.75	+++

10	288.78	289.07	2.86	2.79	3.13

11	274.75	275.11	2.58	2.43	3.02

12	316.79	317.09	2.50	2.43	2.88

13	366.81	367.09	2.96	2.88	3.45

14	352.82	353.22	3.00	2.88	3.31	+

15	390.80	391.26	3.39	3.32	3.76	++

16	415.90	416.29	2.48	2.42	2.81	+

17	412.88	413.22	2.82	2.75	3.22	+

18	396.88	397.14	2.85	2.71	3.31	++

19	260.73	261.06	2.35	2.28	2.65	+

20	328.82	329.14	2.92	2.85	3.22	+

21	288.78	289.07	2.82	2.73	3.22	+

22	382.85	383.14	2.78	2.68	3.19	+

23	340.79	341.06	3.09	3.00	3.52	+

24	391.69	393.02	3.51	3.36	3.88	+

25	323.78	324.28	1.97	1.89	2.20

26	274.75	275.11	2.58	2.49	2.91	+

27	314.82	315.03	3.11	3.00	3.42

28	362.86	363.34	3.41	3.29	3.71

29	342.87	343.19	3.45	3.32	3.99

30	340.82	341.09	1.97	1.87	2.29

31	398.90	399.10	3.58	3.51	4.09

32	329.83	330.18	2.02	1.83	2.50	+

33	370.93	371.16	3.99	3.78	4.45	++++

34	405.93	406.30	2.32	2.23	2.66	+++

35	345.83	346.12	1.95	1.85	2.39

36	300.79	301.21	2.89	2.79	3.13

37	272.74	273.07	2.38	2.30	2.75	+

38	323.78	324.28	1.97	1.89	2.39

39	375.86	376.16	3.08	2.90	3.49	+

40	344.85	345.09	1.83	1.72	1.98

41	366.89	367.17	3.72	3.63	4.22	+

42	329.83	330.19	1.96	1.85	2.20

43	326.79	327.19	1.95	1.83	2.23	+

44	365.87	366.21	2.13	2.05	2.39

45	364.88	365.09	3.58	3.48	4.37	++

46	303.37	304.16	1.77	1.73	1.97

47	240.31	241.08	1.72	1.66	1.92	+

48	254.33	255.36	1.99	1.92	2.28

49	280.37	281.33	2.20	2.09	2.46

50	318.38	319.05	2.22	2.16	2.49

51	278.31	279.19	1.89	1.83	2.16

52	242.28	243.25	1.35	1.27	1.46	+

53	331.42	332.28	1.55	1.43	1.73

54	338.41	339.21	2.56	2.46	2.88	+++

55	328.42	329.24	2.53	2.48	2.66

56	308.43	309.39	2.60	2.46	2.95

57	289.34	290.22	1.37	1.26	1.49	+

58	306.37	307.29	1.42	1.28	1.50	++

59	364.45	365.09	2.75	2.68	3.00	+

60	295.39	296.28	1.43	1.29	1.50	+

61	268.36	269.31	2.25	2.16	2.45

62	336.82	337.33	2.46	2.29	2.76

63	336.48	337.48	2.99	2.89	3.33	++++

64	288.35	289.11	2.13	2.06	2.48	+

65	254.33	255.36	1.99	1.87	2.33

66	240.31	241.08	1.76	1.69	1.99

67	274.33	275.17	2.19	2.09	2.39

68	304.35	305.27	2.19	2.10	2.58	++

69	292.32	293.07	2.29	2.23	2.59

70	275.31	276.27	1.55	1.42	1.79

71	275.31	276.26	1.55	1.40	1.77

72	264.29	265.06	1.55	1.48	1.85	++

73	281.34	282.27	1.98	1.92	2.12

74	282.35	283.34	1.67	1.60	1.98

75	371.49	372.19	1.79	1.65	2.08

76	332.36	333.22	2.16	2.03	2.36

77	311.39	312.23	1.39	1.29	1.52	+

78	337.31	338.29	2.59	2.52	2.75

79	319.32

80	318.38	319.05	2.20	2.09	2.52

81	356.35	357.09	2.59	2.52	2.89	++

82	381.46	382.17	1.79	1.72	2.07	+

83	378.43	379.32	2.05	1.99	2.36	+

84	266.35	267.07	2.02	1.92	2.26

85	238.29	239.06	1.60	1.50	1.96

86	289.34	290.22	1.40	1.17	1.50	+

87	362.43	363.38	2.10	2.05	2.43	+

88	270.33	271.29	1.62	1.59	1.81

89	316.41

90	341.42	342.22	2.32	2.16	2.58

91	290.32	291.07	2.04	1.95	2.21

92	310.40

93	226.28	227.25	1.56	1.36	1.77

94	294.38	295.18	2.05	1.99	2.29	++

95	299.33	300.21	2.05	1.97	2.26	++++

96	317.39	318.13	1.68	1.63	1.88

97	345.45	346.24	1.76	1.69	1.92	+

98	308.77	309.28	2.58	2.50	2.79	++

99	304.35	305.28	2.31	2.22	2.57	++++

100	316.41	317.19	2.53	2.48	2.76

101	304.35	305.30	2.26	2.20	2.72	+

102	359.43	360.29	1.79	1.72	1.97

103	308.77	309.28	2.35	2.16	2.65

104	289.34	290.18	1.66	1.62	1.83

105	332.45	333.27	2.73	2.65	2.89

106	254.33	255.36	1.96	1.86	2.23	++

107	295.39	296.34	1.40	1.24	1.57	+

108	343.22	343.09	2.72	2.62	2.98

109	348.41	349.27	2.00	1.93	2.28	+

110	306.34	307.25	2.25	2.13	2.39

111	357.24	357.09	2.66	2.54	2.96	+++

112	338.80	339.09	2.45	2.38	2.69

113	342.32	343.10	2.82	2.72	3.23	++

114	343.22	343.09	2.69	2.55	3.00	+

115	376.77	377.26	3.01	2.92	3.28

116	353.22	353.09	2.63	2.55	2.85	+

117	299.33	300.24	2.26	2.19	2.53	++++

118	318.38	319.08	1.97	1.90	2.15	++++

119	330.43	331.26	2.93	2.86	3.26

120	316.41	317.19	2.80	2.69	3.05	+

121	330.43	331.27	3.06	2.93	3.41	++++

122	356.47	357.21	3.28	3.18	3.49

123	372.35	373.10	2.86	2.79	3.52

124	302.38	303.08	2.38	2.29	2.68	+

125	332.36	333.22	2.19	2.10	2.35

126	313.36	314.14	2.09	2.02	2.28

127	276.30

128	289.34	290.19	1.55	1.50	1.79

129	354.21	356.09	2.43	2.28	2.59	+

130	343.31	344.13	2.65	2.58	2.75

131	377.76	378.18	2.42	2.32	2.52

132	303.37	304.17	1.80	1.70	2.06

133	268.36	269.31	2.19	1.79	2.65

134	309.76	310.28	1.92	1.85	2.16

135	309.76	310.28	2.15	2.09	2.42	++++

136	305.34	306.28	1.89	1.85	2.20	+

137	309.76	310.68	0.82	0.67	0.83

138	318.34	319.05	2.16	2.12	2.49	+

139	304.35	305.35	1.55	1.42	1.83

140	325.37	326.28	1.73	1.69	1.93

141	391.48	392.27	2.93	2.88	3.25

142	292.34	293.07	1.42	1.29	1.47	++

143	289.34	290.18	1.39	1.30	1.50	++++

144	362.44	363.36	2.93	2.88	3.13	+++

145	319.32	320.12	2.48	2.32	2.65	++

146	324.39	325.29	2.70	2.48	3.06	+

147	330.43	331.26	2.75	2.63	3.13	+

148	317.39	318.12	1.79	1.72	2.12

149	254.33	255.35	1.80	1.65	2.16	++

150	268.36	269.31	2.06	1.99	2.46	++

151	294.40	295.28	2.25	2.17	2.65

152	332.41	333.25	2.26	2.17	2.66	++

153	292.34	293.07	1.95	1.86	2.36

154	256.31	257.30	1.46	1.37	1.79	+

155	345.45	346.20	1.60	1.45	1.93	++

156	352.44	353.28	2.56	2.45	2.91	+

157	342.44	343.22	2.53	2.45	2.83	+

158	322.45	323.25	2.60	2.46	2.96	++

159	303.37	304.14	1.46	1.40	1.60	++

160	320.40	321.25	1.48	1.41	1.71	+++

161	378.48	379.33	2.73	2.58	3.06	++

162	309.41	310.42	1.49	1.36	1.65

163	282.39	283.38	2.28	2.16	2.75	+

164	350.85	351.30	2.49	2.25	2.85	+

165	350.51	351.43	2.99	2.86	3.31

166	302.38	303.08	2.18	2.08	2.48

167	268.36	269.31	2.05	1.99	2.35

168	254.33	255.35	1.86	1.78	1.98

169	288.35	289.11	2.23	2.12	2.53

170	318.38	319.08	2.22	2.12	2.38

171	306.34	307.25	2.33	2.17	2.69

172	289.34	290.20	1.65	1.49	1.87

173	289.34	290.21	1.56	1.50	1.86	++

174	278.32	279.20	1.69	1.55	1.95	++

175	295.37	296.26	2.02	1.95	2.26	+++

176	296.37	297.35	1.76	1.65	2.06	++

177	385.51	386.26	1.82	1.72	2.09

178	346.39	347.14	2.18	2.09	2.46

179	325.41	326.28	1.46	1.16	1.85	++

180	351.34	352.29	2.52	2.40	2.79

181	333.35	334.18	2.46	2.40	2.69	+

182	332.41	333.25	2.22	2.15	2.69	+

183	370.38	371.10	2.63	2.59	2.78

184	395.48	396.11	1.86	1.80	2.05

185	392.46	393.25	2.09	2.00	2.46

186	280.37	281.31	2.06	1.98	2.39

187	252.32	253.16	1.70	1.59	2.00	+

188	303.37	304.16	1.47	1.40	1.92	++

189	376.46	377.34	2.13	2.06	2.43	+

190	284.36	285.24	1.72	1.59	1.97

191	330.43	331.26	2.43	2.35	2.71	++

192	355.44	356.21	2.33	2.25	2.65	++

193	304.35	305.29	2.09	2.03	2.40	++

194	324.43

195	240.31	241.06	1.66	1.46	2.03	+++

196	308.41	309.30	2.12	2.03	2.45	++++

197	313.36	314.12	2.06	1.93	2.25

198	331.42	332.28	1.72	1.59	1.97	+++

199	359.48	360.29	1.79	1.70	2.15

200	322.80	323.14	2.56	2.42	2.85	++++

201	318.38	319.06	2.32	2.19	2.72

202	330.43	331.26	2.55	2.32	2.90	+

203	318.38	319.06	2.29	2.13	2.79

204	373.46	374.22	1.85	1.69	1.93

205	322.80	323.14	2.33	2.19	2.65	+

206	303.37	304.15	1.67	1.56	2.00

207	346.48	347.25	2.72	2.62	3.11

208	268.36	269.30	2.02	1.92	2.32	+

209	309.41	310.43	1.46	1.24	1.59	+++

210	357.24	357.09	2.65	2.55	2.95	+

211	362.43	363.37	2.05	1.90	2.38

212	320.37	321.22	2.26	2.18	2.63	++

213	371.27	371.05	2.66	2.58	2.92	+

214	352.82	353.22	2.46	2.39	2.86	++

215	356.35	357.09	2.78	2.70	3.05	+

216	357.24	357.09	2.59	2.49	2.99	+

217	390.80	391.26	2.93	2.72	3.21	+

218	367.25	369.04	2.61	2.51	2.98	++++

219	313.36	314.12	2.28	2.19	2.60	++++

220	332.41	333.25	2.04	1.86	2.41	+

221	344.46	345.09	2.92	2.82	3.28	++

222	330.43	331.26	2.79	2.73	3.13	++

223	344.46	345.09	3.04	2.82	3.36	++++

224	370.50	371.18	3.25	3.13	3.55

225	386.38	387.20	2.82	2.72	3.12

226	316.41	317.20	2.41	2.31	2.74	+

227	346.39	347.14	2.20	1.96	2.55	+++

228	327.39	328.28	2.13	2.00	2.46	+++

229	290.33

230	303.37	304.16	1.65	1.55	2.08	+

231	368.24	370.07	2.43	2.23	2.65	+

