HK1142335B

HK1142335B - Nicotinic acetylcholine receptor modulators

Info

Publication number: HK1142335B
Application number: HK10108947.5A
Authority: HK
Inventors: Chiara Ghiron; Arianna Nencini; Iolanda Micco; Riccardo Zanaletti; Laura Maccari; Hendrick Bothmann; Simon Haydar; Maurizio Varrone; Carmela Pratelli; Boyd Harrison
Original assignee: Siena Biotech S.P.A.; Wyeth
Priority date: 2007-01-16
Filing date: 2008-01-16
Publication date: 2014-01-03

Description

Nicotinic acetylcholine receptor modulators

Related patent application

This patent application claims priority from U.S. patent application serial No. 60/880,629, filed on 16/1/2007, the entire contents of which are incorporated herein by reference.

Technical Field

The present invention relates to compounds having alpha 7 nicotinic acetylcholine receptor (alpha 7 nAChR) agonist activity, methods for their preparation, pharmaceutical compositions containing them and their use for the treatment of neurological, psychiatric and inflammatory disorders.

Background

Active agents that bind to nicotinic acetylcholine receptors are clinically useful for the treatment and/or prevention of various diseases and disorders, particularly psychiatric disorders, neurodegenerative disorders involving dysfunction of the cholinergic system, and conditions of impaired memory and/or cognition, including, for example, schizophrenia, anxiety, mania, depression, manic depression, Gilles de la Tourette syndrome, Parkinson's disease, Huntington's disease, cognitive disorders (e.g., Alzheimer's disease, dementia with Lewy bodies, amyotrophic lateral sclerosis, impaired memory, memory loss, cognitive deficits, attention deficit hyperactivity disorder); and other uses such as treating nicotine addiction, inducing smoking cessation, treating pain (e.g., analgesic use), providing neuroprotection, and treating jet lag. See, e.g., WO 97/30998; WO 99/03850; WO 00/42044; WO 01/36417; holladay et al, j.med.chem., 40: 26, 4169-94 (1997); schmitt et al, Annual Reports Med. chem., Chapter 5, 41-51 (2000); stevens et al, Psychopharmacology, (1998) 136: 320-27; and Shytle et al, Molecular Psychiatry, (2002), 7, pp.525-535.

Various heterocyclic compounds which carry a basic nitrogen and have affinity for nicotinic and muscarinic acetylcholine receptors or are claimed to be useful in the treatment of alzheimer's disease have been reported, such as 1H-pyrazole and pyrrole-azabicyclic compounds (WO 2004013137); nicotinic acetylcholine receptor agonists (WO 2004039366); ureido-pyrazole derivatives (WO 0112188); oxadiazole derivatives having acetylcholinesterase inhibitory activity and muscarinic receptor agonistic activity (WO 9313083); pyrazole-3-carboxylic acid amide derivatives as pharmaceutical compounds (WO 2006077428); aryl piperidines (WO 2004006924); ureidoalkylpiperidines (US 6605623); compounds having activity at muscarinic receptors (WO 9950247). In addition, WO06008133 by the present applicant also discloses modulators of α 7 nicotinic acetylcholine receptors.

Summary of The Invention

The present invention provides novel compounds that are full agonists or partial agonists of the α 7 nicotinic acetylcholine receptor (α 7 nAChR), pharmaceutical compositions containing the compounds, and their use for the treatment of diseases in which activation of the α 7 nicotinic acetylcholine receptor produces a beneficial effect, such as neurological disorders, neurodegenerative disorders, psychiatric disorders, cognitive disorders, immune disorders, inflammatory disorders, metabolic disorders, addiction, nociceptive disorders, and sexual dysfunction, particularly alzheimer's disease, schizophrenia, and/or other disorders.

Brief Description of Drawings

FIG. 1: x-ray diffraction patterns of various crystalline forms of the hydrochloride salt.

FIG. 2: DSC scans of various crystalline forms of the hydrochloride salt.

FIG. 3: TGA profile of various crystalline forms of the hydrochloride salt.

FIG. 4: DVS analysis of the monohydrochloride salt (no change in crystal form after DVS analysis).

FIG. 5: DVS analysis of the hydrochloride salt (form II) (no change in form after DVS analysis).

FIG. 6: DVS analysis of the hydrochloride salt (form III) results (data immediately prior to selection).

FIG. 7: DVS as the hydrochloride salt (form V).

FIG. 8: influence of pH and HCl equivalents on HCl salt formation.

FIG. 9: influence of pH and HCl equivalents on HCl salt formation.

FIG. 10: conversion of the polyhydrochloride salt to the monohydrochloride salt form I. 259mg of dihydrochloride was added to 4 volumes of acetone +0.5 volumes of ethanol (ASDQ) and mixed at room temperature to give a slurry. The pH of the resulting slurry was about 2. 0.02mL NaOH 30% was added to raise the pH to 5-5.5. The slurry was stirred overnight and converted to the monohydrochloride salt. 173mg of the monohydrochloride were obtained.

FIG. 11: the pH was lowered to convert the monohydrochloride to form II (the slurry was stirred overnight).

FIG. 12: DSC scan of crystalline form I of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride.

FIG. 13: a TGA thermogram of 5- (4-acetyl-1, 4-diazacycloheptan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride crystalline form I.

FIG. 14: an X-ray diffraction pattern of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride form I.

FIG. 15: DVS isothermal analysis of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride form I.

FIG. 16: a DSC scan of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride form II.

FIG. 17: a TGA thermogram of 5- (4-acetyl-1, 4-diazacycloheptan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride crystalline form II.

FIG. 18: an X-ray diffraction pattern of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride form II.

FIG. 19: DVS isothermal analysis of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide hydrochloride form II.

Description of the preferred embodiments

Compound (I)

In a first aspect the present invention provides a compound of formula (I):

wherein

T is (C3-C5) an-alpha, omega-diyl or an-alpha, omega-diyl alkene, optionally carrying an oxo group and optionally substituted by one or more of the following groups: halogen; a hydroxyl group; (C1-C5) alkyl, alkoxy, fluoroalkyl, hydroxyalkyl, alkylene, fluoroalkylene; (C3-C6) cycloalkane-1, 1-diyl, oxacycloalkane-1, 1-diyl; (C3-C6) cycloalkane-1, 2-diyl, oxacycloalkane-1, 2-diyl, wherein the bond of the 1, 2-diyl forms a condensed ring with the T chain; provided that when T bears an oxo group, it is not part of an amide bond;

z is CH₂N, O, S, S (═ O) or S (═ O) 2;

q and q 'are independently an integer from 1 to 4, provided that the sum of q + q' is not more than 6;

p is 0, 1 or 2;

when p ═ 2, R' are independently selected from mono-or di- [ linear, branched or cyclic (C1-C6) alkyl ] aminocarbonyl; linear, branched or cyclic (C1-C6) alkyl, alkoxy, acyl;

q is a group of the formula

R' is C1-C3 alkyl;

j is 0 or 1;

r is a 5 to 10 membered aromatic or heteroaromatic ring;

m is 0, 1, 2 or 3;

when m is greater than 1, Y is independently from each other halogen; a hydroxyl group; a mercapto group; a cyano group; a nitro group; an amino group; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl, di-or trihaloalkoxy, alkoxy or alkylcarbonyl; (C3-C6) cycloalkyl- (C1-C6) alkoxy; (C3-C6) cycloalkyl- (C1-C6) alkyl; a linear, branched or cyclic (C1-C6) alkylcarbonylamino group; mono-or di-linear, branched or cyclic (C1-C6) alkylaminocarbonyl; a carbamoyl group; a linear, branched or cyclic (C1-C6) alkylsulfonylamino group; a linear, branched or cyclic (C1-C6) alkylsulfonyl group; mono-or di-linear, branched or cyclic (C1-C6) alkylsulfamoyl; linear, branched or cyclic (C1-C6) alkoxy- (C1-C6) alkyl; or when m ═ 2, two Y substituents may form a ring together with the atom of the R group to which they are attached.

In a first preferred embodiment, the present invention provides a compound of formula (I) wherein:

t is butane-1, 4-diyl, optionally substituted by one or more (C1-C3) alkyl, halogen;

z is N or O;

q is

p, q ', R', j, R, Y and m have the definitions stated in formula (I);

in this embodiment, particular preference is given to compounds of the formula (I) in which:

t is butane-1, 4-diyl;

z is N or O;

r' is selected from linear, branched or cyclic (C1-C6) alkyl, alkoxy, acyl;

p is 0 or 1;

q is

j is 0;

r is a 5 to 10 membered aromatic or heteroaromatic ring;

q, q', R, Y and m have the meanings indicated for the compounds of formula (I);

in another particularly preferred group of compounds:

t is butane-1, 4-diyl;

z is N;

p is 1;

r' is (C1-C6) acyl;

q is

j is 0;

r is phenyl, pyridyl, thienyl; an indolyl group;

m is 0, 1 or 2;

when m is greater than 1, Y is independently from each other halogen; a hydroxyl group; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl, di-or trihaloalkoxy, alkoxy; (C3-C6) cycloalkyl- (C1-C6) alkyl;

q, q' have the definitions stated for the compounds of the formula (I);

in another preferred embodiment, the present invention provides a compound, hereinafter G1, which is referred to as a compound of formula (I), wherein: ,

t is propane-1, 3-diyl, optionally substituted by (C1-C3) alkyl, halogen;

z is CH₂、N、O；

Q is a group of the formula:

r ', p, q ', R ', j, R, Y and m have the meanings indicated for the compounds of formula (I);

among the compounds of formula (I) which are particularly preferred in G1

T is propane-1, 3-diyl, optionally substituted by (C1-C3) alkyl, halogen;

z is CH₂；

Q is

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl, alkoxy, acyl;

j is 0;

r, Y and m have the meanings indicated for the compounds of formula (I);

in G1, another particularly preferred group of compounds,

t is propane-1, 3-diyl;

z is CH₂；

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl;

q is

j is 0;

r is phenyl, pyridyl, naphthyl;

m is 1 or 2;

when m is greater than 1, Y is independently from each other halogen; a hydroxyl group; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl, di-or trihaloalkoxy, alkoxy; (C3-C6) cycloalkyl- (C1-C6) alkoxy.

In this group of compounds, certain of the newly invented compounds are those in which Q-R is

There is another group of preferred compounds of formula (I) in G1, wherein

T is propane-1, 3-diyl, optionally substituted by (C1-C3) alkyl, halogen;

z is CH₂；

Q is

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl, alkoxy, acyl;

j is 0;

r, Y and m have the meanings indicated for the compounds of formula (I);

in a fourth group of preferred compounds in G1:

t is propane-1, 3-diyl;

z is CH₂；

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl;

q is

j is 0;

r is phenyl, pyridyl, naphthyl;

m is 1 or 2;

Of this group of compounds, the most preferred compounds are those in which Q-R is

It is readily understood by those skilled in the art that, as in the compounds of the present invention, the unsubstituted ring nitrogen atoms of pyrazoles and imidazoles are known to rapidly reach equilibrium in solution as a mixture of tautomers:

thus, in the following description, when a compound of formula (I) is represented in only one tautomeric form, another tautomer is also naturally included within the scope of the present invention.

The compounds of the invention may be in the form of the free base or of an acid addition salt, preferably in the form of a salt with a pharmaceutically acceptable acid. The invention also provides isolated isomers and diastereomers of the compounds of formula (I), or mixtures thereof (e.g., racemic and diastereomeric mixtures), as well as isotopic mixtures thereof.

The pharmacological activity of a representative group of compounds of formula (I) was demonstrated in an in vitro experiment. Cells stably transfected with the α 7 nicotinic acetylcholine receptor were used for this experiment, and cells expressing both the α 1 and α 3 nicotinic acetylcholine receptors and the 5HT3 receptor were used as controls for the selectivity study.

The compounds of formula (I) of the present invention may be in any of a variety of useful forms, such as pharmaceutically acceptable salts, particular crystalline forms, and the like. In some embodiments, the present invention provides prodrugs of one or more compounds of formula (I). Various prodrug forms are known in the art, for example, Bundgaard (ed.), Design of produgs, Elsevier (1985); widder et al (ed.), Methods in enzymology, vol.4, Academic Press (1985); Kgrogsgaard-Larsen et al (eds.); "Design and Application of primers", Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); bundgaard et al, Journal of drug Delivery Reviews, 8: 1-38 (1992); bundgaard et al, j.pharmaceutical sciences, 77: 285 et seq (1988); and prodrug forms as described in Higuchi & Stella (eds.), Prodrugs asNovel Drug Delivery Systems, American Chemical Society (1975).

Use of

Active agents that bind to nicotinic acetylcholine receptors are clinically useful for the treatment and/or prevention of various diseases and disorders, particularly psychiatric disorders, neurodegenerative disorders involving dysfunction of the cholinergic system, and memory and/or cognitive impairment disorders, including, for example, schizophrenia, anxiety, mania, depression, manic depression, gilles de la tourette's syndrome, parkinson's disease, huntington's disease, cognitive disorders (e.g., alzheimer's disease, dementia with lewy bodies, amyotrophic lateral sclerosis, memory impairment, memory loss, cognitive deficits, attention deficit hyperactivity disorder); and other uses such as treating nicotine addiction, inducing smoking cessation, treating pain (e.g., analgesic use), providing neuroprotection, and treating jet lag. See, e.g., WO 97/30998; WO 99/03850; WO 00/42044; WO 01/36417; holladay et al, j.med.chem., 40: 26, 4169-94 (1997); schmitt et al, Annual Reports Med. chem., Chapter 5, 41-51 (2000); stevens et al, Psychopharmacology, (1998) 136: 320-27; and Shytle et al, Molecular Psychiatry, (2002), 7, pp.525-535.

Accordingly, the present invention provides a method of treating a patient, particularly a human, suffering from any of the following diseases: psychotic disorders, neurodegenerative disorders involving dysfunction of the cholinergic system and conditions of impaired memory and/or cognition including, for example, schizophrenia, anxiety, mania, depression, manic depression, gilles de la tourette's syndrome, parkinson's disease, huntington's disease and/or cognitive disorders (e.g., alzheimer's disease, lewy body dementia, amyotrophic lateral sclerosis, impaired memory, memory loss, cognitive deficit, attention deficit hyperactivity disorder) comprising administering to said patient an effective amount of a compound of formula (I).

Neurodegenerative disorders that can be treated by the methods of the invention include, but are not limited to, the treatment and/or prevention of Alzheimer's disease, pick's disease (Friedland, Dementia, (1993) 192. sup. 203; Procter, Dement Geriator Cogn disease (1999) 80-4; Sparks, ArchNeurol.1991) 796-9; Mizukami, Acta Neuropthalol.1989) 52-6; Hansen, Am J Pathol.1988) 507-18), diffuse Lewy body disease, progressive supranuclear palsy (Steel-Richardson syndrome, see Whitehos, J Neural Transpl.1987) 24: 175-82; Whitehosue 722, Arch Neurol.1988 (45: Whitehol. 4; Whiten Neurol.9; Whiten.9; Mah-3. sup. 9) 239-9; Maher-11. sup. 9; Mah-128. sup. 9; Mah-3. sup. 9; Mah-11. sup. 9; Mah-9, including amyotrophic lateral sclerosis (Nakamizo, Biochem Biophys Res Commun. (2005)330(4), 1285-9; Messi, FEBS Lett. (1997)411 (1): 32-8; Mohammadi, Muscle Nerve. (2002) Oct. (26) (4): 539-45; Hanagasi, Brain Reg Brain Bran. (2002)14 (2): 234-44; Crochemor, Neurochem Int. (2005)46 (5): 357-68), degenerative ataxia, corticobasal degeneration 2006, ALS combined with Parkinson's disease and dementia, subacute sclerosing panencephalitis, Huntington's disease (Kanazawa, J Neurol Sci. (1985) 151-65; Manurol. (J Neurol. (3114) 1990, Lauram J.281; Neurol et. (2003-4; Moummy. 2003; Moummy. J. (2003) 1518; Smith. (2003-9; Smith et. (2003) 1518; Moummy. J.103; Smith et. (2003-9; Smith. (2003, Mah Sanger. (1518), neurology. (2006)1268-71), Parkinson's disease, synucleinopathy, primary progressive aphasia, striatal substantia nigra degeneration, Mayas disease/spinocerebellar ataxia type 3, olivopontocerebellar degeneration, Gilles de la Tourette syndrome, bulbar paralysis, pseudobulbar paralysis, spinal muscular atrophy, spinal bulbar muscular atrophy (Kennedy's disease), primary lateral sclerosis, familial spastic paraplegia, Werdnig-Hoffmann's disease, Kugelberg-Welander disease, Tay-Sach disease, dhoff disease, familial spasticity, Wohlfart-gelberg-Welander disease, spastic paraplegia, progressive multifocal encephalopathy, prion diseases (e.g., Creutzldt-Jakob disease, Gehtmann-Strauss-Afeffenfex disease, kuhlung disease and cerebral infarction), ischemic and ischemic stroke, including any type of cerebral hemorrhage including but not limited to intracranial hemorrhage, cerebral infarction, cerebral embolism, and ischemic stroke including any type of cerebral hemorrhage including but not limited to intracranial hemorrhage, Subdural hemorrhage, subarachnoid hemorrhage, and cerebral hemorrhage), intracranial and intravertebral injuries (including but not limited to contusions, punch-throughs, shear-cuts, crush-wounds, and lacerations).

Furthermore, α 7nACh receptor agonists such as the compounds of the present invention are useful in the treatment of age-related dementia and other dementias and conditions associated with memory loss, including age-related memory loss, aging, vascular dementia, diffuse white matter disease (Binswanger's disease), endocrine or metabolic dementia, head traumatic dementia and diffuse brain injury, dementia pugilistica, alcoholic dementia (Korsakoff syndrome) and frontal dementia. See, e.g., WO 99/62505, Tomimoto comment Geriatr Cogn disorder (2005), 282-8; Tohgi-J neuroltram, (1996), 1211-20; casamnti, Neuroscience (1993)465-71, Kopelman, Br J Psychiatry (1995) 154-73; cochrane, Alcohol. (2005) 151-4).

Amyloid Precursor Protein (APP) and A β peptides derived from APP, such as A β 1-42 and other fragments, are known to be associated with the pathology of Alzheimer's disease. The a β 1-42 peptide is not only associated with neurotoxicity but also inhibits cholinergic neurotransmitter function. In addition, it has been detected that a β peptides bind to the α 7nACh receptor. The inflammatory reflex is the response of the autonomic nervous system to inflammatory signals. Upon sensing inflammatory stimuli, the autonomic nervous system responds by releasing acetylcholine from the vagus nerve and activating α 7 nicotinic receptors on macrophages. These macrophages then release cytokines. This pathway dysfunction is known to be associated with inflammatory diseases in humans, including rheumatoid arthritis, diabetes and sepsis. Macrophages express α 7 nicotinic receptors, which may mediate cholinergic anti-inflammatory responses. See, e.g., Czura, C J, et al, j.lnn.med., (2005)257(2), 156-66; wang, H. et al Nature (2003) 421: 384-; de Journal of Pharaology (2007)151, 915-. Mammalian spermatozoa acrosome reaction is an exocytosis process that plays an important role in the process of fertilization of an egg by sperm. Activation of the α 7nAChR on sperm cells has been shown to be essential for acrosome reactions (Son, J. -H. and Meizel, S.biol. Reproduct.68: 1348-1353, 2003). In addition, nicotinic receptors have been found to play a role in the body's response to alcohol intake. Thus, α 7nACh receptor agonists, such as the compounds of the present invention, are also useful in the treatment of these disorders, diseases and conditions.

For example, agonists of the α 7nACh receptor subtype are also useful in the treatment of nicotine addiction, in inducing smoking cessation, in the treatment of pain, jet lag, obesity, diabetes, sexual dysfunction and fertility disorders (e.g. premature ejaculation or vaginal dryness, see US6448276), in drugs of abuse (Solinas, Journal of neuroscience (2007)27(21), 5615-.

Recently, some studies found that nicotine has potential neuroprotective effects in neurodegenerative models in various animals and cell cultures, including excitotoxic injury (Prendergast, M.A., et al Med. Sci. unit. (2001), 7, 1153-1160; Garrido, R., et al (2001), J.Neurochem.76, 1395-1403; Semba, J., et al (1996) BrainRes.735, 335-338; Shimohama, S.et al (1996), an.N.Y.Acad.Sci.777, 356-361; Akaike, A. et al (1994) Brain Res.644, 181-187), nutritional deficiencies (Yamahata shita, H., Nakamura, S. (S.) Neurosche.Le213, 145-147), ischemia (Shimohaoha. S. (1998), Neurosche. J. 10, J. 1997, J. 35-121; Brasshaki, J. 35-121; Brasshaya J. 35, J. 35-35, 35-35; Brasshaya. J. 35, 1996; Brasshaoha. J. 35, J. E. A. 35, 1997, J. 35, J. A, t, et al (2001) J.biol.chem.276, 13541-containing 13546) and protein aggregation-mediated neuronal degeneration (Kelton, M.C. et al (2000) Brain Cogn 43, 274-containing 282). In many cases where nicotine exhibits neuroprotective effects, direct involvement of receptors containing the α 7 subtype was found (Shimohama, S. et al (1998) Brain Res.779, 359-363; Kihara, T. et al (2001) J.biol. chem.276, 13541-13546; Kelton, M.C. et al (2000) Brain Cogn 43, 274-282; Kem, W.R. (2000) Brain Brain Res.113, 169-181; Dajas-Bailador, F.A. et al (2000) Neuropharmacology 39, 2799-2807; Strahendorf, J.C. et al (2001) Brain Brares.901, 71-78), suggesting that activation of nicotinic acetylcholine receptors containing the α 7 subtype may play an important role in mediating neuroprotection in nicotine. The present data indicate that α 7 nicotinic acetylcholine receptors can serve as potent molecular targets in the development of neuroprotective molecules with agonist/positive modulator activity. Indeed, researchers have recognized that α 7 nicotinic receptor agonists could be a clue for the development of neuroprotective drugs and have begun to evaluate their value in this regard (Jonnala, R.R., et al (2002) Life Sci.70, 1543-1554; Bencherif, M.et al (2000) Eur.J.Pharmacol.409, 45-55; Donnelly-Roberts, D.L. et al (1996) Brain Res.719, 36-44; Meyer, E.M. et al (1998) J.Pharmacol.Exp.Ther.284, 1026-1032; Stevens, T.R. et al (2003) J.Neuroscience 23, 10093-10099). The compounds described herein are useful in the treatment of such diseases.

The present invention provides a method of treating a patient, particularly a human, suffering from age-related dementia and other dementias and disorders associated with memory loss, comprising administering to said patient an effective amount of a compound of formula (I).

The present invention includes a method of treating a patient suffering from memory impairment due to, for example, mild cognitive impairment caused by aging, alzheimer's disease, schizophrenia, parkinson's disease, huntington's disease, pick's disease, Creutzfeldt-Jakob disease, depression, aging, head trauma, stroke, central nervous system hypoxia, brain aging, multi-infarct dementia and other neurological disorders as well as HIV and cardiovascular disease, comprising administering to the patient an effective amount of a compound of formula (I).

One embodiment of the present invention provides a method of treating and/or preventing dementia in a patient with alzheimer's disease comprising administering to said patient a therapeutically effective amount of a compound of formula (I) to inhibit binding of an Α β peptide (preferably Α β 1-42) to nACh receptors, preferably α 7nACh receptors, most preferably human α 7nACh receptors (this embodiment also provides a method of treating and/or preventing other clinical manifestations of alzheimer's disease including but not limited to cognitive and speech deficits, akinesia, depression, delusional disorders and other neuropsychiatric symptoms and manifestations and dyskinesias and gait).

The invention also provides methods for treating other amyloidosis cases such as hereditary cerebrovascular disease, non-neuropathic hereditary amyloidosis, down's syndrome, macroglobulinemia, secondary familial mediterranean fever, Muckle-Wells syndrome, multiple myeloma, amyloidosis associated with the pancreas and heart, chronic dialysis osteoarthropathy, and Finnish-and Chlamydomonas amyloidosis.

In addition, nicotinic receptors have been found to play a role in the body's response to alcohol intake. Therefore, agonists of the α 7nACh receptor are useful for the treatment of alcohol withdrawal syndrome and for anti-intoxication therapy. Accordingly, one embodiment of the present invention provides a method of treating a patient having symptoms of alcohol withdrawal or treating a patient receiving anti-intoxication therapy comprising administering to said patient an effective amount of a compound of formula (I).

Agonists of the α 7nACh subtype receptor may also be useful in protecting nerves in injuries associated with stroke, ischemia, and glutamate-induced excitotoxicity. Accordingly, one embodiment of the present invention provides a method of providing neuroprotection in injury associated with stroke, ischemia, and glutamate induced excitotoxicity to treat a patient comprising administering to said patient an effective amount of a compound of formula (I).

