Benzoisoxazole-substituted compounds as MGLUR4 allosteric potentiators, compositions, and methods of treating neurological dysfunction

Compounds which are useful as allosteric potentiators/positive allosteric modulators of the metabotropic glutamate receptor subtype 4 (mGluR4); synthetic methods for making the compounds; pharmaceutical compositions comprising the compounds; and methods of using the compounds, for example, in treating neurological and psychiatric disorders or other disease state associated with glutamate dysfunction.

BACKGROUND

The amino acid L-glutamate (referred to herein simply as glutamate) is the principal excitatory neurotransmitter in the mammalian central nervous system (CNS). Within the CNS, glutamate plays a key role in synaptic plasticity (e.g., long term potentiation (the basis of learning and memory)), motor control and sensory perception. It is now well understood that a variety of neurological and psychiatric disorders, including, but not limited to, schizophrenia general psychosis and cognitive deficits, are associated with dysfunctions in the glutamatergic system. Thus, modulation of the glutamaterigic system is an important therapeutic goal. Glutamate acts through two distinct receptors: ionotropic and metabotropic glutamate receptors. The first class, the ionotropic glutamate receptors, is comprised of multi-subunit ligand-gated ion channels that mediate excitatory post-synaptic currents. Three subtypes of ionotropic glutamate receptors have been identified, and despite glutamate serving as agonist for all three receptor subtypes, selective ligands have been discovered that activate each subtype. The ionotropic glutamate receptors are named after their respective selective ligands: kainate receptors, AMPA receptors and NMDA receptors.

The second class of glutamate receptor, termed metabotropic glutamate receptors, (mGluRs), are G-protein coupled receptors (GPCRs) that modulate neurotransmitter release or the strength of synaptic transmission, based on their location (pre- or postsynaptic). The mGluRs are family C GPCR, characterized by a large (˜560 amino acid) “venus fly trap” agonist binding domain in the amino-terminal domain of the receptor. This unique agonist binding domain distinguishes family C GPCRs from family A and B GPCRs wherein the agonist binding domains are located within the 7-strand transmembrane spanning (7TM) region or within the extracellular loops that connect the strands to this region. To date, eight distinct mGluRs have been identified, cloned and sequenced. Based on structural similarity, primary coupling to intracellular signaling pathways and pharmacology, the mGluRs have been assigned to three groups; Group I (mGluR1 and mGluR5), Group II (mGluR2 and mGluR3) and Group III (mGluR4, mGluR6, mGluR7 and mGluR8). Group I mGluRs are coupled through Gαq/11 to increase inositol phosphate and metabolism and resultant increases in intracellular calcium. Group I mGluRs are primarily located post-synaptically and have a modulatory effect on ion channel activity and neuronal excitability. Group II (mGluR2 and mGluR3) and Group III (mGluR4, mGluR6, mGluR7 and mGluR8) mGluRs are primarily located pre-synaptically where they regulate the release of neurotransmitters, such as glutamate. Group II and Group III mGluRs are coupled to Gαi and its associated effectors such as adenylate cyclase.

mGluR4 belongs to the group III mGluR subfamily and is located in predominantly presynaptic locations in the central nervous system (Benitez et at., 2000; Bradley et al., 1996; Bradley et al., 1999; Mateos et al., 1998; Phillips et al., 1997) where it is functions as an auto- and heteroreceptor to regulate the release of both GABA and glutamate. mGluR4 has also been shown to be expressed at a low level in some postsynaptic locations (Benitez et al., 2000). Numerous reports indicate that mGluR4 is expressed in most brain regions, particularly in neurons known to play key roles in functions of the basal ganglia (Bradley et al., 1999; Corti et al., 2002; Kuramoto et al. 2007; Marino et al., 2003a), learning and memory (Bradley et al., 1996), vision (Akazawa et al., 1994; Koulen et al., 1996; Quraishi et al., 2007), cerebellar functions (Makoff et al., 1996), feeding and the regulation of hypothalamic hormones (Flor et al., 1995), sleep and wakefulness (Noriega et al., 2007) as well as many others. There are now a number of literature reports describing a role for mGluR4 modulation in Parkinson's disease (Battaglia et al., 2006; Lopez et al., 2007; Marino et al., 2005; Marino et al., 2003b; Ossowska et al., 2007; Valenti et al., 2003), anxiety (Stachowicz et al., 2006; Stachowicz et al., 2004), motor effects after alcohol consumption (Blednov et al., 2004), neurogenic fate commitment and neuronal survival (Saxe et al., 2007), epilepsy (Chapman et al., 2001; Pitsch et al., 2007; Snead et al., 2000; Wang et al., 2005) and cancer, particularly medulloblastoma (Iacovelli et al., 2004).

In addition, there is evidence that activation of mGluR4 receptors (expressed in islets of Langerhans) would inhibit glucagon secretion (Uehara et al., 2004). Thus, activation of mGluR4 may be an effective treatment for disorders involving detects in glucose metabolism such ashypoglycemia. Type 2 diabetes, and obesity.

Also, there are reports that activation of Group III mGluRs, specifically mGluR4, may be an effective treatment for neuroinflammatory diseases, such as multiple sclerosis and related disorders (Besong et al., 2002).

Also, that activation of Group III mGluRs, specifically mGlu4 positive allosteric modulators (PAMs), may be an effective treatment for neuroinflammatory diseases, such as multiple sclerosis and related disorders (Besong et al., 2002; Taylor et al., 2003; Fallarino et al., 2010).

There are two variants of the mGluR4 receptor which are expressed in taste tissues; and thus activation of mGluR4 may be used as taste enhancers, blockade of certain tastes, or taste agents, flavoring agents or other food additives (Kurihara, 2009; Chaudhari et al., 2009).

Despite advances in mGluR4 research, there is still a scarcity of compounds that effectively potentiate mGluR4 which are also effective in the treatment of neurological and psychiatric disorders associated with glutamatergic neurotransmission dysfunction and diseases. As well as inflammatory central nervous system disorders, medulloblastomas, metabolic disorders and taste enhancing associated with glutamatergic dysfunction and diseases in which mGluR4 receptor is involved. Further, conventional mGluR4 receptor modulators typically lack satisfactory aqueous solubility and exhibit poor oral bioavailability. These needs and other needs are satisfied by the present invention.

SUMMARY

In accordance with the purpose(s) of the invention, as embodied and broadly described herein, the invention, in one aspect, relates to compounds useful as allosteric modulators of mGluR4 receptor activity, methods of making same, pharmaceutical compositions comprising same, and methods of treating neurological and psychiatric disorders associated with glutamate dysfunction, for example Parkinson's disease, using same. Further disclosed are methods and pharmaceutical compositions useful for treating a disease related to mGluR4 activity. In one aspect, the disclosed compounds can affect the sensitivity of mGluR4 receptors to agonists without binding to the orthosteric agonist binding site or acting as orthosteric agonists themselves. A “receptor allosteric agonist”, as used herein, is generally defined as a ligand that functions as both an allosteric modulator and as an agonist on its own (though the latter is usually only at higher concentrations). The presence of a the receptor allosteric agonist (Ago-PAM) activity may offer advantages in various CNS and neurological disorders where the basal glutamatergic tone is low in given brain regions.

Disclosed are methods for the treatment of a neurotransmission dysfunction or other disease state associated with mGluR4 activity in a mammal comprising the step of administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula (I):

Also disclosed are methods for potentiating mGluR4 activity in a subject comprising the step of administering to the subject at least one compound at least one compound having a structure represented by formula (I):

Also disclosed are methods of potentiating mGluR4 activity in at least one cell comprising the step of contacting at least one cell with at least one compound having a structure represented by formula (I);

Also disclosed are compounds having a structure represented by formula (I):

Also disclosed pharmaceutical compositions comprising a compound having a structure represented by formula (I):

Also disclosed are methods for potentiating mGluR4 activity in at least one cell comprising the step of contacting at least one cell with at least one disclosed compound in an amount effective to potentiate mGluR4 receptor activity in at least one cell.

Also disclosed are methods for potentiating mGluR4 activity in a subject comprising the step of administering to the subject a therapeutically effective amount of at least one disclosed compound in a dosage and amount effective to potentiate mGluR4 receptor activity in the subject.

Also disclosed are methods for the treatment of a disorder associated with mGluR4 neurotransmission dysfunction or other mGluR4 mediated disease states in a mammal comprising the step of administering to the mammal at least one disclosed compound in a dosage and amount effective to treat the disorder in the mammal.

Also disclosed are methods for making a compound comprising the steps of providing an amine compound having a structure represented by formula (I):

Also disclosed are the products of the disclosed methods of making.

Also disclosed are methods for the manufacture of a medicament for potentiating mGluR4 receptor activity in a mammal comprising combining a compound having a structure represented by formula (I) :

Also disclosed are the products of the disclosed methods for the manufacture of a medicament.

Also disclosed are uses of a compound for potentiating mGluR4 receptor activity in a mammal, wherein the compound has a structure represented by formula (I):

Also disclosed are methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula (I):

Also disclosed are methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula (I):

Also disclosed are methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by a Also disclosed are methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula (I):

Also disclosed are kits comprising a compound of the present invention.

Also disclosed are methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula (I):

DESCRIPTION

As used herein, the term “receptor positive allosteric modulator” refers to any exogenously administered compound or agent that directly or indirectly augments the activity of any form (for example, homomeric, heteromeric, oligomeric, or interacting, with other proteins or cellular components) of the receptor in the presence or in the absence of the endogenous ligand (such as glutamate, L-serine O-phosphate (L-SOP), other endogenous ligands, other neurotransmitters, etc.) in an animal, in particular a mammal, for example a human. The term “receptor positive allosteric modulator” includes a compound that is a “receptor allosteric potentiator” or a “receptor allosteric agonist,” as well as a compound that has mixed activity as both a “receptor allosteric potentiator” and an “mGluR receptor allosteric agonist.”

As used herein, the term “receptor allosteric potentiator” refers to any exogenously administered compound or agent that directly or indirectly augments the response of any form (for example, homomeric, heteromeric, oligomeric, or interacting with other proteins or cellular components) of the receptor produced by the endogenous ligand (such as glutamate, L-serine O-phosphate (L-SOP), other endogenous ligands, other neurotransmitters, etc.) when it binds to an allosteric site of any form of receptor in an animal, in particular a mammal, for example a human. The receptor allosteric potentiator binds to a site other than the orthosteric site (an allosteric site) and positively augments the response of the receptor to an agonist. As it is predicted to induce less desensitization of the receptor, activity of a compound as a receptor allosteric potentiator provides advantages over the use of a pure receptor allosteric agonist. Such advantages can include, for example, increased safety margin, higher tolerability, diminished potential for abuse, and reduced toxicity.

As used herein, the term “receptor allosteric agonist” refers to any exogenously administered compound or agent that directly augments the activity of any form (for example, homomeric, heteromeric, oligomeric, or interacting with other proteins or cellular components) of the receptor in the absence of the endogenous ligand (such as glutamate, L-serine O-phosphate (L-SOP), other endogenous ligands, other neurotransmitters, etc.) in an animal, in particular a mammal, for example a human. The receptor allosteric agonist binds to the allosteric glutamate site of the receptor and directly influences the orthosteric site of the receptor. The term “receptor positive allosteric modulator” refers to any exogenonsly administered compound or agent that directly or indirectly augments the activity of the receptor in the presence or in the absence of the endogenous ligand (such as glutamate, L-serine O-phosphate (L-SOP), other endogenous ligands, other neurotransmitters, etc.) in an animal, in particular a mammal, for example a human. The term “receptor positive allosteric modulator” includes a compound that is a “receptor allosteric potentiator” or a “receptor allosteric agonist,” as well as a compound that has mixed activity as both a “receptor allosteric potentiator” and an “mGluR receptor allosteric agonist.”

In some aspects of the disclosed methods, the subject has been diagnosed with a need for treatment of one or more neurological and/or psychiatric disorder and/or any other disease state associated with glutamate dysfunction prior to the administering step. In some aspects of the disclosed method, the subject has been diagnosed with a need for potentiation of metabotropic glutamate receptor activity prior to the administering step. In some aspects of the disclosed method, the subject has been diagnosed with a need for partial agonism of metabotropic glutamate receptor activity prior to the administering step. In some aspects, the disclosed methods can further comprise a step of identifying a subject having a need for treatment of a disclosed disorder.