232	357.34	358.17	2.62	2.48	2.82	+

233	391.78	392.10	2.44	2.36	2.65	+

234	317.39	318.12	1.82	1.76	2.05

235	282.39	283.38	2.23	2.16	2.52

236	323.78	324.28	1.96	1.86	2.06

237	323.78	324.28	2.16	2.05	2.46	+

238	319.37	320.14	1.95	1.88	2.20

239	323.78	324.28	2.16	2.07	2.49	+

240	332.36	333.22	2.20	2.09	2.63	+

241	318.38	319.08	1.65	1.57	1.92

242	339.40	340.18	1.78	1.72	2.02	+

243	405.50	406.35	2.93	2.79	3.19	++

244	306.37	307.28	1.47	1.39	1.59	+

245	303.37	304.15	1.47	1.37	1.82	+

246	376.46	377.34	2.93	2.88	3.12

247	333.35	334.18	2.48	2.33	2.72

248	338.41	339.20	2.68	2.60	2.88	+

249	344.46	345.09	2.75	2.66	3.12

250	303.37	304.16	1.70	1.65	1.92

251	240.31	241.07	1.72	1.67	1.82

252	254.33	255.40	1.99	1.89	2.25

253	280.37	281.35	2.19	2.10	2.53

254	318.38	319.08	2.19	2.10	2.56

255	278.31	279.21	1.83	1.76	2.11	+

256	242.28	243.27	1.32	1.19	1.42	++

257	331.42	332.28	1.50	1.42	1.75	++

258	338.41	339.22	2.49	2.40	2.88

259	328.42	329.29	2.45	2.35	2.79

260	308.43	309.43	2.50	2.38	2.90	+

261	289.34	290.27	1.30	1.09	1.42	+

262	306.37

263	364.45	365.15	2.69	2.56	2.92	+

264	295.39	296.31	1.35	1.23	1.67	+

265	268.36	269.33	2.22	2.05	2.60

266	336.82	337.37	2.42	2.33	2.69	+++

267	336.48	337.49	2.96	2.80	3.32

268	288.35	289.14	2.11	1.98	2.41

269	254.33	255.40	1.98	1.89	2.26

270	240.31	241.10	1.73	1.63	1.95

271	274.33	275.18	2.18	2.09	2.39

272	304.35	305.31	2.17	2.10	2.33

273	292.32	293.07	2.28	2.10	2.50

274	275.31	276.27	1.50	1.40	1.73

275	275.31	276.25	1.47	1.40	1.66	+

276	264.29	265.06	1.57	1.45	1.85	+

277	281.34	282.27	1.95	1.87	2.12	+++

278	282.35	283.38	1.65	1.56	1.96	+

279	371.49	372.23	1.73	1.63	2.00

280	332.36	333.25	2.09	2.02	2.52

281	311.39	312.28	1.32	1.09	1.43	+

282	337.31	338.29	2.46	2.40	2.65	+

283	319.32	320.12	2.40	2.35	2.58

284	318.38	319.08	2.12	2.08	2.51	+

285	356.35	357.09	2.52	2.42	2.88	+

286	381.46	382.24	1.75	1.60	1.92	+

287	378.43	379.34	2.00	1.93	2.30

288	266.35	267.10	1.98	1.92	2.31

289	238.29	239.06	1.56	1.50	1.79

290	289.34	290.27	1.32	1.23	1.62	+

291	362.43	363.42	2.06	1.99	2.39	+

292	270.33	271.33	1.59	1.53	1.81	+

293	316.41	317.22	2.36	2.26	2.65	+

294	341.42	342.25	2.25	2.07	2.59

295	290.32	291.07	2.01	1.91	2.33

296	310.40

297	226.28	227.26	1.53	1.42	1.77	+

298	294.38	295.20	2.00	1.93	2.31	+++

299	299.33	300.26	2.02	1.95	2.35	+

300	317.39	318.13	1.62	1.56	1.95	+

301	345.45	346.22	1.69	1.60	2.15	++

302	308.77	309.30	2.52	2.40	2.72	+

303	304.35	305.31	2.25	2.13	2.59	+

304	316.41	317.22	2.48	2.36	2.86

305	304.35	305.30	2.24	2.14	2.55	+

306	359.43	360.29	0.72	0.63	0.86

307	308.77	309.29	2.26	2.10	2.58

308	289.34	290.21	1.59	1.55	1.81

309	332.45	333.30	2.65	2.59	2.92

310	254.33	255.40	1.93	1.85	2.16	+

311	295.39	296.33	1.30	1.00	1.57	+

312	343.22	343.09	2.62	2.52	2.80	+

313	348.41	349.30	1.99	1.93	2.25	+

314	306.34	307.27	2.19	2.12	2.45	+

315	357.24	357.09	2.58	2.49	2.91	+

316	338.80	339.13	2.39	2.29	2.72	++++

317	342.32	343.09	2.73	2.62	3.09

318	343.22	343.09	2.56	2.45	2.95	+

319	376.77	377.27	2.88	2.82	3.18	+

320	353.22	355.12	2.58	2.49	2.85

321	299.33	300.24	2.22	2.12	2.39

322	318.38	319.08	1.96	1.88	2.29	+

323	330.43	331.27	2.88	2.80	3.16

324	316.41	317.22	2.75	2.66	3.09	+

325	330.43	331.28	3.00	2.75	3.26	+

326	356.47	357.22	3.21	2.95	3.38	+

327	372.35	373.10	2.78	2.68	3.05

328	302.38	303.08	2.36	2.26	2.69	+

329	332.36	333.24	2.13	2.08	2.46

330	313.36	314.14	2.05	1.95	2.28	+

331	276.30	277.12	1.50	1.40	1.72

332	289.34	290.22	1.50	1.45	1.73

333	354.21	354.11	2.39	2.32	2.53	+

334	343.31	344.13	2.65	2.45	2.85

335	377.76

336	303.37	304.15	1.72	1.67	2.03

337	268.36	269.34	2.16	2.09	2.40	++

338	309.76	310.28	1.86	1.70	1.99

339	309.76	310.28	2.11	1.98	2.31	+

340	305.34	306.28	1.86	1.81	2.12

341	309.76	310.28	2.08	1.99	2.22	+

342	318.34	319.04	2.13	2.03	2.46	+

343	304.35	305.31	1.50	1.43	1.73

344	325.37	326.28	1.67	1.62	1.87	+

345	391.48	392.29	2.87	2.77	3.19

346	292.34	293.07	1.35	1.04	1.62	+

347	289.34	290.24	1.35	1.26	1.45	+

348	362.44	363.38	2.90	2.85	3.06

349	319.32	320.12	2.42	2.35	2.60

350	324.39	325.32	2.62	2.49	2.90	+

351	330.43	331.26	2.66	2.56	3.14

352	371.37	372.09	2.82	2.65	3.08	+

353	308.31	309.31	3.13	3.02	3.38

354	322.33	323.19	3.49	3.36	3.73	++

355	348.37	349.28	3.61	3.48	3.92	+

356	386.38	387.19	3.55	3.46	3.92

357	346.31	347.09	3.29	3.16	3.52

358	310.28	311.29	2.50	2.40	2.76	+

359	399.42	400.17	2.58	2.45	2.79

360	406.41	407.23	3.81	3.69	4.19

361	396.42	397.16	3.88	3.75	4.18	+

362	376.43	377.35	3.96	3.63	4.41	+

363	357.34	358.15	2.42	2.36	2.58	+

364	374.37	375.19	2.41	2.32	2.68

365	432.45	433.25	3.95	3.86	4.39	+

366	363.39	364.29	2.44	2.36	2.68

367	336.36	337.40	3.65	3.55	3.93

368	404.82	405.29	3.78	3.68	4.02	+

369	404.48	405.40	4.38	4.21	4.64

370	356.35	357.09	3.48	3.37	3.68

371	322.33	323.18	3.42	3.26	3.73

372	308.31	309.31	3.16	3.01	3.38

373	342.32	343.10	3.62	3.53	3.82

374	372.35	373.10	3.55	3.45	3.73

375	360.31	361.24	3.65	3.59	3.88

376	343.31	344.10	2.63	2.56	2.76

377	343.31	344.12	2.55	2.48	2.73

378	332.29	333.21	2.78	2.71	3.14	+

379	349.34	350.24	3.32	3.23	3.56

380	350.34	351.30	2.98	2.82	3.21

381	439.48	440.32	2.73	2.66	2.98

382	400.36	401.15	3.42	3.31	3.64

383	379.39	380.28	2.39	2.30	2.60	+

384	405.31	406.21	4.01	3.88	4.09	+

385	387.32	388.23	4.02	3.88	4.29

386	386.38	387.19	3.46	3.38	3.72

387	424.35	425.11	3.82	3.71	4.06

388	449.46	450.12	2.96	2.83	3.16	+

389	446.43	447.13	3.28	3.18	3.56

390	334.34	335.32	3.42	3.33	3.71

391	306.29	307.23	2.98	2.85	3.22

392	357.34	358.16	2.42	2.30	2.71

393	430.43	431.33	3.32	3.22	3.66	+

394	338.33	339.19	2.68	2.59	2.83	+

395	384.40	385.24	3.75	3.66	4.02

396	409.41	410.25	3.49	3.41	3.76	++

397	358.32	359.11	3.43	3.36	3.61	+

398	378.40

399	294.28	295.21	2.95	2.85	3.28

400	362.38	363.31	3.41	3.32	3.68

401	367.33	368.09	3.41	3.31	3.77	+

402	385.39	386.19	2.71	2.64	3.04	+

403	413.45	414.20	2.82	2.68	3.05	+

404	376.77	377.28	3.93	3.86	4.22	++

405	372.35	373.10	3.85	3.74	4.12

406	384.40	385.25	3.91	3.81	4.11	+

407	372.35	373.10	3.65	3.55	3.88

408	427.43	428.17	2.89	2.78	3.15

409	376.77	377.24	3.89	3.82	4.15

410	357.34	358.17	2.69	2.59	3.05	+

411	400.45	401.24	4.21	4.08	4.49	+

412	322.33	323.18	3.35	3.18	3.58

413	363.39	364.29	2.39	2.30	2.62

414	411.21	411.02	4.24	4.08	4.41

415	416.40	417.33	3.28	3.18	3.55

416	374.34	375.13	3.55	3.42	3.79

417	425.24	425.02	3.95	3.78	4.18

418	406.79	407.15	3.77	3.71	3.92	+

419	410.32	411.18	4.08	3.97	4.22	+

420	411.21	410.98	4.19	4.06	4.32	+

421	444.77	445.18	4.29	4.15	4.49	+

422	421.22	422.95	4.02	3.92	4.39	++

423	367.33	368.13	3.63	3.48	3.81	+

424	386.38	387.17	3.28	3.19	3.58

425	398.43	399.17	4.25	4.11	4.54

426	384.40	385.25	4.11	3.99	4.49	+

427	398.43	399.15	4.35	4.21	4.49	+++

428	424.47	425.18	4.54	4.45	4.94	+++

429	440.35	441.20	4.12	4.04	4.28

430	370.38	371.10	3.72	3.63	4.09

431	400.36	401.14	3.52	3.43	3.76

432	381.36	382.18	3.36	3.25	3.63

433	344.30	345.06	2.88	2.78	3.02

434	357.34	358.12	2.62	2.56	2.82

435	422.21	424.00	3.88	3.82	4.25

436	411.31	412.10	3.98	3.91	4.19

437	445.75	446.12	3.74	3.68	3.86

438	371.37	372.12	2.85	2.65	3.22

439	336.36	337.40	3.67	3.57	3.82	+

440	377.76	378.18	3.49	3.36	3.68

441	377.76	378.21	3.51	3.42	3.69

442	373.34	374.14	3.19	3.01	3.44

443	377.76

444	386.33	387.13	3.52	3.38	3.74	+

445	372.35	373.10	2.71	2.65	2.89	+

446	393.37	394.15	2.76	2.69	3.04

447	459.48	460.32	4.15	4.07	4.47

448	360.34	361.27	2.36	2.32	2.59	+

449	357.34	358.14	2.40	2.33	2.70

450	430.43

451	387.32

452	392.38	393.17	3.98	3.85	4.16

453	398.43	399.16	3.99	3.79	4.22	+

454	292.74	292.94	2.85	2.78	3.16	+

455	332.81	333.20	3.42	3.29	3.71	++