Agonists of the α 7nACh subtype receptor may also be useful in the treatment of nicotine addiction, the induction of smoking cessation, the treatment of pain, the treatment of jet lag, obesity, diabetes, sexual dysfunction and fertility disorders (e.g. premature ejaculation or vaginal dryness, see US6448276), drug abuse (Solinas, Journal of neuroscience (2007)27(21), 5615-. Accordingly one embodiment of the present invention provides a method of treating a patient suffering from nicotine addiction, pain, jet lag, obesity and/or diabetes, or a method of inducing smoking cessation in a patient, comprising administering to the patient an effective amount of a compound of formula (I).

The inflammatory reflex is the response of the autonomic nervous system to inflammatory signals. Upon sensing inflammatory stimuli, the autonomic nervous system responds by releasing acetylcholine from the vagus nerve and activating α 7 nicotinic receptors on macrophages. These macrophages then release cytokines. This pathway dysfunction is known to be associated with human inflammatory diseases including rheumatoid arthritis, diabetes and sepsis. Macrophages express α 7 nicotinic receptors, which may mediate cholinergic anti-inflammatory responses. Compounds having affinity for the α 7nACh receptor of macrophages are therefore useful in human inflammatory diseases including rheumatoid arthritis, diabetes and sepsis. See, e.g., Czura, C J et al, j.lntern.med., (2005)257(2), 156-66, Wang, h. et al, Nature (2003) 421: 384-; de Journal of Pharmacology (2007)151, 915-.

Accordingly one embodiment of the present invention provides a method of treating a patient (e.g. a mammal such as a human) suffering from an inflammatory disease such as, but not limited to, rheumatoid arthritis, diabetes or sepsis, comprising administering to said patient an effective amount of a compound of formula (I).

Mammalian spermatozoa acrosome reaction is an exocytosis process that plays an important role in the process of fertilization of an egg by sperm. Activation of the α 7nAChR on sperm cells has been shown to be essential for acrosome reactions (Son, J. -H. and Meizel, S.biol. Reproduct.68: 1348-1353, 2003). Thus, selective α 7 active agents have utility in the treatment of fertility disorders.

Furthermore, due to the affinity of the compounds of formula (I) for the α 7nACh receptor, labelled derivatives of the compounds of formula (I) (e.g. C11 or F18 labelled derivatives) are useful for neuroimaging of receptors within, for example, the brain. Thus, in vivo receptor imaging using such labeled active agents can be performed using, for example, PET imaging techniques.

Memory impairment manifests as a failure to learn new information and/or an inability to recall previously learned information. Memory disorders are the main symptoms of dementia and may also be symptoms associated with e.g. alzheimer's disease, schizophrenia, parkinson's disease, huntington's disease, pick's disease, Creutzfeldt-Jakob disease, aids, cardiovascular diseases and head trauma and age-related cognitive decline.

Accordingly, one embodiment of the present invention provides a method of treating a patient suffering from, for example, Mild Cognitive Impairment (MCI), vascular dementia (VaD), age-related cognitive decline (AACD), amnesia associated with open chest surgery, cardiac arrest and/or general anesthesia, memory deficits resulting from narcotics received early in life, sleep-deficit-induced cognitive impairment, chronic fatigue syndrome, narcolepsy, aids-related dementia, cognitive dysfunction associated with epilepsy, down syndrome, alcohol-related dementia (Korsakoff syndrome), drug/drug-induced memory impairment, boxer dementia (boxer syndrome), and animal dementia (e.g., dog, cat, horse, etc.), comprising administering to the patient an effective amount of a compound of formula (I).

The dosage of the compounds used in the treatment may vary depending, for example, on the route of administration, the nature and severity of the disease. In general, the daily dose to achieve a pharmaceutically acceptable effect in humans may be from 0.01 to 200 mg/kg.

In some embodiments of the invention, one or more compounds of formula (I) are administered in combination with one or more other pharmaceutically active agents. The phrase "in combination" as used herein refers to the administration of multiple active agents to an individual simultaneously. It is understood that two (or more) active agents are considered to be administered "in combination" if an individual is exposed to the two (or more) active agents simultaneously. Each of the two or more active agents may be administered according to a different schedule; it is not required that each dose of the different active agents be administered simultaneously or in the same composition. Conversely, two (or more) active agents are considered to be administered "in combination" as long as they are still in the individual.

For example, a compound of formula (I) in the form described herein may be administered in combination with one or more other alpha 7 nicotinic acetylcholine receptor modulators. Alternatively or additionally, a compound of formula (I) in the form described herein may be administered in combination with one or more antipsychotic, analgesic, anti-inflammatory or other pharmaceutically active agents.

Effective amounts of various other pharmaceutically active agents are well known to those skilled in the art. However, determining the optimal effective amount range of such other pharmaceutically active agents is well within the skill of one of ordinary skill in the art. The compound of formula (I) and the other pharmaceutically active agent may have an additive effect or, in some embodiments, a synergistic effect. In some embodiments of the invention, the effective amount of the compound of formula (I) is lower when another pharmaceutically active agent is administered to an animal than when the other pharmaceutically active agent is not administered. In this case, without being bound by any theory, it is believed that the compound of formula (I) has a synergistic effect with said another pharmaceutically active agent. In some cases, a patient in need of treatment is treated with one or more additional pharmaceutically active agents. In some cases, a patient in need of treatment is treated with at least two other pharmaceutically active agents.

In some embodiments, the additional pharmaceutically active agent is selected from one or more antidepressants, anxiolytics, antipsychotics, or cognitive function-enhancing agents. Examples of classes of antidepressant drugs that may be used in combination with the active compounds of the present invention include norepinephrine reuptake inhibitors, Selective Serotonin Reuptake Inhibitors (SSRIs), NK-1 receptor antagonists, monoamine oxidase (MAO) inhibitors, reversible inhibitors of monoamine oxidase (RIMA), Serotonin and Norepinephrine Reuptake Inhibitors (SNRIs), Corticotropin Releasing Factor (CRF) antagonists, alpha-adrenoceptor antagonists and atypical antidepressants. Suitable norepinephrine reuptake inhibitors include tertiary amine tricyclic antidepressants and secondary amine tricyclic antidepressants. Suitable tertiary amine tricyclic antidepressants and secondary amine tricyclic antidepressants include amitriptyline, clomipramine, doxepin, imipramine, trifluoropropyline, dothiepin, butiline, iprindole, lofepramine, nortriptyline, protriptyline, amoxapine, desipramine, and meprotiline. Suitable selective serotonin reuptake inhibitors include fluoxetine, citalopram, escitalopram, fluvoxamine, paroxetine and sertraline. Examples of monoamine oxidase inhibitors include isocarboxazid, phenelzine and tranylcypromine. Suitable reversible inhibitors of monoamine oxidase include moclobemide. Serotonin and norepinephrine reuptake inhibitors suitable for use in the present invention include venlafaxine, nefazodone, milnacipran, and duloxetine. Suitable CRF antagonists include compounds described in international patents publication nos. WO 94/13643, WO 94/13644, WO 94/13661, WO 94/13676 and WO 94/13677. Suitable atypical antidepressants include bupropion, lithium, nefazodone, trazodone, and viloxazine. Suitable NK-1 receptor antagonists include the compounds described in International patent WO 01/77100.

Anxiolytics that may be used in combination with the compounds of formula (I) include, but are not limited to, benzodiazepinesSteroid and serotonin 1A (5-HT)_1A) Agonists or antagonists, especially 5-HT_1APartial agonists and Corticotropin Releasing Factor (CRF) antagonists. BenzodiazepinesSuitable examples of classes include alprazolam, chlordiazepoxide, clonazepam, dipotassium chlordiazepoxide, diazepam, halazepam, lorazepam, oxazepam and pramipepam. Examples of suitable 5-HT1A receptor agonists or antagonists include buspirone, fluoroxingcron, gepirone and ixabepilone.

Antipsychotic agents that may be used in combination with the compounds of formula (I) include, but are not limited to, aliphatic phenothiazines, piperazine phenothiazines, phenylbutanone, substituted benzamides, and thioxanthenes. Other examples of such drugs include, but are not limited to, haloperidol, olanzapine, clozapine, risperidone, pimozide, aripiprazole, and ziprasidone. In some cases, the drug is an anticonvulsant drug, such as phenobarbital, phenytoin, primidone, or carbamazepine.

Drugs that enhance cognitive function that may be used in combination with a compound of formula (I) include, but are not limited to, drugs that modulate neurotransmitter levels (acetylcholinesterase or cholinesterase inhibitors, cholinergic receptor agonists or serotonin receptor antagonists), drugs that modulate soluble a β levels, amyloid fibril formation, or amyloid plaque burden (e.g., gamma-secretase inhibitors, β -secretase inhibitors, antibody therapies, and degradative enzymes), and drugs that protect neuronal integrity(e.g., antioxidants, kinase inhibitors, cystatins and hormones). Other representative drugs that may be selected for co-administration with the compounds of the invention include cholinesterase inhibitors (e.g., tacrine)DonepezilRivastigmineGalanthamineMetrofosfamid, physostigmine, and huperzine A), N-methyl-D-aspartate (NMDA) antagonists and agonists (e.g., dextromethorphan, memantine, diloxetine maleate (MK-801), clomiphene, limamin, eliprodil, amantadine, D-cycloserine, felbamate, ifenprodil, CP-101606(Pfizer), Delucemine, and the compounds described in U.S. Pat. Nos. 6,821,985 and 6,635,270), glutamate receptor modulators (ampathines) (e.g., cyclothiazine, aniracetam, CX-516, and huperzine A), and N-methyl-D-aspartate (NMDA) antagonists and agonists (e.g., dextromethorphan, memantine, mexican, dilCX-717, CX-516, CX-614 and CX-691(Cortex Pharmaceuticals, Inc. Irvine, CA), 7-chloro-3-methyl-3-4-dihydro-2H-1, 2, 4-benzothiadiazine S, S-dioxide (see Zivkovic et al, 1995, J.Pharmacol. Exp. Therap., 272: 300-]Hept-5-en-2-yl-6-chloro-3, 4-dihydro-2H-1, 2, 4-benzothiadiazine-7-sulfonamide-1, 1-dioxide (Yamada et al, 1993, J.Neurosc.13: 3904-; 7-fluoro-3-methyl-5-ethyl-1, 2, 4-benzothiadiazine-S, S-dioxide; compounds described in us patent 6,620,808 and international patent applications WO 94/02475, WO 96/38414, WO 97/36907, WO 99/51240 and WO 99/42456), benzodiazepines(BZD)/GABA receptor complex modulators (e.g., halogat, gengabane, zaleplon, and the drugs described in U.S. patent nos. 5,538,956, 5,260,331, and 5,422,355); serotonin antagonists (e.g. 5HT receptor modulators, 5HT_1AAntagonists or agonists (including but not limited to Lecozotan and the compounds described in U.S. Pat. Nos. 6,465,482, 6,127,357, 6,469,007 and 6,586,436 and PCT patent WO 97/03982) and 5-HT₆Antagonists (including but not limited to the compounds described in U.S. patent nos. 6,727,236, 6,825,212, 6,995,176, and 7,041,695)); nicotinic agents (e.g., niacin); muscarinic agents (e.g., xanomeline, CDD-0102, cevimeline, tasalidine, oxybutynin, tolterodine, propiverine, trospium chloride (tropsiumchloride), and darifenacin); monoamine oxidase type B (MAO B) inhibitors (e.g., rasagiline, selegiline (deprenyl), lazabemide, safinamide (safinamide), clorgoline, pargyline, N- (2-aminoethyl) -4-chlorobenzamide hydrochloride and N- (2-aminoethyl) -5- (3-fluorophenyl) -4-thiazolecarboxamide hydrochloride); phosphodiesterase (PDE) IV inhibitors (e.g., roflumilast, alotheophylline, cilomilast, rolipram, RO-20-1724, theophylline, denbufylline, ARIFLO, roflumilast, CDP-840 (a triarylethane), CP80633 (a pyrimidinone), RP 73401(Rhone-Poulenc Rorer), denbufylline (SmithKlineBeecham), arotheophylline (Almirall), CP-77,059(Pfizer), pyrido [2, 3d ] theophylline]Pyridazin-5-ones (Syntex), EP-685479(Bayer), T-440(Tanabe Seiyaku) and SDZ-ISQ-844 (Novartis)); a G protein; a channel modulator; immunotherapeutics (e.g., the compounds described in US 2005/0197356 and US 2005/0197379); anti-amyloid or amyloid level lowering agents (e.g., Bapineuzumab and compounds described in U.S. patent 6,878,742 or U.S. patent US 2005/0282825 or US 2005/0282826); statins and peroxisome proliferator-activated receptor (PPARS) modulators (e.g., gemfibrozil)FenofibrateRosiglitazone maleatePioglitazone (Actos)^TM) Rosiglitazone (Avandia)^TM) Clofibrate and bezafibrate); a cysteine protease inhibitor; advanced glycation end product receptor inhibitors (RAGE) (e.g., aminoguanidine, pyridoxamine, carnosine, phenazine diamine, OPB-9195, and tenacitan); direct or indirect neuroprotective agents (e.g.Piperacillin, oxiracetam, AIT-082(Emilieu, 2000, Arch. Neurol.57: 454)); beta-secretase (BACE) inhibitors, alpha-secretase, immunophilins, cysteine protease-3 inhibitors, Src kinase inhibitors, Tissue Plasminogen Activator (TPA) activators, AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) modulators, M4 agonists, JNK3 inhibitors, LXR agonists, H3 antagonists, and angiotensin IV antagonists. Other cognitive enhancers include, but are not limited to, acetyl-1-carnitine, brain phosphatidylcholine, huperzine, DMAE (dimethylaminoethanol), Bacopa moneirii extract, sage extract, L-alpha glycerophosphorylcholine, ginkgo biloba and ginkgo biloba extracts, vinpocetine, DHA, nootropic drugs including benztropine, Pikatropin (manufacturer CreativeCompounds, LLC, Scott City, MO), besiflozin, enopridine, siberidine, estrogens and estrogens, idebenone, T-588(Toyama Chemical, Japan) and FK960(Fujisawa Pharmaceutical Co. Ltd.). The compounds described in us patents 5,219,857, 4,904,658, 4,624,954 and 4,665,183 may also be useful as cognitive enhancers described herein. Mechanism of action cognitive enhancers that fall within one or more of the above-described mechanisms are included within the scope of the invention.

In some embodiments, the compounds of formula (I) have an additive effect with the cognitive enhancer, or, in some embodiments, they have a synergistic effect. In some embodiments, when a cognitive enhancer is co-administered to an animal with a compound of formula (I) of the present invention, the effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof is less than the effective amount when the cognitive enhancer is not administered. In some embodiments, when a cognitive enhancer is co-administered to an animal with a compound of formula (I), the effective amount of the cognitive enhancer is less than the effective amount when the compound of the invention or a pharmaceutically acceptable salt thereof is not administered. In some embodiments, when a cognitive enhancer is co-administered to an animal with a compound of formula (I) of the present invention, the dosage of both is lower than if not co-administered. In these cases, without being bound by any theory, it is believed that the compounds of formula (I) have a synergistic effect with cognitive enhancers.

In some embodiments, the other pharmaceutically active agent is an agent useful for the treatment of alzheimer's disease or a related disorder thereof, such as dementia. Examples of active agents useful for treating alzheimer's disease include, but are not limited to, donepezil, rivastigmine, galantamine, memantine, and tacrine.

In some embodiments, the compound of formula (I) is administered with another pharmaceutically active agent or in the same composition.

In some embodiments, an effective amount of a compound of formula (I) and an effective amount of another pharmaceutically active agent may be administered in the same composition.

In another embodiment, a composition containing an effective amount of a compound of formula (I) is administered simultaneously with another composition containing an effective amount of another pharmaceutically active agent. In another embodiment, an effective amount of a compound of formula (I) is administered before or after an effective amount of another pharmaceutically active agent. In this embodiment, the compound of formula (I) is administered at the time the other pharmaceutically active agent exerts its therapeutic effect, or the other pharmaceutically active agent is administered at the time the compound of formula (I) exerts its prophylactic or therapeutic effect.

Thus, in some embodiments, the present invention provides a composition comprising an effective amount of a compound of formula (I) of the present invention and a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a second pharmaceutically active agent.

In another embodiment, the composition further comprises another pharmaceutically active agent selected from one or more antidepressants, anxiolytics, antipsychotics or cognitive enhancers. Antidepressants, anxiolytics, antipsychotics or cognitive enhancers suitable for use in the compositions of the present invention include the aforementioned antidepressants, anxiolytics, antipsychotics and cognitive enhancers.

In another embodiment, the pharmaceutically acceptable carrier is suitable for oral administration and the composition comprises an oral dosage form.

In some embodiments, one or more compounds of formula (I) are administered in combination with antidepressant drug therapy, antipsychotic drug therapy, and/or anticonvulsant drug therapy.

In some embodiments, the compounds of formula (I) are administered in combination with one or more Selective Serotonin Reuptake Inhibitors (SSRIs) (e.g., fluoxetine, citalopram, escitalopram oxalate, fluvoxamine maleate, paroxetine or sertraline), tricyclic antidepressants (e.g., desipramine, amitriptyline, Amoxipine, clomipramine, doxepin, imipramine, nortriptyline, protriptyline, trifluoroperazine, duloxetine, butiline, iprindole or lofepramine), aminoketones (e.g., butoxypropiophenone); in some embodiments, the compounds of formula (I) are combined with monoamine oxidase (MAO) inhibitors (e.g., phenelzine, isocarboxazid, or tranylcypromine), Serotonin and Norepinephrine Reuptake Inhibitors (SNRI) (e.g., venlafaxine, nefazodone, milnacipran, duloxetine), Norepinephrine Reuptake Inhibitors (NRI) (e.g., reboxetine), 5-HT_1APartial agonists (e.g. buspirone), 5-HT_2AReceptor antagonists (e.g., nefazodone), typical or atypical antipsychotics. Examples of such antipsychotic agents include aliphatic phenothiazines, piperazine phenothiazines, phenylbutanone, substituted benzamides, and thioxanthenes. Other examples of such drugs include haloperidol, olanzapine, clozapine, risperidonePimozide, aripiprazole, ziprasidone. In some cases, the drug is an anticonvulsant drug such as phenobarbital, phenytoin, primidone, or carbamazepine. In some cases, the compound of formula (I) is administered with at least two drugs that are antidepressants, antipsychotics, anticonvulsants, or a combination thereof.

Pharmaceutical composition

Another aspect of the invention relates to pharmaceutical compositions containing one or more compounds of formula (I) together with pharmaceutically acceptable carriers and excipients. The pharmaceutical compositions may be in the form of solid, semi-solid or liquid preparations, preferably in the form of solutions, suspensions, powders, granules, tablets, capsules, syrups, suppositories, aerosols or controlled release systems. The compositions may be administered in a variety of routes including oral, transdermal, subcutaneous, intravenous, intramuscular, rectal and intranasal routes, and are preferably prepared in unit dosage form containing from about 1 to about 1000mg, preferably 1 to 600mg, of the active ingredient per dose. The compounds of the invention may be in the form of the free base or of an acid addition salt, preferably with a pharmaceutically acceptable acid. The invention also includes isolated isomers and diastereomers of the compounds of formula I, or mixtures (e.g., racemic mixtures) thereof. Principles and methods for preparing pharmaceutical compositions are described, for example, in Remington's pharmaceutical Science, Mack Publishing Company, Easton (PA).

When administered to an animal, one or more compounds of formula (I) in any desired form (e.g., salt, crystalline form, etc.) may be administered as a compound or as a component of a pharmaceutical composition containing a physiologically acceptable carrier or excipient. Such pharmaceutical compositions of the invention may be prepared according to standard procedures, for example, by mixing the compound or compounds with a physiologically acceptable carrier, excipient or diluent. The compounds of formula (I) may be mixed with physiologically acceptable carriers, excipients or diluents according to known methods.

The pharmaceutical compositions provided herein (i.e., compositions comprising one or more compounds of formula (I) in an appropriate form) may be administered by the oral route. Alternatively, the pharmaceutical compositions provided herein may be administered by any other convenient route, for example, by parenteral routes (e.g., subcutaneous, intravenous, etc. infusion or bolus injection), by absorption through epithelial or mucosal layers (e.g., oral, rectal, vaginal, small intestinal mucosa, etc.), and the like. Administration may be systemic or topical. Various known delivery systems may be used, including, for example, liposome encapsulation, microparticles, microcapsules, capsules.

Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intracerebral, intravaginal, transdermal, rectal, by inhalation methods, or topical administration, particularly to the ear, nose, eye, or skin. In some cases, the compound (and/or one or more metabolites thereof) will be released into the blood following administration. The mode of administration may be determined by a physician.

In some embodiments, the pharmaceutical compositions provided herein are administered by the oral route; in some embodiments, the pharmaceutical compositions provided herein are administered via the intravenous route.

In some embodiments, it may be desirable to administer the pharmaceutical composition topically. Topical application can be by means such as local infusion during surgery, topical application such as by postoperative binding to wound gauze, injection, intubation, by suppository or enema, or implants which are porous or non-porous substances or gel materials including thin films such as silicone rubber membranes or fibers.

In some embodiments, it may be desirable to introduce the compounds of formula (I) into the central nervous system, blood circulation system, or gastrointestinal tract by any suitable route, including intraventricular injection, intrathecal injection, periradicular injection, epidural injection, enema, peripheral nerve side injection, and the like. Intraventricular injections may be assisted by an intraventricular cannula, which may be connected to a reservoir, for example, to an Ommaya sac.

It can also be administered pulmonary, for example, using an inhaler or nebulizer, prepared with an aerosolizing agent, or infused with a fluorocarbon or synthetic pulmonary surfactant. In some embodiments, the compounds of formula (I) may be prepared as suppositories using conventional binders and excipients such as triglycerides.

In some embodiments, one or more compounds of formula (I) may be delivered in vesicles, particularly in Liposomes (see Langer, Science 249: 1527-.

In some embodiments, one or more compounds of formula (I) may be delivered in a Controlled Release system or a delayed Release system (see, e.g., Goodson, Medical Applications of Controlled Release, vol.2, pp.115-138, 1984). Other discussions of controlled or delayed release systems can be found in review of Langer, Science 249: 1527-1533, 1990. In some embodiments, pumps may be utilized (Langer, Science 249: 1527-. In another embodiment, polymeric materials may be utilized (see medical applications of Controlled Release (Langer and Wise eds., 1974); Controlled Drug Bioavailability, Drug Product Design and Performance (Smolen and Ball eds., 1984); Range and Peppas, J.Macromol.Sci.Rev.Macromol.Chem.2: 61, 1983; Levy et al, Science 228: 190, 1935; During et al, an.Neural.25: 351, 1989; and Howard et al, J.Neurosurg.71: 105, 1989).

As noted above, the pharmaceutical compositions provided herein may optionally comprise a suitable amount of a physiologically acceptable excipient. Examples of physiologically acceptable excipients may be liquids such as water and oils, including petroleum, animal, vegetable or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. For example, usable physiologically acceptable excipients may be saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. Or adjuvants, stabilizers, thickeners, lubricants and colorants may be used.

In some embodiments, the physiologically acceptable excipient is sterile when administered to an animal. Such physiologically acceptable excipients are desirably stable under the conditions of manufacture and storage, and in particular can prevent contamination by microorganisms. Water is a particularly useful excipient when the compound of formula (I) is administered by the intravenous route. Saline solutions and aqueous dextrose and glycerol solutions are all useful as liquid vehicles, particularly for injectable solutions. Suitable physiologically acceptable excipients also include starch, glucose, lactose, sucrose, gelatin, maltose, rice starch, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. If desired, the pharmaceutical compositions provided herein may also contain minor amounts of wetting or emulsifying agents, pH buffering agents.

Liquid carriers can be used in the preparation of solutions, suspensions, emulsions, syrups and elixirs. The compounds of formula (I) may be dissolved or suspended in a pharmaceutically acceptable liquid carrier, such as water, an organic solvent or a mixture of both, or a pharmaceutically acceptable oil or fat. Such liquid carriers may contain other suitable pharmaceutical additives including cosolvents, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, coloring agents, viscosity modifiers, stabilizers or osmo-modifiers. Suitable examples of liquid carriers for oral or parenteral administration include water (particularly water containing additives as described above, e.g., cellulose derivatives including sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols such as glycols) and their derivatives and oils (e.g., fractionated coconut oil and peanut oil). For parenteral administration, the carrier may also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are used in sterile liquid compositions for parenteral administration. The liquid carrier of the compressed composition may be a halogenated hydrocarbon or other pharmaceutically acceptable propellant.

The pharmaceutical compositions provided herein may be in the form of solutions, suspensions, emulsions, tablets, pills, pellets, capsules, liquid-containing capsules, powders, sustained release formulations, suppositories, emulsions, aerosols, sprays, suspensions, or any other form suitable for use. In some embodiments, the present invention provides pharmaceutical compositions in the form of a capsule. Other examples of suitable physiologically acceptable excipients can be found in Remington's Pharmaceutical Sciences 1447-.