As used herein, the term “prevent” or “preventing” refers to precluding, averting, obviating, forestalling, stopping, or hindering something from happening, especially by advance action. It is understood that where reduce, inhibitor prevent are used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed.

As used herein, the term “diagnosed” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by the compounds, compositions, or methods disclosed herein. For example, “diagnosed with a disorder treatable by potentiation of mGluR4 activity” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by a compound or composition that can favorably potentiate mGluR4 activity. As a further example, “diagnosed with a need for potentiation of mGluR4 activity” refers to having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition characterized by abnormal mGluR4 activity. Such a diagnosis can be in reference to a disorder, such as Parkinson's disease, and the like, as discussed herein.

As used herein, the phrase “identified to be in need of treatment for a disorder,” or the like, refers to selection of a subject based upon need for treatment of the disorder. For example, a subject can be identified as having a need for treatment of a disorder (e.g., a disorder related to mGluR4 activity) based upon an earlier diagnosis by a person of skill and thereafter subjected to treatment for the disorder. It is contemplated that the identification can, in one aspect, be performed by a person different from the person making the diagnosis. It is also contemplated, in a further aspect, that the administration can be performed by one who subsequently performed the administration.

As used herein, the term “receptor allosteric agonist”, as used herein, is generally defined as a ligand that functions as both an allosteric modulator and as an agonist on its own (though the latter is usually only at higher concentrations).

As used herein, the term “diagnosed with a need for potentiation of metabotropic glutamate receptor activity” refers to having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by potentiation of metabotropic glutamate receptor activity.

As used herein, “diagnosed with a need for partial agonism of metabotropic glutamate receptor activity” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by partial agonism of metabotropic glutamate receptor activity.

As used herein, “diagnosed with a need for treatment of one or more neurological and/or psychiatric disorder or any disease state associated with glutamate dysfunction” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have one or more neurological and/or psychiatric disorder associated with glutamate dysfunction.

A residue of a chemical species, as used in the specification and concluding claims, refers to the moiety that is the resulting product of the chemical species in a particular reaction scheme or subsequent formulation or chemical product, regardless of whether the moiety is actually obtained from the chemical species. Thus, an ethylene glycol residue in a polyester refers to one or more —OCH2CH2O— units in the polyester regardless of whether ethylene glycol was used to prepare the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more —CO(CH2)8CO— moieties in the polyester, regardless of whether the residue is obtained by reacting sebacic acid or an ester thereof to obtain the polyester.

As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. The permissible substituents can be one or more and the same or different for appropriate organic compounds. In other words, all substitutions are “independent.” For example, “substituted with at least one R” means that each R group is independent from the other one, and as such may be the same or different. For purposes of this disclosure, the heteroatoms, such, as nitrogen, can have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. Also, the terms “substitution” or “substituted with” include the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

The term “alkyl” as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. The alkyl group can be cyclic or acyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted with one or more groups including, but not limited to, optionally substituted alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein. A “lower alkyl” group is an alkyl group containing from one to six (e.g., from one to four) carbon atoms.

The term “polyalkylene group” as used herein is a group having two or more CH2groups linked to one another. The polyalkylene group can be represented by a formula —(CH2)a—, where “a” is an integer of from 2 to 500.

The term “aldehyde” as used herein is represented by a formula —C(O)H. Throughout this specification “C(O)” is a short hand notation for a carbonyl group, i.e., C═O.

The terms “amine” or “amino” as used herein are represented by a formula NA1A2A3, where A1, A2, and A3can be, independently, hydrogen or optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

The term “carboxylic acid” as used herein is represented by a formula —C(O)OH.

The term “ester” as used herein is represented by a formula —OC(O)A1or —C(O)OA1, where A1can be an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “polyester” as used herein is represented by a formula -(A1O(O)C-A2-C(O)O)a— or -(A1O(O)C-A2OC(O))a—, where A1and A2can be, independently, an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group described herein and “a” is an integer from 1 to 500. “Polyester” is as the term used to describe a group that is produced by the reaction between a compound having at least two carboxylic acid groups with a compound having at least two hydroxyl groups.

The term “halide” as used herein refers to the halogens fluorine, chlorine, bromine, and iodine.

The term “hydroxyl” as used herein is represented by a formula —OH.

The term “ketone” as used herein is represented by a formula A1C(O)A2, where A1and A2can be, independently, an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein.

The term “azide” as used herein is represented by a formula —N3.

The term “nitro” as used herein is represented by a formula —NO2.

The term “nitrile” as used herein is represented by a formula —CN.

The term “silyl” as used herein is represented by a formula —SiA1A2A3, where A1, A2and A3can be, independently, hydrogen or an optionally substituted alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

The term “sulfo-oxo” as used herein is represented by a formulas —S(O)A1, —S(O)2A1, —OS(O)2A1, or —OS(O)2OA1, where A1can be hydrogen or an optionally substituted alkyl, cycloalkyl alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. Throughout this specification “S(O)” is a short hand notation for S═O. The term “sulfonyl” is used herein to refer to the sulfo-oxo group represented by a formula —S(O)2A1, where A1can be hydrogen or an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl or heteroaryl group as described herein. The term “sulfone” as used herein is represented by a formula A1S(O)2A2, where A1and A2can be, independently, an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “sulfoxide” as used herein is represented by a formula A1S(O)A2; where A1and A2can be, independently, an optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

The term “thiol” as used herein is represented by a formula —SH.

The term “organic residue” defines a carbon containing residue, i.e., a residue comprising at least one carbon atom, and includes but is not limited to the carbon-containing groups, residues, or radicals defined hereinabove. Organic residues can contain various heteroatoms, or be bonded to another molecule through a heteroatom, including oxygen, nitrogen, sulfur, phosphorus, or the like. Examples of organic residues include but are not limited alkyl or substituted alkyls, alkoxy or substituted alkoxy, mono or di-substituted amino, amide groups, etc. Organic residues can preferably comprise 1 to 18 carbon atoms, 1 to 15, carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In a further aspect, an organic residue can comprise 2 to 18 carbon atoms, 2 to 15, carbon atoms, 2 to 12 carbon atoms, 2 to 8 carbon atoms, 2 to 4 carbon atoms, or 2 to 4 carbon atoms

A very close synonym of the term “residue” is the term “radical,” which as used in the specification and concluding claims, refers to a fragment, group, or substructure of a molecule described herein, regardless of how the molecule is prepared. For example, a 2,4-thiazolidinedione radical in a particular compound has the structure

regardless of whether thiazolidinedione is used to prepare the compound. In some aspects the radical (for example an alkyl) can be further modified (i.e., substituted alkyl) by having bonded thereto one or more “substituent radicals.” The number of atoms in a given radical is not critical to the present invention unless it is indicated to the contrary elsewhere herein.

“Organic radicals,” as the term is defined and used herein, contain one or more carbon atoms. An organic radical can have, for example, 1-26 carbon atoms, 1-18 carbon atoms, 1-12 carbon atoms, 1-8 carbon atoms, 1-6 carbon atoms, or 1-4 carbon atoms. In a further aspect, an organic radical can have 2-26 carbon atoms, 2-18 carbon atoms, 2-12 carbon atoms, 2-8 carbon atoms, 2-6 carbon atoms, or 2-4 carbon atoms. Organic radicals often have hydrogen bound to at least some of the carbon atoms of the organic radical. One example, of an organic radical that comprises no inorganic atoms is a 5,6,7,8-tetrahydro-2-naphthyl radical. In some aspects, an organic radical can contain 1-10 inorganic heteroatoms bound thereto or therein, including halogens, oxygen, sulfur, nitrogen, phosphorus, and the like. Examples of organic radicals include but are not limited to an alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, mono-substituted amino, di-substituted amino, acyloxy, cyano, carboxy, carboalkoxy, alkylcarboxamide, substituted alkylcarboxamide, dialkylcarboxamide, substituted dialkylcarboxamide, alkylsulfonyl, alkylsulfinyl, thioalkyl, thiohaloalkyl, alkoxy, substituted alkoxy, haloalkyl haloalkoxy, aryl, substituted aryl, heteroaryl heterocyclic, or substituted heterocyclic radicals, wherein the terms are defined elsewhere herein. A few non-limiting examples of organic radicals that include heteroatoms include alkoxy radicals, trifluoromethoxy radicals, acetoxy radicals, dimethylamino radicals and the like.

“Inorganic radicals” as the term is defined and used herein, contain no carbon atoms and therefore comprise only atoms other than carbon. Inorganic radicals comprise bonded combinations of atoms selected from hydrogen, nitrogen, oxygen, silicon, phosphorus, sulfur, selenium, and halogens such as fluorine, chlorine, bromine, and iodine, which can be present individually or bonded together in their chemically stable combinations. Inorganic radicals have 10 or fewer, or preferably one to six or one to four inorganic atoms as listed above bonded together. Examples of inorganic radicals include, but not limited to, amino, hydroxy, halogens, nitro, thiol, sulfate, phosphate, and like commonly known inorganic radicals. The inorganic radicals do not have bonded therein the metallic elements of the periodic table (such as the alkali metals, alkaline earth metals, transition metals, lanthanide metals, or actinide metals), although such metal ions can sometimes serve as a pharmaceutically acceptable cation for anionic inorganic radicals such as a sulfate, phosphate, or like anionic inorganic radical. Typically, inorganic radicals do not comprise metalloids elements such as boron, aluminum, gallium, germanium, arsenic, tin, lead, or tellurium, or the noble gas elements, unless otherwise specifically indicated elsewhere herein.

As used herein, the term “derivative” refers to a compound having a structure derived from the structure of a parent compound (e.g., a compounds disclosed herein) and whose structure is sufficiently similar to those disclosed herein and based upon that similarity, would be expected by one skilled in the art to exhibit the same or similar activities and utilities as the claimed compounds, or to induce, as a precursor, the same or similar activities and utilities as the claimed compounds. Exemplary derivatives include salts, esters, amides, salts of esters or amides, and N-oxides of a parent compound.

The term “hydrolysable residue” is meant to refer to a functional group capable of undergoing hydrolysis, e.g., under basic or acidic conditions. Examples of hydrolysable residues include, without limitation, residues of acid halides or activated carboxylic acids, residues of trialkylsilyl halides, residues of alkyloxymethyl halides, and various other protecting groups known in the art (see, for example, “Protective Groups in Organic Synthesis,” T. W. Greene, P. G. M. Wuts, Wiley-Interscience, 1999).

The term “leaving group” refers to an atom (or a group of atoms) with electron withdrawing ability that can be displaced as a stable species, taking with it the bonding electrons. Examples of suitable leaving groups include sulfonate esters, including, but not limited to, triflate, mesylate, tosylate, brosylate, and halides.

which is understood to be equivalent to a formula:

wherein n is typically an integer. That is, Rnis understood to represent five independent substituents, Rn(a), Rn(b), Rn(c), Rn(d), Rn(e). By “independent substituents,” it is meant that each R substituent can be independently defined. For example, if in one instance Rn(a)is halogen, then Rn(b)is not necessarily halogen in that instance. Likewise, when a group R is defined as four substituents, R is understood to represent four independent substituents, Ra, Rb, Rc, and Rd. Unless indicated to the contrary, the substituents are not limited to any particular order or arrangement.

In one aspect, the invention relates to compounds, or pharmaceutically acceptable derivatives thereof, useful as potentiators of mGluR4 activity. In general, it is contemplated that each disclosed derivative can be optionally further substituted. It is also contemplated that any one or more derivative can be optionally omitted from the invention. It is understood that a disclosed compound can be provided by the disclosed methods. It is also understood that the disclosed compounds can be employed in the disclosed methods of using. It is also understood that the disclosed compounds can all be employed as corresponding pharmaceutical compositions.

In one aspect, the invention relates to compounds having a structure represented by formula (I):

X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof.

Also disclosed are compounds of the following formula:

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof.

Also disclosed are compounds of the following formula:

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof.

Also disclosed are compounds of the following formula:

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof.