456	370.81	371.00	3.35	3.28	3.77

457	330.75	331.01	2.99	2.89	3.36	+

458	294.72	295.05	2.14	2.05	2.51

459	383.86	384.11	2.26	2.10	2.60	+

460	390.85	391.26	3.59	3.42	4.01	+

461	380.85	381.20	3.66	3.56	3.93

462	360.86	361.26	3.76	3.59	4.48	+

463	341.78	342.07	2.06	1.96	2.35	+

464	358.81	359.18	2.07	1.92	2.40

465	416.89	417.29	3.78	3.69	4.14	+

466	347.82	348.15	2.08	1.81	2.51	++

467	320.80	321.14	3.41	3.35	3.81

468	389.26	389.25	3.56	3.45	3.95	+

469	388.92	389.35	4.28	4.15	4.61	+

470	340.79	340.98	3.23	3.09	3.58

471	306.77	307.20	3.15	3.05	3.49

472	292.74	292.93	2.89	2.79	3.36	+

473	344.75

474	316.72

475	333.77

476	334.78	335.28	2.70	2.62	3.08	+

477	423.92	424.11	2.40	2.18	2.75	+

478	384.80	385.13	3.16	3.09	3.46

479	363.82	364.29	2.03	1.86	2.33	+

480	370.81	370.99	3.22	3.14	3.48	+

481	408.79	409.02	3.56	3.41	3.89	+

482	433.89	434.10	2.65	2.50	3.03	+

483	430.87	431.29	3.01	2.91	3.33

484	318.78	318.94	3.18	3.11	3.42

485	290.73	291.01	2.64	2.52	2.91	+

486	341.78	342.06	2.08	1.99	2.48

487	414.87	415.26	3.09	3.00	3.42	+

488	322.77

489	393.85	394.10	3.26	3.13	3.65

490	362.84	367.09	3.79	3.42	3.81

491	278.72	279.01	2.61	2.51	2.97

492	346.81	346.99	3.15	3.05	3.42	+

493	384.88	385.25	4.11	4.04	4.66	+

494	306.77	307.22	3.12	3.03	3.51	++

495	347.82	348.14	2.02	1.83	2.38	+

496	400.84	401.06	2.98	2.89	3.38	+

497	358.78	359.13	3.29	3.15	3.65	+

498	409.68	408.97	3.71	3.61	4.38

499	443.91

500	344.78

501	341.78	342.06	2.06	1.93	2.30

502	371.76

503	382.87	383.18	3.78	3.68	4.05	++++

504	388.92	389.37	3.98	3.81	4.52	+

505	290.73	291.07	2.53	2.43	2.85	+

506	370.81	371.07	3.17	2.99	3.94	+

507	383.86	384.13	2.25	2.16	2.49

508	340.79	341.04	3.11	2.99	3.56	++

509	384.80	385.19	3.05	2.96	3.51	+

510	370.81	371.05	3.11	3.02	3.61	+

511	408.79	409.12	3.44	3.28	3.84

512	430.87	431.31	2.93	2.80	3.38	+

513	346.81

514	400.84	401.14	2.93	2.78	3.36

515	358.78	359.14	3.16	2.99	3.48	+++

516	409.68	409.00	3.53	3.42	4.04	+

517	408.91

518	382.87	383.21	3.62	3.45	4.19

519	396.64

520	386.22

521	372.81	373.08	3.27	3.18	3.58	+

522	400.84	401.12	3.12	3.02	3.45	++++

523	386.81	387.15	2.69	2.62	3.01

524	354.81	355.22	3.21	3.12	3.71

525	400.95	401.27	3.71	3.55	4.11

526	302.76	303.08	2.29	2.22	2.71	+

527	382.85	383.15	2.92	2.78	3.22

528	395.89	396.11	2.08	2.03	2.39

529	352.82	353.22	2.85	2.75	3.38	+++

530	396.83	397.05	2.90	2.76	3.38

531	382.85	383.14	2.85	2.69	3.29

532	420.82	421.07	3.22	3.06	3.66

533	442.90	443.32	2.68	2.50	3.00

534	358.85	359.10	2.76	2.65	3.13	+

535	412.88	413.18	2.66	2.53	3.15	+

536	370.81	371.05	2.92	2.81	3.37	+

537	421.71	421.00	3.32	3.16	3.74	+

538	407.78

539	328.76

540	394.90	395.14	3.39	3.31	3.72

541	384.82

542	408.67

543	384.84	385.22	3.05	2.89	3.38	+

544	412.88	413.22	2.89	2.78	3.45	+++

545	398.85	399.10	2.43	2.28	2.82

546	366.85	367.09	2.96	2.82	3.45

547	343.22

548	378.65

549	368.23

550	354.81	355.21	3.12	2.96	335

551	382.85	383.16	2.98	2.83	3.45	++++

552	368.82	369.10	2.49	2.33	2.86	+

553	336.82	337.34	3.05	2.92	3.62	+

554	259.37	260.12	3.39	3.29	3.82

555	341.52	342.16	4.21	4.12	4.37

556	243.33

557	323.42

558	336.46	337.40	2.50	2.43	2.72

559	293.39

560	337.40

561	323.42

562	361.39

563	383.47

564	321.44	322.20	3.64	3.34	3.98	+

565	299.42

566	353.44

567	311.38

568	362.28

569	279.36

570	309.39

571	297.35

572	304.37	305.19	3.31	3.24	3.75	++

573	313.81

574	309.39	310.28	3.72	3.62	4.07

575	321.44	322.20	3.78	3.56	4.28

576	309.39	310.28	3.52	3.35	4.04

577	313.81	314.05	3.85	3.76	3.98

578	348.25	348.23	4.15	3.92	4.25

579	348.25	347.69	4.39	4.22	4.48

580	358.26

581	304.37	305.16	3.68	3.59	3.93	+

582	335.47	336.43	4.16	3.95	4.79

583	321.44	322.20	3.95	3.88	4.41	+

584	335.47	336.35	4.01	3.85	4.22	+++

585	361.51	362.30	4.39	4.27	4.59	+

586	307.42	308.28	3.65	3.49	4.22	+

587	337.40	338.24	3.43	3.32	3.71	+

588	323.37	324.27	3.43	3.36	3.93	+

589	324.36

590	342.35

591	324.36

592	343.83	344.09	3.72	3.61	4.08	+

593	347.36					+

594	381.81

595	323.42

596	377.39	378.23	4.04	3.93	4.39	+

597	308.41	309.30	2.72	2.63	3.00	++

598	280.35

599	280.35	281.22	2.45	2.40	2.66

600	322.43	323.16	2.59	2.45	3.19	+

601	350.49	351.31	2.78	2.66	2.96

602	364.47	365.09	2.82	2.73	3.05

603	294.38

604	318.40	318.95	3.29	3.19	3.53	+

605	281.34

606	359.25	359.09	3.85	3.72	4.22

607	348.35	349.20	3.91	3.84	4.54	+

608	382.79	383.07	3.68	3.53	4.11	+

609	308.41

610	314.80	314.87	3.42	3.36	3.61	+

611	310.38	311.24	3.08	3.00	3.38

612	314.80

613	309.39

614	330.41	331.12	2.68	2.56	2.91

615	396.51

616	269.33

617	286.38	287.07	3.22	3.16	3.55

618	329.42	330.10	3.86	3.83	3.98

619	335.47	336.38	3.88	3.71	4.15	+

620	385.49

621	351.43	352.29	3.69	3.58	3.85

622	422.55	423.07	2.86	2.65	3.49	+

623	339.42	340.11	3.31	3.19	3.53

624	323.37	324.25	3.11	3.06	3.38

625	357.82

626	351.43

627	336.42

628	313.81

629	325.39

630	349.24	349.09	3.96	3.91	4.19

631	369.44	370.13	3.38	3.25	3.65	+

632	319.39	320.10	3.08	2.99	3.27	+

633	338.82

634	337.44	338.28	3.83	3.73	3.99

635	347.36	348.13	3.92	3.85	4.19	+

636	363.36

637	363.36	364.29	4.07	3.82	4.09	+

638	325.41

639	337.44					+

640	315.34	316.10	3.98	3.82	4.12	+

641	353.44

642	323.42	324.28	3.71	3.59	4.06

643	323.42	324.28	3.62	3.55	3.85	++

644	335.47	336.39	4.12	3.88	4.69

645	377.39	378.30	3.79	3.75	3.92

646	339.42

647	307.42	308.25	3.46	3.38	3.79

648	398.91	399.10	3.72	3.62	4.32

649	346.41

650	393.87

651	348.47

652	341.42	342.17	2.25	1.96	2.86

653	304.35	305.27	1.85	1.72	2.38

654	382.26	384.07	2.60	2.36	2.85	+

655	371.37

656	405.81	406.19	2.59	2.53	2.91	+

657	331.42	332.28	1.96	1.90	2.33

658	337.81

659	333.39

660	337.81	338.27	2.35	2.29	2.45

661	332.41	333.25	1.83	1.63	2.12

662	353.43	354.25	1.85	1.73	2.06

663	419.53

664	292.34	293.06	1.80	1.70	2.13

665	309.39	310.28	2.16	2.03	2.59

666	352.44	353.27	2.78	2.69	3.23

667	358.49	359.29	2.82	2.65	3.39

668	408.50

669	374.44	375.22	2.72	2.63	3.11

670	445.57	446.26	2.07	1.99	2.45

671	362.43	363.37	2.21	2.12	2.69

672	360.46	361.32	2.78	2.69	3.14

673	370.38	371.10	2.85	2.69	3.32

674	386.38	387.18	2.98	2.81	3.22	++

675	386.38	387.20	2.95	2.81	3.38	+

676	348.42	349.29	2.48	2.39	2.98	+

677	360.46	361.32	2.75	2.65	3.23

678	338.36	339.18	2.62	2.52	3.05

679	376.46	377.34	2.38	2.18	2.91

680	346.43	347.22	2.59	2.52	3.06

681	346.43	347.21	2.54	2.48	2.91

682	358.49	359.29	3.08	2.98	3.67

683	400.40	401.21	2.81	2.65	3.46

684	362.43	363.37	1.99	1.90	2.23

685	330.43	331.26	2.42	2.33	2.80	+++

686	421.93	422.03	2.58	2.50	2.96	+

687	369.43	370.17	2.42	2.16	2.82

688	416.89

689	371.49	372.18	2.00	1.95	2.42

690	372.26

691	392.46	393.26	2.29	2.15	2.73

692	342.40	343.20	2.12	1.93	2.72

693	361.83	362.29	2.75	2.69	3.08

694	336.82	337.35	2.71	2.55	3.09

695	359.43	360.22	2.00	1.93	2.32

696	374.44	375.22	2.68	2.58	3.15	++++

697	380.83

698	346.39	347.15	2.15	1.98	2.48

699	348.41	349.29	2.05	1.97	2.78

700	289.77

701	371.91	372.09	4.25	4.09	4.79

702	273.72	274.06	2.52	2.33	2.96

703	353.81	354.12	3.25	2.96	3.69	+

704	366.85	367.02	2.19	1.87	2.43

705	323.78	324.16	3.13	2.89	3.61

706	367.79	368.06	3.08	2.86	3.33

707	353.81	354.12	3.11	2.92	3.46

708	391.78

709	413.86	414.08	2.89	2.79	3.18

710	329.81	327.05	5.28	4.88	5.69	+

711	383.84	384.12	2.88	2.75	3.22	++

712	341.78	342.08	3.18	3.08	3.31	++++

713	392.67	391.93	3.61	3.52	3.88

714	378.65

715	378.65

716	354.76

717	372.75

718	354.76

719	348.79

720	389.64

721	365.87	366.12	3.75	3.61	4.15	+

722	353.77

723	388.21

724	344.20

725	379.64

726	369.21

727	355.80	356.12	3.33	3.21	3.71

728	383.84	384.09	3.15	2.89	3.45

729	369.81	370.10	2.62	2.55	2.83	++++

730	337.81

731	376.81