In some embodiments, the compound of formula (I) (in appropriate form) is prepared according to conventional methods as a composition suitable for oral administration to a human. Compositions for oral use may be in the form of, for example, tablets, troches, lozenges, aqueous or oily suspensions or solutions, granules, powders, emulsions, capsules, syrups or elixirs. Compositions for oral administration may comprise one or more excipients such as sweetening agents, for example fructose, aspartame or saccharin; flavoring agents such as peppermint, oil of wintergreen, or cherry flavoring; a colorant; and a preservative to prepare a pharmaceutically palatable preparation. In powders, the carrier may be a finely divided solid which may be admixed with the finely divided compound or a pharmaceutically acceptable salt thereof. In tablets, the compound of the present invention or a pharmaceutically acceptable salt thereof is mixed with a carrier having necessary tabletting properties in an appropriate ratio and then compressed into tablets of a desired shape and size. Powders and tablets may contain up to 99% of a compound of the invention or a pharmaceutically acceptable salt thereof.

Capsules may contain mixtures of one or more compounds of formula (I) with inert fillers and/or diluents such as pharmaceutically acceptable starches (e.g., corn, potato or tapioca starch), sugars, artificial sweeteners, cellulose powders (e.g., microcrystalline cellulose), flours, gelatins, gums, and the like.

May be prepared according to conventional tableting, wet granulation or dry granulation techniques using pharmaceutically acceptable diluents, binders, lubricants, disintegrants, surface modifying agents (including surfactants), suspending or stabilizing agents (including, but not limited to, magnesium stearate, stearic acid, sodium lauryl sulfate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, microcrystalline cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, polyvinyl pyrrolidone, alginic acid, acacia gum, tragacanth gum, sodium citrate, complex silicates, calcium carbonate, glycine, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, low melting waxes and ion exchange resins). Surface modifiers include nonionic and anionic surface modifiers. Representative examples of surface modifying agents include, but are not limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidal silica, phosphate esters, sodium lauryl sulfate, magnesium aluminum silicate, and triethanolamine.

In addition, when the pharmaceutical composition provided by the present invention is in the form of a tablet or pill, it may be coated to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a prolonged period of time. Selectively permeable membranes surrounding osmotically active motile compounds are also suitable for oral compositions. In the latter, liquid in the environment surrounding the capsule can be absorbed by the driver compound, which then swells to displace the active agent or active agent composition from the pores. The release kinetics of this delivery form are essentially zero order, unlike the peaked release kinetics of immediate release dosage forms. Time delay materials such as glyceryl monostearate or glyceryl stearate may also be used. Oral compositions may also contain standard excipients such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose and magnesium carbonate. In some embodiments, the excipient is a pharmaceutical grade excipient.

In some embodiments, one or more compounds of formula (I) (in appropriate form) may be prepared for intravenous administration. Typically, the composition for intravenous administration comprises a sterile isotonic buffered aqueous solution. If desired, the composition may contain a co-solvent. Compositions for intravenous administration may optionally include a local anesthetic such as lignocaine to alleviate injection local pain. Typically, the ingredients are provided separately or mixed together in a unit dosage form, e.g., as a lyophilized powder or non-aqueous concentrate, enclosed in a sealed container such as an ampoule or sachet indicating the active agent content. If the compound of formula (I) is to be administered by infusion, it may be dispensed into injection vials containing sterile pharmaceutical grade water or saline. If the compound of formula (I) is to be administered by injection, an ampoule of sterile water for injection or saline may be provided, into which the ingredients are mixed prior to administration.

In some embodiments, one or more compounds of formula (I) (in appropriate form) may be administered by a transdermal route using a transdermal patch. Transdermal administration includes methods of administration through body surfaces and lining of body passageways, including epithelial and mucosal tissues. Such methods of administration may employ the compounds of the present invention in the form of lotions, creams, foams, patches, suspensions, solutions and suppositories (e.g., rectal or vaginal).

Transdermal administration may employ a transdermal patch containing, in appropriate form, one or more compounds of formula (I) and a carrier which is inert to the compound or a pharmaceutically acceptable salt thereof, which is non-toxic to the skin and through which the active agent is delivered to the systemic blood circulation. The carrier may be in a variety of forms, such as a cream or ointment, a paste, a gel, or an occlusive device. The cream or ointment may be a viscous liquid or a semisolid emulsion of the oil-in-water or water-in-oil type. Pastes include absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be used. Various occlusive devices may be used to release the compounds of the invention or pharmaceutically acceptable salts thereof into the blood stream, such as a semi-permeable membrane covering a reservoir containing the compound of formula (I) with or without a carrier, or a matrix containing the active ingredient.

One or more compounds of formula (I) (in appropriate form) may be administered rectally or vaginally in the form of conventional suppositories. Suppositories can be prepared from conventional materials, including cocoa butter (with or without the addition of waxes to alter the suppository's melting point) and glycerin. Water-soluble suppository bases such as polyethylene glycols of various molecular weights may also be used.

One or more compounds of formula (I) (in appropriate form) may be delivered by controlled or sustained release means or delivery devices known to those skilled in the art. Such dosage forms may provide controlled or sustained release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof, to provide a desired release profile and release ratio. Controlled release dosage forms or sustained release dosage forms known to those skilled in the art, including those described herein, can be readily selected for use with the active ingredients of the present invention. Thus, the present invention includes single dosage unit forms suitable for oral administration, including, but not limited to, tablets, capsules, gelcaps, and caplets suitable for controlled or sustained release.

In some embodiments, the controlled or sustained release compositions comprise a minimal amount of a compound of formula (I) to treat or prevent one or more disorders, diseases, or conditions associated with α 7 nicotinic acetylcholine receptor activity. Advantages of controlled or sustained release compositions include prolonged drug action, reduced frequency of administration, and enhanced compliance of the treated animal. In addition, controlled or sustained release compositions can have a beneficial effect on the onset time of action or other characteristics of the compounds of the invention or pharmaceutically acceptable salts thereof, such as blood levels, thereby reducing the occurrence of adverse side effects.

Controlled or sustained release compositions may first release an amount of one or more compounds of formula (I) to immediately elicit a desired therapeutic or prophylactic effect, and then gradually and continuously release a greater amount of the compound over a period of time to maintain the desired level of the compound for its therapeutic or prophylactic effect. To maintain a constant level of the compound in the body, the rate at which the compound is released from the dosage form may be balanced with the rate at which the body metabolizes or excretes the compound. Controlled or sustained release of the active ingredient can be stimulated by a variety of conditions, including but not limited to changes in pH, changes in temperature, concentration of the enzyme or the presence of the enzyme, concentration of water or the presence of water, or other physiological conditions or compounds.

In some embodiments, the pharmaceutical compositions provided herein deliver an amount of a compound of formula (I) effective to treat one or more disorders, diseases, or conditions associated with the activity (or inactivation) of α 7 nicotinic acetylcholine receptors. In accordance with the present invention, in vitro or in vivo assays may optionally be used to assist in determining the optimal dosage range. The precise dosage employed will also depend upon the route of administration, the nature and severity of the condition to be treated, various physical factors of the individual to be treated, and may be decided upon at the discretion of a physician. Equivalent doses may be administered over various periods of time, including, but not limited to, about every 2 hours, about every 6 hours, about every 8 hours, about every 12 hours, about every 24 hours, about every 36 hours, about every 48 hours, about every 72 hours, about every week, about every 2 weeks, about every 3 weeks, about every month, about every 2 months. The size and frequency of doses administered during a complete course of treatment will be at the discretion of the physician. An effective dose as described herein generally refers to the total amount administered, that is, if more than one compound of formula (I) is administered, the effective dose corresponds to the total amount administered.

An effective amount of a compound of formula (I) for use herein is typically from about 0.001mg/kg to about 600mg/kg body weight per day, in some embodiments, from about 1mg/kg to about 600mg/kg body weight per day, in another embodiment, from about 10mg/kg to about 400mg/kg body weight per day, in another embodiment, from about 10mg/kg to about 200mg/kg body weight per day, in another embodiment, from about 10mg/kg to about 100mg/kg body weight per day, in another embodiment, from about 1mg/kg to about 10mg/kg body weight per day, in another embodiment, from about 0.001mg/kg to about 100mg/kg body weight per day, in another embodiment, from about 0.001mg/kg to about 10mg/kg body weight per day, in another embodiment, about 0.001mg/kg to about 1mg/kg body weight per day.

In some embodiments, the pharmaceutical composition is provided in dosage unit form, for example in the form of a tablet, capsule, powder, solution, suspension, emulsion, granule, or suppository. In this form, the composition is subdivided into unit doses containing appropriate quantities of the active ingredient; unit dosage forms may be packaged compositions such as powders, vials, ampoules, prefilled syringes or sachets enclosing a liquid. The unit dosage form may be, for example, a capsule or tablet itself, or any number of packaged forms of any such compositions. Such unit dosage forms may contain, for example, from about 0.01mg/kg to about 250mg/kg of the active ingredient, and may be administered in a single dose or in two or three doses. The dosage of the drug will necessarily fluctuate depending on the species, weight, condition of the patient to be treated and the individual response of the patient to the drug itself.

In some embodiments, the unit dosage form is about 0.01 to about 1000 mg. In another embodiment, the unit dosage form is from about 0.01 to about 500 mg; in another embodiment, the unit dosage form is from about 0.01 to about 250 mg; in another embodiment, the unit dosage form is from about 0.01 to about 100 mg; in another embodiment, the unit dosage form is from about 0.01 to about 50 mg; in another embodiment, the unit dosage form is from about 0.01 to about 25 mg; in another embodiment, the unit dosage form is from about 0.01 to about 10 mg; in another embodiment, the unit dosage form is from about 0.01 to about 5 mg; in another embodiment, the unit dosage form is about 0.01 to about 10 mg.

The desired therapeutic or prophylactic activity of a compound of formula (I) may be assayed in vitro or in vivo prior to use in humans. Animal model systems can be used to demonstrate safety and efficacy.

Synthesis and preparation

The compounds of formula (I) or precursors thereof can be prepared by a variety of synthetic routes, several of which are illustrated in schemes 1-5 below:

scheme 1

According to scheme 1, omega-haloalkanochlorides 1 (here, omega-bromoalkanoyl chlorides are exemplified) are reacted with a suitable heterocyclic amine 2 in the presence of a base such as but not limited to triethylamine, Hunig's base (diisopropylethylamine) or an inorganic base such as potassium carbonate in a solvent such as but not limited to dichloromethane, dimethylformamide, dimethylacetamide, tetrahydrofuran, ethyl acetate or mixtures thereof to provide the coupled amide product 3, which product 3 may or may not be isolated and purified. Amide 3 is then reacted with amine X in a suitable solvent such as, but not limited to, dichloromethane, dimethylformamide or dimethylacetamide, with or without additional base such as triethylamine or Hunig's base, to give the title compound of formula (I), wherein amine X may or may not be in excess.

Scheme 2

According to scheme 2, an omega-haloalkanoic acid amide 3 is suitably activated and reacted with a suitable heterocyclic amine using, for example, but not limited to, 1' -carbonyldiimidazole in a solvent such as dichloromethane, dimethylformamide or mixtures thereof to afford an intermediate omega-haloalkanoic acid amide 3, which intermediate 3 may or may not be isolated and purified. Amide 3 is then reacted with amine X in a suitable solvent such as, but not limited to, dichloromethane, dimethylformamide or dimethylacetamide, with or without additional base such as triethylamine or Hunig's base, to give the title compound of formula (I), wherein amine X may or may not be in excess.

Scheme 3

According to scheme 3, the omega-aminoalkanoic acids are suitably activated using, for example, but not limited to, 1' -carbonyldiimidazole in a solvent such as dichloromethane, dimethylformamide or mixtures thereof and reacted with the appropriate heterocyclic amine to give the title compounds of formula (I).

Scheme 4

According to scheme 4, an omega-aminoalkanoic acid 5 is suitably activated and reacted with a suitable brominated heterocyclic amine using, for example, but not limited to, 1' -carbonyldiimidazole in a solvent such as dichloromethane, dimethylformamide or mixtures thereof to give the brominated heteroaryl amide of formula 7, which is then further reacted with 7 under cross-coupling conditions such as Suzuki conditions to give the title compound of formula (I).

Scheme 5 shows one possible synthetic route to the compound precursor of formula (I), i.e. chain substituted acid 5.

According to scheme 5, an alkyl substituted malonic acid diester is treated with a base such as, but not limited to, sodium hydride in a solvent such as tetrahydrofuran or dimethylformamide and reacted with an α, ω -dihaloalkane. The di-substituted malonic acid diester thus obtained is treated with a strong acid such as hydrobromic acid to hydrolyze it and remove one of the carboxyl groups. The esterification reaction is then carried out by treatment with, for example, methanol and a catalytic amount of acid. Omega-halogens can be substituted by heating with a suitable amine in a suitable solvent such as, but not limited to, toluene. Finally hydrolysis of the ester with a water soluble base gives an intermediate of formula 5 which can be activated as described herein to give a compound of formula (I).

The compounds of formula (I) and their optical isomers or diastereomers may be purified or isolated according to known procedures including, but not limited to, chiral chromatography and fractional crystallization.

Experimental part

Experimental procedure-Compound Synthesis

General procedure

Unless otherwise stated, all nuclear magnetic resonance spectra were recorded using a Varian Mercury Plus 400MHz spectrometer equipped with a PFG ATB broadband probe.

The HPLC-MS analytical instrument was a Waters 2795 separation unit, equipped with Waters micro ZQ (ES ionization) and Waters PDA 2996, and the column was Waters XTerra MS C183.5 μm 2.1x50mm.

Preparative HLPC was performed on a Waters 2767 system equipped with a binary gradient module Waters2525 pump, connected to Waters micro ZQ (ES) or Waters 2487 DAD on a Supelco Discovery HS C185.0 μm 10X21.2mm column.

Gradient elution was with 0.1% formic acid/water and 0.1% formic acid/acetonitrile, gradient change was 5/95 to 95/5, run time see examples.

All column chromatography methods were according to Still, c.; method of org Chem 43, 2923 (1978). All TLC analyses were carried out on silica gel (Merck 60F 254), with spots observed at 254nm under UV light or with KMnO₄Or ninhydrin dyeing

When indicated for use in the continuous synthesis, heating was in BuchiIs carried out in the system.

All microwave reactions were carried out in a CEM Discover microwave oven.

Abbreviations used during the experiments

AcOEt Ethyl acetate

DCM dichloromethane

DCE 1, 2-dichloroethane

DMEA N, N-dimethylethylamine

DMF N, N-dimethylformamide

DMSO, DMSO dimethyl sulfoxide

DAM N, N-Dimethylacetamide

SCX strong cation exchanger

TEA Triethylamine

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

LC-MS liquid chromatography-mass spectrometry

HPLC high performance liquid chromatography

General Synthesis of 3-amino-5-aryl/heteroaryl pyrazoles

The 3-amino-5-aryl/heteroaryl pyrazoles used in the examples can be obtained commercially or synthesized according to the route in the following scheme:

general procedure for the synthesis of aryl/heteroaryl β -ketonitriles (a 1):

methyl aryl or heteroaryl carboxylates are commercially available or synthesized according to the following standard methods: aryl or heteroaryl carboxylic acid (32mmol) was dissolved in MeOH (40mL) and sulfuric acid (1mL) was added. The mixture was refluxed overnight, then the solvent was evaporated under reduced pressure; the crude product was dissolved in DCM and saturated NaHCO₃And (4) washing the solution. The organic phase was dried and evaporated under reduced pressure and the crude product was used in the next step without purification.

To a solution of methyl aryl or heteroaryl formate (6.5mmol) in dry toluene (6mL) under nitrogen was carefully added NaH (50-60% dispersion in mineral oil, 624mg, 13 mmol). The mixture was heated at 80 ℃ and then dried CH was added dropwise₃CN (1.6mL, 30.8 mmol). The reaction was heated for 18 hours and the product precipitated from the reaction mixture, typically as the sodium salt.

The reaction was then cooled to room temperature, the solid formed was filtered off, dissolved in water, acidified with 2N HCl solution to pH 2-6 (depending on the ring substitution in the aryl/heteroaryl system), the product precipitated out and filtered. If no precipitate separated out, the product was extracted with DCM.

After work-up, the product is generally used in the subsequent steps without purification. The yield is generally 40-80%.

General synthetic procedure for aryl/heteroaryl β -ketonitriles (scheme A1 bis):

aryl-or heteroaryl-carboxylic acid methyl esters are commercially available or synthesized according to standard methods described below in general procedure a 1.

A solution of dry alkanenitrile in toluene (1mmol/mL, 5 eq.) was cooled to-78 ℃ under nitrogen, then a solution of N-butyllithium in N-hexane (1.6N, 3.5 eq.) was added dropwise, stirred at-78 ℃ for 20 minutes, and then methyl arylcarboxylate or methyl heteroarylformate was addedBenzene solution (0.75mmol/mL, 1eq.) was allowed to warm to room temperature. After about 20 minutes the reaction was complete, the mixture was cooled to 0 ℃ and HCl 2N was added to pH 2. Recovering the organic phase with Na₂SO₄Drying and concentration under reduced pressure gave the title compound, which was used in the next step usually without further purification.

General procedure for the synthesis of arylaminopyrazoles (scheme A2):

to a solution of β -ketonitrile (7.5mmoL) in anhydrous EtOH (15mL) was added hydrazine monohydrate (0.44mL, 9.0mmoL), heated at reflux for 18 h, cooled to room temperature, and the solvent was evaporated under reduced pressure. The residue was dissolved in DCM and washed with water.

Evaporating organic phase under reduced pressure to obtain crude product, and treating with SiO₂Column purification or from Et₂And precipitating in O for purification. The yield is usually 65-90%.

General Process for preparing methyl esters from hydroxy-aryl-or hydroxy-heteroaryl-carboxylic acids

4-hydroxy-benzoic acid (typically 24.0mmol) was dissolved in MeOH (50mL) and sulfuric acid (1mL/g substrate) was added. The mixture was refluxed overnight, then the solvent was evaporated under reduced pressure, the crude product was dissolved in DCM and washed with saturated NaHCO₃Washed until the pH is basic. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification. The yield is 80-90%.

Preparation of F from hydroxy-aryl-or hydroxy-heteroaryl-carboxylic acid methyl ester₂General procedure for CHO-aryl-or heteroarylmethyl carboxylates

In a two-necked round bottom flask under nitrogen atmosphere, 4-hydroxy-benzoic acid methyl or ethyl ester (1.0eq) and sodium difluorochloroacetate (1.2eq) were dissolved in DMF (20-25mL), potassium carbonate (1.2eq) was added and the mixture was heated at 125 ℃ to LC-MS to confirm complete conversion of the starting material. The reaction mixture was then diluted with water, extracted with DCM, the organic phase dried, evaporated under reduced pressure and the crude product purified on a Si column to give the product (yield 20-70%).

Table 1 below gives a series of F prepared according to the general procedure described above₂CHO-aryl-or F₂Yield and analytical data for CHO-heteroaryl-methyl formate

TABLE 1

Raw materials	Methyl ester-OH	Methyl ester-OCHF₂
			3-fluoro-4-hydroxy-benzoic acid	C₈H₇FO₃The yield is 85 percent₁H NMR(DMSO-d₆)δ3.78(3H，s)，7.00-7.05(1H，m)，7.60-7.65(2H，m)	C₉H₇F₃O₃Yield 66% 1H NMR (DMSO-d)₆)δ3.78(3H，s)，6.24(1H，m)，7.61(1H，m)，7.64(1H，m)，10.89(1H，bs)
2, 6-difluoro-4-hydroxy-benzoic acid	C₈H₆F₂O₃Yield 85% 1H NMR (DMSO-d)₆)δ3.79(s，3H，s)，6.53(2H，d，J＝10.8Hz)，11.13(1H，s)	C₉H₆F₄O₃Yield 34% 1H NMR (DMSO-d)₆)δ3.86(3H，s)，7.18-7.24(2H，m)，7.42(1H，t，J＝72.4Hz)。
			3, 5-dichloro-4-hydroxy-benzoic acid	Are commercially available	C₉H₆Cl₂F₂O₃Yield 74% 1H NMR (DMSO-d)₆)δ3.31(3H，s)，7.22(1H，t，J＝71.6Hz)，8.05(2H，s)。
3-chloro-4-hydroxy-benzoic acid	Are commercially available	C₉H₇ClF₂O₃Yield 85% 1H NMR (DMSO-d)₆)δ3.85(3H，s)，7.39(1H，t，J＝72.4Hz)，7.50(1H，t，J＝8.4Hz)，7.82-7.89(2H，m)。

4-hydroxy-3-methoxy-benzoic acid	Are commercially available	C₁₀H₁₀F₂O₄Yield 85% 1H NMR (DMSO-d)₆)3.84(3H，s)，3.87(3H，s)；7.22(1H，t，J＝73.6Hz)，7.29(1H，d，J＝8.4Hz)，7.57-7.60(2H，m)。
			4-hydroxy-2-methyl-benzoic acid	C₉H₁₀O₃Yield 95% 1H NMR (DMSO-d)₆)2.43(3H，br s)，3.72(3H，s)；6.61-6.64(2H，m)；7.71-7.73(1H，m)，10.10(1H，s)	C₁₀H₁₀F₂O₃Yield 85% 1H NMR (DMSO-d)₆)2.52(3H，br s)，3.80(3H，s)；7.07-7.13(2H，m)；7.34(1H，t，J＝73.6Hz)，7.89(1H，d，J＝8.8Hz)。

3-imidazo [1, 2-a ] pyridin-6-yl-3-oxo-propionitrile

The product was obtained according to general procedure a1 starting from imidazo [1, 2-a ] pyridine-6-carboxylic acid methyl ester.

The yield thereof was found to be 39%

C₁₀H₇N₃O Mass (calculated) [185 ]](ii) a (found value) [ M + H +]＝186[M-H]＝184

LC Rt ═ 0.23, 100% (3 min method)

¹H-NMR：(DMSO-d₆)：4.72(2H，s)，7.61-7.65(2H，m)，7.70(1H，m)，8.07(1H，s)，9.40(s，1H)。

5-imidazo [1, 2-a ] pyridin-6-yl-1H-pyrazol-3-ylamine

The title compound was synthesized according to general procedure a2 starting from 3-imidazo [1, 2-a ] pyridin-6-yl-3-oxo-propionitrile.

Yield: 84 percent

C₁₀H₉N₅Mass (calculated) [199](ii) a (found value) [ M +1]＝200

LCMS, (5 min method, RT ═ 0.21min),

NMR(¹H，400MHz，MeOH-d₄)3,34(s，2H)，5,90(br s，1H)，7,57(s，1H)，7,63(br s，1H)，7,86(s，1H)，8,73(s，1H)

synthesis of chlorocinnamonitrile (route B1)

Adding POCl₃(2 equivalents relative to aryl/heteroaryl acetophenone) was added dropwise to 4 molar equivalents of anhydrous DMF cooled to 0 ℃ at a rate such that the temperature did not exceed 10 ℃. Acetophenone (1eq) was then added dropwise and the reaction was allowed to warm to room temperature.

Stirring was then continued for 30 minutes and 0.4mmol of hydroxylamine hydrochloride was added. The reaction solution was then heated to 50 ℃ and the heat source removed and an additional 4 equivalents of hydroxylamine hydrochloride were added in portions (at a rate such that the temperature did not exceed 120 ℃). Stirring was continued until the temperature of the reaction mixture naturally dropped to 25 ℃. Water (100mL) was added, and the mixture was extracted with diethyl ether. With Na₂SO₄The organic phase was dried and evaporated under reduced pressure. The crude product was used in the next step without further purification.

Synthesis of arylaminopyrazoles (route B2)

To a solution of chlorocinnamonitrile (0.5mmol/mL, 1eq) in anhydrous EtOH was added 2 equivalents of hydrazine monohydrate and the reaction was heated to reflux for 4 hours. The reaction mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. With Et₂The residue was triturated to recover the title compound, which was used in the next step generally without further purification.

5- (2-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

a) 3-oxo-3- (2-trifluoromethyl-phenyl) -propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a1) starting from 2-trifluoromethyl-benzoic acid methyl ester (3.1g, 14.0mmol, 1.0 eq). The crude product was precipitated from HCl to give the title compound as a yellow solid (2.8g, yield: 94%).

C₁₀H₆F₃NO

¹H-NMR(CD₃OD)：4.90(2H，br s)；7.52-7.86(4H，m)。

b)5- (2-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a Si column (eluent DCM) and dried to give the title compound (0.6g, 20% yield).

C₁₀H₈F₃N₃

5- (2, 6-dimethyl-phenyl) -2H-pyrazol-3-ylamine

a)3- (2, 6-dimethyl-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A1), and the mixture was refluxed overnight and then heated at 110 ℃ under reflux for 2 h. The crude product was extracted with DCM and used in the next step without further purification (2.2g, yield: 76%).

C₁₁H₁₁NO

b)5- (2, 6-dimethyl-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a Si column (eluent DCM), washed with water, extracted and dried to give the product (0.25g, yield 10%).

C₁₁H₁₃N₃

¹H-NMR(CD₃OD)：2.09-2.23(6H，m)；7.04-7.12(2H，m)；7.18-7.26(2H，m)。

5- (2-chloro-4-fluoro-phenyl) -2H-pyrazol-3-ylamine

a)3- (2-chloro-4-fluoro-phenyl) -3-oxo-propionitrile

Prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a1) starting from 2-chloro-4-fluoro-benzoic acid methyl ester (0.7g, 3.7mmol, 1.0 eq). The crude product was extracted with DCM and used in the next step without further purification (0.4g, yield: 60%).