Also disclosed are compounds of the following formula:

The compounds disclosed herein can include all salt forms, for example, salts of both basic groups, inter alia, amines, as well as salts of acidic groups, inter alia, carboxylic acids. The following are non-limiting examples of anions that can form salts with protonated basic groups: chloride, bromide, iodide, sulfate, bisulfate, carbonate, bicarbonate, phosphate, formate, acetate, propionate, butyrate, pyruvate, lactate, oxalate, malonate, maleate, succinate, tartrate, fumarate, citrate, and the like. The following are non-limiting examples of cations that can form salts of acidic groups: ammonium, sodium, lithium, potassium, calcium, magnesium, bismuth, lysine, and the like.

The analogs (compounds) of the present disclosure are arranged into several categories to assist the formulator in applying a rational synthetic strategy for the preparation of analogs which are not expressly exampled herein. The arrangement into categories does not imply increased or decreased efficacy for any of the compositions of matter described herein.

In one aspect, the invention relates to pharmaceutical compositions comprising the disclosed compounds. That is, a pharmaceutical composition can be provided comprising a therapeutically effective amount of at least one disclosed compound or at least one product of a disclosed method and a pharmaceutically acceptable carrier.

In practice, the compounds of the invention, or pharmaceutically acceptable salts thereof, of this invention can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compounds of the invention, and/or pharmaceutically acceptable salt(s) thereof, can also be administered by controlled release means and/or delivery devices. The compositions can be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

Thus, the pharmaceutical compositions of this invention can include a pharmaceutically acceptable carrier and a compound or a pharmaceutically acceptable salt of the compounds of the invention. The compounds of the invention, or pharmaceutically acceptable salts thereof, can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds. The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

In preparing the compositions for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques

The pharmaceutical compositions of the present invention can comprise a compound of the invention (or pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents or adjuvants. The instant compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

Pharmaceutical compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, mouth washes, gargles, and the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations can be prepared, utilizing a compound of the invention, or pharmaceutically acceptable salts thereof, via conventional processing methods. As an example, a cream or ointment is prepared by mixing hydrophilic material and water, together with, about 5 wt % to about 10 wt % of the compound, to produce a cream or ointment having a desired consistency.

A potentiated amount of an mGluR agonist to be administered in combination with an effective amount of a disclosed compound is expected to vary from about 0.1 milligram per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day and is expected to be less than the amount that is required to provide the same effect when administered without an effective amount of a disclosed compound. Preferred amounts of a co-administered mGluR agonist are able to be determined by one skilled in the art.

In the treatment conditions which require potentiation of metabotropic glutamate receptor activity an appropriate dosage level will generally be about 0.01 to 500 mg per kg patient body weight per day and can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg/kg per day; more preferably 0.5 to 100 mg/kg per day. A suitable dosage level can be about 0.01 to 250 mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kg per day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5.0 or 5.0 to 50 mg/kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient for the symptomatic adjustment of the dosage of the patient to be treated. The compound can be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosing regimen can be adjusted to provide the optimal therapeutic response.

It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors. Such factors include the age, body weight, general health, sex, and diet of the patient. Other factors include the time and route of administration, rate of excretion, drug combination, and the type and severity of the particular disease undergoing therapy.

The disclosed pharmaceutical compositions can further comprise other therapeutically active compounds, which are usually applied in the treatment of the above mentioned pathological conditions.

It is understood that the disclosed compositions can be prepared from the disclosed compounds. It is also understood that the disclosed compositions can be employed in the disclosed methods of using.

Further disclosed herein are pharmaceutical compositions comprising one or more of the disclosed mGluR4 potentiators and a pharmaceutically acceptable carrier.

Accordingly, the pharmaceutical compositions of the present invention include those that contain one or more other active ingredients, in addition to a compound of the present invention.

The above combinations include combinations of a disclosed compound not only with one other active compound, but also with two or more other active compounds. Likewise, disclosed compounds may be used in combination with other drugs that are used in the prevention, treatment, control, amelioration, or reduction of risk of the diseases or conditions for which disclosed compounds are useful. Such other drugs may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of the present invention. When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of the present invention.

In such combinations the compound of the present invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element can be prior to, concurrent to, or subsequent to the administration of other agent(s).

Accordingly, the subject compounds can be used alone or in combination with other agents which are known to be beneficial in the subject indications or other drugs that affect receptors or enzymes that either increase the efficacy, safety, convenience, or reduce unwanted side effects or toxicity of the disclosed compounds. The subject compound and the other agent may be coadministered, either in concomitant therapy or in a fixed combination.

In a further aspect, the compound can be employed in combination with levodopa (with or without a selective extracerebral decarboxylase inhibitor such as carbidopa or benserazide), anticholinergics such as biperiden (optionally as its hydrochloride or lactate salt) and trihexyphenidyl (benzhexol) hydrochloride, COMT inhibitors such as entacapone, MOA-B inhibitors, antioxidants, A2a adenosine receptor antagonists, cholinergic agonists, NMDA receptor antagonists, serotonin receptor antagonists and dopamine receptor agonists such as alentemol, bromocriptine, fenoldopam, lisuride, naxagolide, pergolide and pramipexole. It will be appreciated that the dopamine agonist may be in the form of a pharmaceutically acceptable salt, for example, alentemol hydrobromide, bromocriptine mesylate, fenoldopam mesylate, naxagolide hydrochloride and pergolide mesylate. Lisuride and pramipexol are commonly used in a non-salt form.

In a further aspect, the compound can be employed in combination with a compound from the phenothiazine, thioxanthene, heterocyclic dibenzazepine, butyrophenone, diphenylbutylpiperidine and indolone classes of neuroleptic agent. Suitable examples of phenothiazines include chlorpromazine, mesoridazine, thioridazine, acetophenazine, fluphenazine, perphenazine and trifluoperazine. Suitable examples of thioxanthenes include chlorprothixene and thiothixene. An example of a dibenzazepine is clozapine. An example of a butyrophenone is haloperidol. An example of a diphenylbutylpiperidine is pimozide. An example of an indolone is molindolone. Other neuroleptic agents include loxapine, sulpiride and risperidone. It will be appreciated that the neuroleptic agents when used in combination with the subject compound may be in the form of a pharmaceutically acceptable salt, for example, chlorpromazine hydrochloride, mesoridazine besylate, thioridazine hydrochloride, acetophenazine maleate, fluphenazine hydrochloride, flurphenazine enathate, fluphenazine decanoate, trifluoperazine hydrochloride, thiothixene hydrochloride, haloperidol decanoate, loxapine succinate and molindone hydrochloride. Perphenazine, chlorprothixene, clozapine, haloperidol, pimozide and risperidone are commonly used in a non-salt form. Thus, the subject compound may be employed in combination with acetophenazine, alentemol, aripiprazole, amisulpride, benzhexol, bromocriptine, biperiden, chlorpromazine, chlorprothixene, clozapine, diazepam, fenoldopam, fluphenazine, haloperidol, levodopa, levodopa with benserazide, levodopa with carbidopa, lisuride, loxapine, mesoridazine, molindolone, naxagolide, olanzapine, pergolide, perphenazine, pimozide, pramipexole, quetiapine, risperidone, sulpiride, tetrabenazine, trihexyphenidyl, thioridazine, thiothixene, trifluoperazine or ziprasidone.

In the treatment of conditions which require potentiation of mGluR4 activity an appropriate dosage level will generally be about 0.01 to 500 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg/kg per day; more preferably about 0.5 to about 100 mg/kg per day. A suitable dosage level may be about 0.01 to 250 mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, and 1000 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosage regimen may be adjusted to provide the optimal therapeutic response. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

In one aspect, the invention relates to a pharmaceutical composition comprising a compound disclosed herein or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof; and a pharmaceutically acceptable carrier.

In one aspect, the invention relates to pharmaceutical compositions comprising a compound having a structure represented by a compound of the following formula:

X is CH, C-alkyl, M, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof, and a pharmaceutically acceptable carrier.

Also disclosed are pharmaceutical compositions comprising a compound of the following formula:

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof; and a pharmaceutically acceptable carrier.

Also disclosed are pharmaceutical compositions comprising a compound of the following formula:

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof; and a pharmaceutically acceptable carrier.

Also disclosed are pharmaceutical compositions comprising a compound of the following formula:

Also disclosed are pharmaceutical compositions wherein the compound is of the following formula:

D. Methods of Using the Compounds and Compositions

mGluR4 belongs to the group III mGluR subfamily and is located in predominantly in presynaptic locations in the central nervous system where it is functions as an auto- and heteroreceptor to regulate the release of both GABA and glutamate. In addition, mGluR4 is also expressed at a low level in some postsynaptic locations, mGluR4 is expressed in most brain regions, particularly in neurons known to play key roles in the following functions of the CNS:

a) learning and memory;

b) regulation of voluntary movement and other motor functions

c) motor learning

d) emotional responses

e) habit formation, including repetitive tasks and perseverative thought processes

f) reward systems

g) vision and olfaction

i) feeding and the regulation of hypothalamic hormones; and

j) sleep and wakefulness.

As such, mGluR4 plays a major role in the modulation of CNS-related diseases, syndromes and non-CNS related diseases or conditions the like, for example:

c) Huntington's diseases and other disorders involving involuntary movements and dyskinesias

d) Tourette's syndrome and related ticking disorders

e) Obsessive/compulsive disorder and other perseverative behavioral disorders

g) Schizophrenia and other psychotic disorders

j) motor effects after alcohol consumption or other drug-induced motor disorders;

k) neurogenic fate commitment and neuronal survival;

The disclosed compounds can act as potentiators of the metabotropic glutamate receptor activity (mGluR4). Therefore, in one aspect, the disclosed compounds can be used to treat one or more mGluR4 associated disorders that result in dysfunction in a mammal.

The disclosed compounds can be used as single agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of risk of the aforementioned diseases, disorders and conditions for which compounds of formula I or the other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. The other drug(s) can be administered by a route and in an amount commonly used therefore, contemporaneously or sequentially with a disclosed compound. When a disclosed compound is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compound is preferred. However, the combination therapy can also be administered on overlapping schedules. It is also envisioned that the combination of one or more active ingredients and a disclosed compound will be more efficacious than either as a single agent.

1. Treatment Methods

The compounds disclosed herein are useful for treating, preventing, ameliorating, controlling, or reducing the risk of a variety of neurological and psychiatric disorders associated with glutamate dysfunction. Thus, provided is a method of treating or preventing a disorder in a subject comprising the step of administering to the subject at least one disclosed compound; at least one disclosed pharmaceutical composition; and/or at least one disclosed product in a dosage and amount effective to treat the disorder in the subject.

Also provided is a method for the treatment of one or more neurological and/or psychiatric disorders associated with glutamate dysfunction in a subject comprising the step of administering to the subject at least one disclosed compound; at least one disclosed pharmaceutical composition; and/or at least one disclosed product in a dosage and amount effective to treat the disorder in the subject.

Anxiety disorders that can be treated or prevented by the compositions disclosed herein include generalized anxiety disorder, panic disorder, and obsessive compulsive disorder. Addictive behaviors include addiction to substances (including opiates, nicotine, tobacco products, alcohol, benzodiazepines, cocaine, sedatives, hypnotics, etc.), withdrawal from such addictive substances (including substances such, as opiates, nicotine, tobacco products, alcohol, benzodiazepines, cocaine, sedatives, hypnotics, etc.) and substance tolerance.

Also provided is a method for treating or prevention anxiety, comprising: administering to a subject at least one disclosed compound; at least one disclosed pharmaceutical composition; and/or at least one disclosed product in a dosage and amount effective to treat the disorder in the subject. At present, the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) (1994, American Psychiatric Association, Washington, D.C.), provides a diagnostic tool including anxiety and related disorders. These include: panic disorder with or without agoraphobia, agoraphobia without history of panic disorder, specific phobia, social phobia, obsessive-compulsive disorder, post-traumatic stress disorder, acute stress disorder, generalized anxiety disorder, anxiety disorder due to a general medical condition, substance-induced anxiety disorder and anxiety disorder not otherwise specified.