732	372.35

733	443.47	444.39	2.51	2.39	2.88

734	402.38	403.31	3.18	2.88	3.69

735	457.46	458.32	2.56	2.42	3.00

736	406.79	407.14	3.44	3.18	3.81

737	436.82	437.12	3.33	3.22	3.72

738	440.35	441.20	3.59	3.39	3.96

739	474.79	475.01	3.78	3.58	4.21

740	428.46	429.28	3.75	3.48	4.09

741	414.43	415.29	3.68	3.43	4.15

742	428.46	429.29	3.86	3.58	4.24

743	470.37	471.33	3.64	3.45	4.01

744	430.39	431.31	3.09	2.95	3.59

745	452.23	451.96	3.39	3.19	3.51

746	441.34	442.22	3.52	3.28	3.81

747	407.78	408.11	3.09	2.93	3.53

748	403.36	404.29	2.79	2.52	3.28

749	407.78	408.12	3.08	2.95	3.62

750	440.35	441.21	3.56	3.18	4.04

751	456.35	457.24	3.61	3.23	3.91

752	428.46	429.29	3.81	3.55	4.18

753	470.37	471.32	3.49	3.18	3.86

754	425.41	426.12	3.19	2.92	3.59

755	322.34	323.15	2.46	2.09	2.75

756	393.47	394.17	1.95	1.88	2.33

757	352.37	353.24	2.52	2.36	2.78

758	407.45	408.17	1.98	1.89	2.31

759	356.79	357.09	2.80	2.66	3.13

760	386.81	387.12	2.69	2.53	3.03

761	390.34	391.27	2.99	2.86	3.38

762	424.79	425.00	3.19	3.06	3.53

763	378.45	379.30	3.15	2.93	3.43

764	364.42	365.09	2.98	2.85	3.55

765	378.45	379.30	3.24	3.09	3.56

766	420.37	421.09	3.05	2.89	3.28

767	380.38	381.22	2.43	2.32	2.75

768	402.23	404.19	2.68	2.60	2.79

769	391.33	392.14	2.86	2.78	2.99

770	357.77	358.10	2.39	2.30	2.63

771	353.36	354.15	2.13	2.02	2.39

772	357.77

773	390.34	391.26	2.96	2.81	3.24

774	406.34	407.18	3.02	2.88	3.32

775	378.45	379.30	3.21	3.06	3.48

776	420.37	421.10	2.89	2.75	3.12

777	375.41	376.17	2.55	2.33	2.96

778	304.35	305.26	2.25	2.12	2.52

779	375.47	376.28	1.83	1.69	1.93

780	334.38	335.31	2.33	2.19	2.59

781	389.46	390.41	1.83	1.73	1.99

782	338.80	339.09	2.60	2.50	2.99

783	368.82	369.09	2.49	2.40	2.73

784	372.35	373.10	2.81	2.68	2.98

785	406.79	407.14	3.01	2.86	3.23

786	360.46	361.32	2.95	2.82	3.12

787	346.43	347.18	2.83	2.68	3.19

788	360.46	361.30	3.08	2.92	3.52

789	402.38	403.31	2.88	2.72	3.12

790	362.39	363.32	2.26	2.16	2.52

791	384.24

792	373.34

793	339.78	340.09	2.19	2.09	2.49

794	335.37	336.35	1.96	1.87	2.30

795	339.78	340.09	2.22	2.08	2.43

796	372.35	373.10	2.76	2.63	3.12

797	388.35	389.30	2.82	2.71	3.09

798	360.46	361.30	2.99	2.85	3.36

799	402.38	403.31	2.70	2.53	2.99

800	357.42	358.19	2.39	2.25	2.52

801	413.91	414.13	3.75	3.51	4.04

802	485.03	485.36	2.99	2.85	3.32

803	443.94	444.32	3.76	3.49	4.15

804	499.02	499.19	3.08	2.91	3.56

805	448.36	448.07	4.04	3.89	4.41

806	481.91	482.15	4.12	3.79	4.45

807	516.35	516.14	4.31	4.02	4.62

808	470.02	470.30	4.28	4.18	4.85

809	455.99	456.26	4.18	4.06	4.68

810	470.02	470.31	4.39	4.19	4.87

811	511.94	512.34	4.15	3.99	4.42

812	471.95	472.32	3.63	3.43	4.05

813	482.90	483.25	4.06	3.88	4.31

814	449.34	448.93	3.71	3.51	3.99

815	444.92	445.27	3.41	3.16	3.71

816	497.91	498.33	4.11	3.96	4.52

817	470.02	470.34	4.32	4.21	4.71

818	511.94	512.34	3.99	3.78	4.38

819	466.97	467.21	3.72	3.48	4.19

820	337.81	338.29	3.15	2.96	3.71

821	408.93	409.26	2.49	2.32	2.70

822	367.84	368.14	3.19	2.99	3.76

823	422.92	423.05	2.52	2.35	2.75

824	372.26	371.98	3.51	3.16	3.99

825	402.28	402.14	3.39	3.06	3.96

826	440.26	440.13	3.92	3.82	4.32

827	393.92	394.16	3.81	3.65	4.45

828	379.89	380.24	3.68	3.41	3.96

829	393.92	394.16	3.96	3.56	4.58

830	435.84	436.24	3.72	3.61	4.04

831	395.85	396.11	3.05	2.91	3.38

832	406.80	407.17	3.61	3.32	3.99

833	373.24	373.02	3.12	2.86	3.51

834	368.83	369.09	2.75	2.58	2.95

835	405.81	406.21	3.69	3.49	4.14

836	421.81	421.98	3.71	3.49	4.21

837	393.92	394.19	3.89	3.53	4.12

838	435.84	436.22	3.55	3.16	4.01

839	390.88	391.31	3.13	2.78	3.79

840	376.77	377.24	3.41	3.26	3.71

841	447.89	448.10	2.66	2.32	3.12

842	406.79	407.14	3.45	3.00	3.83

843	461.87	462.22	2.78	2.63	3.22

844	411.21	410.98	3.68	3.36	4.11

845	441.24	441.08	3.56	3.16	3.85

846	479.21	478.96	3.99	3.62	4.34

847	432.88	433.22	3.96	3.72	4.44

848	418.85	419.13	3.85	3.58	4.25

849	432.88	433.22	4.12	3.78	4.61

850	474.79	475.01	3.83	3.59	4.19

851	434.81	435.12	3.32	2.92	3.82

852	456.65	458.13	3.63	3.35	4.15

853	445.75	446.10	3.71	3.48	4.06

854	412.20	412.09	3.32	3.18	3.63

855	407.78	408.11	3.05	2.90	3.36

856	444.77	445.16	3.78	3.34	4.18

857	460.77	461.12	3.85	3.55	4.14

858	432.88	433.22	4.05	3.65	4.58

859	474.79	475.01	3.68	3.42	3.98

860	429.83	430.18	3.41	3.13	3.95

861	334.38	335.32	2.30	2.13	2.78

862	405.50	406.38	1.85	1.63	2.09

863	364.40	365.09	2.36	2.17	2.78

864	419.48	420.22	1.87	1.80	2.15

865	368.82	369.10	2.65	2.49	2.93

866	398.85	399.10	2.50	2.36	2.70

867	402.38	403.31	2.80	2.69	3.09

868	436.82	437.12	2.99	2.70	3.25

869	390.49	391.35	2.95	2.69	3.15

870	376.46	377.35	2.82	2.66	3.08

871	390.49	391.35	3.02	2.93	3.29

872	432.40	433.25	2.86	2.69	3.16

873	392.42	393.21	2.28	2.10	2.46

874	414.26	414.09	2.52	2.41	2.82

875	403.36	404.34	2.63	2.49	2.88

876	369.81	370.11	2.23	2.10	2.49

877	365.39	366.18	2.02	1.66	2.23

878	369.81	370.12	2.25	2.02	2.43

879	402.38	403.32	2.79	2.65	2.99

880	418.38	419.17	2.83	2.69	3.09

881	390.49	391.35	3.02	2.88	3.26

882	432.40	433.23	2.72	2.59	2.99

883	387.44	388.28	2.42	2.16	2.82

884	381.87	382.19	3.15	2.93	3.52

885	452.99	453.12	2.52	2.35	2.92

886	411.89	412.12	3.18	2.96	3.86

887	466.97

888	416.31	416.25	3.46	3.23	3.86

889	446.34	446.15	3.34	3.21	3.96

890	449.86	450.13	3.62	3.42	4.06

891	484.31	484.14	3.79	3.59	4.15

892	437.97

893	423.95	424.16	3.66	3.45	4.04

894	437.97	438.20	3.88	3.73	4.19

895	479.89	480.09	3.66	3.46	4.05

896	439.90	440.22	3.09	2.93	3.49

897	461.75

898	450.85

899	417.30	417.27	3.06	2.93	3.52

900	412.88	413.24	2.78	2.65	3.25

901	417.30

902	449.86	450.12	3.61	3.36	4.12

903	465.86	466.25	3.62	3.43	4.11

904	437.97	438.18	3.79	3.49	4.22

905	479.89	480.08	3.52	3.38	3.98

906	434.93	435.28	3.18	3.03	3.59

907	421.89

908	323.78

909	394.91	395.15	2.21	2.12	2.52

910	353.81

911	408.89

912	358.23	358.11	3.18	2.99	3.36

913	388.26	388.17	3.01	2.93	3.29

914	391.78	392.10	3.35	3.15	3.58

915	426.23	426.03	3.54	3.42	3.87

916	379.89

917	365.87	366.14	3.35	3.25	3.63

918	379.89	380.30	3.59	3.51	3.78

919	421.81	421.98	3.41	3.32	3.71

920	381.82	382.13	2.73	2.59	2.99

921	403.67

922	392.77

923	359.22	359.09	2.72	2.65	3.05

924	354.80	355.16	2.38	2.29	2.48

925	359.22

926	391.78	392.11	3.31	3.22	3.62

927	407.78	408.12	3.33	3.22	3.49

928	379.89	380.25	3.53	3.35	3.79

929	421.81	421.97	3.19	2.99	3.39

930	376.85	377.27	2.83	2.75	3.12

931	338.80	339.10	2.55

932	409.92	410.23	2.03

933	368.82	369.10	2.62

934	423.90	424.11	2.05

935	373.24	373.01	2.88

936	403.27	403.27	2.73

937	406.79	407.14	3.05

938	441.24	441.10	3.25

939	394.90	395.14	3.19

940	380.88	381.24	3.06

941	394.90	395.16	3.33

942	436.82	437.12	3.12

943	396.83	397.08	2.52

944	418.68	420.02	2.76

945	407.78	407.51	3.25

946	374.23	374.08	2.45

947	369.81	370.10	2.22

948	374.23	374.08	2.51

949	406.79	407.13	3.02

950	422.79	422.96	3.08

951	394.90	395.17	3.26

952	436.82	437.12	2.95

953	391.86

954	378.82

955	386.86

956	440.83

957	375.82

958	332.79

959	392.77

960	428.94

961	448.47

962	462.49

963	449.46

964	415.90

965	440.95

966	415.90

967	380.88

968	394.90

969	421.93

970	437.97

971	407.90

972	393.88

973	377.76

974	400.89

975	414.91

976	378.74

977	442.81

978	420.41	421.10	3.05	2.95	3.23

979	360.31	361.23	3.43	3.33	3.72

980	363.44	364.33	2.26	2.16	2.69

981	423.54	424.16	1.98	1.76	2.34

982	322.34	323.15	2.98	2.85	3.28

983	382.44	383.12	2.58	2.18	2.78

984	377.42	378.28	2.32	2.19	2.72

985	437.52	438.15	1.98	1.79	2.19

986	326.76	327.10	3.26	3.15	3.54

987	386.86	387.09	2.85	2.73	3.12

988	348.42	349.28	3.59	3.36	4.12

989	408.52	409.25	3.22	3.09	3.53

990	350.35	351.27	2.88	2.72	3.29