C₉H₅ClFNO

b)5- (2-chloro-4-fluoro-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was dissolved in DCM, washed with saturated sodium bicarbonate solution, extracted and dried to give the title compound (0.12g, yield 26%).

C₉H₇ClFN₃

¹H-NMR(DMSO-d₆)：7.03-7.53(4H，m)。

5- (5-tert-butyl-thiophen-2-yl) -2H-pyrazol-3-ylamine

a)3- (5-tert-butyl-thiophen-2-yl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole (scheme A1) starting from 5-tert-butyl-thiophene-2-carboxylic acid methyl ester (3.0g, 15.0mmol, 1.0 eq). The crude product was extracted with DCM and used in the next step without further purification (2.7g, yield: 86%).

C₁₁H₁₃NOS

b)5- (5-tert-butyl-thiophen-2-yl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was washed with water and precipitated to give the title compound (2.7g, 91% yield).

C₁₁H₁₅N₃S

Quality (meter)Calculated value) [221](ii) a (found value) [ M + H [)⁺]＝222。

LC Rt 2.53min, 94% (10 min method)

¹H-NMR(DMSO-d₆)：1.26-1.29(9H，m)；4.87(2H，br s)；5.47(1H，brs)；6.66-6.79(1H，m)；6.97-7.02(1H，m)

5- (3-chloro-2-methyl-phenyl) -2H-pyrazol-3-ylamine

a) 2-Ethyl-benzoic acid methyl ester

2-Ethyl-benzoic acid (3.0g, 17.6mmol) was dissolved in MeOH (20mL), sulfuric acid (1mL) was added, refluxed overnight, then the solvent was evaporated under reduced pressure and the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to alkaline pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification (3.1g, 96% yield).

C₉H₉ClO₂

¹H-NMR(DMSO-d₆)：2.48(3H，br s)；3.82(3H，s)；7.31(1H，t，J＝7.6Hz)；7.63-7.67(2H，m)。

b)3- (3-chloro-2-methyl-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole (scheme a1) starting from 3-chloro-2-methyl-benzoic acid methyl ester (3.1g, 16.8mmol, 1.0 eq). The crude product was precipitated from water and used in the next step without further purification (2.4g, yield: 74%).

C₁₀H₈ClNO

¹H-NMR(DMSO-d₆)：2.31(3H，br s)；4.64(2H，br s)；7.27-7.36(2H，m)；7.54-7.77(1H，m)。

c)5- (3-chloro-2-methyl-phenyl) -2H-pyrazol-3-ylamine

According to the general procedure for the synthesis of aminopyrazoles (scheme A)2) And (3) preparing a product. The crude product is passed through SiO₂Column (20g) purified, eluting with a gradient from 100% EtOAc to EtOAc-MeOH 80: 20. The title compound (1.3g, yield 50%) was obtained.

C₁₀h₁₀ClN₃

Mass (calculated value) [207](ii) a (found value) [ M + H [)⁺]＝208。

LC Rt 1.96min, 85% (10 min method)

¹H-NMR(CDCl₃)：2.41(3H，s)；5.74(1H，s)；7.16(1H，t，J＝8.0Hz)；7.20-7.26(1H，m)；7.38-7.40(1H，m)。

5- (2-ethyl-phenyl) -2H-pyrazol-3-yl-amines

a) 2-Ethyl-benzoic acid methyl ester

2-Ethyl-benzoic acid (3.0g, 20.0mmol) was dissolved in MeOH (20mL), a catalytic amount of sulfuric acid (1mL) was added, refluxed overnight, the solvent was evaporated under reduced pressure, the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to a basic pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification (2.9g, 88% yield).

C₁₀H₁₂O₂

¹H-NMR(DMSO-d₆)：1.12(3H，t，J＝7.2Hz)；2.86(2H，q，J＝7.2Hz)；3.81(3H，s)；7.27-7.34(2H，m)；7.46-7.51(1H，m)；7.73-7.75(1H，m)。

b)3- (2-ethyl-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a1) starting from methyl 2-ethyl-benzoate (2.9g, 17.6mmol, 1.0 eq). The crude product was extracted with DCM as a yellow oil and used in the next step without further purification (2.8g, yield: 92%).

C₁₁H₁₁NO

¹H-NMR(DMSO-d₆)：1.10-1.18(3H，m)；2.78(2H，q，J＝7.2Hz)；4.67(1H，s)；7.23-7.53(3H，m)；7.73-7.78(1H，m)。

c)5- (2-ethyl-phenyl) -2H-pyrazol-3-yl-amines

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column (20g) purification, gradient elution from 100% EtOAc to EtOAc-MeOH 80: 20, afforded the title compound (1.2g, 40% yield).

C₁₁H₁₃N₃

Mass (calculated value) [187](ii) a (found value) [ M + H [)⁺]＝188。

LC Rt 1.58min, 90% (10 min method)

¹H-NMR(CDCl₃)：1.15(3H，t，J＝7.6Hz)；2.71(2H，q，J＝7.6Hz)；5.72(1H，s)；7.20-7.26(1H，m)；7.29-7.35(3H，m)。

5- (4-methoxy-phenyl) -4-methyl-2H-pyrazol-3-ylamine

a)3- (4-methoxy-phenyl) -2-methyl-3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a1) starting from 4-methoxy-benzoic acid methyl ester (3.0mL, 18.0mmol, 1.0eq), NaH (1.4g, 36.0mmol, 2.0eq) and propionitrile (6.1mL, 84.9mmol, 4.7 eq). The crude product was purified by Si column (eluent hexane/ethyl acetate) to obtain 2.1g of the title product (yield: 62%).

C₁₁H₁₁NO₂

b)5- (4-methoxy-phenyl) -4-methyl-2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a2), the crude product was washed with basic water, dried and used in the next step without further purification (1.8g, yield 80%).

C₁₁H₁₃N₃O

Mass (calculated value) [203](ii) a (found value) [ M + H [)⁺]＝204。

LC Rt ═ 1.34min, 91% (10 min method)

¹H-NMR(CDCl₃)：2.03(3H，s)；3.84(3H，s)；6.96-6.98(2H，m)；7.37-7.39(2H，m)。

4-methyl-5- (4-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

a) 2-methyl-3-oxo-3- (4-trifluoromethyl-phenyl) -propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a1) starting from 4-trifluoromethyl-benzoic acid methyl ester (3.0g, 14.7mmol, 1.0eq), NaH (1.2g, 29.4mmol, 2.0eq) and propionitrile (4.9mL, 69.4mmol, 4.7 eq). The crude product was extracted with DCM and used in the next step without further purification (3.2g, yield: 96%).

C₁₁H₈F₃NO

b) 4-methyl-5- (4-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was washed with basic water, dried and the title compound was used in the next step without further purification (2.8g, yield 84%).

C₁₁H₁₀F₃N₃

Mass (calculated value) [ 241%](ii) a (found value) [ M + H [)⁺]＝242。

LC Rt 2.34min, 92% (10 min method)

¹H-NMR(CDCl₃)：2.05(3H，s)；7.56(2H，d，J＝8.4Hz)；7.64(2H，d，J＝8.4Hz)。

5- (4-cyclopropylmethoxy-2-methyl-phenyl) -2H-pyrazol-3-ylamine

a) 4-hydroxy-2-methyl-benzoic acid methyl ester

4-hydroxy-2-methyl-benzoic acid (4.8g, 32.0mmol) was dissolved in MeOH (40mL), a catalytic amount of sulfuric acid (1mL) was added, refluxed overnight, then the solvent was evaporated under reduced pressure, the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to alkaline pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification (5.0g, yield 95%).

C₉H₁₀O₃

¹H-NMR(DMSO-d₆)：2.43(3H，s)；3.72(3H，s)；6.62-6.64(2H，m)；7.71-7.73(1H，m)；10.10(1H，s)。

b) 4-Cyclopropylmethoxy-2-methyl-benzoic acid methyl ester

4-hydroxy-2-methyl-benzoic acid methyl ester (1.0g, 6.0mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.45g, 3.0mmol, 0.5eq) and K were added₂CO₃(1.66g, 12.0mmol, 2.0eq), and stirred at room temperature for 20 minutes. (bromomethyl) cyclopropane (0.53mL, 5.4mmol, 0.9eq) was added and refluxed for 2 days. The solvent was then evaporated under reduced pressure, 10% NaOH was added and the crude product was extracted with DCM and dried. 0.42g of the title compound (yield 32%) was obtained and used in the next step without further purification.

C₁₃H₁₆O₃

¹H-NMR(CDCl₃)：0.23-0.34(2H，m)；0.52-0.64(2H，m)；1.15-1.24(1H，m)；2.52(3H，s)；3.75(2H，d，J＝7.2Hz)；3.77(3H，s)；6.64-6.66(1H，m)；7.83-7.85(2H，m)。

c)3- (4-Cyclopropylmethoxy-2-methyl-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole starting from 4-cyclopropylmethoxy-2-methyl-benzoic acid methyl ester (scheme A1 bis). 0.54g of the title compound was extracted from water, dried and used in the next step without further purification (yield 69%).

C₁₄H₁₅NO₂

d)5- (4-cyclopropylmethoxy-2-methyl-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification, gradient elution from 100% EtOAc to EtOAc-MeOH 90: 10. The title compound (206mg, yield 36%) was obtained.

C₁₄H₁₇N₃O

¹H-NMR(CD₃OD)：0.29-0.36(2H，m)；0.54-0.63(2H，m)；1.18-1.28(1H，m)；2.33(3H，s)；3.81(2H，d，J＝7.2Hz)；5.67(1H，s)；6.74-6.80(2H，m)；7.25(1H，d，J＝8.8Hz)。

5- (3-chloro-4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine

a) 3-chloro-4-cyclopropylmethoxy-benzoic acid methyl ester

3-chloro-4-hydroxy-benzoic acid methyl ester (1.1g, 6.0mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.45g, 3.0mmol, 0.5eq) and K were added₂CO₃(1.66g, 12.0mmol, 2.0eq), and stirred at room temperature for 20 minutes. (bromomethyl) cyclopropane (0.53mL, 5.4mmol, 0.9eq) was added and refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, the crude product was extracted with DCM and dried and the title compound was used in the next step without further purification (0.88g, yield 32%).

C₁₂H₁₃ClO₃

¹H-NMR(DMSO-d₆)：0.33-0.37(2H，m)；0.55-0.60(2H，m)；1.25-1.27(1H，m)；3.80(3H，s)；3.99(2H，d，J＝7.2Hz)；7.21(1H，s，J＝8.8Hz)；7.85-7.91(2H，m)。

b)3- (3-chloro-4-cyclopropylmethoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to a general synthetic procedure starting from 3-chloro-4-cyclopropylmethoxy-benzoic acid methyl ester (scheme A1 bis). 0.74g of the title compound was extracted from water and dried and used directly in the next step (yield 81%).

C₁₃H₁₂ClNO₂

c)5- (3-chloro-4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification (gradient elution from 100% EtOAc to EtOAc-MeOH 90: 10). 521mg of the title compound (yield 67%) was obtained.

C₁₃H₁₄ClN₃O

Mass (calculated) [263](ii) a (found value) [ M + H [)⁺]＝264。

LC Rt 2.51min, 90% (10 min method)

¹H-NMR(CD₃OD)：0.25-0.29(2H，m)；0.52-0.55(2H，m)；1.10-1.18(1H，m)；3.81(2H，d，J＝6.8Hz)；5.74(1H，s)；6.95-6.99(1H，m)；7.24-7.30(2H，m)。

5- (4-Cyclopropylmethoxy-2-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

a) 4-hydroxy-2-trifluoromethyl-benzoic acid methyl ester

4-hydroxy-2-trifluoromethyl-benzoic acid (5.0g, 24.0mmol) was dissolved in MeOH (50mL), a catalytic amount of sulfuric acid was added, refluxed overnight, then the solvent was evaporated under reduced pressure, the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to alkaline pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification.

C₉H₇F₃O₃

b) 4-Cyclopropylmethoxy-2-trifluoromethyl-benzoic acid methyl ester

4-hydroxy-2-trifluoromethyl-benzoic acid methyl ester (1.1g, 4.8mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.5eq) and K were added₂CO₃(1.04g, 2.0eq), and stirred at room temperature for 30 minutes. (bromomethyl) cyclopropane (0.42mL, 4.3mmol, 0.9eq) was added and the mixture was refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, extracted with DCM and dried. The title compound was obtained and used in the next step without further purification (1.21g, yield 92%).

C₁₃H₁₃F₃O₃

c)3- (4-Cyclopropylmethoxy-2-trifluoromethyl-phenyl) -3-oxo-propionitrile

The product was prepared according to the general synthetic procedure (scheme A1 bis). The mixture was acidified with HCl1M, the organic phase was separated and dried to yield 1.2g of the title compound, which was used directly in the next step (yield 94%).

C₁₄H₁₂F₃NO₂

Mass (calculated) [283](ii) a (found value) [ M + H [)⁺]＝284

LC Rt 3.86min, 98% (10 min method)

d)5- (4-Cyclopropylmethoxy-2-trifluoromethyl-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification (gradient elution from ethyl acetate-cyclohexane 1: 1 to ethyl acetate-MeOH 90: 10). 650mg of the title compound (yield 52%) was obtained.

C₁₄H₁₄F₃N₃O

Mass (calculated) [ 297)](ii) a (found value) [ M + H [)⁺]＝298。

LC Rt 2.78min, 59% (10 min method)

¹H-NMR(CDCl₃)：032-0.44(2H，m)；0.64-0.62(2H，m)；1.22-1.37(1H，m)；3.80-3.92(2H，m)；5.78(1H，s)；7.04-7.07(1H，m)；7.24-7.26(1H，m)；7.38-7.40(1H，m)

5- (4-cyclopropylmethoxy-2, 3-difluoro-phenyl) -2H-pyrazol-3-ylamine

a) 4-hydroxy-2, 3-difluoro-benzoic acid methyl ester

4-hydroxy-2, 3-difluoro-benzoic acid (2.0g, 11.5mmol) was dissolved in MeOH (20mL), a catalytic amount of sulfuric acid was added, reflux was overnight, then the solvent was evaporated under reduced pressure and the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to alkaline pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification.

C₈H₆F₂O₃

b) 4-Cyclopropylmethoxy-2, 3-difluoro-benzoic acid methyl ester

4-hydroxy-2, 3-difluoro-benzoic acid methyl ester (0.9g, 4.8mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.5eq) and K were added₂CO₃(1.03g, 2.0eq), and stirred at room temperature for 30 minutes. (bromomethyl) cyclopropane (0.42mL, 0.9eq) was added and the mixture refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, extracted with DCM and dried. The title compound was obtained and used in the next step without further purification (0.97g, yield 84%). C₁₂H₁₂F₂O₃

c)3- (4-cyclopropylmethoxy-2, 3-difluoro-phenyl) -3-oxo-propionitrile

The product was prepared according to the general synthetic procedure (scheme A1 bis). The mixture was acidified with HCl1M, the organic phase was separated and dried to yield 0.79g of the title compound, which was used directly in the next step (yield 79%).

C₁₃H₁₁F₂NO₂

Mass (calculated) [251 ]](ii) a (found value) [ M + H [)⁺]＝252。

LC Rt 3.53min, 82% (10 min method)

d)5- (4-cyclopropylmethoxy-2, 3-difluoro-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification (gradient elution from 100% EtOAc-cyclohexane to EtOAc-MeOH 90: 10). Yield 810mg of the title compound (yield 97%).

C₁₃H₁₃F₂N₃O

Mass (calculated) [265 ]](ii) a (found value) [ M + H [)⁺]＝266。

LC Rt 2.59min, 75% (10 min method)

¹H-NMR(CDCl₃)：032-0.47(2H，m)；0.64-0.75(2H，m)；1.19-1.38(1H，m)；3.67-4.15(4H，m)；5.95(1H，s)；6.74-6.88(1H，m)；7.17-7.26(1H，m)；

5- (3, 5-dichloro-4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine

a)3, 5-dichloro-4-cyclopropylmethoxy-benzoic acid methyl ester

3, 5-dichloro-4-hydroxy-benzoic acid ethyl ester (1.0g, 4.5mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.5eq) and K were added₂CO₃(0.98g, 9.0mmol, 2.0eq), and stirred at room temperature for 30 minutes. (bromomethyl) cyclopropane (0.39mL, 4.1mmol, 0.9eq) was added and the mixture was refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, extracted with DCM and dried. The title compound was obtained and used in the next step without further purification (0.98g, yield 79%).

C₁₂H₁₂Cl₂O₃

b)3- (3, 5-dichloro-4-cyclopropylmethoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to the general synthetic procedure (scheme A1 bis). The mixture was acidified with HCl1M, the organic phase was separated and dried to yield 0.91g of the title compound, which was used directly in the next step (yield 90%).

C₁₃H₁₃Cl₂N₃O

Mass (calculated) [283](ii) a (found value) [ M + H [)⁺]＝284。

LC Rt 4.06min, 99% (10 min method)

c)5- (3, 5-dichloro-4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification (eluent gradient 100% EtOAc-cyclohexane 1: 1 to ethyl acetate: MeOH 90: 10). 750mg of the title compound were obtained (yield 79%).

C₁₃H₁₃Cl₂N₃O

Mass (calculated) [ 297)](ii) a (found value) [ M + H [)⁺]＝298。

LC Rt 3.23min, 93% (10 min method)

¹H-NMR(CDCl₃)：023-0.46(2H，m)；0.64-0.74(2H，m)；1.30-1.48(1H，m)；3.60-4.04(4H，m)；5.86(1H，s)；7.48(2H，s)

5- (4-Cyclopropylmethoxy-3-methoxy-phenyl) -2H-pyrazol-3-ylamine

a) 4-Cyclopropylmethoxy-3-methoxy-benzoic acid methyl ester

4-hydroxy-3-methoxy-benzoic acid methyl ester (1.0g, 5.5mmol, 1.0eq) was dissolved in acetone (14mL) and addedNaI (0.5eq) and K₂CO₃(1.0g, 2.0eq), and stirred at room temperature for 30 minutes. (bromomethyl) cyclopropane (0.53mL, 0.9eq) was added and the mixture refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, extracted with DCM and dried. The title compound was obtained and used in the next step without further purification (1.21g, yield 93%).

C₁₃H₁₆O₄

b)3- (4-Cyclopropylmethoxy-3-methoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to the general synthetic procedure (scheme A1 bis). The mixture was acidified with HCl1M, the organic phase was separated and dried to yield 1.24g of the title compound, which was used directly in the next step (yield 99%).

C₁₄H₁₅NO₃

Mass (calculated value) [245](ii) a (found value) [ M + H [)⁺]＝246。

LC Rt 3.03min, 100% (10 min method)

c)5- (4-Cyclopropylmethoxy-3-methoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product is passed through SiO₂Column purification (eluent gradient 100% EtOAc-cyclohexane 1: 1 to ethyl acetate: MeOH 90: 10). 220mg of the title compound (yield 50%) were obtained.

C₁₄H₁₇N₃O₂

Mass (calculated value) [ 259)](ii) a (found value) [ M + H [)⁺]＝260。

LC Rt 1.86min, 93% (10 min method)

¹H-NMR(CDCl₃)：027-0.43(2H，m)；0.56-0.72(2H，m)；1.23-1.40(1H，m)；348(2H，m)；3.87(3H，s)；3.98(2H，br s)；5.82(1H，s)；6.85-6.89(1H，m)；7.05-7.10(2H，m)；

3-amino-5- (3-fluoro-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

3-amino-5- (3-fluoro-phenyl) -pyrazole (5.0g, 28.0mmol, 1.0eq) and KOH4.5M (50mL, 226mmol, 8eq) were dissolved in DCM (200mL), di-tert-butyl dicarbonate (6.5g, 30.0mmol, 1.1eq) was added and stirred at room temperature until LC-MS confirmed complete conversion. The organic phase was washed with saturated brine, then the solvent was evaporated and the crude product was crystallized from MeOH to yield 7.4g of the title product (95% yield).

C₁₄H₁₆FN₃O₂

¹H-NMR(DMSO-d₆)：1.57(9H，s)，5.80(1H，s)，6.43(2H，br s)，7.16-7.21(1H，m)，7.41-7.47(1H，m)；7.50-7.54(1H，m)；7.58-7.60(1H，m)。

3-amino-5-o-tolyl-pyrazole-1-carboxylic acid tert-butyl ester

3-amino-5-o-tolyl-pyrazole (0.5g, 2.89mmol, 1.0eq) and KOH4.5M (5.1mL, 23.1mmol, 8.0eq) were dissolved in DCM (20mL), di-tert-butyl dicarbonate (0.66g, 3.0mmol, 1.1eq) was added and stirred at room temperature until LC-MS confirmed complete conversion. The organic phase was washed with saturated brine and the solvent was evaporated to give 0.6g of the title product (yield 76%).

C₁₅H₁₉N₃O₂

Mass (calculated value) [273](ii) a (found value) [ M + H [)⁺]＝274。

LC Rt 2.34min, 96% (5 min method)

3-amino-5- (4-trifluoromethyl-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

3-amino-5- (4-trifluoromethyl-phenyl) -pyrazole (2.0g, 8.8mmol, 1.0eq) and KOH4.5M (15.7mL, 70.5mmol, 8.0eq) were dissolved in DCM (70mL), di-tert-butyl dicarbonate (2.02g, 9.2mmol, 1.1eq) was added and stirred at room temperature until LC-MS confirmed conversionAnd (4) completing. The organic phase was washed with saturated brine, the solvent was evaporated and the crude product was in CH₃CN to yield 1.9g of the title product (69%).

C₁₅H₁₆F₃N₃O₂

Mass (calculated) [327](ii) a (found value) [ M + H [)⁺]＝328。

LC Rt 2.59min, 100% (5 min method)

¹H-NMR(DMSO-d₆)：1.57(9H，s)，5.83(1H，s)，6.46(2H，s)，7.74(2H，d，J＝8.4Hz)，7.95(2H，d，J＝8.8Hz)

5-pyridin-2-yl-2H-pyrazol-3-ylamine

a) Oxo-pyridin-2-yl-acetonitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole (scheme A1) starting from pyridine-2-carboxylic acid methyl ester (3.0g, 21.9mmol, 1.0 eq). The crude product was precipitated from HCl to give the title product as a solid which was used directly in the next step (2.2g, yield: 69%).

C₈H₆N₂O

b) 5-pyridin-2-yl-2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was dissolved in EtOAc and washed with NaHCO₃Washed, dried and then evaporated. NMR analysis showed that most of the crude mixture was still in the ring-opened state. Dissolving the mixture in CH₃COOH, heated at 80 ℃ overnight, to allow ring closure of the ring-opened form. The acylated form was then recovered and treated with HCl 6N at 60 ℃ overnight to deacylate it to give the title compound (0.816g, 60% yield).

C₈H₈N₄

¹H-NMR(DMSO-d₆)：4.81(2H，bs)，5.92(1H，s)，7.21-7.24(1H，m)，7.76(2H，d)，8.51(1H，d)，11.96(1H，bs)

5- (3-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine

a) 3-Difluoromethoxybenzoic acid methyl ester

Difluoromethoxy-benzoic acid (2.0g, 10.6mmol, 1.0eq) was dissolved in MeOH (15mL) with addition of catalytic amount of sulfuric acid, refluxed overnight, then the solvent was evaporated under reduced pressure, the crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to alkaline pH. The organic phase was dried and evaporated under reduced pressure and the product used in the next step without further purification (1.9g, yield 90%).

C₉H₈F₂O₃

¹H-NMR(DMSO-d₆)：3.86(3H，s)，7.33(1H，t，J＝73.6Hz)，7.46-7.50(1H，m)，7.59(1H，t，J＝8.0Hz)，7.67(1H，s)；7.82(1H，d，J＝7.6Hz)。

b)3- (3-difluoromethoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A1bis) starting from 3-difluoromethoxy-benzoic acid methyl ester (1.5g, 7.4mmol, 1.0 eq). The crude product was precipitated by addition of aqueous HCl to give the product, which was used directly in the next step.

C₁₀H₇F₂NO₂

c)5- (3-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a Si column (eluent gradient 100% EtOAc to ethyl acetate: MeOH 90: 10). 1.45g of the title product are obtained (yield 87%).