In one aspect, the invention relates to methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of administering to the mammal at least one compound of the present invention in a dosage and amount effective to treat the dysfunction in the mammal. In certain embodiments, the compound has a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

In one aspect, the invention relates to methods for potentiating mGluR4 activity in a subject comprising the step of administering to the subject at least one compound of the present invention in a dosage and amount effective to treat the dysfunction in the mammal. In certain embodiments, the compound has a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

In one aspect, the invention relates to methods of potentiating mGluR4 activity in at least one cell comprising the step of contacting the at least one cell with at least one compound having a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

In certain aspects, a subject, for example a mammal or a human, has been diagnosed with the dysfunction prior to the administering step. In further aspects, a disclosed method can further comprise the step of identifying a subject, for example a mammal or a human, having a need for treatment of a dysfunction. In further aspects, a subject for example a mammal or a human, has been diagnosed with a need for potentiation of mGluR4 receptor activity prior to the administering step. In further aspects, a disclosed method can further comprise the step of identifying a subject, for example a mammal or a human, having a need for potentiation of mGluR4 receptor activity. In further aspects, a cell (e.g., a mammalian cell or a human cell) has been isolated from a subject, for example a mammal or a human, prior to the contacting step. In further aspects, contacting is via administration to a subject, for example a mammal or a human.

In one aspect, the invention relates to methods for potentiating mGluR4 activity in at least one cell comprising the step of contacting the at least one cell with at least one disclosed compound in an amount effective to potentiate mGluR4 receptor activity in the at least one cell.

In one aspect, the invention relates to methods for potentiating mGluR4 activity in a subject comprising the step of administering to the subject a therapeutically effective amount of at least one disclosed compound in a dosage and amount effective to potentiate mGluR4 receptor activity in the subject.

In one aspect, the invention relates to methods for the treatment of a disorder associated with mGluR4 neurotransmission dysfunction or other disease state in a mammal comprising the step of administering to the mammal at least one disclosed compound in a dosage and amount effective to treat the disorder in the mammal.

The disclosed compounds can be used to treat a wide range of neurological and psychiatric disorders and other disease states associated with glutamate dysfunction. Non-limiting examples of these diseases includes movement disorders, including akinesias and akinetic-rigid syndromes (including Parkinson's disease), dystonia, epilepsy, chorea, neurogenerative diseases such as dementia, Huntington's disease, Amyotrophic Lateral Sclerosis, Alzheimer's disease, Pick's disease, Creutzfeldt-Jakob disease, pain, migraines, diabetes, obesity and eating disorders, sleep disorders including narcolepsy, and anxiety or affective disorders, including generalized anxiety disorder, panic attacks, unipolar depression, bipolar disorder, psychotic depression, and related disorders, cognitive disorders including dementia (associated with Alzheimer's disease, ischemia, trauma, stroke, HIV disease, Parkinson's disease, Huntington's disease and other general medical conditions or substance abuse), delirium, amnestic disorders, age-related cognitive decline, schizophrenia or psychosis including schizophrenia (paranoid, disorganized, catatonic or undifferentiated), schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, substance-related disorder, cancer and inflammation (including MS). Of the disorders above, the treatment of Parkinson's disease, movement disorders, cognitive disorders, neurodegenerative diseases, obesity and pain are of particular importance.

In one aspect, the disclosed compounds can be used to treat, or can be a component of a pharmaceutical composition used to treat movement disorders. As such, disclosed herein in a method for treating a movement disorder, comprising the step of administering to a mammal in need of treatment at least one compound in a dosage and amount effective to treat the disorder in the mammal, wherein the disorder is selected from Parkinson's disease, Huntington's disease, dystonia, Wilson's disease, chorea, ataxia, ballism, akathesia, athetosis, bradykinesia, rigidity, postural instability, inherited ataxias such as Friedreich's ataxia, Machado-Joseph disease, spinocerebellar ataxias, Tourette syndrome and other tic disorders, essential tremor, cerebral palsy, stroke, encephalopathies, and intoxication.

In a further aspect, the disclosed compounds can be used to treat, or can be a component of a pharmaceutical composition used to treat cognitive disorders. As such, disclosed herein in a method for treating a cognitive disorder, comprising the step of administering to a mammal in need of treatment at least one compound in a dosage and amount effective to treat the disorder in the mammal, wherein the disorder is selected from dementia (associated with Alzheimer's disease, ischemia, trauma, stroke, HIV disease, Parkinson's disease, Huntington's disease and other general medical conditions or substance abuse), delirium, amnestic disorders and age-related cognitive decline. The fourth edition (Revised) of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR.) (2000, American Psychiatric Association, Washington D.C.) provides a diagnostic tool for cognitive disorders including dementia (associated with Alzheimer's disease, ischemia, trauma, stroke, HIV disease, Parkinson's disease, Huntington's disease and other general medical conditions or substance abuse), delirium, amnestic disorders and age-related cognitive decline.

In a further aspect, the disclosed compounds can be used to treat, or can be a component of a pharmaceutical composition used to neurodegenerative disorders. As such, disclosed herein in a method for treating a neurodegenerative disorder, comprising the step of administering to a mammal in need of treatment at least one compound in a dosage and amount effective to treat a neurodegenerative disorder in the mammal.

In a still further aspect, the disclosed compounds provide a method for treating schizophrenia or psychosis. As such, disclosed herein in a method for treating a disorder related to schizophrenia or psychosis, comprising the step of administering to a mammal in need of treatment at least one compound in a dosage and amount effective to treat the disorder in the mammal, wherein the disorder related to schizophrenia or psychosis is selected from paranoid, disorganized, catatonic or undifferentiated, schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, substance-induced psychotic disorder. The fourth edition (Revised) of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR) (2000, American Psychiatric Association, Washington D.C.) provides a diagnostic tool for c include paranoid, disorganized, catatonic or undifferentiated, schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, substance-induced psychotic disorder.

The subject compounds are further useful in the prevention, treatment, control, amelioration or reduction of risk of the aforementioned diseases, disorders and conditions in combination with other agents, including an mGluR agonist.

The disclosed compounds may be used as single agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of risk of the aforementioned diseases, disorders and conditions for which compounds of formula I or the other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. The other drug(s) may be administered by a route and in an amount commonly used therefore, contemporaneously or sequentially with a disclosed compound. When a disclosed compound is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the compound is preferred. However, the combination therapy can also be administered on overlapping schedules. It is also envisioned that the combination of one or more active ingredients and a disclosed compound can be more efficacious than either as a single agent.

In a further aspect, the subject compound may be used in combination with levodopa (with or without a selective extracerebral decarboxylase inhibitor), anitcholinergics such as biperiden, COMT inhibitors such as entacapone, A2a adenosine antagonists, cholinergic agonists, NMDA receptor antagonists and dopamine agonists.

In one aspect, the invention relates to methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

In one aspect, the invention relates to methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal, the compound having a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

In one aspect, the invention relates to methods for the treatment of a neurotransmission dysfunction and other disease states associated with mGluR4 activity in a mammal comprising the step of co-administering to the mammal at least one compound in a dosage and amount effective to treat the dysfunction in the mammal the compound having a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

The disclosed compounds and compositions can be evaluated for their ability to act as a potentiator of metabotropic glutamate receptor activity, in particular mGluR4 activity, by any suitable known methodology known in the art. For example, Chinese Hamster Ovary (CHO) cells transacted with human mGluR4 or HEK cells co-transfected with rat mGluR4 and the G-protein regulated Inwardly Rectifying Potassium channel (GIRK) were plated in clear bottom assay plates for assay in a Hamamatsu FDSS Fluorometric Plate Reader. The cells were loaded with either a Ca2+-sensitive fluorescent dye or the thallium responsive dye and the plates were washed and placed into a suitable kinetic plate reader. For human mGluR4 assays, a fluorescence baseline was established for 3-5 seconds, the disclosed compounds were then added to the cells, and the response in cells was measured. Approximately two and a half minutes later, a concentration of mGluR4 orthosteric agonist (e.g. glutamate or L-AP4) eliciting approximately 20% (EC20) of the maximal agonist response was added to the cells, and the response was measured. Two minutes later, a concentration of mGluR4 agonist (e.g. glutamate or L-AP4) eliciting 80% (EC80) of the maximal agonist response was added to the cells, and the response was measured. For rat mGluR4/GIRK experiments, a baseline was established for approximately five seconds, disclosed compounds were added, and either an EC20 or EC80 concentration of agonist was added approximately two and one half minutes later. Potentiation of the agonist response of mGluR4 by the disclosed compounds was observed as an increase in response to the EC20 concentration of agonist in the presence of compound compared to the response to agonist in the absence of compound. Similarly, antagonism of the agonist response of mGluR4 by the disclosed compounds was observed as a decrease in response to the EC80 concentration of agonist in the presence of compound compared to the response to agonist in the absence of compound.

The above described assay operated in two modes. In the first mode, a range of concentrations of the disclosed compounds are added to cells, followed by a single fixed concentration of agonist. If the compound acts as a potentiator, an EC50so value for potentiation and a maximum extent of potentiation by the compound at this concentration of agonist is determined by non-linear curve fitting. If the compound acts as a noncompetitive antagonist, an IC50value is determined by non-linear curve fitting. In the second mode, several fixed concentrations of the disclosed compounds are added to various wells on a plate, followed by a range in concentrations of agonist for each concentration of disclosed compound. The EC50values for the agonist at each concentration of compound are determined by non-linear curve fitting. A decrease in the EC50value of the agonist with increasing concentrations of the sample compound (a leftward shift of the agonist concentration-response curve) is an indication of the degree of mGluR4 potentiation at a given concentration of the sample compound. A decrease in the maximal response of the agonist with increasing concentrations of the sample compounds, with or without a rightward shift in agonist potency, is an indication of the degree of noncompetitive antagonism at mGluR4. The second mode also indicates whether the sample compounds also affect the maximum response to mGluR4 to agonists.

In particular, the compounds of the following examples were found to have activity in potentiating the mGluR4 receptor in the aforementioned assays, generally with an EC50for potentiation of less than about 10 μM. One aspect of the disclosed compounds have activity in potentiating rat and human mGluR4 receptors with an EC50for potentiation of less than about 500 nM. These compounds further caused a leftward shift of the agonist EC50by greater than 3-fold. These compounds are positive allosteric modulators (potentiators) of human and rat mGluR4 and were selective for mGluR4 compared to the other seven subtypes of metabotropic glutamate receptors.

F. Manufacture of a Medicament

In one aspect, the invention relates to methods for the manufacture of a medicament for potentiating mGluR4 receptor activity in a mammal comprising combining a compound having a structure represented by formula:

X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

Thus, the disclosed compounds and compositions can be further directed to a method for the manufacture of a medicament for potentiating glutamate receptor activity (e.g., treatment of one or more neurological and/or psychiatric disorder and other disease states associated with glutamate dysfunction) in mammals (e.g., humans) comprising combining one or more disclosed compounds, products, or compositions with a pharmaceutically acceptable carrier or diluent.

G. Uses of Compounds

In one aspect, the invention relates to uses of a compound for potentiating mGluR4 receptor activity in a mammal, wherein the compound has a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

The disclosed uses for potentiating mGluR4 receptor activity in a mammal can further be directed for use in treating one or more disorders, for example neurological and psychiatric disorders and other disease states associated with glutamate dysfunction (e.g., Parkinson's disease) in a subject, for example a mammal or a human.

In one aspect, the invention relates to kits comprising a compound having a structure represented by formula:

wherein X is CH, C-alkyl, N, NH, N-alkyl, alkoxy, or may form a 6-membered ring with Y optionally substituted with at least one R;

Y is CH, C-alkyl, N, or may form a 6-membered ring with X optionally substituted with at least one R;

X1is O or C—R1;

X2is O or C—R2;

or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable derivative thereof, and one or more of a drug having a known side-effect of increasing metabotropic glutamate receptor activity, a drug known to treat a disorder associated with increasing metabotropic glutamate receptor activity, and/or a drug known to treat the neurotransmission dysfunction and other disease states.

In various aspects, the kits can comprise disclosed compounds, compositions, and/or products co-packaged, co-formulated, and/or co-delivered with other components. For example, a drug manufacturer, a drug reseller, a physician, or a pharmacist can provide a kit comprising a disclosed oral dosage forms and another component for delivery to a patient.