991	410.45	411.14	2.46	2.29	2.76

992	372.20

993	432.30

994	361.30	362.22	3.35	3.22	3.68

995	421.40	421.97	2.88	2.73	3.09

996	327.75	328.23	2.86	2.68	3.15

997	387.85	388.15	2.42	2.29	2.75

998	360.31	361.23	3.41	3.18	3.69

999	420.41	421.08	3.02	2.78	3.25

1000	376.31	377.27	3.44	3.32	3.67

1001	436.41	437.12	3.08	2.86	3.36

1002	350.40	351.30	3.31	3.07	3.62

1003	410.50	411.20	2.93	2.75	3.28

1004	345.38	346.10	2.96	2.76	3.33

1005	405.48

1006	371.39

1007	431.49

1008	361.30	362.21	3.12	2.96	3.51

1009	421.40	421.96	2.71	2.52	2.98

1010	424.38	425.02	3.49	3.39	3.64

1011	484.48	485.21	3.11	2.99	3.28

1012	370.41	371.04	2.66	2.56	2.96

1013	430.50	431.29	2.23	2.15	2.33

1014	384.43	385.22	2.79	2.69	3.15

1015	444.53	445.23	2.40	2.32	2.85

1016	398.46	399.10	2.99	2.80	3.25

1017	458.56	459.23	2.59	2.35	2.95

1018	366.81	367.09	3.04	2.95	3.62	++

1019	338.80	339.09	3.18	3.09	3.59	+

1020	308.77	309.28	3.12	3.05	3.58	+

1021	347.81	348.15	2.90	2.83	3.18	++++

1022	338.80	339.11	3.05	2.92	3.43	++

1023	326.76	327.11	3.19	3.12	3.47

1024	309.76	310.28	2.21	2.09	2.61	++++

1025	309.76	310.28	2.09	2.02	2.46	++++

1026	298.73	299.17	2.29	2.19	2.53	+

1027	324.77	325.23	2.93	2.85	3.18

1028	333.78	334.18	2.93	2.78	3.32	+

1029	343.22	343.09	3.52	3.42	3.81	+

1030	338.80	339.12	3.34	3.22	3.68	++++

1031	350.85	351.29	3.45	3.34	3.91

1032	343.22	343.09	3.41	3.33	3.81	+

1033	377.66	377.16	3.79	3.71	4.14	+

1034	373.24	373.03	3.33	3.22	3.66	+

1035	376.77	377.26	3.67	3.54	3.89	++++

1036	377.66	377.17	3.74	3.61	4.05

1037	411.21	411.00	3.89	3.78	4.24	+++

1038	387.67	389.18	3.57	3.48	3.91	+

1039	333.78	334.15	3.16	3.09	3.48	+

1040	352.82	353.22	2.78	2.71	3.15	+++

1041	364.88	365.09	3.85	3.75	4.31	++++

1042	350.85	351.29	3.71	3.62	4.05	++++

1043	364.88	365.09	3.96	3.85	4.68	++

1044	390.92	391.32	4.19	3.99	4.57	++++

1045	406.79	407.15	3.73	3.65	4.14	++

1046	336.82	337.34	3.29	3.16	3.59	+

1047	344.20	344.08	2.90	2.83	3.32	++

1048	344.20	344.08	3.02	2.92	3.31	+

1049	339.78	340.10	2.66	2.58	2.98	+

1050	344.20	344.08	3.05	2.88	3.36	++++

1051	352.78	353.15	3.05	2.90	3.48	+

1052	338.80	339.13	2.29	2.22	2.51	+

1053	396.88	397.10	3.83	3.53	4.24	++

1054	337.81	338.29	2.40	2.25	2.80	+

1055	315.78	316.06	2.82	2.72	3.16	+

1056	371.76	371.96	3.56	3.48	3.83

1057	353.77	354.12	3.53	3.46	3.78

1058	304.78	305.24	2.33	2.29	2.61	+

1059	350.85	351.30	3.29	3.21	3.66

1060	351.84	352.29	2.28	2.16	2.62	++++

1061	379.89	380.27	2.39	2.19	2.82	++++

1062	393.88	394.15	2.43	2.36	2.89	+

1063	323.78	324.28	2.28	2.17	2.59

1064	310.75	311.25	2.35	2.23	2.53

1065	323.78	324.28	2.19	2.12	2.39	++

1066	388.65	390.14	3.42	3.35	3.63	++

1067	377.76	378.19	3.56	3.51	3.82	++

1068	412.20	412.11	3.32	3.13	3.63

1069	337.81	338.29	2.42	2.36	2.64

1070	302.81	303.08	3.15	3.06	3.59	+

1071	359.82	360.14	2.33	2.22	2.59	+

1072	425.92	426.12	3.78	3.68	4.49	++

1073	353.77	354.15	3.39	3.31	3.56	++++

1074	358.83	359.15	3.58	3.48	3.92	++++

1075	355.80	356.12	2.50	2.38	2.93

1076	306.77	307.22	3.25	3.18	3.46

1077	326.76	327.10	3.35	3.28	3.59

1078	356.79	357.01	3.28	3.19	3.75

1079	327.75	328.19	2.25	2.13	2.58	+

1080	327.75	328.21	2.18	2.12	2.53	+

1081	389.75	390.15	3.82	3.66	4.21

1082	371.76	371.94	3.78	3.71	4.16

1083	368.84	369.07	3.56	3.46	3.96	+

1084	342.76	343.09	3.18	3.08	3.49	+

1085	351.77	352.15	3.31	3.12	3.61

1086	369.83	370.11	2.39	2.29	2.76	+

1087	397.88	398.10	2.48	2.26	2.99

1088	361.21	361.02	3.69	3.58	3.96	+

1089	356.79	357.00	3.76	3.69	4.05

1090	368.84	369.04	3.83	3.66	4.26

1091	356.79	357.00	3.38	3.29	3.85	+

1092	411.87	412.10	2.61	2.46	3.19

1093	361.21

1094	341.78	342.07	2.40	2.26	2.73

1095	395.65	394.97	4.45	4.31	4.51

1096	391.23	391.22	3.52	3.41	3.81	++

1097	394.76	395.00	3.81	3.68	4.05

1098	395.65	396.95	4.34	4.24	4.61

1099	429.20	428.97	4.02	3.88	4.31

1100	405.66	406.99	3.75	3.66	4.09	++++

1101	351.77	352.14	3.31	3.22	3.59	++++

1102	370.81	371.01	2.89	2.82	3.22

1103	382.87	383.16	4.05	3.95	4.26	+

1104	368.84	369.07	3.91	3.82	4.30

1105	382.87

1106	408.91	409.21	4.44	4.31	4.85

1107	424.79	424.96	3.86	3.76	4.21

1108	354.81	355.19	3.63	3.56	3.95

1109	384.80	385.11	3.22	3.15	3.55

1110	365.80	366.10	3.09	3.02	3.59

1111	328.74	329.16	2.60	2.50	2.85

1112	341.78	342.06	2.32	2.23	2.70	+

1113	406.64	408.06	3.76	3.69	3.99

1114	395.75	396.06	3.84	3.71	4.07	+

1115	430.19	430.12	3.55	3.49	3.88	+

1116	355.80	356.13	2.58	2.46	2.76

1117	320.80	321.12	3.46	3.36	4.02

1118	362.19	362.15	3.44	3.11	3.62	+

1119	362.19	362.14	3.18	3.11	3.45

1120	357.77	358.10	2.88	2.79	3.23	+

1121	362.19

1122	370.77	370.99	3.23	3.15	3.49

1123	356.79	357.07	2.40	2.29	2.73	+++

1124	377.81	378.17	2.40	2.32	2.78	++

1125	414.87	415.24	4.05	3.89	4.38

1126	376.82	377.27	3.75	3.66	4.11	++

1127	306.77	307.22	3.06	2.95	3.46	+

1128	368.84	369.10	3.36	3.28	3.73

1129	326.76	327.10	3.21	3.12	3.52

1130	356.79	357.09	3.13	3.03	3.63

1131	344.75	344.97	3.28	3.12	3.53	+

1132	351.77	352.22	2.99	2.86	3.29

1133	361.21	361.13	3.53	3.41	3.88	+

1134	356.79	357.04	3.35	3.09	3.73

1135	368.84	369.10	3.49	3.35	4.02	+

1136	356.79	357.09	3.24	2.89	3.79	+

1137	361.21	361.09	3.39	3.12	3.79

1138	395.65	394.98	3.74	3.49	4.12

1139	395.65	394.97	3.66	3.46	4.06	++++

1140	405.66	407.03	3.61	3.48	4.09

1141	351.77	352.18	3.21	3.13	3.42	++++

1142	382.87	383.17	3.87	3.77	4.24	++++

1143	368.84	369.08	3.72	3.48	4.11

1144	382.87	383.18	3.99	3.79	4.56	++++

1145	354.81	355.20	3.33	3.22	3.79	++++

1146	384.80	385.18	3.12	3.03	3.49

1147	370.77	370.99	3.14	2.99	3.59

1148	371.76	371.94	3.42	3.33	3.69

1149	389.75	390.22	3.49	3.41	3.83	++++

1150	371.76	372.00	3.49	3.41	3.92

1151	391.23	391.22	3.38	3.21	3.73	++++

1152	394.76	395.03	3.68	3.51	3.91	++

1153	429.20	428.99	3.91	3.79	4.19

1154	370.81	371.04	2.90	2.78	3.19

1155	424.79	425.01	3.76	3.63	3.95

1156	355.80	356.13	2.45	2.28	2.82	+

1157	327.75	328.21	2.28	2.15	2.59	++++

1158	327.75	328.23	2.17	2.07	2.49

1159	369.83	370.13	2.39	2.25	2.63

1160	397.88	398.14	2.43	2.30	2.99

1161	411.87	412.11	2.56	2.35	2.98

1162	341.78	342.08	2.33	2.15	2.60	++++

1163	365.80	366.10	2.99	2.86	3.53

1164	328.74	329.18	2.46	2.39	2.78

1165	406.64	408.06	3.46	3.38	3.81

1166	395.75	396.10	3.58	3.48	3.85	++

1167	430.19	430.13	3.35	3.15	3.75

1168	355.80	356.14	2.48	2.26	2.85

1169	362.19	362.16	3.12	2.95	3.46	+

1170	357.77	358.11	2.79	2.65	3.18	++

1171	362.19	362.19	3.12	3.07	3.45

1172	356.79	357.09	2.36	2.30	2.55

1173	377.81	378.19	2.40	2.12	2.78	++++

1174	443.91

1175	316.72	317.08	2.42	2.33	2.83	+

1176	333.77	334.13	2.91	2.84	3.29

1177	376.82	377.26	3.61	3.45	4.12	++++

1178	432.89	433.22	3.72	3.48	4.02

1179	398.82	399.10	3.51	3.31	3.82

1180	469.95	470.32	2.66	2.56	2.98

1181	386.81	387.15	2.98	2.80	3.38

1182	370.77	371.04	2.85	2.78	3.21	+

1183	405.22	405.21	2.99	2.80	3.26	++++

1184	398.82	399.10	3.45	3.36	3.91

1185	383.81	384.11	2.65	2.52	3.03

1186	361.21	361.12	3.52	3.41	3.78

1187	372.79	373.05	2.78	2.68	3.06	++

1188	416.84	417.30	3.08	2.96	3.61

1189	366.79	367.07	2.79	2.66	3.31	+++

1190	384.84	385.21	3.56	3.48	3.92	++

1191	394.76	395.06	3.65	3.51	3.96

1192	410.76	411.04	3.71	3.62	3.96	++

1193	410.76	411.01	3.73	3.62	4.12	+

1194	384.84	385.22	3.58	3.46	3.82	+

1195	362.74	363.29	3.43	3.33	3.72	+++

1196	370.81	371.10	3.42	3.31	3.76

1197	370.81	371.06	3.32	3.21	3.71

1198	382.87	383.20	3.91	3.75	4.14

1199	424.79	425.00	3.55	3.46	3.72

1200	446.31	446.12	3.38	3.28	3.61	++++

1201	393.81	394.10	3.16	3.08	3.48

1202	441.27	441.01	2.26	2.20	2.56

1203	395.87	396.11	2.61	2.11	3.01	+

1204	318.81	319.00	2.76	2.68	3.26	+

1205	380.88	381.24	3.11	2.98	3.46

1206	338.80	339.12	2.95	2.79	3.33	+

1207	368.82	369.09	2.89	2.75	3.36	+