C₁₀H₉F₂N₃O

¹H-NMR(DMSO-d₆)：4.89(2H，br s)，5.75(1H，s)，7.02(1H，d)，7.25(1H，t，J＝74.0Hz)，7.36-7.42(2H，m)，7.48-7.50(1H，d)，11.76(1H，br s)

5-pyrazolo [1, 5-a ] pyridin-3-yl-2H-pyrazol-3-ylamine

a) 3-oxo-3-pyrazolo [1, 5-a ] pyridin-3-yl-propionitrile

A solution of dry acetonitrile in toluene (0.66mL, 13mmol, 5eq) was cooled to-78 deg.C under nitrogen, and a solution of n-butyllithium in n-hexane (5.2mL, 13mmol, 5eq) was then added dropwise. The mixture was stirred at-78 ℃ for 20 minutes, and then pyrazolo [1, 5-a ] was added]Pyridine-3-carboxylic acid methyl ester (0.46g, 2.6mmol, 1eq, prepared according to the reported method (Anderson et al Journal of heterocyclic Chemistry 1981, 18, 1149-1152)) in toluene and the reaction was allowed to warm to room temperature. After about 20 minutes the reaction was complete, the mixture was cooled to 0 ℃ and HCl 2N was added to pH 2. Recovering the organic phase with Na₂SO₄Drying and concentration under reduced pressure gave the title compound, which was used in the next step without further purification.

C₁₀H₇N₃O

b) 5-pyrazolo [1, 5-a ] pyridin-3-yl-2H-pyrazol-3-ylamine

To a solution of 3-oxo-3-pyrazolo [1, 5-a ] pyridin-3-yl-propionitrile (0.66g, 3.6mmol) in anhydrous EtOH (25mL) was added hydrazine monohydrate (0.44mL, 9.0mmol), heated to reflux for 18 hours and cooled to room temperature, the solvent was evaporated under reduced pressure, and the residue was dissolved in DCM and washed with water.

Evaporating the organic phase under reduced pressure to obtain a crude product, passing the crude product through SiO₂Column purification (gradient elution of DCM to DCM: MeOH 95: 5 to 85: 15) gave the title compound in 41% yield (0.29g, 1.48 mmol).

C₁₀H₉N₅

¹H-NMR(DMSO-d₆)：8.68(s，1H)；8.21(s，1H)；7.92(s，1H)；7.28(s，1H)；6.90(s，1H)；5.75(s，1H)；5.10(s，2H)。

Mass (calculated) [199](ii) a (found value) [ M + H [)⁺]＝200。

LC Rt 0.86min, 92% (5 min method).

General synthesis of omega-bromo-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides

A solution of ω -bromoalkanoyl chloride (15.7mmol, 1eq) in dry DMA (35mL) was cooled to-10 ℃ (ice water bath) under nitrogen; a solution of 5-aryl/heteroaryl-1H-pyrazol-3-ylamine (15.7mmol, 1eq) and diisopropylethylamine (15.7mmol, 1eq) in dry DMA (15mL) was added over 30 minutes. After 2 hours at-10 ℃ the end of the reaction is generally confirmed by monitoring by LC-MS (acylation of the pyrazole ring is also detected). Then H is added₂The reaction was quenched with O (about 50mL) and the large amount of white precipitate formed upon addition of water was filtered off. With Et₂An O (3X10mL) wash is generally more effective in removing acylated by-products on the pyrazole ring.

General synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides

Omega-bromo-alkanoic acid [ 5-aryl-1H-pyrazol-3-yl ] -amide (0.6mmol, 1eq) was dissolved in DMF (4mL), sodium iodide (0.6mmol, 1.0eq) was added followed by secondary amine (1.5mmol, 2.5eq) and diisopropylethylamine (0.6mmol, 1 eq). The reaction was then stirred at +50 ℃ for 18 hours under nitrogen.

After the reaction was complete (LC-MS monitoring), the solvent was removed under reduced pressure and the resulting oily residue was dissolved in DCM (20mL) and saturated Na was used₂CO₃(2X20mL) and saturated NaCl (2X20mL) and washed with Na₂SO₄The organic phase was dried and the solvent was removed under reduced pressure. The title compound was purified by silica gel column or preparative HPLC.

General procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides: acylation-nucleophilic substitution

A solution of omega-bromoalkanoyl chloride (0.94mmol, 1eq) in DMA (1mL) was cooled to 0 deg.C, a solution of 3-amino-5-aryl/heteroaryl pyrazole (0.94mmol, 1eq) and diisopropylethylamine (1.88mmol, 2eq) in DMA (2mL) was added and the reaction was stirred at 0 deg.C for 1 hour. Secondary amine (2.35mmol, 2.5eq) and NaI (0.94mmol, 1eq) were then added. For 3 carbon chain derivatives, the reaction is usually complete after 2 hours at room temperature; for 4 carbon chain derivatives, the reaction mixture is typically heated at 60 ℃ for 24 to 48 hours. When the bromine intermediate was completely converted (monitored by LC-MS), the solvent was removed under reduced pressure. The residue was dissolved in DCM (2mL) and saturated Na₂CO₃And (4) washing with an aqueous solution. The organic phase was concentrated under reduced pressure and the crude product was isolated from CH₃Recrystallized from CN or via SiO₂Column purification (gradient elution: 100% DCM to DCM-NH)₃MeOH 2N solution 8: 2) or by preparative HPLC (standard acidic conditions).

General procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides via the amino acid route

General Synthesis of omega-amino esters (scheme C1)

To a solution of amine X (65mmol) in toluene (15mL) was added omega-bromoalkanoic acid ethyl ester (26mmol) and the reaction mixture was refluxed for 10 hours. The mixture was cooled to room temperature, then the solid was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give the omega-amino ester, which was used in the next step without further purification.

General Synthesis of omega-amino acids (scheme C2)

To a suspension of the crude omega-aminoalkanoic acid ethyl ester obtained in the previous step (ca. 25mmol) in 15mL of water, NaOH (1.4g, 25mmol) was added and the mixture was heated at reflux for 16 h. The reaction was then cooled to room temperature, the solution was acidified with HCl 6N at 0 ℃ and then concentrated under reduced pressure. The residue was treated with EtOH and NaCl and the precipitate was filtered off. Evaporation of the solvent under reduced pressure gave the omega-amino acid as a white solid or as a colorless oil.

4- (2-methyl-piperidin-1-yl) -butyric acid

a)4- (2-methyl-piperidin-1-yl) -butyric acid ethyl ester

The title compound is prepared according to the general synthesis of omega-amino esters (scheme C1). After filtering off the excess 2-methylpiperidine, the organic phase is concentrated under reduced pressure to give 4.6g of amino ester (yield 99%), which is used in the next step without further purification.

C₁₂H₂₃NO₂

¹H-NMR(DMSO-d₆)：0.94(3H，d，J＝6.0Hz)；1.11-1.19(4H，m)；1.31-1.40(1H，m)；1.46-1.62(5H，m)；1.97-2.02(1H，m)；2.12-2.28(5H，m)；2.52-2.59(1H，m)；2.68-2.73(1H，m)；4.02(2H，q，J＝7.2Hz)。

b)4- (2-methyl-piperidin-1-yl) -butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Evaporation of water under reduced pressure gave 4.1g of the title compound (99% yield).

C₁₀H₁₉NO₂

¹H-NMR(DMSO-d₆)：1.01(3H，d，J＝6.4Hz)；1.19-1.27(2H，m)；1.40-1.49(2H，m)；1.54-1.61(4H，m)；2.10-2.13(2H，m)；2.18-2.25(1H，m)；2.28-2.35(1H，m)；2.42-2.48(1H，m)；2.62-2.69(1H，m)；2.69-2.84(1H，m)。

4- (2-methyl-pyrrolidin-1-yl) -butyric acid

a)4- (2-methyl-pyrrolidin-1-yl) -butyric acid ethyl ester

The product was prepared according to the general synthesis of omega-amino esters (scheme C1). After filtering off the excess 2-methylpyrrolidine, the organic phase is concentrated under reduced pressure to give 4.1g of amino ester (yield 99%), which is used in the next step without further purification.

C₁₁H₂₁NO₂

¹H-NMR(CDCl₃)：1.09-1.11(3H，m)；1.23(3H，t，J＝6.8Hz)；1.41-1.48(2H，m)；1.63-1.95(6H，m)；2.10-2.14(2H，m)；2.78-2.81(1H，m)；3.17-3.21(2H，m)；4.10(2H，q，J＝7.2Hz)

b)4- (2-methyl-pyrrolidin-1-yl) -butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Water was evaporated under reduced pressure and crystallized from acetone to give 1.4g of the title compound (49% yield).

C₉H₁₇NO₂

¹H-NMR(DMSO-d₆)：1.31(3H，d，J＝6.4Hz)；1.51-1.60(1H，m)；1.81-1.91(4H，m)；2.03-2.17(1H，m)；2.24-2.37(2H，m)；2.82-2.95(1H，m)；2.97-3.02(1H，m)；3.19-3.32(2H，m)；3.49-3.57(1H，m)；10.06(1H，br s)。

4((S) -2-methyl-piperidin-1-yl) -butyric acid

a)4- ((S) - (2-methyl-piperidin-1-yl) -butyric acid ethyl ester

The product was prepared according to the general synthesis of omega-amino esters (scheme C1). After filtering off excess (S) -2-methylpiperidine, the organic phase is concentrated under reduced pressure to give 2.4g of an amino ester (yield 92%), which is used in the next step without further purification.

C₁₂H₂₃NO₂

¹H-NMR(CDCl₃)：0.93(3H，d，J＝6.0Hz)；1.10-1.21(5H，m)；1.31-1.39(1H，m)；1.44-1.64(5H，m)；1.97-2.03(1H，m)；2.11-2.25(4H，m)；2.53-2.59(1H，m)；2.68-2.72(1H，m)；4.01(2H，q，J＝6.8Hz)。

b)4((S) -2-methyl-piperidin-1-yl) -butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Evaporation of water under reduced pressure gave 1.9g of the title compound (85% yield).

C₁₀H₁₉NO₂

¹H-NMR(DMSO-d₆)：1.22(3H，d，J＝6.4Hz)；1.40-1.43(1H，m)；1.50-1.70(4H，m)；1.76-1.83(3H，m)；2.26-2.33(2H，m)；2.80-2.89(2H，m)；2.95-3.00(1H，m)；3.11-3.19(2H，m)。

4- ((R) -2-methyl-pyrrolidin-1-yl) -butyric acid

a)4- ((R) -2-methyl-pyrrolidin-1-yl) -butyric acid ethyl ester

(R) -2-methyl-pyrrolidine hydrochloride (1.0g, 8.2mmol, 1.1eq) was dissolved in 2-butanone (25mL) and potassium carbonate (2.2g, 15.7mmol, 2.1eq) was added. Ethyl 4-bromobutyrate (1.07mL, 7.5mmol, 1.0eq) was added and the reaction mixture was refluxed for 2 days. The mixture was cooled to room temperature, the solid filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give 1.5g of the title compound (yield 99%), which was used in the next step without further purification.

C₁₁H₂₁NO₂

¹H-NMR(DMSO-d₆)：0.95(3H，d，J＝6.0Hz)；1.15(3H，t，J＝7.2Hz)；1.20-1.27(1H，m)；1.56-1.64(4H，m)；1.77-1.86(1H，m)；1.91-1.99(2H，m)；2.15-2.22(1H，m)；2.25-2.30(2H，m)；2.62-2.69(1H，m)；2.97-3.01(1H，m)；4.01(2H，q，J＝7.2Hz)。

b)4- ((R) -2-methyl-pyrrolidin-1-yl) -butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Evaporation of water under reduced pressure gave 1.4g of the hydrochloride salt of the title compound (88% yield).

C₉H₁₇NO₂

¹H-NMR(DMSO-d₆Hydrochloride salt): 1.34(3H, d, J ═ 6.4 Hz); 1.56-1.61(1H, m); 1.83-1.92(3H, m); 2.11-2.14(1H, m); 2.31-2.39(2H, m); 2.81-2.90(1H, m); 2.95-3.04(1H, m); 3.19-3.44(3H, m); 3.51-3.58(1H, m); 10.20(1H, brs); 12.29(1H, br s).

2-methyl-4- (pyrrolidin-1-yl) -2-butanoic acid

a) 4-bromo-2-methyl-butyryl bromide

2-methylbutyrolactone (50mmol, 5.0g) and phosphorus tribromide (41mmol, 3.7mL) were heated at 140 ℃ for 2.5 hours. The reaction mixture was transferred to a Kugelrohr distiller and distilled under reduced pressure (40mmHg, T ═ 128 ℃) to give 6.21g (yield: 51%) of 4-bromo-2-methyl-butyryl bromide as a colorless transparent oil.

C₅H₈Br₂O

¹H-NMR(CDCl₃)：3.45(2H，t，J＝6.8Hz)；3.22-3.18(1H，m)；2.42-2.36(1H，m)；1.99-1.94(1H，m)；1.32(3H，d，J＝7.2Hz)。

b) 4-bromo-2-methyl-butyric acid methyl ester

4-bromo-2-methyl-butyryl bromide (6.2g, 43.0mmol, 1.0eq) in CHCl₃The solution in (10mL) was cooled to 0 ℃. MeOH (10mL) was added slowly and stirred at room temperature for 16 h. The solvent was evaporated and the residue was dissolved in CHCl₃And washed with water and brine. Collecting the organic layer, adding Na₂SO₄And (5) drying. Evaporation of the solvent gave methyl 4-bromo-2-methyl-butyrate as a viscous oil (4.3g, 51% yield).

C₆H₁₁BrO₂

¹H-NMR(DMSO-d₆)：1.19(3H，d，J＝7.2Hz)；1.94-1.89(2H，m)；2.29-2.23(2H，m)；3.43-3.40(1H，m)；3.69(3H，s)。

c) 2-methyl-4- (pyrrolidin-1-yl) -2-butanoic acid

Pyrrolidine (5.4mL, 66mmol) was dissolved in toluene (40 mL). 4-bromo-2-methyl-butyric acid methyl ester (4.3g, 22.0mmol) was added and the reaction was stirred at reflux for 2.5 hours. The solvent and excess amine were removed under reduced pressure to give methyl 2-methyl-4- (pyrrolidin-1-yl) -butyrate as a viscous oil. The crude product was diluted with MeOH (3mL), 1.0M aqueous NaOH (22mL) was added and the reaction was stirred at reflux for 18 h.

After cooling to room temperature, the mixture was concentrated under reduced pressure to remove the organic solvent and water. Adding HCl 6N to pH 4.5; then EtOH was added to precipitate NaCl. After filtration the solvent was evaporated under reduced pressure (keeping the temperature of the water bath at room temperature to prevent esterification) to give 4-pyrrolidine-2-methyl-butyric acid as a yellow oil (3.58g, yield 90%).

C₉H₁₇NO₂

Mass (calculated) [199](ii) a (found value) [ M + H [)⁺]＝200。

LC Rt 1.12 min; 90% (5 min method):

¹H-NMR(DMSO-d₆)：2.79(4H，m)；2.73(2H，m)；2.37(1H，m)；1.84(2H，m)；1.81-1.75(3H，br m)；1.57(1H，m)；1.5(3H，d，J＝7.2Hz)

2-methyl-4-piperidin-1-yl-butyric acid

Piperidine (1.1mL, 20.0mmol, 3.0eq) was dissolved in toluene (15 mL). 4-bromo-2-methyl-butyric acid methyl ester (1.3g, 6.6mmol, 1.0eq) was added and the reaction was stirred at reflux for 3 hours. The solvent and excess amine were removed under reduced pressure to give 4-pyrrolidine-2-methyl-butyric acid methyl ester as a viscous oil. The crude product was diluted with MeOH (2mL), 1.0M aqueous NaOH (14mL, 7.0eq) was added and the reaction was stirred at reflux for 16 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to remove the organic solvent and water. Adding HCl 6N to pH 4.5; then EtOH was added to precipitate NaCl. After filtration the solvent was evaporated under reduced pressure (keeping the temperature of the water bath at room temperature to prevent esterification) to give 4-pyrrolidine-2-methyl-butyric acid as a yellow oil (0.9g, yield 66%).

C₁₀H₁₉NO₂

Mass (calculated value) [171](ii) a (found value) [ M + H [)⁺]＝172。

LC Rt ═ 0.22 min; 90% (5 min method).

¹H-NMR(CDCl₃)：3.66(m，1H)；3.59(m，1H)；3.53(m，2H)；3.45(m，2H)；2.93(m，1H)；1.62-1.51(br m，8H)；1.10(d，3H，J＝7.2)

5- [1, 4] -oxazepan-4-yl-butyric acid

Homomorpholine (1.0g, 7.3mmol, 1.2eq) was dissolved in toluene (15mL), 4-bromo-2-methyl-butyric acid methyl ester (0.9g, 6.1mmol, 1.0eq) was added and the reaction was stirred at reflux for 3 hours. The solvent and excess amine were removed under reduced pressure to give the methyl ester as an oil. The crude product is treated with H₂O (10mL) and MeOH (2mL) were diluted, 1.0M aqueous NaOH (0.3g, 7.0eq) was added, and the reaction was stirred at reflux for 18 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to remove the organic solvent and water. Adding HCl 6N to pH 4; then EtOH was added to precipitate NaCl. After filtration, the solvent was evaporated under reduced pressure at room temperature to give 4-pyrrolidine-2-methyl-butyric acid as a yellow oil (0.9g, yield 66%).

C₉H₁₇NO₃

¹H-NMR(DMSO-d₆)：3.73(m，2H)；3.68(m，2H)；3.16-3.11(m，2H)；2.93(m，2H)；2.28(m，2H)；2.23(m，2H)；1.96(m，2H)；1.79(m，2H)。

4-pyrrolidin-1-yl-butyric acid

a) 4-Pyrrolidin-1-yl-butyric acid ethyl ester

To a solution of pyrrolidine (8.42mL, 102mmol, 4.0eq) in toluene (30mL) was added ethyl 4-bromobutyrate (3.8mL, 26mmol, 1.0eq) and the reaction was refluxed for 10 hours. The mixture was cooled to room temperature and the white precipitate was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give the title compound, which was used in the next step without further purification.

b) 4-pyrrolidin-1-yl-butyric acid hydrochloride

4-pyrrolidin-1-yl-butyric acid ethyl ester (about 25mmol) was suspended in 100mL NaOH 10%, heated to reflux for 10 hours, then cooled to room temperature and washed with AcOEt. The aqueous layer was recovered by extraction, adjusted to pH 4 with HCl 37% at 0 ℃ and then concentrated under reduced pressure. The residue was treated with EtOH and then filtered to remove the sodium chloride precipitate. Treating the crude product with diethyl ether and filtering; evaporation of the solvent under reduced pressure gave 2.5g of the title compound as a white solid in 61% overall yield of steps a) and b).

C₈H₁₅NO₂

Mass (calculated value) [157](ii) a (found value) [ M + H [)⁺]＝158。

LC Rt 0.21min, 100% (5 min method)

¹H-NMR(DMSO-d₆HCl salt): 1.80-1.93(6H, m); 2.31(2H, t, J ═ 14.8); 3.03-3.11(2H, m); 3.18-3.32(4H, m, broad peak)

4-morpholin-4-yl-butyric acid

a) 4-Morpholin-4-yl-butyric acid ethyl ester

To a solution of morpholine (8.96mL, 102mmol, 4.0eq) in toluene (30mL) was added ethyl 4-bromobutyrate (3.8mL, 26mmol, 1.0eq) and the reaction mixture was heated under reflux for 10h, then cooled to room temperature, the white precipitate was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give the title compound, which was used in the next step without further purification.

b) 4-morpholin-4-yl-butyric acid

4-morpholin-4-yl-butyric acid ethyl ester (about 25mmol) is suspended in 100mL NaOH 10%. The reaction mixture was heated to reflux for 10 hours, then cooled to room temperature and washed with AcOEt. The aqueous layer was recovered by extraction, adjusted to pH 4 with HCl 37% at 0 ℃ and then concentrated under reduced pressure. The residue was treated with EtOH and the precipitated NaCl was then filtered off. The crude product was treated with acetone, filtered and the solvent was evaporated under reduced pressure to give 3.2g of the title compound as a white solid in 72% total yield from steps a) and b).

C₈H₁₅NO₃

Mass (calculated value) [173](ii) a (found value) [ M + H [)⁺]＝174。

LC Rt 0.30min, 100% (5 min method)

¹H-NMR(DMSO-d₆Hydrochloride salt): 1.86-1.95(2H, m); 2.29-2.34(2H, m); 2.94-3.08(4H, m); 3.34-3.38(2H, m); 3.74-3.83(2H, m); 3.88-3.91(2H, m); 11.24(1H, s)

General procedure for amide coupling

To a suspension of omega-amino acid (7.93mmol) in 1, 2-dichloroethane (20mL) was added N, N' -carbonyldiimidazole (1.2g, 7.4mmol) and the mixture was stirred at room temperature for 2 hours (the suspension was usually seen to dissolve when the amino acid was fully activated). 3-amino-5-aryl/heteroaryl pyrazole (5.29mmol) was then added and the reaction mixture was stirred for a further 10 hours. At the end of the reaction (LC-MS monitoring), if two dissimilarities are formedStructure, the mixture was heated at 50 ℃ to the less stable isomer and converted to the title compound (monitored by LC-MS). The solvent was washed with a saturated aqueous sodium carbonate solution, extracted, and the solvent was removed under reduced pressure. The crude product is in CH₃Recrystallized from CN or via SiO₂Column purification or purification by preparative HPLC.

4- (4-trifluoromethoxy-phenyl) -1H-imidazol-2-ylamine

a) N- [4- (4-trifluoromethoxy-phenyl) -1H-imidazol-2-yl ] -acetamide

Acetylguanidine (2.6g, 25.7mmol, 3.0eq) was dissolved in anhydrous DMF (40mL), 2-bromo-1- (4-trifluoromethoxy-phenyl) -ethanone (2.4g, 8.6mmol, 1.0eq) was added and the mixture was stirred at room temperature for 4 days. DMF was removed under reduced pressure, the residue was washed with water, filtered and dried over sodium sulfate; crystallization from MeOH gave 0.7g of the title compound (30% yield).

C₁₂H₁₀F₃N₃O₂

¹H-NMR(DMSO-d₆)：2.14(3H，s)；7.37-7.40(3H，m)；7.88-7.91(2H，m)；11.33(1H，s)；11.78(1H，br s)。

b)4- (4-trifluoromethoxy-phenyl) -1H-imidazol-2-ylamine

N- [4- (4-trifluoromethoxy-phenyl) -1H-imidazol-2-yl ] -acetamide (0.7g, 2.6mmol, 1.0eq) was dissolved in water (18mL) and methanol (18mL), 20 drops of sulfuric acid were added. The reaction was refluxed for 2 days, the mixture was dried, the residue was diluted with water, pH adjusted to 8 with NaOH 2N, the product was extracted with DCM and concentrated under reduced pressure to give 0.6g of the title compound (yield 98%).

C₁₀H₈F₃N₃O

¹H-NMR(DMSO-d₆)：5.73(2H，br s)；7.10(1H，s)；7.26(2H，d，J＝8.0Hz)；7.67-7.69(2H，m)。

Example 1

5-azepan-1-yl-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide

5- (4-methoxy-phenyl) -1H-pyrazol-3-yl-amine (0.089g, 0.45mmol) was dissolved in DCE: DMF 4: 1(2.5mL), 5-bromo-n-valeryl chloride (0.057mL, 0.43mmol) was added, followed by diisopropylethylamine (0.078mL, 0.45 mmol). The reaction was stirred at 0 ℃ for 1 hour under nitrogen. Then azepane (0.152mL, 1.35mmol) and more diisopropylethylamine (0.078mL, 0.45mmol) were added together. The reaction was stirred at +50 ℃ for 18 hours. At the end of the reaction (monitored by LC-MS), the solvent was removed under reduced pressure and the resulting oily residue was dissolved in DCM (20mL) and saturated Na₂CO₃(2X20mL) and saturated NaCl (2X20mL) washes; na for organic layer₂SO₄And (5) drying.

Purification by preparative HPLC (standard acidic conditions) gave 0.046g of the title compound as the formate salt (0.11mmol, 25% yield).

C₂₁H₃₀N₄O₂Mass (calculated value) [370.50](ii) a (found value) [ M + H [)⁺]＝371

LC Rt ═ 1.97, 96% (10 min method)

NMR(400MHz，DMSO-d₆)：1.79-1.71(6H，m)；1.89(6H，m)；3.17(2H，t)；3.34(2H，m)；3.82(3H，s)；6.7(1H，s)；6.98(2H，d)；7.58(2H，d)；8.26(1H，HCOOH，s)；10.21(1H，s)。

Example 2

5- (4-methyl-piperidin-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide

5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide (0.106g, 0.6mmol) was dissolved in DMF (2mL), sodium iodide (0.045g, 0.6mmol) was added, followed by 4-methylpiperidine (0.054mL, 1.5mmol) and diisopropylethylamine (0.052mL, 0.6mmol, 1 eq). The reaction was stirred under nitrogen at +50 ℃ for 18 hours.

At the end of the reaction (monitored by LC-MS), the solvent was removed under reduced pressure and the resulting oily residue was dissolved in DCM (20mL) and saturated Na₂CO₃(2X20mL) and saturated NaCl (2X20mL) washes; na for organic layer₂SO₄And (5) drying.

Purification by preparative HPLC (standard acidic conditions) gave 0.057g of the title compound as the formate salt (0.14mmol, 45% yield).