In further aspects, the kits can comprise one or more other components (e.g., one or more of a drug having a known side-effect of increasing metabotropic glutamate receptor activity, a drug known to treat a disorder associated with increasing metabotropic glutamate receptor activity, and/or a drug known to treat the neurotransmission dysfunction and other disease states) and instructions for coadministration to a patient with one or more disclosed compounds, compositions, and/or products. For example, a drug manufacturer, a drug reseller, a physician, or a pharmacist can provide a kit comprising one or more other components (e.g., one or more of a drug having a known side-effect of increasing metabotropic glutamate receptor activity, a drug known to treat a disorder associated with increasing metabotropic glutamate receptor activity, and/or a drug known to treat the neurotransmission dysfunction and other disease states) and instructions for coadministration to a patient with one or more disclosed compounds, compositions, and/or products.

Several methods for preparing the compounds of this invention are illustrated in the following Examples. Starting materials and the requisite intermediates are in some cases commercially available, or can be prepared according to literature procedures or as illustrated herein. All NMR spectra were recorded on either a Varian Inova 400 (400 MHz) or Varian Inova 500 (500 MHz) spectrophotometer.1H chemical shifts are reported in δ values in ppm downfield from Me4Si as the internal standard in CDCl3. Data are reported as follows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, br=broad, m=multiplet), integration, coupling constant (Hz).13C chemical shifts are reported in δ values in ppm with the CDCl3carbon peak set to 77.23 ppm. Low resolution mass spectra were obtained on an HP1100 MSD with electrospray ionization. High resolution mass spectra were recorded on a Bruker Daltonics 3T Fourier transform ion cyclotron resonance mass spectrometer (FT/ICR) with electrospray ionization. Analytical thin layer chromatography was performed on EM Reagent 0.25 mm silica gel 60-F plates. Analytical HPLC was performed on an HP1100 with UV detection at 214 and 254 nm along with ELSD detection, LC/MS (J-Sphere80-C18, 3.0×50 mm, 4.1 min gradient, 5% [0.05% TFA/CH3CN]: 95% [0.05% TFA/H2O] to 100% [0.05% TFA/CH3CN]. Preparative purification was performed on a custom HP1100 purification system (reference 16) with collection triggered by mass detection. Solvents for extraction, washing and chromatography were HPLC grade. N-Boc-p-phenylenediamine was purchased from Fluka and 1,2-benzenedisulfonyl dichloride was purchased from TCI America. All other reagents were purchased from Aldrich Chemical Co. and were used without purification.

(2). To a solution of 3-bromophenol (1) (1.0 eq) in Dichloroethane (1.2 M) was added AlCl3(1.5 eq) in three equal portions over 5 min period. After 5 min, the acid chloride (1.02 eq) was added dropwise and the rxn was heated to reflux. After 12 h, the heat was removed. The rxn was poured onto ice/1N HCl (1:1, 50 mL). The mixture was extracted with EtOAc (3×) and the combined organic layers were washed with brine, dried (MgSO4) and filtered. After the solvent was removed, the crude residue was purified by normal phase chromatography (Biotage, 0-10% EtOAc/hexanes) to afford the desired product.

(3). A solution of 7M NH3in MeOH (5 eq) was added to (2) (1 eq) at rt. After 30 min, a bright yellow precipitate was observed. After an additional 30 min, the rxn was filtered and the yellow precipitate was washed with cold MeOH. After the solid was dried overnight, the desired hydroxyl imine was obtained and taken through without any further purification.

(4&5). To a solution of (3) (1 eq) and K2CO3(2 eq) in THF (0.15 M) at rt was added N-chlorosuccinimide (1.5 eq). After 12 h at rt, LCMS confirmed product. The rxn was added to EtOAc:water (1:1) and the organic layer was separated. The organic layer was further washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was purified by normal phase chromatography (Biotage, 100% DCM) to afford pure material (4) and when observed the chlorinated (5).

(6) In a microwave vial, was added 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1 eq), NaOtBu (2.1 eq),tBuXPhos (1 mol %) andtBuXPhos Palladacycle (1 mol %). The vial was sealed and evacuated and purged with Argon (3×). A solution of (4) (1 eq) intBuOH was added and the rxn was heated to 60° C. After 4 h, the rxn was added to EtOAc:NH4Cl (aq) (1:1). The organic layer was separated and washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was redissolved in TFA:toluene (1:1) and subjected to microwave irradiation at 140° C. for 20 min. The solvent was removed and the crude residue was purified by reverse phase chromatography (Gilson, 20-80% acetonitrile:water w/0.1% TFA). The collected fractions were neutralized with NaHCO3(aq); EtOAc (1:1) and the organic layers were concentrated to provide desired product.

(7). A solution of 7M NH3in MeOH (5 eq) was added to hydroxyketone (1 eq) at rt. After 30 min, a bright yellow precipitate was observed. After 12 h, the rxn was filtered and the yellow precipitate was washed with cold MeOH. After the solid was dried overnight in a vacuum dessicator, the desired hydroxyl imine was obtained and taken through without any further purification.

(8). To a solution of (7) (1 eq) and K2CO3(2 eq) in THF (0.15 M) at rt was added N-chlorosuccinimide (1.5 eq). After 12 h at rt, LCMS confirmed product. The rxn was added to EtOAc:water (1:1) and the organic layer was separated. The organic layer was further washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was purified by normal phase chromatography (Biotage, 100% DCM) to afford pure material (8).

(9) In a microwave vial, was added 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1 eq), NaOtBu (2.1 eq),tBuXPhos (1 mol %) andtBuXPhos Palladacycle (1 mol %). The vial was sealed and evacuated and purged with Argon (3×). A solution of (8) (1 eq) intBuOH was added and the rxn was heated to 60° C. After 4h, the rxn was added to EtOAc:NH4Cl (aq) (1:1). The organic layer was separated and washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was redissolved in TFA:toluene (1:1) and subjected to microwave irradiation at 140° C. for 20 min. The solvent was removed and the crude residue was purified by reverse phase chromatography (Gilson, 20-80% acetonitrile:water w/0.1% TFA). The collected fractions were neutralized with NaHCO3(aq):EtOAc (1:1) and the organic layers were concentrated to provide desired product.

(10) To a solution of 4-bromo-2-hydroxybenzoic acid (1 eq) in Methanol (0.2 M) at rt was added sulfuric acid (1 eq). The dark red/brown solution was heated to 75° C. for 12 h. LCMS confirms product. The rxn was concentrated and crude residue was partitioned between EtOAc:water (1:1). The organic layer was washed with NaHCO3(aq), water and dried (MgSO4). After filtration, the organic layer was concentrated to give a tan solid (58%).

(11) To a solution of hydroxylamine hydrochloride (1.5 eq) and NaOH (3.5 eq) in water (0.25 M) was added Methyl 4-bromo-2-hydroxybenzoate (1 eq) in 1,4-dioxane (2.0 M) dropwise. The reaction turns light yellow shortly after addition. The solution is stirred at rt for 18 hrs. 1,4-dioxane was removed in vacuo and HCl (2N) was added until a precipitate was formed. The heterogenous mixture was filtered and washed with water. The solid was dried overnight in a vacuum oven to give the title compound.

(12) A solution of 1,1′-carbonyldiimidazole (2.0 eq) in THF (0.1 M) was added dropwise to a refluxing solution of 4-bromo-2-hydroxy-benzenecarbohydroxamic acid (1.0 eq) in THF (0.15 M) and then stirred for 2 h at refluxing conditions. The THF was removed under vacuum and the residue was suspended in water. HCl (2N) was added dropwise at rt. A precipitate formed and was collected by filtration, washed with water and dried overnight in a vacuum oven to afford 6-bromo-1,2-benzoxazol-3-ol (95% yield).

(13) 6-bromo-1,2-benzoxazol-3-ol (1.0 eq), potassium carbonate (1.5 eq) were added to a rbf, followed by DMF (0.2 M). 4-Methoxybenyl chloride (1.1 eq) was added and the rxn was stirred overnight at rt. LCMS shows both product regioisomers. Rxn is added to water (0.1 M) and extracted with EtOAc (2×). The combined extracts are washed with water (3×), brine, and dried over MgSO4. Solvent was removed in vacuo and the residue was purified by normal phase chromatography (15-30% EtOAc:hexanes). The two regioisomers were collected separately. The major product, 6-bromo-3-[(4-methoxyphenyl)methoxy]-1,2-benzoxazole (39% yield) and the minor product, 6-bromo-2-[(4-methoxyphenyl)methyl]-1,2-benzoxazol-3-one (15% yield).

(1.4) To a rxn vial, was added 6-bromo-3-[(4-methoxyphenyl)methoxy]-1,2-benzoxazole (1.0 eq), cesium carbonate (2.0 eq), 1-[(4-methoxyphenyl)methyl]pyrazolo[4,3-b]pyridin-3-amine (1.1 eq), XPhos (0.15 eq), Pd2(dba)3(0.075 eq). The rxn was evacuated and purged with Nitrogen (3×), followed by the addition of 1,4-Dioxane (0.07 M). The heterogenous rxn was heated to 100° C. After 24 h, the rxn was added to NH4Cl (aq):EtOAc, the organic layer was collected, washed with water, brine, dried over MgSO4and the solvent was removed. Residue was purified by normal phase chromatography (10-65% EtOAc:hexanes). The product was dissolved in Trifluoroacetic acid (7.5 mL) and Toluene (5 mL) and the solution were transferred to a 20 mL MW vial. The rxn was subjected to 150° C. in the MW for 45 min. After LCMS confirms complete loss of the PMB protecting group, the solvent was removed. The material was carried through without further purification.

(15) & (16) Alkyl halide (1.0 eq) and potassium carbonate (3.0 eq) were added to a 2 mL μW vial. A solution of 6-(1H-pyrazolo[4,3-b]pyridin-3-ylamino)-1,2-benzoxazol-3-ol; 2,2,2-trifluoroacetic acid (1.0 eq) in DMF (1.5 mL) was added to the MW vial. The mixture was subjected to 120° C. in the MW for 15 min. LCMS shows two products formed. Rxn was syringe filtered and purified by Gilson HPLC (15-85% acetonitrile:water with 0.1% TFA). The two products were collected separately, free based in NaHCO3(aq), extracted with EtOAc and washed with water (2×). The solvent was removed to afford the products (15 and 16).

(17) To a suspension of pyridine (3.0 eq) and 6-bromo-1,2-benzoxazol-3-ol (1.0 eq) in DCM (0.05 M) at 0° C. under nitrogen was added trifluoromethanesulfonic anhdride (1.5 eq, 1M in DCM), dropwise. After 10 min, the ice bath was removed. It was stirred for an additional 45 min at rt. Rxn was added to DCM:Water (1:1). The organic layer was separated with the hydrophobic phase separator, solvent was removed and the product (6-bromo-1,2-benzoxazol-3-yl)trifluoromethanesulfonate was carried forward without further purification.

(18) To a 2 mL μW vial was added amine (5.0 eq), (6-bromo-1,2-benzoxazol-3-yl) trifluoromethanesulfonate (1.0 eq), and 1-methyl-2pyrrolidinone (0.5 M). The rxn was heated to 130° C. for 15 min in the MW. LCMS confirmed product and the rxn was added to EtOAc:NH4Cl (1:1). The organic layer was collected and the aq. layer re-extracted with EtOAc (2×). The combined organic extracts were washed with water, brine, and dried over MgSO4, filtered and concentrated. The crude residue was purified by normal phase chromatography (25-85% EtOAc:hexanes) to yield desired product (9-20% yield).

(19) In a microwave vial, was added 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1 eq), NaOtBu (2.1 eq),tBuXPhos (1 mol %) andtBuXPhos Palladacycle (1 mol %). The vial was sealed and evacuated and purged with Argon (3×). A solution of (18) (1 eq) intBuOH was added and the rxn was heated to 90° C. After 4 h, the rxn was added to EtOAc:NH4Cl (aq) (1:1). The organic layer was separated and washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was redissolved In TFA:toluene (1:1) and subjected to microwave irradiation at 140° C. for 20 min. The solvent was removed and the crude residue was purified by reverse phase chromatography (Gilson, 20-80% acetonitrile:water w/0.1% TFA). The collected fractions were neutralized with NaHCO3(aq):EtOAc (1:1) and the organic layers were concentrated to provide desired product.