1208	356.79	357.09	3.02	2.95	3.26

1209	363.81	364.29	2.76	2.65	2.90	+

1210	373.24	373.02	3.31	3.19	3.69	+

1211	368.82	369.10	3.09	2.96	3.51	++

1212	380.88	381.25	3.23	3.11	3.75	++

1213	368.82	369.09	2.98	2.86	3.35	++

1214	373.24	373.01	3.14	2.95	3.65

1215	407.69	407.09	3.49	3.39	3.93

1216	407.69	409.01	3.42	3.36	3.83	++++

1217	417.69	419.04	3.35	3.19	3.85

1218	363.81	364.29	2.99	2.92	3.32	++++

1219	394.90	395.14	3.61	3.46	4.08

1220	380.88	381.26	3.49	3.38	3.95

1221	394.90	395.14	3.73	3.58	4.15	++

1222	420.94	421.17	3.96	3.73	4.35

1223	366.85	367.09	3.09	2.95	3.51	++++

1224	396.83	397.07	2.88	2.72	3.26

1225	382.81	383.10	2.90	2.69	3.39

1226	383.79	384.08	3.19	3.12	3.41

1227	401.78	402.12	3.31	3.16	3.65	++++

1228	383.79	384.09	3.29	3.18	3.56

1229	403.27	403.27	3.15	3.06	3.45	+++

1230	406.79	407.15	3.46	3.36	3.93	++

1231	441.24	441.09	3.69	3.56	4.01	+

1232	382.85	383.14	2.65	2.49	3.01

1233	436.82	437.12	3.54	3.42	3.89

1234	367.84	368.13	2.36	2.28	2.70	+

1235	339.78	340.09	2.16	2.03	2.55	++++

1236	339.78	340.11	2.09	2.03	2.42	+

1237	381.87	382.16	2.22	2.16	2.63

1238	409.92	410.21	2.32	2.26	2.69

1239	423.90	424.12	2.36	2.29	2.69

1240	353.81	354.15	2.23	2.13	2.51	++++

1241	377.83	378.20	2.76	2.68	3.31

1242	340.77	340.96	2.32	2.13	2.70	++

1243	418.68	420.00	3.23	3.15	3.63	+

1244	442.23	442.16	3.15	2.96	3.39

1245	367.84	368.15	2.39	2.13	2.66

1246	374.23	374.08	2.86	2.76	3.25

1247	369.81	370.12	2.56	2.49	2.93	++

1248	374.23	374.09	2.89	2.70	3.38	+

1249	368.83	369.14	2.23	2.12	2.46	+

1250	389.84	390.25	2.26	2.20	2.83	++++

1251	455.95	456.21	3.58	3.43	3.95	+

1252	345.81	345.95	2.70	2.63	2.98	+

1253	388.86	389.31	3.35	3.23	3.79	++++

1254	444.92	445.29	3.49	3.32	3.99

1255	410.86	411.14	3.26	3.16	3.45

1256	481.98	482.22	2.50	2.39	2.66

1257	398.85	399.10	2.73	2.55	3.16	+

1258	382.81	383.10	2.63	2.52	3.03	+

1259	417.25	417.27	2.79	2.69	3.29	++++

1260	410.86	411.14	3.22	3.13	3.61

1261	395.85	396.09	2.46	2.39	2.83

1262	373.24	373.04	3.28	3.13	3.75

1263	428.88	429.25	2.85	2.73	3.19	++++

1264	378.82	379.25	2.59	2.36	3.06	++

1265	398.25	398.03	3.31	3.22	3.72	++++

1266	396.88	397.14	3.33	3.22	3.92

1267	406.79	407.15	3.44	3.21	3.82	++

1268	422.79	422.95	3.48	3.39	3.85

1269	422.79	422.95	3.52	3.42	3.92	+

1270	396.88	397.14	3.32	3.22	3.82	+

1271	374.78	375.08	3.19	3.08	3.59	+++

1272	382.85	383.14	3.18	3.09	3.62

1273	382.85	383.15	3.09	2.96	3.56	+

1274	394.90	395.15	3.68	3.58	4.18

1275	436.82	437.12	3.35	3.25	3.79	++

1276	458.35	458.26	3.18	3.03	3.63	+

1277	405.84	406.22	2.90	2.55	3.25	+++

1278	453.30	452.96	2.09	1.94	2.38	+

1279	407.90	408.13	2.32	2.13	2.39

1280	414.90	415.27	3.59	3.51	4.22

1281	380.83	381.22	3.41	3.25	4.02

1282	451.96	452.15	2.53	2.36	2.88	+

1283	368.82	369.10	2.83	2.72	3.34

1284	352.78	353.17	2.71	2.63	2.83

1285	387.23	387.06	2.85	2.79	3.06

1286	380.83	381.22	3.33	3.23	3.99	+

1287	365.82	366.14	2.52	2.45	2.89

1288	354.80	355.20	2.61	2.51	2.84	+

1289	398.85	399.10	2.93	2.82	3.39

1290	348.79	349.24	2.66	2.52	3.03	++

1291	366.85	367.09	3.45	3.35	4.01	+

1292	376.77	377.26	3.55	3.42	4.18

1293	392.77	393.09	3.58	3.48	4.01	+

1294	392.77	393.06	3.64	3.37	4.20	+

1295	366.85	367.10	3.45	3.25	4.08	+

1296	344.75	345.01	3.32	3.08	3.86	+

1297	352.82	353.23	3.31	3.11	3.75

1298	352.82	353.21	3.16	3.09	3.36	+

1299	364.88	365.09	3.81	3.66	4.34

1300	406.79	407.17	3.46	3.35	3.84

1301	428.32	428.12	3.26	3.16	4.17	+++

1302	375.82	376.16	3.02	2.95	3.29

1303	423.28	422.93	2.10	2.03	2.42

1304	377.88	378.24	2.29	2.07	2.49	+

1305	351.84	352.18	3.51	3.44	3.72

1306	309.76	310.14	3.35	3.26	3.54	++++

1307	339.78	340.07	3.31	3.21	3.39

1308	327.75	328.18	3.41	3.29	3.65	+

1309	334.77	335.24	3.41	3.25	3.75	+

1310	344.20	344.05	3.72	3.66	3.91

1311	339.78	340.07	3.58	3.45	3.75

1312	351.84	352.16	3.98	3.88	4.48

1313	339.78	340.08	3.38	3.29	3.63

1314	344.20	344.04	3.95	3.86	4.22	++++

1315	388.65	390.11	3.79	3.71	4.01

1316	334.77	335.24	3.28	3.19	3.51	++++

1317	365.87

1318	351.84	352.15	3.98	3.78	4.28

1319	365.87	366.11	4.11	3.99	4.44	+++

1320	391.90	392.13	4.52	4.36	4.89	++++

1321	337.81	338.21	3.73	3.68	3.95

1322	367.79	368.07	3.23	3.16	3.53

1323	353.77

1324	374.23	374.05	3.53	3.41	3.73

1325	377.76	378.12	3.82	3.75	4.02	+

1326	412.20	414.31	3.56	3.51	3.78

1327	353.81

1328	407.78	408.13	3.86	3.79	4.06	+

1329	338.80	339.10	2.43	2.15	2.79	+

1330	310.75	311.21	2.15	2.10	2.26	+++

1331	310.75	311.20	2.09	2.00	2.45

1332	352.83	353.20	2.36	2.19	2.63

1333	380.88	381.22	2.48	2.36	2.66

1334	394.86	395.04	2.55	2.35	2.86	+

1335	324.77	325.23	2.30	2.23	2.70	+

1336	311.73	312.08	2.64	2.52	2.79	++++

1337	378.74	379.16	3.92	3.84	4.11

1338	413.19	412.97	3.58	3.49	3.89

1339	338.80	339.06	2.51	2.25	2.63

1340	345.19	344.94	3.12	2.86	3.36

1341	340.77	340.95	2.82	2.61	3.23	++

1342	345.19	344.92	3.18	2.98	3.41	+

1343	339.79	340.08	2.36	2.13	2.79	+

1344	360.81	361.08	2.38	2.07	2.66	+

1345	426.91	426.98	4.08	3.99	4.49	++++

1346	299.72	300.05	2.35	2.22	2.70

1347	316.77	316.95	2.86	2.76	3.11

1348	359.82	360.09	3.85	3.71	4.24	++

1349	415.88	416.23	3.88	3.78	4.28	+

1350	381.82	382.10	3.65	3.55	3.86	+

1351	452.94	453.04	2.69	2.52	2.78

1352	369.81	370.08	3.08	2.88	3.79

1353	381.82	382.11	3.59	3.48	3.88

1354	366.81	366.98	2.69	2.49	3.12

1355	355.78	356.10	2.82	2.55	3.12	++

1356	399.84	400.09	3.15	2.89	3.36	+

1357	349.78	350.17	2.89	2.55	3.13	++++

1358	367.84	368.10	3.75	3.58	4.08	+

1359	377.76	378.12	3.79	3.51	4.02

1360	393.76	394.08	3.83	3.73	4.05	+

1361	393.76	394.06	3.86	3.73	4.24	++

1362	367.84	368.08	3.74	3.45	4.04	+

1363	345.74	345.86	3.55	3.41	3.79	++++

1364	353.81	354.12	3.61	3.45	3.95	++++

1365	353.81	354.11	3.51	3.36	3.85

1366	365.87	366.11	4.16	3.95	4.51

1367	407.78	408.09	3.66	3.43	3.95

1368	429.31	429.06	3.58	3.39	3.79

1369	424.26	423.90	2.20	2.12	2.39

1370	378.86	379.24	2.68	2.28	3.03

1371	342.32	343.10	3.19	2.99	3.58

1372	413.45	414.14	2.49	2.20	2.96

1373	372.35	373.06	3.22	3.01	3.75

1374	427.43	428.13	2.53	2.38	2.93

1375	376.77	377.25	3.51	3.25	3.92

1376	406.79	407.16	3.38	3.21	3.88

1377	410.32	411.07	3.65	3.51	3.95

1378	444.77	445.18	3.83	3.56	4.26

1379	398.43	399.18	3.79	3.59	4.15

1380	384.40	385.26	3.68	3.55	4.15

1381	398.43	399.17	3.91	3.78	4.35

1382	440.35	441.22	3.70	3.55	4.07

1383	400.36	401.10	3.13	3.00	3.66

1384	422.21	421.94	3.43	3.19	3.82

1385	411.31	412.10	3.53	3.31	3.91

1386	377.76	378.16	3.11	2.98	3.52

1387	373.34	374.11	2.78	2.55	3.19

1388	377.76	378.15	3.15	2.93	3.65

1389	410.32	411.07	3.63	3.42	3.96

1390	426.32	426.99	3.67	3.52	4.01

1391	398.43	399.16	3.86	3.55	4.38

1392	440.35	441.20	3.53	3.33	3.99

1393	395.39

1394	360.31	361.22	3.31	3.11	3.62

1395	431.44	432.27	2.59	2.17	2.99

1396	390.34	391.27	3.35	3.23	3.53

1397	445.42	446.16	2.66	2.53	3.08

1398	394.76	395.03	3.58	3.45	3.91

1399	424.79	424.99	3.45	3.11	3.92

1400	428.31	429.13	3.72	3.33	4.01

1401	462.76	463.12	3.93	3.66	4.19

1402	416.42	417.32	3.86	3.65	4.42

1403	402.40	403.31	3.78	3.62	4.26

1404	416.42	417.32	4.01	3.85	4.68

1405	458.34	459.23	3.78	3.59	4.21

1406	418.35	419.13	3.21	3.08	3.42

1407	440.20	442.12	3.55	3.24	3.87

1408	429.30	430.15	3.63	3.26	4.05

1409	395.75	396.07	3.22	3.05	3.56

1410	391.33	392.11	2.93	2.63	3.33

1411	395.75	396.07	3.22	3.02	3.69

1412	428.31	429.12	3.68	3.55	4.04

1413	444.31	445.19	3.73	3.61	4.18

1414	416.42	417.31	3.93	3.66	4.55

1415	458.34	459.22	3.58	3.32	3.99

1416	413.38	414.12	3.34	3.18	3.46

1417	441.94

1418	457.94

1419	459.91

1420	441.48

For purposes of the Table 1, activity of each compound is expressed as follows:
“+” compound exhibited 0-25% inhibition of calcium ion influx induced by capsaicin stimulation.