C₂₁H₃₀N₄O₂Mass (calculated value) [370.50](ii) a (found value) [ M + H [)⁺]＝371.26

LC Rt ═ 1.73, 100% (10 min method)

NMR(400MHz，DMSO-d₆)：0.84(3H，d，J＝6.23Hz)；1.13-1.07(2H，m)；1.33-1.27(4H，m)；1.45(1H，m)；1.50(2H，m)；1.96(2H，m)；2.26(2H，m)；2.35(2H，m)；2.88(2H，m)；3.14(3H，s)；6.71(1H，s)；6.96(2H，d)；7.6(2H，d)；8.17(1H，s，HCOOH)；10.13(1H，s)。

Example 3

5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid (5-thiophen-2-yl-1H-pyrazol-3-yl) -amide

Bromovaleryl chloride (1.62mL, 12.12mmol) was dissolved in DMA (50 mL). To this was added a solution of 5-thiophen-2-yl-2H-pyrazol-3-ylamine (2g, 12.12mmol) and DIEA (2.1mL, 12.12mmol) in portions at 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 hour, then at room temperature for 2 hours. After a total of 3 hours, triphosphonylmethylamine (PS-Trisamine) (1g, ca. 4mmol/g) was added to the mixture and stirred for 2 hours. N-acetyl homopiperazine (4.3g, 30.3mmol) was then added and the mixture was stirred at room temperature for a further 60 hours. After evaporation of DMA under reduced pressure, water (50mL) was added and extracted with ethyl acetate (3 × 30 mL). The aqueous layer was basified with solid NaOH, extracted with ethyl acetate at pH 10 and then re-extracted at pH 11. All organic layers were combined, dried and evaporated. The residue was purified by chromatography on silica gel (gradient eluent: ethyl acetate/methanol 9: 1 to ethyl acetate/methanol 8: 2 to give the title compound as a yellow oil (800mg, 17%).

C₁₉H₂₇N₅O₂S Mass (calculated) [389.52 ]](ii) a (found value) [ M + H [)⁺]＝390.11

NMR(400MHz，CDCl₃)：1.52(2H，m)；1.77(2H，m)；1.82(2H，m)；2.13+2.09(3H，s)；2.44(2H，m)；2.56(2H，m)；2.62(1H，m)；2.76-2.70(3H，m)；3.51(2H，m)；3.61(1H，m)；3.64(1H，m)；6.48(1H，s)；6.56(1H，s)；7.05-7.02(2H，m)；6.9-7.26(2H，m)；8.94(1H，s)；9.53(1H，s)。

A solution of HCl in ether (1.05mL, 2N) was added to a suspension of (5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid (5-thiophen-2-yl-2H-pyrazol-3-yl) -amide (800mg, 2.05mmol) in MeOH (10mL) to convert the title compound to the hydrochloride salt the solution was stirred at room temperature for 1 hour and then evaporated to dryness to give the title compound as a yellow powder (750mg, 86%).

Example 4

5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide

a) First method

ai) 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide

A solution of 5-bromovaleryl chloride (2.1mL, 15.7mmol, 1eq) in dry DMA (35mL) was cooled to-10 ℃ (ice water bath) under nitrogen and a solution of 5- (4-methoxy-phenyl) -1H-pyrazol-3-ylamine (3.0g, 15.7mmol, 1eq) and diisopropylethylamine (2.74mL, 15.7mmol, 1eq) in dry DMA (15mL) was added over 30 minutes. After 2 hours at-10 deg.C, the reaction was complete by LC-MS and stopped by the addition of water (about 50 mL). The solid precipitate was filtered off and washed with diethyl ether to give 4.68g of 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide as a white powder (13.3mmol, 85% yield).

mp＝149.5-151.5℃。

C₁₅H₁₈BrN₃O₂Mass (calculated value) [352.23](ii) a (found value) [ M + H [)⁺]＝352.09/354.10

LC Rt 2,07, 95% (5 min method)

NMR(400MHz，DMSO-d₆)：1.69-1.63(2H，m)；1.81-1.75(2H，m)；2.29(2H，t)；3.52(2H，t)；3.75(3H，s)；6.75(1H，bs)；6.96(2H，d)；7.6(2H，d)；10.28(1H，s)；12.57(1H，s)

aii)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide

To 750mg (1.96mmol) of 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl]-amide in 7mL DMA solution, adding N-acetyl-diazepine(278mg, 1.96mmol) and NaI (240mg, 1.96mmol) were heated at 60 ℃ for 18 hours. At the end of the conversion (monitored by LC-MS), it was diluted with 20mL of DCM and washed with water. The organic phase is concentrated under reduced pressure and the residue obtained is taken up in SiO₂Column (10g) purification (gradient elution: DCM to DCM-MeOH 90: 10). The pure title compound (380mg) was recovered (yield 46%).

C₂₂H₃₁N₅O₃Mass (calculated value) [413 ]](ii) a (found value) [ M + H [)⁺]＝414

LC Rt ═ 1.91, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆)：1.53-1.75(4H，m)，1.90-2.15(5H，m)，2.28-2.42(2H，m)，2.90-3.26(3H，m)，3.34-3.58(3H，m)，3.71-3.88(7H，m)

b) Second method

bi)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide (monohydrochloride)

To a solution of 5- (4-methoxyphenyl) -1H-pyrazol-3-ylamine (12g, 62.8mmol) and N, N-diisopropylethylamine (10.96mL, 62.8mmol) in dry N, N-dimethylformamide (150mL) was slowly (about 40min) added a solution of 5-bromovaleryl chloride (8.4mL, 62.8mmol) in dry N, N-dimethylformamide (50mL) at-10 ℃ and the reaction mixture was stirred at-10 to 0 ℃ for 8H. Sodium iodide (9.44g, 62.8mmol) was added at 0 ℃ followed by N-acetylhomopiperazine (8.24mL, 62.8mmol) and N, N-diisopropylethylamine (10.96mL, 62.8mmol), and the reaction mixture was stirred at 50 ℃ for 18 h. The solvent was removed in vacuo. The residue was dissolved in dichloromethane (500mL) and saturated aqueous sodium bicarbonate (500mL) and stirred at room temperature for 30 min. The organic layer was separated, dried over sodium sulfate and the solvent removed in vacuo to give 25.8g (99%) of 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide as a viscous, pale yellow oil (crude product).

To a solution of the crude product 5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide (free base) in dichloromethane (270mL) was then slowly added hydrogen chloride (65mL, 1.0M in ether) at room temperature. The resulting suspension was stirred at room temperature for 1 hour and the solvent was removed in vacuo to give 33g of the monohydrochloride as a yellow foam. The foam was dissolved in a solvent (330mL, 33: 1 acetonitrile: methanol) at 60-70 ℃ and seeded. The mixture was slowly cooled to room temperature and stirred at room temperature for 15 hours. The precipitate formed is filtered off to give 20.5g (72%) of the monohydrochloride salt of the title compound as white crystals. MS [ M-H ]]^- m/z 412.3；mp.132-133℃。

c) Third method

ci)3- (4-methoxyphenyl) -3-oxopropanenitrile

The solution of methyl anisate in acetonitrile was cooled to-10 ℃. Lithium bis (trimethylsilyl) amide (1M THF solution) was added dropwise over a minimum of 3 hours. The mixture was kept at-10 to 0 ℃ until the end of the reaction. Water was added to terminate the reaction, and the pH was adjusted to 3-4 with concentrated hydrochloric acid. The mixture was stirred for 1 hour, the product was isolated by filtration, washed with water and dried in a vacuum oven. The yield thereof was found to be 73%.

cii)5- (4-methoxyphenyl) -1H-pyrazol-3-amine

A suspension of 3- (4-methoxyphenyl) -3-oxopropanenitrile in ethanol was heated to 60 ℃. At this temperature, hydrazine hydrate is added dropwise over at least 30 minutes. The resulting solution is kept at 60 ℃ until the end of the reaction, which usually takes 15 to 18 hours. The reaction was quenched by addition of water and distilled to about 5 volumes to remove ethanol. The product was isolated by filtration, washed with water and dried in a vacuum oven. The yield thereof was found to be 88-95%.

ciii) 5-bromo-N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide

A solution of 5- (4-methoxyphenyl) -1H-pyrazol-3-amine and diisopropylethylamine in a 10 volume 9: 1 mixture of acetonitrile: DMF was cooled to-10 ℃. 5-bromovaleryl chloride is added dropwise at-10 ℃ over at least 3 hours. The resulting solution was kept at-10 ℃ until the end of the reaction, which usually took 2 hours. The reaction was stopped by adding water, the product was isolated by filtration, washed with water and TBME and drained. The product wet cake was purified by reslurrying in TBME at 35 ℃ for at least 2 hours. The yield is 70-80%.

civ)5- (4-acetyl-1, 4-diazepan-1-yl) -N- (5- (4-methoxyphenyl) -1H-pyrazol-3-yl) pentanamide

Reacting bromopyrazole with K at room temperature₂CO₃And KI in 10 volumes of acetone, at 1N-acetyl homopiperazine was added over a period of hours. The reaction mixture was stirred until the end of the reaction. The mixture was filtered to remove inorganic material, washed with acetone and distilled to 2 volumes. The free base was extracted into methyl THF/EtOH, with NaCl and NaHCO₃And (6) washing. The solution concentration was determined by replacing the solvent with EtOH and 0.93 equivalents of HCl were added to a mixture of acetone, ethanol and water, depending on the free base concentration in the solution. Careful monitoring of the pH gave the desired crystalline product form 1 in 70% overall yield.

d) Fourth method

di)5- (4-methoxy-phenyl) -1H-pyrazol-3-ylamine

The intermediate 5- (4-methoxy-phenyl) -1H-pyrazol-3-ylamine is commercially available from Sigma-enrich (usa) and can also be prepared according to the following general method:

synthesis of aryl beta-ketonitriles

To a solution of the aromatic ester (6.5mmol) in dry toluene (6mL) under nitrogen was carefully added NaH (50-60% dispersion in mineral oil, 624mg, 13 mmol). Heating the mixture at 80 deg.C, and adding dry CH dropwise₃CN (1.6mL, 30.8 mmol). The reaction solution is heated for 18 hours and the product precipitates from the reaction, usually in the form of a salt. The reaction was cooled to room temperature and the solid formed was filtered off and dissolved in water. The solution was acidified to pH 2-4 with 2N HCl solution and the precipitated product was filtered off. If no precipitate separated out, the product was extracted with DCM. The product purity is usually high enough to be used in the next step without further purification by aqueous workup. The one-step yield is usually 40-80%. Synthesis of arylaminopyrazoles

To a solution of β -ketonitrile (7.5mmol) in anhydrous EtOH (15mL) was added hydrazine monohydrate (0.44mL, 9.0mmol) and the reaction was heated at reflux for 18 h. The reaction mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. The residue was dissolved in 20mL of DCM and washed with water. Concentrating the organic phase to obtain a crude product, passing through SiO₂Purifying by column, or precipitating from diethyl ether. For example, the 2-methoxy derivative is purified by chromatography on silica gel with a DCM/MeOH gradientDe (100% DCM to 90/10 DCM/MeOH); the 3-methoxy derivative was triturated in ether. The yield is usually 65-90%. dii) 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl]Amides of carboxylic acids

A solution of 5-bromovaleryl chloride (2.1mL, 15.7mmol) in dry Dimethylacetamide (DMA) (35mL) was cooled to-10 deg.C (ice water bath) under nitrogen and a solution of 5- (4-methoxy-phenyl) -1H-pyrazol-3-ylamine (3.0g, 15.7mmol) and diisopropylethylamine (2.74mL, 15.7mmol) in dry DMA (15mL) was added over 30 minutes. After 2 hours at-10 ℃ LCMS indicated the end of the reaction (acylation on the pyrazole ring was also detected). The reaction was quenched by the addition of water (about 50mL) and the large amount of white precipitate formed upon the addition of water was filtered off. When the reaction was warmed to room temperature before terminating the reaction, it was concluded that exchange of Br with Cl may occur, which causes reactivity problems in subsequent steps. Washing with diethyl ether (3x10mL) effectively removed the by-product (acylation on the pyrazole ring). 4.68g of the title compound are obtained as a white powder (13.3mmol, 85% yield). Mp 149.5-151.5 ℃.

diii)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide

Reacting 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl]Amide (1.5g, 4.26mmol) was dissolved in DMF (15mL), sodium iodide (0.64g, 4.26mmol) was added, followed by N-acetyl homopiperazine (0.56mL, 4.26mmol) and diisopropylethylamine (0.74mL, 4.26 mmol). Stirring was carried out at 50 ℃ for 18 hours under nitrogen. At the end of the reaction (LCMS monitoring), the solvent was removed under reduced pressure and the resulting oily residue was dissolved in DCM (20mL) and saturated Na₂CO₃(2X20mL) and saturated NaCl (2X20mL) washes with Na₂SO₄And (5) drying. After removal of the solvent, 1.7g of crude product are obtained as a viscous oil. The product is processed by SiO₂Chromatography (10g silica gel column of IST-flash chromatography SI II) eluting with DCM and DCM: MeOH 9: 1 afforded 0.92g of pure product and 0.52g of less pure product. With 5g SiO₂The column again purified the less pure product, eluent as above. To give 1.09g of 5- (4-acetyl- [1, 4)]Diazacyclo ringHeptane-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl]Amide (2.64mmol, 62% yield) as a pale yellow viscous oil. MS (ES +): 414.26(M + H)⁺。

div)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide hydrochloride

Reacting 5- (4-acetyl- [1, 4)]Diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl]Amide (1.05g, 2.54mmol) was dissolved in a minimum amount of DCM (5mL) and cooled to 0 ℃. HCl (2.0M in Et) was added₂O, 1.4mL, 2.89mmol), and the mixture was stirred at room temperature until complete precipitation of the salt (about 10 min.). The solid was filtered off, washed several times with diethyl ether and dried in a desiccator to give 1.09g of hydrochloride (2.42mmol, 95% yield). The melting point was not determined because the sample was very hygroscopic. MS (ES +): 414.26(M + H)⁺。

e) Fifth method

ei)5- (4-acetyl- [1, 4] diazepan-1-yl) -N- [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -pentanamide

To a 3L column reactor equipped with nitrogen inlet, stirrer, condenser/distillation top, temperature control and jacketed shell was added 5-bromo-pentanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] amide (0.15kg, 0.426mol), potassium carbonate (0.059kg, 0.426mol), potassium iodide (0.071kg, 0.426mol) and acetone (1.18kg, 1.5L) (at 20 ℃) to form a white mixture. The mixture was stirred (235rpm) at 25-30 ℃ for at least 15 minutes. N-acetyl homopiperazine (0.062kg, 0.057L, 0.434mol) was added to the reactor via an addition funnel over at least 45 minutes, maintaining the temperature at 25-30 ℃. The addition funnel was washed with 0.05L of acetone and the mixture in the reactor remained white. Stirring (235rpm) at 25-30 ℃ for at least 16 hours resulted in a white/yellow mixture. The reaction was monitored by HPLC and considered complete when the starting material (bromopyrazole) in the reactor was < 2% and iodopyrazole < 2%.

The temperature of the reaction was reduced to 5-15 ℃ with stirring (295rpm) over at least 15 minutes to form a white/yellow mixture, and stirring was continued for at least 1 hour. The mixture was filtered for 1.5 minutes using a laboratory vacuum pump and a buchner funnel with filter paper to remove the minerals. The filter cake was washed twice with acetone at 5-15 deg.C (0.24 kg, 0.30L total). The washing solution was combined with the filtered mother liquor and the reactor was washed with the combined solution. The filtrate was concentrated to about 0.45L to give a clear solution.

eii) aqueous treatment

To a reactor containing the product of step i, a freshly prepared homogeneous solution of 1.5L of methyl THF (1.22kg, 1.42L) and ethanol (0.059kg, 0.075L) was added at 25 ℃ to form a cloudy solution. To this was added 0.45L of a 5% solution of sodium chloride (0.022kg) in water (0.43L) at 25 ℃. The resulting mixture was heated to 30-35 ℃ over 15 minutes with stirring to form a clear biphasic solution. Stopping stirring, standing for layering, and allowing the product to be in the upper layer. The two layers were separated, leaving the emulsion in the upper organic layer. The organic layer was left, washed with a homogeneous solution of 5% sodium bicarbonate (0.03kg) in water (0.57L) at 25 ℃ and stirred at 10-15 ℃ for at least 5 minutes. Stopping stirring, standing for layering, and allowing the product to be in the upper layer. The two layers were separated, leaving the emulsion in the upper organic layer. The organic layer was left to concentrate to 0.35L, forming a cloudy solution. The mixture was treated with ethanol to remove residual water.

eiii)5- (4-acetyl- [1, 4] diazepan-1-yl) -N- [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -pentanamide HCl

To the reactor containing the product of step ii, 0.47kg (0.60L) of acetone was added. The resulting mixture was heated to 25-30 ℃ over at least 10 minutes with stirring to form a cloudy solution. The contents of the reactor were filtered through a polypropylene filter pad to a 2L tared filter flask for clarification, and the contents were maintained at 25-30 ℃ at all times. And keeping the negative pressure until the filtration is finished. The reactor and filter pad were washed with acetone (0.05L) at 20-25 deg.C. The filtrate from the suction flask was transferred to the reactor and rinsed with acetone (0.05L). A solution of 5% HCl (0.042kg, 0.036L) in acetone (0.174L) and ethanol solution (0.0174L ethanol: acetone (91: 9) v/v) was prepared and stirred at 10 ℃ to give a homogeneous solution. 0.05L of water was added to the reactor to form a clear solution. One third of the prepared 5% HCl solution (0.076L) was added to the reactor over at least 20 minutes, maintaining the temperature at 20-25 ℃. Then a second third of 5% HCl solution (0.076L) was added to the reactor over at least 20 minutes, maintaining the temperature at 20-25 ℃.75 mg of 5- (4-acetyl- [1, 4] diazepan-1-yl) -N- [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -pentanamide HCl (e.g. type 1) are seeded in the reactor, and then the last third of a 5% HCl solution (0.076L) is added over a minimum of 20 minutes, maintaining the temperature at 20-25 ℃. A further 0.08 equivalents of 5% HCl solution (0.023L) was then added over at least 30 minutes, maintaining the temperature at 20-25 ℃. The pH was monitored accurately to a pH of 5.2-5.8.

The mixture was stirred at 20-25 ℃ for at least 1 hour to form a thin suspension. Acetone (0.6L) was added over at least 60 minutes, maintaining the temperature at 20-25 ℃. The mixture was stirred at 20-25 ℃ for at least 60 minutes. Acetone (1.5L) was added to the reactor over at least 3 hours, maintaining the temperature at 20-25 ℃ to form a thick suspension. The mixture was stirred at 20-25 ℃ for at least 12 hours. Crystallization is considered to be complete when the product in the mother liquor is equal to or less than 20%.

The mixture was then suction filtered using a buchner funnel (polypropylene filter pad) and a laboratory vacuum pump. A solution of water (0.009L), acetone (0.23L) and 0.06L of alcohol (ethanol: acetone (91: 9) v/v) was stirred to form a homogeneous solution (20% ethanol, 3% water, total 77% acetone). The filter cake was washed twice with this solution (0.15Lx 2). A solution of water (0.009L), acetone (0.171L) and 0.12L of alcohol (ethanol: acetone (91: 9) v/v) was stirred to form a homogeneous solution (40% ethanol, 3% water, 57% acetone total). The filter cake (0.30L) was washed with this solution. After dropping was stopped, the wet cake was pumped under nitrogen for 30 minutes with a laboratory vacuum pump. And detecting the purity of the product by using HPLC, and continuously washing if the total impurities are not less than 2%. The product was dried in a vacuum oven under a stream of nitrogen at a temperature of 38-45 deg.C and the vacuum was maintained at 20 torr for at least 12 hours until the loss on drying was less than 1%. After drying, 0.119kg of the title compound is obtained in 62% yield (67% adjusted yield, considering the aliquot removed in the process; 60% adjusted yield, considering concentration or purity). Melting point 185 ℃; the crystal form is 1; the grain size is D90 < 89.4um, D50 < 19.2 um.

f)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride

This example describes the preparation of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride. The hydrochloride salt form readily forms a solid. In fact, at least 4 different crystalline forms of the hydrochloride salt were observed (i.e. polymorphs, see table below).

Counter ions used	The obtained solid	Start of melting	Moisture absorption property
				Hydrochloric acid	Crystal	About 100 to about 180 from three peaks at 185 ℃ 165 ℃ and 125 ℃ 125 ℃; and about 200 deg.C	Does not absorb moisture and absorbs moisture slightly? Moisture absorption

DSC data for each solid form were measured with a differential scanning calorimeter (TA instruments, model Q1000)The parameters are as follows: 50mL/min purge gas (N)₂) (ii) a The scanning range is 40-200 ℃, and the scanning speed is 10 ℃/min. TGA data was measured using a thermogravimetric analyzer (Mettler Toledo, model TGA/SDTA 851e) with a parameter of 40ml/min purge gas (N)₂) (ii) a The scanning range is 30-250 ℃; the scanning rate was 10 ℃/min. X-ray derivative spectra were obtained using an X-ray powder diffractometer (Bruker-axs, model advanced D8) with the parameters: voltage 40kV, current 40.0mA, scan range (2 θ)5-30 °, scan step 0.01 °, total scan time 33 minutes, VANTEC detector. The anti-scatter slit is 1 mm. Figures 1-7 show the characteristic data for the hydrochloride salt.

The hydrochloride salt had polymorphic forms with DSC endothermic peaks at 119 ℃ (form III), 127 ℃ (form IV), 167 ℃ (form II) and 186 ℃ (form I). Another form may be the form of an ethanol solvate with multiple endothermic peaks corresponding to 1) desolvation at about 100 ℃; 2) form I, about 183 ℃; 3) another polymorph is possible, about 200 ℃. The following crystalline form table illustrates some of the characteristics observed for each crystalline form of the hydrochloride salt.

Crystal form meter

Crystal form I	Crystal form II	Crystal form III	Crystal form IV	Crystal form V
					Monohydrochloride (8% HCl)
Melting temperature: 180 ℃ to 186 DEG C	Melting temperature: 165 deg.C	Melting temperature: 125 deg.C	Melting temperature: 125 deg.C	Three peaks: about 100 deg.C, about 180 deg.C, about 200 deg.C
					Non-hygroscopic (see fig. 4)	Slightly hygroscopic (5% moisture at RH 50%; see FIG. 10)	Having hygroscopicity (10% moisture at RH 50%; see FIG. 11)	Not detected	Having hygroscopicity (7% at RH 50%; see FIG. 12)

Of the various forms of the hydrochloride salt observed, only form I (186 ℃) is relatively non-hygroscopic, absorbing only about 0.5% of water when equilibrium is reached at an RH of less than or equal to 70%. Form I absorbs at least about 12% of water at 70-100% RH, but as RH is lowered it loses absorbed water without significant hysteresis. No evidence of a hydrated hydrochloride salt was observed.

Depending on the amount of hydrochloric acid present in the solution during reactive crystallization, polyhydrochloride salts are also formed. Adjusting the pH of the solution to about 4-5 converts the polyhydrochloride to the monohydrochloride. But continued conditioning may result in the formation of inorganic salts. In some embodiments, the conditions for making a pure monohydrochloride salt are that the hydrochloric acid equivalent and the slurry pH are < 0.95eq. (e.g., 0.93) and ph.5 (see, e.g., fig. 8-11), respectively.

g) Characterization of certain crystalline forms of the hydrochloride salt

This example describes the characterization of two unexpectedly non-hygroscopic crystalline forms (forms I and II, as described above) of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride:

both crystal forms are readily soluble in water. Type I melting point is 185 ℃ (± 2 ℃); form II has a melting point of 166 ℃ (± 2 ℃).

Form I absorbs moisture at a Relative Humidity (RH) of about 50% and eventually absorbs no more than about 2% moisture (90% RH) and is lost as RH decreases (< 50%). The characteristic peaks of the X-ray diffraction pattern of form I have 2 θ angles of 15.3 ° and 21.9 °, which fluctuate by 0.3 ° depending on the instrument and the measurement method.

Form II absorbs moisture at approximately 20% RH and eventually absorbs no more than 7% moisture (90% RH) and retains 2% moisture at very low RH (0%). The 2 theta angles of the characteristic peaks of the X-ray diffraction pattern of form II were 20.2 ° and 24.9 °, which varied by 0.3 ° depending on the instrument and measurement method. DSC data of each crystal form are measured by a differential scanning calorimeter (TA instrument, model Q1000), and parameters are as follows: 50mL/min purge gas (N)₂) (ii) a The scanning range is 40-200 ℃, and the scanning speed is 10 ℃/min.

TGA data were measured using a thermogravimetric analyzer (Mettler Toledo, model TGA/SDTA 851e) with the following parameters: 40ml/min purge gas (N)₂) (ii) a The scanning range is 30-250 ℃, and the scanning speed is 10 ℃/min.

X-ray diffraction patterns were obtained using an X-ray powder diffractometer (Bruker-axs, model No. advanced D8) with the parameters: voltage 40kV, current 40.0mA, scan range (2 θ)3.7-30 °, scan step 0.01 °, total scan time 33 minutes, VANTEC detector, anti-scatter slit 1 mm.