(20). A solution of methyl 2-(4-bromo-2-nitrophenyl)acetate (3.43 g, 12.5 mmol, 1.0 eq.) in ethanol (62.5 mL, 0.2 M) was treated with isoamyl nitrite (2.18 mL, 16.25 mmol, 1.3 eq.) and sodium ethoxide (893 mg, 13.13 mmol, 1.05 eq.). The reaction mixture was stirred at 60° C. After 24 h, the mixture was neutralized with 1M HCl solution and concentrated. The residue was extracted with EtOAc (3×). The combined extracts were washed with brine, dried over Na2SO4, filtered and concentrated. Purification using flash chromatography on silica gel (0-35% EtOAc/hexanes) to provide the title compound as a pale yellow solid (1.73 g, 51% yield); ES-MS [M+1]+: 270.2

(22). To a solution of ethyl 6-((1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazole-3-carboxylate (300 mg, 0.79 mmol, 1.0 eq) in 1,4-dioxane (3.9 mL) at 0° C. was added a solution of 1N LiOH (2.76 mL, 2.76 mmol, 3.5 eq.). The reaction mixture was stirred at rt for 45 min. Solvent was removed. The aqueous layer was adjusted to pH 3-4 with 1M HCl solution. The resulting mixture was extracted with CHCl3/iPA (3:1, v/v, 3×). The combined extracts were concentrated to provide the title compound as a yellow solid (280 mg, 85.4% yield): ES-MS [M+1]+: 416.2. Note: Decarboxylation will occur upon heating the sample.

General Procedure (23): A mixture of 6-((1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazole-3-carboxylic acid (14.45 mg, 0.035 mmol, 1.0 eq.), amine (0.07 mmol. 2.0 eq.) and HATU (19.96 mg, 0.05 mmol, 1.5 eq.) in DMF (1.0 mL) was stirred for 5 min at rt and DIEA (9.55 uL, 0.07 mmol, 2.0 eq.) was added. The reaction mixture was continued to stir at rt for 45 min. Upon completion by LCMS, the reaction mixture was purified using reverse phase Gilson HPLC system with acidic method (ACN/water/0.1% TFA). The Gilson fractions were concentrated. The resulting residue was dissolved in TFA (0.1 M) and heated in a microwave reactor for 30 min at 150° C. Solvent was removed. The crude product was purified using reverse phase Gilson HPLC system with acidic method (ACN/water/0.1% TFA). The sample was concentrated and submitted as a TFA salt.

(24). To a microwave vial 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1.1 equiv), 1-(6-bromo-1,2-benzoxazol-3-yl)ethanone (1 equiv), cesium carbonate (3 equiv), XantPhos (0.05 equiv), and tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct (0.05 equiv) were added. The vial was evacuated and purged with Nitrogen three times. Then, dioxane (10 mL) was added into the vial. The mixture was stirred at room temperature for 3 min, and heated under microwave irradiation at 140° C. for 30 min. The mixture was filtered through celite, and the filtrate was concentrated under reduced pressure. Then, the resulting residue was purified by column chromatography, eluted with 0% -70% ethyl acetate in hexane, to afford the product.(25). To a solution of 1-(6-((1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazol-3-yl)ethan-1-one (1 equiv) in methanol (0.25 M), 4-methylbenzenesulfonohydrazide (1 equiv) was charged. The solution was stirred at room temperature for 1 h or until all starting material was consumed. The mixture was then concentrated under reduced pressure and purified by column chromatography, eluted with 0%-50% ethyl acetate in hexane, to afford the product.(26). To a solution of (E)-N′-(1-(5-((1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazol-3-yl)ethylidene)-4-methylbenzenesulfonohydrazide (1 equiv) in dioxane (0.07 M), boronic acid (1.5 equiv) and potassium carbonate (3 equiv) were charged. The mixture was stirred at 110° C. for overnight. After stirring, the mixture was filtered through syringe filter, and the filtrate was concentrated. The residue was purified by HPLC, eluted with 15%-100% acetonitrile in water with 0.5 mL/L NH4OH. The resulting solid was dissolved in TFA (0.07 M) and stirred under microwave irradiation at 130° C. for 30 min. The solution was concentrated, and the residue was purified by HPLC, eluted with 15% -100% acetonitrile in water with 0.5 mL/L NH4OH, to obtain the final product.

(27). To a solution of6-bromobenzo[d]isoxazole-3-carbaldehyde (1 equiv) in methanol (0.25 M), 4-methylbenzenesulfonohydrazide (1 equiv) was charged. The solution was stirred at room temperature for 1 h or until all starting material was consumed. The mixture was then concentrated under reduced pressure and purified by column chromatography, eluted with 0% -50% ethyl acetate in hexane, to afford the product.(28). To a solution of (Z)-N′-((6-bromobenzo[d]isoxazol-3-yl)methylene)-4-methylbenzenesulfonohydrazide (1 equiv) in dioxane (0.07 M), boronic acid (1.5 equiv) and potassium carbonate (3 equiv) were charged. The mixture was stirred at 110° C. for overnight. After stirring, the mixture was filtered with syringe filter. The filtrate was purified by HPLC, eluted with 15% -100% acetonitrile in water with 0.5 mL/L NH4OH, to obtain the product, I.(29). To a microwave vial, 1-(4-methoxybenzyl)-1H-pyrazolo4,3-pyridin-3-amine (1.1 equiv), I (1 equiv), caesium carbonate (3 equiv), XantPhos (0.05 equiv), and tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct (0.05 equiv) were added. The vial was evacuated and purged with Nitrogen three times. Then, dioxane (0.07M) was added into the vial. The mixture was stirred at room temperature for 3 min, and heated under microwave irradiation at 140° C. for 30 min. The mixture was filtered through syringe filter, and the filtrate was concentrated. Then, the resulting residue was purified by HPLC, eluted with 15% -100% acetonitrile in water with 0.1% TFA. The resulting solid was dissolved in TFA (0.07 M) and stirred under microwave irradiation at 130° C. for 30 min. The solution was concentrated, and the residue was purified by HPLC, eluted with 15% -100% acetonitrile in water with 0.5 mL/L NH4OH, to obtain the final product.

(30). To a solution of 1-(6-bromobenzo[d]isoxazol-3-yl)ethan-1-one (1.0 eq) in THF (0.15M) at −78° C. was added a solution of cycloproylmagnesium bromide (2.0 eq) in THF (0.5M). After 2h at −78° C., NH4Cl (aq) was added dropwise and the cold bath was removed. The rxn was extracted with EtOAc (2×) and the organic layer was washed with water, brine and concentrated. The residue was purified using normal phase Biotage Isolera (0-15% EtOAc/hexanes) to afford 1-(6-bromobenzo[d]isoxazol-3-yl)-1-cyclopropylethan-1-ol (85% yield).

(31). To a solution of 1-(6-bromobenzo[d]isoxazol-3-yl)-1-cyclopropylethan-1-ol (1.0 eq) in DCM (0.1M) was added DAST (3.0 eq) at 0° C. The ice both was removed. After 2 h, NaHCO3(aq) was added dropwise to the rxn. The rxn was extracted with DCM (2×) and the organic layer was passed through a phase separator. After concentration, the residue was purified using normal phase Biotage Isolera (1 g column, liquid loading, gradient: 0-15% EtOAc/hexanes) to provide 6-bromo-3-(1-cyclopropyl-1-fluoroethyl)benzo[d]isoxazole (40% yield).

(32). In a microwave vial was added 6-bromo-3-(1-cyclopropyl-1-fluoroethyl)benzo[d]isoxazole (1.0 eq), Pd2(dba)3(0.18 eq), Xantphos (0.24 eq). Cesium Carbonate (2.1 eq) and 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1.0 eq). The mixture was evacuated and purged with Nitrogen (3×), followed by the addition of 1,4-dioxane (0.1 M). The rxn was heated to 150° C. for 1 h in the microwave. After confirmation of product via LCMS, the mixture was filtered through a pad of Celite and washed with EtOAc. After the solvent, was removed, the crude residue was purified by reversed phase Gilson HPLC (60-90% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure. The residue was dissolved in toluene:TFA (1:1) and heated to 150° C. in the microwave. After the solvent was removed, the material was purified by reversed phase Gilson HPLC (60-90% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure to afford 1-(6-((1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazol-3-yl)-1-cyclopropylethan-1-ol (14% yield).

(33). In a microwave vial was added 6-bromo-3-(2-fluorohexan-2-yl)benzo[d]isoxazole (1.0 eq), Pd2(dba)3(0.18 eq), Xantphos (0.24 eq), Cesium Carbonate (2.1 eq) and tert-butyl carbamate (1.0 eq). The mixture was evacuated and purged with Nitrogen (3×), followed by the addition of 1,4-dioxane (0.1 M). The rxn was heated to 140° C. for 1 h in the microwave. After confirmation of product via LCMS, the mixture was filtered through a pad of Celite and washed with EtOAc. After the solvent was removed, the crude residue was taken forward without further purification.

The above material was dissolved in TFA:DCM (1:5; 0.1M) at rt. The rxn was monitored by LCMS. After complete consumption of starting material, the solvent was removed. The crude residue was purified by reversed phase Gilson HPLC (60-90% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure to afford 3-(2-fluorohexan-2-yl)benzo[d]isoxazol-6-amine.

(34). In a vial was added 3-(2-fluorohexan-2-yl)benzo[d]isoxazol-6-amine (1.0 eq), Pd2(dba)3(0.18 eq), Xantphos (0.24 eq). Cesium Carbonate (2.1 eq.) and 3-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[4,3b]pyridine (1.0 eq). The mixture was evacuated and purged with Nitrogen (3×), followed by the addition of 1,4-dioxane (0.1 M). The rxn was heated to 100° C. After 12 h, the reaction was removed from the heat and cooled to rt. The rxn was added to EtOAc:NH4Cl (aq) (1:1). The organic layer was separated and washed with water (2×), brine and dried (MgSO4). After filtration, the solvent was removed under reduced pressure. The crude residue was purified by reversed phase Gilson HPLC (35-95% acetonitrile/water with 0.1% TFA). The collected fractions were concentrated.

The above material was dissolved in 4N HCl in MeOH (0.1M) at rt. The rxn was heated to 55° C. After complete consumption of starting material, the solvent was removed. The crude residue was purified by reversed phase Gilson HPLC (25-95% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure to afford 3-(2-fluorohexan-2-yl)-N-(1H-pyrazolo[4,3-b]pyridin-3-yl)benzo[d]isoxazol-6-amine.

(35) Acid Chloride method: To a solution of 3-neopentyl-N-(1H-pyrazolo[4,3-b]pyridin-3-yl)benzo[d]isoxazol-6-amine (1.0 eq) in DMA (0.2M ) at rt was added Cs2CO3(2.0 eq). After 15 min, acid chloride (1.5 eq) was added. After an additional 30 min, the rxn mixture was diluted with EtOAc:water (1:1). The organic layer was separated and washed with water (4×), dried (MgSO4), filtered and concentrated. The crude residue was purified by reversed phase Gilson HPLC (60-85% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure to afford the desired amide.(35) Carboxylic Acid Method: To a vial was charged carboxylic acid (3.0 eq), HOBt (3.0 eq), EDC (3.0 eq) and NMP (0.3M) at rt. Once a solution was achieved, a solution of 3-neopentyl-N-(1H-pyrazolo[4,3-b]pyridin-3-yl)benzo[d]isoxazol-6-amine (1.0 eq) in NMP (0.5M was added dropwise. DMAP (3.0 eq) was added. After 30 min, TFA was added until a solution was obtained. The crude rxn was purified by reversed phase Gilson HPLC (30-60% acetonitrile/water with 0.1% TFA). The collected fractions were neutralized with NaHCO3(aq) and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried (MgSO4), filtered and concentrated under reduced pressure to afford the desired amide.