“++” compound exhibited 25-50% inhibition of calcium ion influx induced by capsaicin stimulation.
“+++” compound exhibited 50-75% inhibition of calcium ion influx induced by capsaicin stimulation.
“++++” compound exhibited 75% or greater inhibition of calcium ion influx induced by capsaicin stimulation.

EXAMPLE 3

High Throughput Screening of VR1 Antagonists for Determination of In Vitro Efficacy Using a Calcium Imaging Assay

VR1 protein is a heat-gated cation channel that exchanges about 10 calcium ions for every sodium ion resulting in neuronal membrane depolarization and elevated intracellular calcium ion levels. Therefore the functional activity of compounds at the VR1 receptor may be determined by measuring changes in intracellular calcium levels in 293 cells expressing capsaicin-insensitive VR1 receptor variants. A dual wavelength ratiometric dye, Fura2, was used as an indicator of relative levels of calcium ions in a 96 well format using a bench top scanning fluorometer with integrated fluidics and temperature control (Flex Station, Molecular Devices).
A dual wavelength ratiometric dye, Fura2, was used as an indicator of relative levels of [Ca2+] in a 96 well format using a bench top scanning fluorometer with integrated fluidics and temperature control (Flex Station, Molecular Devices).
293 cells were grown on PDL coated 96-well black-walled plates, in the presence of a DMEM medium containing 5% Penstrep, 5% Glutamax, 200 ug/mL Hygromycin, 5 μg/mL Blasticide and 10% heat inactivated FBS. Prior to assay, the cells were loaded with 5 μg/mL Fura2 in normal saline solution at 37° C. for 40 minutes. Cells were then washed with normal saline to remove the dye.
The assay consists of two stages; a pre-treatment phase followed by a treatment phase.
50 μl of a compound solution was added to the cells (Pre-treatment). Immediately following, 50 μl of the test compound in a saline solution at pH 5.1 was added. Fura2 was excited at 340 and 380 nM to indicate relative calcium concentration. Changes in wavelength measurements were made throughout the course of the experiment in 4 second intervals over a period of 3 minutes. Responses were measured as peak fluorescence ratio after test compound addition minus baseline fluorescence ratio prior to pre-treatment and were calculated using SoftMaxPro softwareData were expressed as percentage inhibition calculated using Excel as follows:
$Percentage Inhibition = \frac{\begin{matrix} (Compound Response) - \\ (Control Response) \end{matrix}}{(\begin{matrix} Agonist Response - \\ Control Response \end{matrix})} \times 100$
All compounds with percentage inhibition values greater than 75% are considered hits and earmarked for further investigation at lower concentrations. The relative strengths of the percentage inhibition values are set forth in Table 1.

EXAMPLE 4

Whole-Cell Patch Clamp Electrophysiology

Dorsal root ganglion (DRG) neurons were recovered from either neonatal or adult rats and plated onto poly-D-lysine coated glass coverslips. The plated neurons were transferred into a chamber to allow drug solutions to be added to the cells using a computer-controlled solenoid-valve based perfusion system. The cells were imaged using standard DIC optics. Cells were patched using finely-pulled glass electrodes. Voltage-clamp electrophysiology experiments were carried out using an Axon Instruments Multiclamp amplified controlled by pCLAMP8 software.
The cells were placed into a whole-cell voltage clamp and help at a voltage of −80 mV while monitoring the membrane current in gap-free recording mode. 500 nM capsaicin was added for 30 seconds as a control. Test compounds at various concentrations at ranging from (10-1000 nM) were added to the cells for 1 minute prior to a 30 second capsaicin application. Differences between control experiments and drug positive capsaicin experiments were used to determine the efficacy of each test compound. All compounds that inhibited capsaicin induced current greater than 50% were considered positives. The data obtained for compounds 24 and 18 are set forth in Table 2, below. FIG. 3 represents a dose response curve demonstrating the increasing effectiveness of compound 18 in inhibiting a capsaicin induced calcium ion influx at higher concentrations tested, namely 50 nM, 100 nM and 250 nM. Likewise, FIG. 4 represents a dose response curve demonstrating the increasing effectiveness of Compound 24 in inhibiting a capsaicin induced calcium ion influx at higher concentrations tested, namely 20 nM, 40 nM, 100 nM and 200 nM.

TABLE 2

			% inhibition of
		Treatment time	capsaicin induced
Compound ID	Concentration	(seconds)	current

24	200 nM	25	100
18	100 nM	20	<75

FIG. 1 demonstrates the activity of the compounds tested in inhibiting the capsaicin induced current.

EXAMPLE 5

Plasma Extravasation Study

A Measure of Neurogenic Inflammation

The density of TRPV1 expression is enhanced during an inflammatory condition. Therefore TRPV1 antagonists have been investigated in three different models of inflammatory pain, namely plasma extravasation, paw lick assay, and thermal hyperalgesia.

Methods

Sprague-Dawley male rats obtained from Charles River, San Diego, Calif. were dosed with compound 18. Two hours later, they were injected with Evan's Blue (10 ml/kg or 150 μl). Thirty minutes after I.V. injection, 10 μl of 25 mM capsaicin in 100% ethanol was applied to the left ear of the animal, followed by 10 μl vehicle (100% EtOH) to the right ear of the mouse. Fifteen minutes later, the animal was sacrificed using CO₂. Each ear was removed, placed into labeled tubes and weighed. The dye was then extracted from the ears by drying the ears at 55-60° overnight. The following day, 250 μl formamide was added and left at 55-60° overnight.
An Evans blue standard curve was first generated (2.5, 5, 10, 20, 40, and 80 mg/μl). Samples (with ears) were spun for at least 1 minute before addition of 100 μl of each sample to appropriate wells. Samples were analyzed in a plate holder by SOFTmax PRO. Standards were graphed to generate a standard curve and then used to extrapolate Evans Blue concentrations of experimental samples.

Results

Compound 18 at a dose of 30 mg/kg significantly blocks capsaicin induced plasma extravasation in rats. FIG. 2 demonstrates the results in μg EB/mg tissue (Evans Blue). The results are represented for the delivery vehicle HPBCD, control compound BCTC, Compound 18 at two concentrations and capsaicin alone in vehicle.

EXAMPLE 6

Paw Lick Assay

This assay was performed to test the ability of Compound 18 to inhibit the response to capsaicin challenge.

Methods

Animals were acclimatized at least 2 days prior to testing by placing them in a behavioral chamber for one hour. On the day of testing, animals were trained for 30-60 minutes prior to dosing. Animals were dosed with 30 mg/kg of Compound 18 at least 30 minutes prior to testing and then placed in behavioral chambers for acclimatization. Animals were then placed into a falcon tube restrainer and injected with a 0.16 mg/ml capsaicin solution (or vehicle) into the plantar surface of their paw. Animals were then returned to the behavior chamber and monitored for paw licking behavior (including paw and leg licking behavior) over the next 5 minute interval.

Results

Compound 18 (30 mg/kg) significantly inhibited pawlick response induced by treatment with capsaicin. FIG. 5 demonstrates the pawlick times per second when a delivery vehicle is administered alone, capsaicin is administered, capsaicin is administered with a control compound, and capsaicin is administered with Compound 18.