Dynamic moisture sorption analysis (DVS) was performed at 26 ℃.

The thermal study results of form I and form II are shown in fig. 12-19.

h) Preparation of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride form I

This example describes the preparation of crystalline form I of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride.

The first method comprises the following steps: 611.7mg of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide in the form of the free base are dissolved in 1.97mL of acetone at 35 ℃. A solution of 5% HCl in acetone-water was prepared by diluting 37.5% aqueous HCl with acetone. 0.6ml of this 5% HCl solution was slowly added to the above solution, followed by 1.2ml of EtOH ASDQ (100: 10 ethanol: methanol). Within a few minutes, the solution turned milky and stirred for about 5 minutes. 0.25ml of 5% HCl was slowly added. After 5 minutes, 0.25ml 5% HCl was slowly added. After 5 minutes, 0.087ml of 5% HCl was slowly added. The mixture was heated to about 40-50 ℃. The mixture was stirred at room temperature overnight. The crystals are filtered off, washed with 2ml of acetone and dried at 45 ℃ for about 7 hours. 505mg of solid were obtained.

The second method comprises the following steps: 377mg of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide in the form of the free base are dissolved in 1.2ml of acetone at 35 ℃. 0.754ml ethanol ASDQ (100: 10 ethanol: methanol) was added. A solution of 5% HCl in acetone-water was prepared by diluting 37.5% aqueous HCl with acetone. 0.18ml of a diluted HCl solution was slowly added to the above solution, followed by slow addition of seed crystals of 5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide hydrochloride form I. Slowly add 0.18ml of dilute HCl solution. After about 2 minutes, 0.18ml of dilute HCl solution was slowly added. After about 2 minutes, 0.18ml of dilute HCl solution was slowly added. The mixture was heated to about 40-50 ℃ and then stirred at room temperature overnight. The crystals were filtered off, washed with 1.5ml of acetone and dried at 45 ℃ for about 6 hours.

Example 5

5-piperidin-1-yl-pentanoic acid [5- (3-bromo-phenyl) -2H-pyrazol-3-yl ] -amide

a)3- (3-bromo-phenyl) -3-chloro-acrylonitrile

To 30.9mL of dry DMF (400mmol) cooled to 0 ℃ was added dropwise 18.3mL of POCl₃(200mmol), the temperature was kept below 10 ℃. 19.9g (100mmol) of 1- (3-bromophenyl) ethanone was added dropwise to the mixture, and the reaction was warmed to room temperature.

After the addition was complete, the reaction was stirred for a further 30 minutes, then 2.7g (40mmol) of hydroxylamine hydrochloride were added and the reaction was heated to 50 ℃. The heat source was removed and 27g (400mmol) of hydroxylamine hydrochloride (to a temperature not exceeding 120 ℃ C.) were added in portions.

After the last portion was added, the reaction was stirred until the temperature of the mixture naturally dropped to 25 ℃. Water (100mL) was then added and the mixture was extracted with ether. Na for organic phase₂SO₄Drying, and concentrating under reduced pressure. The crude product was used in the next step without further purification.

C₉H₅BrClN

¹H-NMR(400MHz，DMSO-d₆)：7.03(s，1H)，7.44-7.54m，1H)，7.72-7.84(m，2H)，8.00(br s，1H)

The yield thereof is 68%

b)5- (3-bromo-phenyl) -2H-pyrazol-3-ylamine

To a solution of 3- (3-bromo-phenyl) -3-chloro-acrylonitrile (10mmoL) in anhydrous EtOH (20mL) was added hydrazine monohydrate (1mL, 20mmoL) and the reaction was heated to reflux for 4 hours. The reaction mixture was then cooled to room temperature and the solvent was evaporated under reduced pressure. The residue was triturated with ether to give 1.8g of the title compound as a pure product (yield 54%).

C₉H₈BrN₃

¹H-NMR(400MHz，DMSO-d₆): 4.58, 5.03(1H, 2 tautomeric peaks), 5.64, 5.84(1H, 2 tautomeric peaks), 7.28(1H, s), 7.35(1H, s), 7.53-7.65(1H, m), 7.77(1H, s), 11.56, 11.97(1H, 2 tautomeric peaks).

c) 5-piperidin-1-yl-pentanoic acid [5- (3-bromo-phenyl) -2H-pyrazol-3-yl ] -amide

To a solution of 5-bromo-valeryl chloride (500 μ L, 3.74mmol) in 5mL DMA cooled to 0 ℃ was added a solution of 5- (3-bromo-phenyl) -2H-pyrazol-3-ylamine (890mg, 3.74mmol) in 3mL DMA and the reaction was stirred at 0 ℃ for 1 hour. After the reaction was complete, the reaction was diluted with 5mL and the product was extracted with 20mL of DCM. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The oily product containing DMA was used in the next step without further purification, assuming 100% yield.

To 5-bromo-pentanoic acid [5- (3-bromo-phenyl) -2H-pyrazol-3-yl]-amide (ca. 3.74mmol) in 10ml DMF, Na was added₂CO₃(1.23g, 7.48mmol), piperidine (738. mu.L, 7.48mmol) and NaI (561mg, 3.74mmol), and the mixture was heated at 60 ℃ for 5 hours. At the end of the reaction, the solvent was removed under reduced pressure and the residue was diluted with DCM and washed with saturated sodium bicarbonate solution. The organic phase was dried over sodium sulfate and then concentrated under reduced pressure. The crude product was purified by column on silica gel (10g) with a gradient eluent of 100% DCM to DCM-NH₃(2N MeOH) 95: 5 to give the title compound (1.2g, 79% yield).

C₁₉H₂₅BrN₄O

Mass (calculated value) [405 ]](ii) a (found value) [ M + H [)⁺]＝405-407

LC Rt 2.48, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆)：1.24-1.70(10H，m)，2.06-2.41(6H，m)，3.15-3.17(2H，m)，6.96(1H，s)，7.29-7.45(1H，m)，7.46-7.57(1H，m)，7.63-7.83(1H，m)，7.94(1H，s)，10.43(1H，s)，12.89(1H，s)。

Example 6

5-piperidin-1-yl-pentanoic acid [5- (1H-indol-5-yl) -2H-pyrazol-3-yl ] -amide

a) 1-Triisopropylsilyl-1H-indole-5-carboxylic acid methyl ester

To a solution of 1g methyl indole-5-carboxylate (5.7mmol) in 10mL dry DMF was added 273mg NaH (50-60% dispersion in mineral oil, 5.7mmol) and the mixture was cooled to 0 ℃. Triisopropylchlorosilane (1.06g, 5.7mmol) was added dropwise, and LC-MS showed complete conversion of the starting material to the title compound after 1 hour. The mixture was diluted with 30mL of DCM and washed with saturated sodium carbonate solution. The organic phase was dried over sodium sulfate and then concentrated under reduced pressure. The crude product was purified by silica gel column eluting with n-hexane to give the title compound (500mg, yield 26%).

C₁₉H₂₉NO₂Si

Mass (calculated value) [331 ]](ii) a (found value) [ M + H [)⁺]＝332

LC Rt ═ 3.39, 100% (5 min method)

¹H-NMR：(DMSO-d₆)：1.06(d，18H，J＝7.52)，1.75(quin，3H，J＝7.52)，6.75(m，1H)，7.48(m，1H)，7.60(m，1H)，7.72(m，1H)，8.25(s，1H)。

b) 3-oxo-3- (1-triisopropylsilyl-1H-indol-5-yl) -propionitrile

393 mu L of anhydrous CH₃A solution of CN (7.5mmol) in 6mL of dry toluene was cooled to-78 deg.C, and 5.35mL of a solution of butyllithium in hexane (1.6N) was added dropwise thereto. The mixture was stirred at-78 ℃ for 20 minutes, then a solution of 500mg of methyl 1-triisopropylsilyl-1H-indole-5-carboxylate (1.5mmol) in 2mL of dry toluene was added and the reaction was warmed to room temperature. After about 20 minutes the reaction was complete, the mixture was cooled to 0 ℃ and adjusted to pH 2 by addition of HCl 2N. The organic phase was separated, dried over sodium sulfate and concentrated under reduced pressure to yield 490mg of the title product, which was used in the next step without further purification (yield 96%).

C₂₀H₂₈N₂OSi

Mass (calculated value) [340](ii) a (found value) [ M + H [)⁺]＝341[M-H⁺]＝339

LC Rt ═ 3.10, 89% (5 min method)

¹H-NMR：(DMSO-d₆)：1.06(18H，d，J＝7.52)，1.76(3H，quin，J＝7.52)，4.76(1H，d)，7.78-7.81(1H，m)，7.48-7.52(1H，m)，7.60-7.73(2H，m)，8.25(s，1H)。

c)5- (1H-indol-5-yl) -2H-pyrazol-3-ylamine

To a solution of 3-oxo-3- (1-triisopropylsilyl-1H-indol-5-yl) -propionitrile (490mg, 1.44mmol) in 15mL of anhydrous EtOH was added 720. mu.L of hydrazine monohydrate (14.4mmol) and the reaction was refluxed for 18 hours. LC-MS showed complete conversion to aminopyrazole and complete deprotection of the silyl group. The reaction mixture was concentrated under reduced pressure and purified by silica gel column (gradient eluent 100% DCM to DCM: MeOH 9: 1) to give the title compound (120mg, yield: 41%).

C₁₁H₁₀N₄

Mass (calculated value) [198](ii) a (found value) [ M + H [)⁺]＝199

LC Rt ═ 0.84, 100% (3 min method)

d) 5-piperidin-1-yl-pentanoic acid [5- (1H-indol-5-yl) -2H-pyrazol-3-yl ] -amide

A solution of 5-bromovaleryl chloride (80. mu.L, 0.60mmol) in DMA (1mL) was cooled to 0 ℃ and a solution of 5- (1H-indol-5-yl) -2H-pyrazol-3-ylamine (120mg, 0.60mmol) and diisopropylethylamine (104. mu.L, 1.20mmol) in DMA (2mL) was added thereto. The reaction was stirred at 0 ℃ for 1h, then piperidine (119 μ L, 1.20mmol) and NaI (90mg, 0.60mmol) were added and the mixture was heated at 60 ℃ for 5h, at which time LC-MS showed complete conversion of the bromo intermediate and the solvent was removed under reduced pressure.

The residue was dissolved in DCM (2mL) and washed with saturated aqueous sodium carbonate solution. The organic phase was concentrated under reduced pressure and the crude product was purified by preparative HPLC.

Yield: 22 percent

C₂₁H₂₇N₅O

Mass (calculated value) [365 ]](ii) a (found value) [ M + H [)⁺]＝366

LC Rt ═ 1.49, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄)：1.47-1.91(10H，m)，2.44-2.56(2H，m)，2.80-3.01(2H，m)，3.07-3.17(2H，m)，3.40-3.60(2H，m)，6.48-6.51(1H，m)，6.76(1H，s)，7.26-7.30(1H，m)，7.40-7.44(2H，m)，7.86(1H，s)，8.28(1H，s，HCOOH)

Example 7

5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid (5-pyridin-3-yl-2H-pyrazol-3-yl) -amide

a) 3-oxo-3-pyridin-3-yl-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A1).

¹H-NMR(400MHz，MeOH-d₄)：9.07(1H，d)，8.81(2H，dd)，8.26(1H，dt)，7.59(1H，dd)，4.79(2H，s)。

b) 5-pyridin-3-yl-2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2).

The crude product was purified over a silica gel column (5g) with gradient eluent: 100% DCM to DCM-NH₃(2N MeOH solution) 95: 5. The title compound was obtained (371mg, 68% yield).

¹H-NMR(400MHz，MeOH-d₄)：8.82(1H，d)，8.41(1H，dd)，7.98(1H，dt)，7.37(1H，dd)，5.82(2H，s)

c)5- (4-acetyl- [1, 4] diazepan-1-yl) -pentanoic acid (5-pyridin-3-yl-2H-pyrazol-3-yl) -amide

The product was prepared according to the general synthetic procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides. The crude product was purified over a silica gel column (5g) with gradient eluent: 100% DCM to DCM-NH₃(2N MeOH solution) 95: 5.

The crude product was further purified by preparative HPLC to give 772mg of pure product (25% yield).

C₂₀H₂₈N₆O₂

Mass (calculated) [384](ii) a (found value) [ M + H [)⁺]＝385

LC Rt ═ 1.91, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄): 8.89(1H, d), 8.49(1H, dd), 8.12(1H, d), 7.48(1H, dd), 6.81(1H, broad peak), 3.60(1H, m), 3.55(3H, m), 2.72(3H, m), 2.63(1H, m), 2.55(2H, m), 2.43(2H, m), 2.07(3H, s), 1.90(1H, m), 1.80(1H, m), 1.70(m, 2H), 1.57(2H, m).

Example 8

5-piperidin-1-yl-pentanoic acid [5- (4-methoxy-phenyl) -4-methyl-2H-pyrazol-3-yl ] -amide

a)3- (4-methoxy-phenyl) -2-methyl-3-oxo-propionitrile

The crude product was purified over a silica gel column (10g) with gradient eluent: 100% hexane to hexane-AcOEt 7: 3 to give 1.43g of pure product (31% yield).

¹H-NMR(400MHz，MeOH-d₄)：7.97(2H，d)，6.98(1H，d)，4.31(1H，q，J＝7.3Hz)，3.89(3H，s)，1.63(3H，d，J＝7.3Hz)。

b)5- (4-methoxy-phenyl) -4-methyl-2H-pyrazol-3-ylamine

The crude product was purified over a silica gel column (10g) with gradient eluent: 100% DCM to DCM-MeOH 8: 2 to give 1.0g of the pure product (65% yield).

¹H-NMR(400MHz，CDCl₃)：7.37(2H，d)，6.97(2H，d)，3.84(3H，s)，2.03(3H，s)。

c) 5-piperidin-1-yl-pentanoic acid [5- (4-methoxy-phenyl) -4-methyl-2H-pyrazol-3-yl ] -amide

The product was prepared according to the general synthetic procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides.

The crude product was purified over a silica gel column (2g) with gradient eluent: 100% DCM to DCM-NH₃(2N MeOH solution) 95: 5.

The crude product was purified again by preparative HPLC to give 54mg of pure product (7% yield).

C₂₁H₃₀N₄O₂

Mass (calculated value) [370 ]](ii) a (found value) [ M + H [)⁺]＝371

LC Rt ═ 1.61, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆): 9.57(1H, s), 8.12(1H, s), 7.47(2H, d), 7.02(2H, d), 3.78(3H, s), 2.41(4H, broad), 2.37(2H, m), 2.29(2H, t), 1.91(3H, s), 1.57(2H, m), 1.50(6H, m), 1.38(2H, m).

Example 9

5-piperidin-1-yl-pentanoic acid (5-furan-2-yl-2H-pyrazol-3-yl) -amide

The crude product was purified by preparative HPLC (yield 15%).

C₁₇H₂₄N₄O₂

Mass (calculated value) [316](ii) a (found value) [ M + H [)⁺]＝317

LC Rt ═ 1.53, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄)：8.48(1H，s)，7.56(1H，s)，6.70(1H，s)，6.66(1H，s)，6.52(1H，m)，5.49(1H，s)，4.88(1H，s)，3.10(2H，m)，2.48(2H，m)，1.77(10，m)。

Example 10

N- [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a) 4-piperidin-1-yl-butyric acid ethyl ester

To a solution of piperidine (5.4g, 65mmol) in toluene (15mL) was added ethyl 4-bromobutyrate (3.8mL, 26mmol) and the reaction mixture was refluxed for 10 hours. The mixture was cooled to room temperature and the white solid (piperidine bromide) was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give the title compound, which was used in the next step without further purification.

C₁₁H₂₁NO₂

Mass (calculated) [199](ii) a (found value) [ M + H [)⁺]＝200

LC Rt ═ 0.2, 100% (5 min method)

¹H-NMR(400MHz，MeOH-d₄): 1.22-1.25(3H, m), 1.46-1.47(2H, m), 1.57-1.63(4H, m), 1.78-1.84(2H, m), 2.30-2.35(4H, m), 2.42(4H, m, broad peak), 4.08-4.14(2H, m).

b) 4-piperidin-1-yl-butyric acid

To a suspension of the crude 4-piperidin-1-yl-butyric acid ethyl ester (ca. 25mmol) obtained in the above step in 15mL of water was added NaOH (1.4g, 25mmol) and the mixture was heated under reflux for 16 hours and then cooled to room temperature. The solution was acidified with HCl 6N at 0 ℃ and then concentrated under reduced pressure. The residue was treated with EtOH and the precipitated sodium chloride was filtered off. Evaporation of the solvent under reduced pressure gave 2.8g of the title compound as a white solid in 58% overall yield from steps a) and b).

C₉H₁₇NO₂

Mass (calculated value) [171](ii) a (found value) [ M + H [)⁺]＝172

LC Rt ═ 0.23, 100% (5 min method)

¹H-NMR(400MHz，DMSO-d₆): 1.44-1.51(2H, m); 1.64-1.80(6H, m); 2.22-2.25(2H, m); 2.75-2.78(2H, m, broad); 2.91-2.94(2H,m, broad peak); 3.30-3.40(2H, m).

c) N- [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

To a suspension of 4-piperidin-1-yl-butyric acid (1.32g, 7.93mmol) in 1, 2-dichloroethane (20mL) was added N, N' -carbonyldiimidazole (1.2g, 7.4mmol) and the mixture was stirred at room temperature for 2 hours (complete dissolution of the suspension was usually observed when all amino acids were activated). Then 3-amino-5- (4-methoxyphenyl) pyrazole (1g, 5.29mmol) was added and stirred for 10 hours. At the end of the reaction (LC-MS monitoring), the formation of two isomers was observed, and the mixture was heated at 50 ℃ until the less stable isomer was converted to the title compound (LC-MS monitoring). With saturated Na₂CO₃The solution was washed with solvent, extracted and then the solvent was removed under reduced pressure. The crude product was crystallized from acetonitrile to yield 1.2g of the title compound (yield: 70%).

C₁₉H₂₆N₄O₂

Mass (calculated value) [342 ]](ii) a (found value) [ M + H [)⁺]＝343

LC Rt ═ 1.54, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆)：1.34-1.40(1H，m)；1.52-1.55(1H，m)；1.62-1.75(6H，m)；1.94-1.98(2H，m)；2.37-2.40(2H，m)；2.81-2.88(2H，m)；2.97-3.03(2H，m)；3.39-3.42(2H，m)；3.77(3H，s)；6.77(1H，s)；6.98(2H，d，J＝8.8Hz)；7.61(2H，d，J＝8.8Hz)；10.47(1H，s)，12.66(1H，s)。

Example 11

N- [5- (3-methoxy-phenyl) -1H-pyrazol-3-yl ] -4-morpholin-4-yl-butyramide

a)3- (3-methoxy-phenyl) -3-oxo-propionitrile

Under nitrogenTo a solution of commercially available ethyl 3-methoxy-benzoate (3.2g, 18mmol) in dry toluene (25mL) was carefully added NaH (50-60% dispersion in mineral oil, 1.44g, 36 mmol). Heating the mixture at 90 deg.C, and adding anhydrous CH dropwise₃CN (4.45mL, 85.2 mmol). After heating the reaction for 18 hours, the product precipitated from the reaction mixture as the sodium salt. The reaction was allowed to cool to room temperature, the resulting solid was filtered off, washed with diethyl ether, then redissolved in water and the solution was acidified to pH 3 with 2N HCl, at which time the title compound was observed to precipitate. The solid was filtered off from the aqueous solution to yield 1.57g of the title product (50% yield).

C₁₀H₉NO₂

Mass (calculated) [175](ii) a (found value) [ M + H [)⁺]＝176

LC Rt ═ 1.69, 94% (5 min method)

b)5- (3-methoxy-phenyl) -2H-pyrazol-3-ylamine

To a solution of 3- (3-methoxy-phenyl) -3-oxo-propionitrile (8.96mmoL) in anhydrous EtOH (20mL) was added hydrazine monohydrate (0.52mL, 15mmoL) and the reaction was heated to reflux for 18 h. The reaction mixture was then cooled to room temperature and the solvent was evaporated under reduced pressure. The crude product was treated with diethyl ether and then filtered to give 1.4g of the title product (83% yield).

C₁₀H₁₁N₃O

Mass (calculated) [ 189)](ii) a (found value) [ M + H [)⁺]＝190

LC Rt ═ 1.13, 100% (5 min method)

¹H-NMR(400MHz，MeOH-d₄)：3.82(3H，s)；5.93(1H，s)；6.86-6.88(1H，m)；7.19-7.31(3H，m)。

c) N- [5- (3-methoxy-phenyl) -1H-pyrazol-3-yl ] -4-morpholin-4-yl-butyramide

Under nitrogen, underA solution of 4-bromobutyryl chloride (0.104mL, 0.9mmol) in dry DMA (1mL) was cooled to-10 deg.C (ice water bath) and a solution of 5- (3-methoxy-phenyl) -2H-pyrazol-3-ylamine (170mg, 0.9mmol) and diisopropylethylamine (0.315mL, 1.8mmol) in dry DMA (1mL) was added. The starting material is completely converted into the intermediate 4-bromo-N- [5- (3-methoxy-phenyl) -1H-pyrazol-3-yl]-butanamide (monitored by LC-MS), morpholine (0.079mL, 0.9mmol) was added and the mixture was heated at 60 ℃ for 16 h. The residue was dissolved in DCM (2mL) and saturated Na₂CO₃And (4) washing the solution. The organic phase was concentrated under reduced pressure and the crude product was purified by column on silica gel (gradient eluent acetonitrile 100% to MeCN/MeOH, NH)₃90/10). Fractions containing the title compound were collected to yield 17mg (5.5% yield).

C₁₈H₂₄N₄O₃

Mass (calculated value) [344 ]](ii) a (found value) [ M + H [)⁺]＝345

LC Rt ═ 1.36, 95% (10 min method)

¹H-NMR(400MHz，MeOH-d₄)：1.77-1.85(2H，m)；2.34-2.40(8H，m)；3.59-3.62(4H，m)；3.76(3H，s)；6.79-6.85(2H，m)；7.15-7.29(3H，m)。

Example 12

4-azepan-1-yl-N- [5- (3-methoxy-phenyl) -1H-pyrazol-3-yl ] -butyramide

A solution of 4-bromobutyryl chloride (0.104mL, 0.9mmol) in dry DMA (1mL) was cooled to-10 deg.C (ice-water bath) under nitrogen and a solution of 5- (3-methoxy-phenyl) -2H-pyrazol-3-ylamine (170mg, 0.9mmol) and diisopropylethylamine (0.315mL, 1.8mmol) in dry DMA (1mL) was added. When the starting material was completely converted to the omega-bromoamide intermediate (monitored by LC-MS), 0.101mL of aza was addedThen stirred at 60 ℃ for 16 hours.

The residue was dissolved in DCM (2mL) and saturated Na₂CO₃And (4) washing the solution. The organic phase was concentrated under reduced pressure and the crude product was purified by column on silica gel (5g) (eluent gradient: acetonitrile 100% to MeCN/MeOH, NH)₃90/10). Fractions containing the title compound were collected and further purified by preparative HPLC to give 20mg of the title compound as formate salt (5.5% yield).

C₂₀H₂₈N₄O₂

Mass (calculated) [356 ]](ii) a (found value) [ M + H [)⁺]＝357

LC Rt ═ 1.71, 99% (10 min method)

¹H-NMR(400MHz，MeOH-d₄): 1.65-1.68(4H, m); 1.80-1.90(4H, m); 1.97-2.04(2H, m); 2.49-2.52(2H, m); 3.12-3.16(2H, m); 3.24-3.30(4H, m, broad peak); 3.75(3H, s); 6.76(1H, s); 6.82-6.85(1H, m); 6.13-6.15(2H, m); 6.23-6.27(1H, m); 8.37(1H, s, formate)

Example 13

4-azepan-1-yl-N- [5- (4-fluoro-phenyl) -2H-pyrazol-3-yl ] -butyramide

The product was prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides. Starting material was commercially available 5- (4-fluoro-phenyl) -2H-pyrazol-3-ylamine which, according to this method, after purification by preparative HPLC gave 25mg of the title compound as its formate salt form (7% yield).

C₁₉H₂₅N₄OF

Mass (calculated value) [344 ]](ii) a (found value) [ M + H [)⁺]＝345

LC Rt 1.69, 100% (10 min method).

¹H-NMR(400MHz，MeOH-d₄): 1.66-1.69(4H, m); 1.80-1.90(4H, m, broad peak); 1.97-2.05(2H, m); 2.52-2.54(2H, m); 3.12-3.18(2H, m); 3.25-3.30(4H, m, broad peak); 6.67(1H, s, broad); 7.08-7.12(2H, m); 7.59-7.63(2H, m); 8.43(1H, s, formate)

Example 14

N- [5- (6-methyl-pyridin-3-yl) -1H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a)3- (6-methyl-pyridin-3-yl) -3-oxo-propionitrile

Oxopropanonitrile is prepared according to the general synthetic procedure for the synthesis of 3-oxopropanenitrile (scheme A1).