(36). A solution of 1-(4-bromo-2-hydroxyphenyl)ethan-1-one (1.0 eq) in toluene (0.5M) was added dropwise to a stirred suspension of sodium hydride (2.8 eq) in toluene (0.5M) at reflux. The mixture was stirred for 10 min and then diethylcarbonate (2.0 eq) was added dropwise over 30 min. After 18 h at reflux, the rxn mixture was removed from heat source. Once at rt, the rxn was cooled to 0° C. and 2N HCl (0.01M) was added dropwise. After an additional 15 min, white solid forms in the aqueous layer. The rxn mixture was extracted with EtOAc (2×). The combined organic layers were washed with water, brine and dried (MgSO4). After nitration, the solvent was removed and the crude residue was carried forward without purification.

The above residue was dissolved in toluene (0.2M) and refluxed. After 12 h, the heat source was removed and upon cooling the resulting precipitate was collected by vacuum filtration. The white solid was washed with ether and dried to provide 7-bromo-4-hydroxy-2H-chromen-2-one (49% yield).

(37). To a solution of NaOEt (2.0 eq) in EtOH (0.8M) was added hydroxylamine hydrochloride (2.0 eq). A solution of 7-bromo-4-hydroxy-2H-chromen-2-one (1.0 eq) in EtOH (1.0M) was added to the rxn. The mixture was heated to reflux. After 4 h at reflux, the solution was cooled to rt. The mixture was poured into a saturated solution of sodium bicarbonate (1.0M). Water:DCM (1:1; 0.5M) were added and the aqueous layer was collected. The aqueous layer was washed with DCM (1.0M) and acidified with 2N HCl, dropwise, until precipitate forms. The precipitate was collected by filtration and dried in the oven to afford 2-(6-bromobenzo[d]isoxazol-3-yl)acetic acid (79% yield).

(38). To a solution of 2-(6-bromobenzo[d]isoxazol-3-yl)acetic acid (1.0 eq) in DMF was added amine (1.3 eq), DIEA (4.0 eq), HOBt (1.0 eq) and HATU (1.3 eq). The solution was heated to 50° C. and monitored by LCMS. After 3 h, the rxn was filtered and purified by reverse phase Gilson HPLC (35-95% acetonitrile/water with 0.05% NH4OH). The solvent was removed to afford, desired product (50-85% yield).

(39). In a microwave vial was added the previously mentioned aryl bromide (1.0 eq), Pd2dba)3(0.09eq), Xantphos (0.12 eq), Cesium Carbonate (2.1 eq) and 1-(4-methoxybenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine (1.0 eq). The mixture was evacuated and purged with Nitrogen (3×), followed by the addition of 1,4-dioxane (0.1 M). The reaction mixture was subjected to a microwave reactor for 45 min at 150° C. After confirmation of product via LCMS, the mixture was filtered through a pad of Celite and washed with EtOAc. After the solvent was removed, the crude residue was dissolved in TFA:toluene (1:1; 0.03M). The reaction mixture was subjected to a microwave reactor for 30 min at 150° C. LCMS confirmed loss of starting material, so solvent was removed. The crude residue was purified by reversed phase Gilson HPLC (60-90% acetonitrile/water with 0.1% TFA). The fractions were concentrated to afford the desired product.

(40). To a solution of 2-(6-((1H-pyrazolo[4,3-b]pyridin-3-yl)amino)benzo[d]isoxazol-3-yl-N-methoxy-N-methylacetamide (1.0 eq) in DMF (0.1M) was added Boc2O (1.5 eq) and Et3N (1.5 eq) at rt. After 12 h, rxn was added to water and extracted with a solution of CHCl3:IPA (3:1). The organic layer was washed with water (2×) and the solvent was removed. The material was carried forward without further purification.

(41). tert-butyl 3-((3-(2-methoxy(methyl)amino-2-oxoethyl)benzo[d]isoxazol-6-yl)amino)-1H-pyrazolo[4,3-b]pyridine-1-carboxylate (1.0 eq) was dissolved in THF (0.2M) and the solution was cooled to 0° C. LiBH4(5.0 eq) was added slowly, followed by MeOH (5.0 eq), dropwise. The ice bath was removed. After 3 h at rt, 1N HCl was added slowly. The rxn was added to EtOAc:water (1:1) and the organic layer was separated. The aqueous layer was further extracted with EtOAc (2×). The combined organic layers were washed with water, brine and dried (MgSO4). The solvent was removed and the residue was carried forward without further purification.

(42). To a solution of tert-butyl 3-((3-(2-hydroxyethyl)benzo[d]isoxazol-6-yl)amino)-1H-pyrazolo[4,3-b]pyridine-1-carboxylate (1.0 eq) in DCM (0.1M) was added (diethylamino)sulfur trifluoride (DAST) (4.0 eq) dropwise. The rxn was added to ice water and the mixture was extracted with EtOAc (3×). The combined organic layers were washed with water, brine and dried (MgSO4). After filtration and concentration, the residue was dissolved in DCM:TFA (2:1) and stirred at rt. After LCMS confirms the loss of starting material, the solvent was removed. The crude residue was purified by reversed phase Gilson. HPLC (15-55% acetonitrile/water with 0.1% TFA) to afford 3-(2-fluoroethyl)-N-(1H-pyrazolo[4,3-b]pyridin-3-yl)benzo-[d]isoxazol-6-amine (22% yield).

(43). To a solution of the starting material (75 mg, 0.3500 mmol) and triethylamine (0.06 mL, 0.42 mmol) in DCM (3 mL, 0.11 M) was added pivaloyl chloride (0.05 mL, 0.42 mmol). After 3 hours at rt, the reaction was heated to 30° C. for 12 hrs, and monitored via LCMS. The reaction was concentrated, and purified using flash chromatography on silica gel (8-30% EtOAc in hexanes) to provide 43 (41.7 mg, 0.14 mmol, 40% yield).(44). In a vial was added 43 (20 mg, 0.07 mmol), 1-[(4-methoxyphenyl)methyl]pyrazolo-[4,3-b]pyridin-3-amine (20 mg, 0.07 mmol), NaOtBu (17 mg, 0.18 mmol), tBuXPhos (0.31 mg, 0.001 mmol), and tBuXPhos-Palladacycle (0.5 mg, 0.001 mmol). The sealed vial was evacuated and backfilled with dry Nitrogen 3×. To the vial was added tBuOH (1 mL, 0.07 M). After 1 hr at 60° C. The rxn was added to a mixture of EtOAc and water. The aqueous layer was extracted with EtOAc 5× (25 mL), and the organic extractions were washed with water 1× (10 mL). The organic extractions were concentrated in vacuo, and the residue was dissolved in TFA and toluene. The solution was heated to 150° C. in μW for 15 min. The solvent was removed in vacuo, and compound was purified with reverse phase chromatography via HPLC (MeCN in 0.1% TFA and water). The fractions were added to water and NaHCO3 solution. The water was extracted with EtOAc. The solvent was removed, and the residue was dissolved in DCM and filtered through a hydrophobic membrane to yield the product. (6.4 mg, 0.018 mmol, 27% yield).

(45). A solution of the starting material in THF (58.5 mL, 0.2 M) was cooled to 0° C. To the reaction was added a solution of LiBH4in THF and MeOH. After 30 min at 0° C., the reaction was quenched with 15% NaOH aqueous solution. The reaction was allowed to warm to room temp, and the filtrate was collected and concentrated. The residue was added to water and EtOAc. The aqueous layer was extracted with EtOAc 3×. The organic extraction was washed with brine 1× and dried over MgSO4. The solution was concentrated to afford the product which was used for future reactions without further purification.(46). To a solution of 45 (2.00 g, 8.77 mmol) in DCM (43.9 mL, 0.2 M) was added Dess-Martin Periodinane (5.58 g, 13.2 mmol). After 90 min at room temp, to the reaction was added water. The water layer was extracted the DCM. The DCM extractions were combined and concentrated. The product was purified by flash chromatography on silica gel (10-45% EtOAc in hexanes) to yield 30 (1.60 g, 7.08 mmol 81% yield).(47). A solution of 46 (60 mg, 0.26 mmol) and 4-methoxypiperdine (0.164 mL, 1.33 mmol) in DCE (2.4 mL, 0.07 mmol) and AcOH (1.6 mL, 0.07 mmol) was heated to 45° C. After 45 min at 45° C., Na(CH3COO)3BH4(84 mg, 0.40 mmol) was added to the solution. After 10 min, the solution was diluted with DCE and quenched with NaHCO3 aqueous solution. The aqueous mixture was extracted with DCE 2×. The organic extractions were combined and filtered through a hydrophobic membrane to yield 47 (61 mg, 0.188 mmol, 71% yield).(48). In a 2 ml vial, was added NaOtBu (28 mg, 0.30 mmol), tBuXPhos-Palladacycle (2.5 mg, 0.02 mmol), tBuXPhos (1.6 mg, 0.02 mmol), 1-[(4-methoxyphenyl)methyl]pyrazolo[4,3-b]pyridin-3-amine (47 mg, 0.18 mmol), and 31 (60 mg, 0.18 mmol). The reaction was evacuated and purged with Nitrogen (3×). To the vial was added tBuOH (2mL). The heterogeneous mixture was heated to 75° C. for 12 hrs. The reaction was added to a mixture of ExOAc and water. The aqueous mixture was extracted with EtOAc 5×. The organic mixture was washed with water 3×. The organic extractions were combined and concentrated. The residue was dissolved in TFA (1 mL) and transferred to a 2 mL MW vial. After 15 min at 150° C. in the MW, the solvent was removed under reduced pressure. The fractions were separated by reverse phase chromatography (ACN/water/0.1% TFA). The combined fractions were added to a mixture of water and NaHCO3. The aqueous layer was extracted with EtOAc 3×, and the organic layer was washed with water 1×. Solvent was removed under reduced pressure, and solids were dissolved in a mixture of DCM:MeOH (1:1) and filtered through a hydrophobic frit to yield the product (7.2 mg, 0.02 mmol, 10.3% yield). ES-MS [M+H]+: 379.4.

(49). To a 1M solution of chloro(2,2-dimethylpropyl)magnesium in THF (9.86 mL, 9.86 mmol) was added 33 (1000 mg, 4.90 mmol) at 0° C. The reaction was removed from the ice bath, and stirred at room tempt for 24 hrs. The reaction was diluted with EtOAc (10 mL). The organic mixture was washed with aqueous NH4Cl solution 1×, and water 2×. The reaction was concentrated in vacuo to yield yellow oil. The residue was dissolved in DCM. To the DCM solution was added DMP (2080 mg, 4.90 mmol). After 1 hr at room temp, the rxn was diluted with DCM. To the reaction was added saturated NaS2O3 and saturated NaHCO3 solution. The organic layers were filtered through a hydrophobic membrane to yield the product which was taken forward without purification and assumed to yield 100% product.(50). To a solution of 49 (1340 mg, 4.9 mmol) in MeOH (50 mL, 0.1 M) was added hydroxylamine hydrochloride (1360 mg, 19.6 mmol) and NaOAc (2009 mg, 24.5 mmol). After 16 hrs at 50° C., the reaction was cooled to room temperature and filtered over a celite pad. The filtrate was added to water and the aqueous mixture was extracted with EtOAc 1× and dried over anhydrous Na2SO4. The solution was concentrated in vacuo to yield 50 as crude product. The product was taken forward without further purification.(51). A solution of NaH (336 mg, 60% w/v) in DMF (5 mL) was cooled to 0° C. in an ice-bath under a nitrogen environment. To the solution was added dropwise a solution of 50 (1420 mg, 4.9 mmol) in DMF (5 mL). The ice bath was removed, and the reaction was stirred for another hour at room temp. After 1 hour, the reaction was cooled to 0° C., and a mixture of EtOH:water (1:1) was added dropwise until no gas evolution was noticed. The solution was added to water, and extracted with EtOAc 2×. The organic extractions were washed with water 3×. The solution was concentrated in vacuo, and fractions were separated by reverse phase HPLC in a gradient of MeCN in water and 0.1% TFA. The fractions were added to water and treated with NaHCO3 solution. The aqueous solution was extracted with EtOAc 2×. The organic extractions were washed with water 3×, brine 1×, and dried over anhydrous Na2SO4. The solution was concentrated to yield 51 (130 mg, 0.48 mmol, 9.8% yield): 1H NMR (400 MHz, CDCl3) δ 8.65 (d, J=1.80 Hz, 1H), 7.99 (d, J=1.8 Hz, 1H), 2.92 (s, 2H), 0.994 (s, 9H).(52). In a 2 mL vial, was added NaOtBu (28 mg, 0.300 mmol), tBuXPhos-Palladacycle (2.5 mg, 0.02 mmol), tBuXPhos (1.6 mg, 0.02 mmol), 1-[(4-methoxyphenyl)methyl]pyrazolo[4,3-b]pyridin-3-amine (57 mg, 0.22 mmol), and 36 (60 mg, 0.22 mmol). The reaction was evacuated and purged with Nitrogen (3×). To the vial was added t-butanol (2mL). The heterogeneous mixture was heated to 75° C. for 12 hrs. The reaction was added to a mixture of ExOAc and water. The aqueous mixture was extracted with EtOAc 5×. The organic mixture was washed with water 3×. The organic extractions were combined and concentrated. The residue was dissolved in TFA (1 mL) and transferred to a 2 mL MW vial. After 15 min at 150° C. in the MW, the solvent was removed under reduced pressure. The fractions were separated by reverse phase chromatography (ACN/water/0.1% TFA). The combined fractions were added to a mixture of water and NaHCO3. The aqueous layer was extracted with EtOAc 3×, and the organic layer was washed with water 1×. Solvent was removed under reduced pressure, and solids were dissolved in a mixture of DCM:MeOH (1:1) and filtered through a hydrophobic frit to yield 52 (8.1 mg, 0.03 mmol, 11.3% yield). ES-MS [M+1]+: 323.9.