EXAMPLE 7

Thermal Hyperalgesia

Sprague-Dawley male rats obtained from Charles River, San Diego, Calif. were purchased at 150-175 g, and held for at least one week before testing. Pain was induced by injecting 100 μl of 2% carrageenan in 0.9% saline sub-Q into the right ventral hindpaw while the animals were under isofluorane anesthesia. Animals were then dosed one hour after with different concentrations of Compound 18 (3, 10, and 30 mg/kg). Two hours later, after acclimatizing in testing chambers for 20-30 minutes, animals were tested on both hindpaws for latency of paw withdrawal using a thermal testing apparatus. 2-3 trials were conducted with 10 minutes between trials. As demonstrated in FIG. 6, a dose of Compound 18 at 30 mg/kg significantly increased latency of paw withdrawal demonstrating reversal of thermal hyperalagesia. The figure depicts the time in seconds until animals withdraw from thermal stimulation at baseline and two hours after administration of delivery vehicle, control compound, and three concentrations of Compound 18.

EXAMPLE 8

Pharmacokinetic Profile

The pharmacokinetic profile of Compound 18 was evaluated following intravenous and oral administration in rats. Sprague-Dawley male rats obtained from Charles River, San Diego, Calif. were acclimated for 24 hours.
Compound 18 was formulated at a concentration of 0.5 mg/mL for IV administration at a 1 mg/kg dose and 1 mg/mL for oral administration at a 5 mg/kg dose. All animals were weighed before dosing. The body weight was used to calculate the actual dose for each animal. The intravenous dose was administered through the jugular vein catheter in less than 1 minute. The oral dose volume was 1.5 mL for all PO rats administered through oral lavage.
For IV dosing, blood samples were collected using a pre-heparinized syringe via the carotid artery catheter before dosing and at t=2, 5, 15, 30, 60, 120, 180, 360, and 480 minutes post dosing. For PO dosing, blood samples were collected using a pre-heparinized syringe via the carotid artery catheter before dosing and at t=5, 15, 30, 60, 120, 180, 360, and 480 minutes post dosing. 250 uL of blood was obtained at each time point from each animal. Equal volumes of 0.9% normal saline were replaced to prevent dehydration. The whole blood samples were maintained on ice until centrifugation. Blood samples were centrifuged at 14,000 rpm for 10 minutes at 4° C., and the upper plasma layer was transferred into a clean vial and store at −80° C.
The plasma was analyzed. Compound 18 demonstrates Oral Bioavailability (% F) of 30.06%, a half life (t½) of 3.71 hours, Clearance (Cl) of 0.53 L/h/Kg, a Volume of distribution (Vd) of 2.82 L/Kg, a Tmax of 180 minutes, and a Cmax of 1.75 mM.

EXAMPLE 9

Aqueous Solubility

Equilibrium solubility was measured in a pH 2.0 isotonic solution of NaCl/HCl and a pH 7.4 aqueous buffer. The pH 2.0 solution was prepared by adjusting the saline solution to a pH of 2.0 using HCl. The pH 7.4 buffer was prepared by adjusting the pH of a 0.07 M solution of NaH₂PO₄to pH 7.4 with 10 N NaOH. Each buffer had an ionic strength of 0.15. At least 1 mg of powder was combined with 1 ml of buffer to make 1 mg/ml mixture. This sample was shaken for 2 hours and left to stand overnight at room temperature. The samples were then filtered through a 0.45-μm Nylon syringe filter that was first saturated with the sample. The filtrate was sampled twice, consecutively. All samples were assayed by LC/MS using electrospray ionization. The typical range of the assay is greater than 1 mg/mL to less than 0.0002 mg/mL, depending on analytical sensitivity. The results demonstrated solubility of <0.0002 mg/ml at pH 2.0 and low solubility at pH 7.4.
The partition coefficient, Log(D), between water-saturated 1-octanol and pH 7.4 buffer was determined for Compound 18. The pH 7.4 buffer was prepared by adjusting the pH of a 0.07 M solution of NaH₂PO₄to pH 7.4 with 10 N NaOH. 15 μL of a 10 mM stock solution of test article were pipetted in duplicate, into test tubes containing 750 μL each of 1-octanol and pH 7.4 buffer. Then 3 μL of 50 mM testosterone was also added to each tube. Then the tubes were rotated for about one hour using a benchtop rotator. Following the rotation, the tubes sat on the bench top for about 1 hour to allow the layers to separate.
Thereafter, 400 μL of the octanol (top) layer was removed and placed into a test tube. Next 400 μL of the aqueous (bottom) layer was removed and placed into a test tube. The following serial dilutions were then made of each layer using 50% methanol as the diluent: Octanol—100×, 1000×, and 10,000× Aqueous—1×, 10×, 100×. The 100×, 1000× and 10,000× diluted octanol samples and the undiluted, 10× and 100× diluted aqueous samples were then aliquoted into appropriate vials.
A standard curve of test article and testosterone was prepared using 50% methanol at the following concentrations: 2, 0.6, 0.2, 0.06, 0.02, 0.006, and 0.002 μM. The samples were analyzed by LC/MS monitoring both test article and testosterone. Log (D) was calculated for each duplicate sample by taking the calculated concentration of the least diluted sample for each phase that fell within the standard curve using the following equation: Log(D)=Log 10(Calc. Conc. in Organic Phase/Calc. Conc. in Aqueous Phase). The results demonstrated Log (D) values at pH 7.4 of less than 4.7 for Compound 18.

EXAMPLE 10

Analysis of Plasma Protein Binding

Membranes from Harvard/Amika with a molecular weight cutoff of 5,000 were rinsed with dH₂O then placed in pH 7.4 PBS supplied by Gibco. The membranes were allowed to soak for 1 hour. A stock of the test article was pooled with Warfarin, Atropine at 2 mM in DMSO. The test article was then dosed into human plasma in sodium citrate, Rat Plasma, and Mouse Plasma to a final 10 μM concentration (0.5% DMSO v/v). The pre-soaked membranes were then placed into dialysis chambers. 500 μL of PBS was added to one side of the chamber, and 500 μL of the Matrix containing the test article was added to the other side of the chamber.
The chambers were then placed into an enclosed, heated rocker, which was pre-warmed to 37° C. and allowed to reach equilibrium for at least 22 hours. After 22 hours both sides were sampled. 100 μL of the donor side was added to 500 μL of PBS. 100 μL of the PBS side was added to 20 μL of fresh matrix. Samples then were crashed with 1:1 Acetonitrile and centrifuged at 10,000 RPM for 10 minutes. 100 μL of supernatant was placed into LC/MS vials for analysis.
Standards were prepared in a 1:5 plasma:PBS mixture at 5, 1.5, 0.5, 0.15, 0.05, 0.015 and 0.005 μM concentrations. The samples and standards were placed into HPLC vials and assayed by LC/MS. Protein binding values were calculated as follows:
% Bound=[(Concentration in Donor−Concentration in Receiver)/(Concentration in Donor)]×100.
% Recovery=[(Concentration in donor+Concentration in Receiver)]/(Concentration in Normal Initial)]×100
Compound 18 had high human plasma protein binding of more than 99.8%. Compound 18 protein binding in rat and mouse plasma could not be established due to low recovery of the test article.

EXAMPLE 11

Cytochrome p450 Inhibition Assessment

The ability of test compounds to inhibit five major human cytochrome p450 isozymes was evaluated. Cytochrome p450 inhibition assays were performed on 96-well microtiter assay plates according to the protocols described in Gentest P450 inhibition instruction manuals. The assays were performed in duplicate at 8 concentrations with the upper concentration of 100 μM followed by a 1:3 serial dilution. Reactions were initiated by the addition of 100 μL of enzyme/substrate mix to 100 μL of cofactor/serial dilution mix, and terminated by addition of 75 μL of a 4:1 acetonitrile: 0.5 M Tris base solution or by 2N NaOH for CYP3A4/DBF. Fluorescence was measured using a fluorescence plate reader (FLUOstar model 403, BMG Lab Technologies, Durham, N.C.). For reactions with inhibition greater than 50%, IC₅₀values were determined by fitting the data to the Hill equation using software GraphPad Prism (Version 4.02, GraphPad Software, San Diego, Calif.).
Known CYP inhibitors inhibited respective CYP enzymes in the expected manners, indicating that CYP enzymes were active and responsive. Compound 18 did not significantly inhibit 2C9, CYP2D6 and CYP3A4 activities in the concentration range tested. It inhibited CYP2C19 activity. The IC₅₀value was estimated to be 26.85 μM. It also inhibited CYP1A2 activity with an estimated IC₅₀of 97.45 μM.
From the foregoing description, various modifications and changes in the compositions and methods of this invention will occur to those skilled in the art. All such modifications coming within the scope of the appended claims are intended to be included therein.
All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

Claims

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44. A method for treating or managing a disease or condition which comprises administering to a patient in need of such treatment or management a therapeutically effective amount of a compound of the formula I′:

or a pharmaceutically acceptable salt, or stereoisomers, or tautomers thereof, wherein:

each of W, X, Y and Z is independently CR⁴;

R^1′ is aryl, heteroaryl, aralkyl, heteroaralkyl, amino or substituted amino;

each R⁴is independently hydrogen, substituted or unsubstituted alkyl, hydroxyalkyl, haloalkyl, amino, alkylamino, dialkylamino, arylamino, alkylarylamino, alkoxy, aryloxy, heteroaryloxy, aminoalkoxy, alkoxy, cycloalkylalkoxy, alkoxycarbonyl, arylalkyloxy, aryl, heteroaryl, arylalkyl, sulfo, sulfonyl, sulfanyl, aminosulfonyl, arylsulfonyl, carboxy, carbamoyl, cyano, cycloheteroalkyl, halo, heteroalkyl, hydroxyl, or thiol; and

the subscript n and the subscript m are independently 0, 1, 2, 3, or 4.

45. The method of claim 44 wherein the disease or condition is a pain condition.

46. The method of claim 44 wherein the disease or condition is an autoimmune disease.

47. The method of claim 44 wherein the disease or condition is an inflammatory disease or condition.

48. The method of claim 44 wherein the disease or condition is a neurological or neurodegenerative disease or condition.

49. A method for treating, ameliorating or managing a disease or condition, which comprises administering to a patient in need of such prevention, treatment or management a therapeutically acceptable amount of a compound of formula I′

each of W, X, Y and Z is independently CR⁴;

R^1′ is aryl, heteroaryl, aralkyl, heteroaralkyl, amino or substituted amino;

the subscript n and the subscript m are independently 0, 1, 2, 3, or 4; and

wherein the disease or condition is: pain including acute, inflammatory and neuropathic pain; chronic pain; dental pain; headache including migraine, cluster headache and tension headache; Parkinson's disease; Alzheimer's disease; multiple sclerosis; diseases and disorders mediated by or result in neuroinflammation, traumatic brain injury, stroke, or encephalitis; centrally-mediated neuropsychiatric diseases and disorders including depression, mania, bipolar disease, anxiety, schizophrenia, eating disorders, sleep disorders and cognition disorders; epilepsy and seizure disorders; prostate, bladder and bowel dysfunction, urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway disease and disorders including allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders mediated by or result in inflammation including arthritis, rheumatoid arthritis and osteoarthritis; myocardial infarction; autoimmune diseases and disorders; uveitis and atherosclerosis; itch/pruritus, psoriasis; alopecia (hair loss); obesity; lipid disorders; cancer; high blood pressure; spinal cord injury; or a renal disorder.

50. The method of claim 49 wherein the disease or condition is Parkinson's disease.

51. The method of claim 49 wherein the disease or condition is migraine.

52. The method of claim 49 wherein the disease or condition is overactive bladder.

53. The method of claim 49 wherein the disease or condition is osteoarthritis.

54. The method of claim 49 wherein the disease or condition is pain.

55. The method of claim 49 wherein the disease or condition is inflammatory pain

56. The method of claim 49 wherein the disease or condition is neuropathic pain.

57. The method of claim 49 wherein the disease or condition is dental pain