(56). To a solution of 6-bromo-1,2-benzoxazole-3-carbaldehyde (678.09 mg, 3.0 mmol, 1.0 eq.) in THF (30 mL) at −78° C. was added a solution of tert-butylmagnesium chloride in THF (2.0 M, 2.25 mL, 4.5 mmol, 1.5 eq). The resulting mixture was stirred at −78° C. After 2 h, sat. soln NH4Cl was added. The mixture was warmed to rt and extracted with EtOAc (3×). The combined extracts were washed with brine, dried over Na2SO4, filtered and concentrated. Purification using flash chromatography on silica gel (0-30% EtOAc/hexanes) to provide the title compound (600 mg, 70.4% yield) as a viscous oil. ES-MS [M+1]+: 284.2, RT=1.128 min, >80% at 254 nm.

(57). To a solution of 1-(6-bromo-1,2-benzoxazol-3-yl)-2,2-dimethyl-propan-1-ol (600 mg, 2.11 mmol, 1.0 eq.) in DCM (10.6 mL) was added Dess-Martin periodinane (1.34 g, 3.2 mmol, 1.5 eq.). After stirring at rt for 1 h, water was added slowly. The mixture was extracted with DCM (3×). The combined extracts were washed with brine, dried over Na2SO4, filtered and concentrated. Purification using flash chromatography on silica gel (0-30% EtOAc/hexanes) to provide the title compound (380 mg, 63.8% yield) as a white crystalline solid, ES-MS [M+1]+: 284.2, RT=1.371 min, >98% at 215 and 254 nm).

(60). To a solution of 6-bromo-1,2-benzoxazole-3-carbaldehyde (750 mg, 3.32 mmol, 1.0 eq.) in THF (16.6 mL) at −78° C. was added a solution, of n-butylmagnesium chloride in THF (2.0 M, 4.98 mL, 9.95 mmol, 3.0 eq). The resulting mixture was stirred at −78° C. After 2 h, sat. soln NH4Cl was added. The mixture was warmed to rt and extracted with EtOAc (3×). The combined extracts were washed with brine, dried over Na2SO4, filtered and concentrated. Purification using flash chromatography on silica gel (0-30% EtOAc/hexanes) to provide the title compound (360.2 mg, 38.2% yield) as a viscous oil. ES-MS [M+1]+: 284.2, RT=1.147 min, >98% at 215 and 254 nm.

(61). To a solution of 1-(6-bromo-1,2-benzoxazol-3-yl)pentan-1-ol (100 mg, 0.35 mmol, 1.0 eq.) in DCE (2.64 mL) was added dropwise DAST (0.86 mL, 1.41 mmol, 4.0 eq.). After stirring at rt for 1 h, the mixture was poured over an ice/water mixture and extracted with DCM. The combined extracts were washed with brine, dried over Na2SO4, filtered and concentrated. Purification using flash chromatography on silica gel (0-30% EtOAc/hexanes) to provide the title compound (85.8 mg, 85.2% yield) as an oil.

(68). To a solution of methyl 6-[(1-tert-butoxycarbonylpyrazolo[4,3-b]pyridin-3-yl)amino]-1,2-benzoxazole-3-carboxylate (119.0 mg, 0.29 mmol, 1.0 eq.) in THF (1.45 mL) at 0° C. was added a solution of lithium borohydride in THF (2.0 M, 0.298 mL, 0.58 mmol, 2.0 eq.) and methanol (23.55 uL, 0.58 mmol, 2.0 eq.). The resulting mixture was stirred at 0° C. for 30 min. Water (10 mL) was added. The mixture was stirred for 20 min at rt and diluted with EtOAc. The mixture was extracted with EtOAc (2×). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrate. Purification using flash column chromatography on silica gel (10-60% EtOAc/hexanes) to provide the title compound. ES-MS [M+1]+: 382.3, RT=1.038 min, >98% at 215 and 254 nm

(69). To a suspension of tert-butyl 3-[[3-(hydroxymethyl)-1,2-benzoxazol-6-yl]amino]pyrazolo[4,3-b]pyridine-1-carboxylate (9.0 mg, 0.02 mmol, 1.0 eq.), 5-hydro-2-methylpyridine (5.15 mg, 0.05 mmol, 2.5 eq.) and triphenylphosphine (13.63 mg, 0.05 mmol, 2.5 eq.) in DCM (0.12 mL) at 0° C. was added di-tert-butyl azodicarboxylate (8.7 mg, 0.04 mmol, 1.6 eq.). The mixture was warmed to rt and stirred for 15 min. TFA (0.10 mL) was added and the reaction was allowed to stir at rt for 2 h. The reaction was concentrated and purified using reverse phase Gilson HPLC system with acidic method (ACN/water/0.1% TFA), gradient: 11-42%. The desired fraction was concentrated to provide the title compound as a TFA salt (3.6 mg mg, 25.4% yield). ES-MS [M+1]+: 373.3, RT=0.668 min, >98% at 215 and 254 nm

(74). 2-methyl-2-propanesulfinamide (1.07g, 8.849 mmol) was added to a solution of 6-bromo-1,2-benzoxazole-3-carbaldehyde (2 g, 8.848 mmol) in toluene (50 mL). After 15 min at room temp, NaOH (353.9 mg, 8.848 mmol) was added to the solution. After stirring overnight at room temp, the reaction was diluted with EtOAc. The organic mixture was washed with water, dried over MgSO4, and concentrated. Fractions were separated by automated normal phase flash chromatography on silica gel in a gradient of EtOAc in hexanes. The fractions were combined and concentrated to yield (NE)-N-[(6-bromo-1,2-benzoxazol-3-yl)methylene]-2-methyl-propan-2-sulfinamide (2.24 g, 6.804 mmol, 77% yield).

(75). A mixture of TBAT (2.71 g, 5.01 mmol) and (NE)-N-[(6-bromo-1,2-benzoxazol-3-yl)methylene]-2-methyl-propane-2-sulfinamide (1.5 g, 4.56 mmol) was sealed the vessel was evacuated and backfilled with dry nitrogen 3×. The vessel was cooled to 0° C. and THF (30 mL) and trimethyl(trifluoromethyl)silane (81290-20-2) (0.876 mL, 5.92 mmol) was added. The reaction was maintained at 0° C. After 30 min, the reaction was quenched with saturated NH4Cl solution, and diluted with EtOAc. The organic layer was extracted, dried over MgSO4, and concentrated. Fractions were separated by automated normal phase flash chromatography on silica gel in a gradient of EtOAc in hexanes. The fractions were concentrated to yield N-[1-(6-bromo-1,2-benzoxazol-3-yl)-2,2,2-trifluoroethyl]-2-methyl-propane-2-sulfinamide (1.57 g, 3.93 mmol, 86% yield). ES-MS [M+1]+: 401.0, RT=1.138 min, >98% at 254 nm.

To a solution of N-[1-(6-bromo-1,2-benzoxazol-3-yl)-2,2-trifluoro-ethyl]-2-methyl-propane-2-sulfinamide (1.57 g, 3.93 mmol) in methanol (15 mL) was added HCl (4M in dioxanes, 3.93 mL, 15.73mmol). After 4 hrs at room temp, the reaction was concentrated. The crude residue was sonicated with EtOAc, and the product was collected as solids by filtration to yield 1-(6-bromo-1,2-benzoxazol-3-yl)-2,2,2-trifluoro-ethanamine hydrochloride (1.03 g, 3.107 mmol, 79% yield). ES-MS [M+1]+: 297.2, RT=0.875 min, >90% at 254 nm.

(77). In a 2 mL vial, was addedtBuONa (865-48-5) (21.9 mg, 0.230 mmol), Pd-tBuXPhos (2 mg, 0.003 mmol),tBuXPhos (1.2 mg, 0.003 mmol), 1-[(4-methoxyphenyl)methyl]pyrazolo[4,3-b]pyridin-3-amine (39.8, 0.157 mmol), and N-[1-(6-bromo-1,2-benzoxazol-3-yl)-2,2,2-trifluoro-ethyl]butan-1-amine (50 mg, 0.142 mmol). The reaction was evacuated and purged with Nitrogen 3×. To the vial was addedtBuOH (2 mL). The heterogeneous mixture was heated to 60° C. for 3 hrs. The reaction was added to water, and extracted with EtOAc. The organic extractions were combined and concentrated in vacuo. The residue was dissolved in DMSO, and fractions were separated via automated reverse phase HPLC in a gradient of MeCN in water (0.1% TFA in water). The fractions were concentrated, and the residue was suspended in TFA (2 mL) and transferred to a 2 mL MW vial. The reaction was heated to 130° C. for 30 min in MW. The solvent was removed under reduced pressure, and residue was dissolved in DMSO (5 mL). Fractions were separated by reverse phase HPLC in a gradient of MeCN in water (MeCN/0.1% TFA in water). The fractions were concentrated and extracted with DCM to yield 3-[1-(butylamino)-2,2,2-trifluoro-ethyl]-N-(1H-pyrazolo[4,3-b]pyridin-3-yl)-1,2-benzoxazol-6-amine; 2,2,2-trifluoroacetic acid (3.2 mg, 0.006 mmol, 4% yield). ES-MS [M+1]+: 405.3, RT=1.034 min, >98% at 215 and 254 nm.

(79). A mixture of N-[1-(6-bromo-1,2-benzoxazol-3-yl)-2,2,2-trifluoro-ethyl]-2,2-dimethyl-propanamide (40 mg, 0.11 mmol), Pd2(dba)3CHCl3(5 mg, 0.005 mmol), Xantphos (161265-03-8) (3 mg, 0.005 mmol), CS2CO3(103 mg, 0.317 mmol), and 1-[(4-methoxyphenyl)methyl]pyrazolo[4,3-b]pyridin-3-amine (30 mg, 0.116 mmol) was sealed in a vial and purged and backfilled with argon. To the vial was added 1,4-dioxane (2 mL). After 3 min at rt, the reaction was heated to 140° C. in MW for 30 min. The solution was concentrated and fractions were separated via automated reverse phase HPLC in a gradient of MeCN in water (0.5 mL NH4OH/1 L water). The fractions were concentrated and suspended in TFA. The TFA solution was heated to 130° C. in MW for 30 min. The reaction was concentrated, and fractious were separated by automated reverse phase HPLC in a gradient of MeCN in water (0.5 mL NH4OH/1 L water). The fractions were concentrated to yield 2,2-dimethyl-N-[2,2,2-trifluoro-1-[6-(1H-pyrazolo[4,3-b]pyridin-3-ylamino)-1,2-benzoxazol-3-yl]ethyl]propanamide.

Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as reaction conditions, and so forth used herein are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the herein are approximations that may vary depending upon the desired properties sought to be determined by the present invention.