C₉H₈N₂O

Mass (calculated value) [160](ii) a (found value) [ M + H [)⁺]＝161

LC Rt ═ 0.63, 100% (5 min method)

¹H-NMR(400MHz，DMSO-d₆)：2.55(3H，s)；4.65(2H，s)；7.43-7.45(m，1)；8.13-8.16(1H，m)；8.94-8.95(1H，m)。

b)5- (6-methyl-pyridin-3-yl) -1H-pyrazol-3-ylamine

Aminopyrazoles are prepared according to the general synthetic procedure described in scheme a 2.

C₉H₁₀N₄

Mass (calculated value) [174](ii) a (found value) [ M + H [)⁺]＝175

LC Rt ═ 0.23, 100% (5 min method)

c) N- [5- (6-methyl-pyridin-3-yl) -1H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

Prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amide, purification by preparative HPLC gave 19mg (6% yield) of the title compound as its formate salt.

C₁₈H₂₅N₅O

Mass (calculated) [327](ii) a (found value) [ M + H [)⁺]＝328

LC Rt ═ 0.33, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄): 1.40-1.90(6H, m); 2.30-2.54(5H, m); 3.05-3.09(4H, m); 3.20-3.24(2H, m); 6.72(1H, s, broad); 7.30(1H, d J ═ 8.0 Hz); 7.92-7.94(1H, m); 8.35(1H, s, formate); 8.67(1H, s).

Example 15

N- [5- (5-methyl-pyridin-3-yl) -1H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a)3- (5-methyl-pyridin-3-yl) -3-oxo-propionitrile

Oxopropanonitrile is prepared according to the general procedure for the synthesis of 3-oxopropanenitrile (scheme A1).

C₉H₈N₂O

Mass (calculated value) [160](ii) a (found value) [ M + H [)⁺]＝161

LC Rt ═ 0.63, 100% (5 min method)

¹H-NMR(400MHz，MeOH-d₄)：2.55(3H，s)；4.65(2H，s)；7.43-7.45(m，1H)；8.13-8.16(1H，m)；8.94-8.95(1H，m)。

b)5- (5-methyl-pyridin-3-yl) -1H-pyrazol-3-ylamine

C₉H₁₀N₄

Mass (calculated value) [174](ii) a (found value) [ M + H [)⁺]＝175

LC Rt ═ 0.23, 100% (5 min method)

c) N- [5- (5-methyl-pyridin-3-yl) -1H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

Prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amide, after preparative HPLC purification yielded 25mg of the title compound as its formate salt (7.4% yield).

C₁₈H₂₅N₅O

Mass (calculated) [327](ii) a (found value) [ M + H [)⁺]＝328

LC Rt ═ 0.33, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄): 1.52-1.70(2H, m, broad); 1.72-1.84(4H, m, broad); 1.98-2.06(2H, m); 2.45(3H, s); 2.48-2.54(2H, m); 3.04-3.10(4H, m); 3.20-3.24(2H, m, broad peak); 6.74(1H, s, broad); 7.88(1H, s); 7.28(1H, s); 8.37(1H, s, formate); 8.67(1H, s).

Example 16

4- (4-acetyl- [1, 4] diazepan-1-yl) -N- [5- (6-methoxy-naphthalen-2-yl) -1H-pyrazol-3-yl ] -butyramide

a) 6-methoxy-naphthalene-2-carboxylic acid methyl ester

To a solution of 6-methoxy-naphthalene-2-carboxylic acid (1.01g, 5mmol) in methanol (10mL) was added a catalytic amount of sulfuric acid. The mixture was then heated at 80 ℃ for 8 hours. After the reaction was complete (LCMS monitoring), the solution was slowly cooled and the product was observed to precipitate. The white solid was filtered off to give 1.01g (94% yield) of the title compound.

C₁₃H₁₂O₃

Mass (calculated) [216 ]](ii) a (found value) [ M + H [)⁺]＝217

LC Rt 2.43, 100% (5 min method)

b)3- (6-methoxy-naphthalen-2-yl) -3-oxo-propionitrile

To a solution of methyl 6-methoxy-naphthalene-2-carboxylate (1.0g, 4.7mmol) in dry toluene (8mL) was added NaH (0.55mg, 9.4mmol) and the mixture was heated to 90 ℃. Acetonitrile (1.2mL) was added dropwise to the hot solution. The reaction was then heated for 18 hours and the product precipitated from the mixture as the sodium salt.

The reaction was cooled to room temperature and the solid formed was first filtered off, washed with diethyl ether, then dissolved in water and acidified to pH 3 with HCl 2N, at which time the title compound was observed to precipitate. The solid was filtered off from the aqueous solution to yield 1.1g of the title compound (100% yield).

C₁₃H₁₂O₃

Mass (calculated value) [225](ii) a (found value) [ M + H [)⁺]＝226

LC Rt 2.13, 90% (5 min method)

c)5- (6-methoxy-naphthalen-2-yl) -1H-pyrazol-3-ylamine

To a solution of 3- (6-methoxy-naphthalen-2-yl) -3-oxo-propionitrile (1.1g, 4.8mmoL) in anhydrous EtOH (10mL) was added hydrazine monohydrate (0.96mL, 19.2mmoL) and the mixture was heated at reflux for 18 h. The reaction mixture was then cooled to room temperature and the solvent was evaporated under reduced pressure. The crude product was treated with diethyl ether and filtered to give 0.95g of the title compound (83% yield).

C₁₄H₁₃N₃O

Mass (calculated value) [239](ii) a (found value) [ M + H [)⁺]＝240

LC Rt ═ 1.49, 90% (5 min method)

d)4- (4-acetyl- [1, 4] diazepan-1-yl) -N- [5- (6-methoxy-naphthalen-2-yl) -1H-pyrazol-3-yl ] -butyramide

Purification by preparative HPLC according to the general procedure for the synthesis of omega-bromo-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amide and general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amide gave 15mg (3% yield) of the title compound as formate salt.

C₂₅H₃₁N₅O₃

Mass (calculated) [ 449)](ii) a (found value) [ M + H [)⁺]＝450

LC Rt ═ 1.91, 100% (10 min method)

¹H-NMR(400MHz，MeOH-d₄): 1.88-2.0(4H, m); 2.06(3H, s); 2.48-2.52(2H, m); 2.94-3.02(2H, m); 3.08-3.18(4H, m); 3.52-3.58(2H, m); 3.64-3.72(2H, m); 3.82(3H, s); 6.78-6.82(1H, m); 7.04-7.10(1H, m); 7.16-7.18(1H, m); 7.62-7.78(3H, m); 7.98-8.02(1H, m); 8.28(1H, s, formate).

Example 17

5-piperidin-1-yl-pentanoic acid [5- (3-fluoro-phenyl) -1H-pyrazol-3-yl ] -amide

a)3- (3-fluoro-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure of scheme a1 with simple modifications. To a solution of methyl 3-fluorobenzoate (3g, 18mmol) in dry toluene (25mL) under nitrogen was carefully added NaH (50-60% dispersion in mineral oil, 1.44g, 36 mmol). The mixture was heated to 90 ℃ and then droppedInto dry CH₃CN (4.45mL, 85.2 mmol). The reaction was heated for 18 hours and the product precipitated from the reaction mixture as the sodium salt. The reaction was cooled to room temperature, the solid formed was filtered off, dissolved again in water and acidified with 2N HCl to pH 5-6, at which time precipitation was observed. The solid was filtered off from the aqueous solution to yield 2.12g of the title compound (72% yield), which was used directly in the next step.

b)5- (3-fluoro-phenyl) -1H-pyrazol-3-yl-amines

The product was prepared according to the general procedure of scheme a2 with simple modifications. To a solution of 3- (3-fluoro-phenyl) -3-oxo-propionitrile (1.92g, 11.77mmoL) in anhydrous EtOH (32mL) was added hydrazine monohydrate (0.685mL, 14.12mmoL) and the reaction was refluxed for 2 hours. The reaction mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. The crude product was treated with diethyl ether and filtered to give 1.71g of the title compound (82% yield).

C₉H₈FN₃

Mass (calculated) [177 ]](ii) a (found value) [ M + H [)⁺]＝190

LC Rt ═ 1.13, 69% (5 min method)

c) 5-piperidin-1-yl-pentanoic acid [5- (3-fluoro-phenyl) -1H-pyrazol-3-yl ] -amide

The product was prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides. A solution of 5-bromovaleryl chloride (0.125mL, 0.94mmol) in dry DMA (1mL) was cooled to-10 deg.C (ice water bath) under nitrogen and a solution of 5- (3-fluoro-phenyl) -2H-pyrazol-3-ylamine (177mg, 0.94mmol) and diisopropylethylamine (0.324mL, 1.88mmol) in dry DMA (1mL) was added.

The reaction was stirred at 0 ℃ for 1h, then piperidine (0.232mL, 2.35mmol) and NaI (141mg, 0.94mmol) were added. The reaction mixture was heated at 60 ℃ until LC-MS showed complete conversion of the bromo intermediate, at which time the reaction was cooled, the solvent was removed under reduced pressure, the residue was dissolved in DCM (2mL) and saturated Na was used₂CO₃And (4) washing the solution. The organic phase was concentrated under reduced pressure and the crude product was purified by silica gel column (gradient eluent: 100% DCM to DCM-NH)₃MeOH 2N solution 8: 2), then purified by preparative HPLC. Fractions containing the title compound were collected to yield 15mg (4.4% yield) of formate salt.

C₁₉H₂₅FN₄O

Mass (calculated value) [344 ]](ii) a (found value) [ M + H [)⁺]＝345

LC Rt ═ 1.64, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆): 1.37-1.58(10H, m); 2.27-2.31(2H, m); 2.35-2.44(6H, m); 6.85(1H, s); 7.14(1H, t, J ═ 8.6 Hz); 7.45(1H, m), 7.53-7.55(2H, m); 8.21(1H, s, formate); 10.47(1H, s).

Example 18

5-azepan-1-yl-pentanoic acid (5-pyridin-4-yl-1H-pyrazol-3-yl) -amide 3-oxo-3-pyridin-4-yl-propionitrile

The product was prepared according to the general procedure of scheme a1 with simple modifications. To a solution of 3g (22mmol) of methyl isonicotinate in dry toluene (30mL) under nitrogen was carefully added NaH (50-60% dispersion in mineral oil, 1.75g, 44 mmol). Heating the mixture to 90 deg.C, and adding dry CH dropwise₃CN (5.39mL, 103 mmol). The reaction was heated for 18 h and the product precipitated from the reaction mixture as sodium salt, the reaction was cooled to room temperature, the solid was filtered off, redissolved in water, adjusted to pH 5-6 with 6N HCl solution and the product was extracted with DCM. The aqueous phase was again adjusted to pH 4-5 and extracted again with DCM to give more product.

The organic phases were combined, dried and evaporated. The product was used directly in the next step with 58% yield of crude product.

b) 5-pyridin-4-yl-1H-pyrazol-3-ylamine

The product was prepared according to the general procedure of scheme a2 with simple modifications. To a solution of 3-oxo-3-pyridin-4-yl-propionitrile (1.86g, 12.74mmoL) in anhydrous EtOH (35mL) was added hydrazine monohydrate (0.74mL, 15.29mmoL) and the reaction was heated at reflux for 2 h. The reaction mixture was then cooled to room temperature and the solvent was evaporated under reduced pressure. The crude product was washed with diethyl ether to give the title compound (yield: 39%).

C₈H₈N₄

Mass (calculated value) [160](ii) a (found value) [ M + H [)⁺]＝161

LC Rt ═ 0.23, 100% (5 min method)

¹H-NMR(400MHz，DMSO-d₆)：5.02(2H，s)；5.85(1H，s)；7.59(2H，d，J＝6Hz)；8.50(2H，d，J＝6Hz)；11.93(1H，s)。

c) 5-azepan-1-yl-pentanoic acid (5-pyridin-4-yl-1H-pyrazol-3-yl) -amide

The product was prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides. A solution of 5-bromovaleryl chloride (0.125mL, 0.94mmol) in dry DMA (1mL) was cooled to-10 deg.C (ice water bath) under nitrogen and a solution of 5-pyridin-4-yl-1H-pyrazol-3-ylamine (151mg, 0.94mmol) and diisopropylethylamine (0.324mL, 1.88mmol) in dry DMA (1mL) was added. The reaction was stirred at 0 ℃ for 1h, then azepane (0.265mL, 2.35mmol) and NaI (0.94mmol, 1eq) were added.

The reaction mixture was heated to 60 ℃ until LC-MS analysis showed complete conversion of the bromo intermediate, at which time the reaction was cooled and the solvent was evaporated under reduced pressure. The residue was dissolved in DCM (2mL) and saturated Na₂CO₃The solution was washed, the organic phase was evaporated under reduced pressure and the crude product was purified by silica gel column (eluent gradient: 100% DCM to DCM-NH)₃MeOH 2N solution 8: 2), the fractions containing the title compound were collected (30mg, 8.8% yield).

C₁₉H₂₇N₅O

Mass (calculated value) [341 ]](ii) a (found value) [ M + H [)⁺]＝342

LC Rt ═ 0.23, 100% (10 min method)

¹H-NMR(400MHz，DMSO-d₆)：1.58-1.75(12H，m)；2.34-2.37(2H，t，J＝6.6Hz)；3.05-3.09(4H，m)；3.31(2H，m)；7.09(1H，s)；7.68(2H，d，J＝4.8Hz)；8.59(2H，d，J＝4Hz)；9.14(1H，s)；10.52(1H，s)；13.17(1H，s)。

Example 19

6- (4-acetyl- [1, 4] diazepan-1-yl) -hexanoic acid [5- (4-methoxy-phenyl) -1H-pyrazol-3-yl ] -amide

The product was prepared according to the general procedure for the one-pot synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides. A solution of 5-bromohexanoyl chloride (0.144mL, 0.94mmol) in dry DMA (1mL) was cooled to-10 deg.C (ice-water bath) under nitrogen and a solution of 5- (4-methoxy-phenyl) -1H-pyrazol-3-ylamine (178mg, 0.94mmol) and diisopropylethylamine (0.324mL, 1.88mmol) in dry DMA (1mL) was added.

The reaction mixture was stirred at 0 ℃ for 1 hour, and then 1- [1, 4] was added]Diazepan-1-yl-ethanone (0.310mL, 2.35mmol) and NaI (0.94mmol, 1 eq). The reaction mixture was heated at 60 ℃ until LC-MS analysis showed complete conversion of the bromo intermediate at which time the reaction was cooled and the solvent removed under reduced pressure. The residue was dissolved in DCM (2mL) and saturated Na₂CO₃And (4) washing the solution.

The organic phase was concentrated under reduced pressure and half of the crude product was purified by silica gel column (eluent gradient: 100% DCM to DCM-NH)₃MeOH 2N solution 8: 2). Fractions containing the title compound (35mg) were collected.

C₂₃H₃₃N₅O₃

Mass (calculated) [427 ]](ii) a (found value) [ M + H [)⁺]＝428

LC Rt ═ 1.61, 96% (10 min method)

¹H-NMR(400MHz，DMSO-d₆)：1.24-1.29(2H，m)；1.36-1.44(2H，m)；1.54-1.58(2H，m)；1.62-1.76(2H，m)；1.94-1.96(3H，m)；2.25-2.28(2H，m)；2.35-2.41(2H，m)；2.51-2.54(2H，m)；2.60-2.62(1H，m)；3.38-3.44(5H，m)；3.77(3H，s)；6.73(1H，s)；6.98(2H，d，J＝8.8Hz)；7.61(2H，d，J＝8.8)；10.32(1H，s)

Example 20

N- [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -2-methyl-4-piperidin-1-yl-butyramide

a) 4-bromo-2-methyl-butyric acid methyl ester

4-bromo-2-methyl-butyric acid (2.16g, 1eq, prepared as described in J.Am.chem.Soc.1990, 112, 2755) was dissolved in MeOH (10mL) and a few drops of concentrated H were added₂SO₄. The reaction was refluxed with stirring for 16 hours. Monitoring by LC-MS showed that after the reaction was complete, the methanol was removed under reduced pressure, the oily residue was diluted with water, the pH was adjusted to 9 with 10% NaOH, the product was extracted with diethyl ether (2X20mL), Na was added₂SO₄And (5) drying. After removal of the solvent the title compound was obtained as a colourless oil (1.29g, 55% yield).

C₆H₁₁BrO₂

NMR(400MHz，CDCl₃)；1.19(3H，d)；1.94-1.89(2H，m)；2.29-2.23(2H，m)；3.43-3.40(1H，m)；3.69(3H，s)。

b) 2-methyl-4-piperidin-1-yl-butyric acid, HCl

Methyl-4-bromo-2-methyl-butyric acid (1.29g, 1eq) was dissolved in toluene (15mL), piperidine (1.07mL, 3eq) was added, and the reaction was stirred for 3 hours. Monitoring by LC-MS indicated the end of the reaction, toluene was removed under reduced pressure and the crude ester was dissolved in 1M NaOH (14mL, 1.1eq) and MeOH (2 mL). The reaction was refluxed for 16 hours with stirring, and after completion of hydrolysis, concentrated under reduced pressure and adjusted to pH 4 with 6N HCl. EtOH was added to promote NaCl precipitation. The organic phase was filtered and the EtOH removed under reduced pressure. The resulting oil was treated with a solution of 2M HCl in ether to give 2-methyl-4-piperidin-1-yl-butyric acid HCl (0.96g, 66% yield).

C₁₀H₁₉NO₂

Mass (calculated value) [185.27](ii) a (found value) [ M + H [)⁺]＝186.27

LC Rt ═ 0.23, 95% (5 min method)

c) N- [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -2-methyl-4-piperidin-1-yl-butyramide

2-methyl-4-piperidin-1-yl-butyric acid HCl (0.45g, 1.2eq) was suspended in 1, 2-DCE (15mL), triethylamine (0.29mL, 1.2eq) was added, 1' -carbonyldiimidazole (0.303g, 1.1eq) was added in one portion, and the reaction was stirred at room temperature for 2 hours. Then 5- (4-methoxy-phenyl) -2H-pyrazol-3-ylamine (0.325g, 1eq) was added and stirring was continued at room temperature for 16 hours. After completion of the reaction, as monitored by LC-MS, the solvent was removed under reduced pressure and the crude amide was purified by column chromatography (Flash-SI 10 g; CH)₃CN∶MeOH 9∶1，CH₃CN∶2N NH₃MeOH 9: 1) to give the title compound as a colorless viscous oil (0.120g, 0.33 mmol).

C₂₀H₂₈N₄O₂

Mass (calculated value) [356.48](ii) a (found value) [ M + H [)⁺]＝357.25

LC Rt ═ 1.67, 97% (10 min method)

NMR(400MHz，DMSO-d₆)；1.18(3H，d)；1.35-1.31(2H，m)；1.46-1.41(4H，m)；1.77-1.72(1H，m)；2.19-2.16(2H，m)；2.27-2.23(4H，m)；2.61-2.58(2H，m)；3.76(3H，s)；6.76(1H，s)；6.92(2H，d)；7.61(2H，d)；10.33(1H，s)。

Example 21

N- [4- (4-methoxy-phenyl) -1H-imidazol-2-yl ] -4-piperidin-1-yl-butyramide

To a suspension of 4-piperidin-1-yl-butyric acid (200mg, 1.17mmol, 1.0eq) in 1, 2-dichloroethane (2mL) was added N, N' -carbonyldiimidazole (179.9mg, 1.11mmol, 0.95eq) and the mixture was stirred at room temperature for 1 hour until the amino acid was fully activated and the suspension was completely dissolved. 4- (4-methoxy-phenyl) -1H-imidazol-2-ylamine (prepared as described in JOC 1994, 59, 24, 7299; 110.5g, 0.58mmol, 0.50eq) was added and stirred at 50 ℃ for 1 day. The slow conversion process was monitored by LC-MS. A further portion of activated acid (4-piperidin-1-yl-butyric acid, 200mg and carbonyldiimidazole, 179.9mg in 2mL of 1, 2-dichloroethane) was added and stirring was continued at 50 ℃ for 2 days.

The solvent was removed under reduced pressure and the crude product mixture was purified by preparative HPLC to give a 9: 1 mixture of product and unreacted 4- (4-methoxy-phenyl) -1H-imidazol-2-ylamine. The crude product was treated with an isocyanate resin and an SCX column to purify it, to obtain 78.0mg (yield: 39%) of the title compound as a white solid.

C₁₉H₂₆N₄O₂Mass (calculated value) [342 ]](ii) a (found value) [ M + H [)⁺]＝343

LC Rt ═ 1.00 (and developing solvent front), 99% (10 min method)

¹H-NMR(400MHz，DMSO)：1.30-1.36(2H，m)；1.43-1.49(4H，m)；1.67-1.75(2H，m)；2.22-2.34(8H，m)；3.73(3H，s，-OCH3)；6.87(2H，d，J＝8.8Hz)；7.10(1H，s)；7.60(2H，d，J＝8.8Hz)；11.26(1H，s，NHCO)，11.52(1H，s，NH)。

¹³C-NMR(400MHz，DMSO)：21.54(1C)；23.63(1C)；24.92(2C)；33.24(1C)；53.6(1C，-OCH₃)；55.02(2C)；57.46(1C)；113.88(2C)；125.18(2C)，141.13(1C)；157.67(1C)；162.33(2C)；163.66(1C)；171.15(1C，CO)。

Example 22

N- (4-methyl-5-o-tolyl-2H-pyrazol-3-yl) -4-pyrrolidin-1-yl-butyramide

a) 2-methyl-3-oxo-3-o-tolyl-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A1). A mixture of methyl 2-methylbenzoate (3.0mL, 20.0mmol, 1.0eq) and NaH (1.6g, 40.0mmol, 2.0eq) in dry toluene (20mL) was heated at 80 deg.C, propionitrile (6.7mL, 94.4mmol, 4.7eq) was added dropwise, and the reaction was heated for 18 hours. The crude product was dissolved in water and extracted with DCM and used in the next step without further purification (3.04g, yield: 88%).

C₁₁H₁₁NO

¹H-NMR(DMSO-d₆)：1.82(3H，s)；2.26(3H，s)；2.48-2.49(1H，m)；7.10-7.42(4H，m)。

b) 4-methyl-5-o-tolyl-2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a silica gel column (20g) with gradient eluent: 100% Ethyl acetate (EtOAc) to EtOAc-MeOH 80: 20. The title product was obtained (1.2g, 37% yield).

C₁₁H₁₃N₃

Mass (calculated value) [187](ii) a (found value) [ M + H [)⁺]＝188。

LC Rt 1.33min, 100% (10 min method)

¹H-NMR(DMSO-d₆)：1.68(3H，s)；2.17(3H，s)；4.36(2H，br s)；7.14(1H，d，J＝7.2Hz)；7.20-7.26(3H，m)；11.24(1H，br s)。

c) N- (4-methyl-5-o-tolyl-2H-pyrazol-3-yl) -4-pyrrolidin-1-yl-butyramide

To the suspension of 4-pyrrolidin-1-yl-butyric acid (118.0mg, 0.8mmol, 1.5eq) in 1, 2-dichloroethane (3mL) was added N, N' -carbonyldiimidazole (113.0mg, 0.7mmol, 1.4eq) and the mixture was stirred at room temperature for 1 hour, then N, N-diisopropylethylamine (87 μ L, 0.5mmol, 1.0eq) was added and the mixture was stirred at room temperature for another 1 hour until the suspension was completely dissolved. 4-methyl-5-o-tolyl-2H-pyrazol-3-ylamine (93.5mg, 0.5mmol, 1.0eq) was added and stirred for 18 hours, then at 50 ℃ for 1 day until the less stable ring nitrogen acylated isomer was converted to the title compound (monitored by LC-MS). The solvent was removed under reduced pressure, and the crude product was purified by means of a silica gel column to give 44.0mg of the title compound (yield: 27%).

C₁₉H₂₆N₄O

Mass (calculated) [ 326)](ii) a (found value) [ M + H [)⁺]＝327，[M+2/2]＝164。

LC Rt ═ 1.56min, 95% (10 min method)

¹H-NMR(CD₃OD)：1.83(3H，s)；2.07-2.11(6H，m)；2.22(3H，s)；2.62(2H，t，J＝7.2Hz)；3.27-3.39(6H，m)；7.22-7.28(2H，m)；7.32-7.34(2H，m)。

Example 23

N- [5- (4-cyclopropylmethoxy-3-fluoro-phenyl) -2H-pyrazol-3-yl ] -4-pyrrolidin-1-yl-butyramide

a) 3-fluoro-4-hydroxy-benzoic acid methyl ester

3-fluoro-4-hydroxy-benzoic acid (5g, 32.0mmol) was dissolved in MeOH (50mL), a catalytic amount of sulfuric acid (1mL) was added and the mixture was refluxed overnight, then the solvent was evaporated under reduced pressure. The crude product was dissolved in DCM and washed with saturated sodium bicarbonate solution to basic pH. The organic phase was dried and evaporated under reduced pressure and the residue was used in the next step without further purification (yield 85%).

C₈H₇FO₃

¹H-NMR(DMSO-d₆)：3.78(3H，s)；7.00-7.02(1H，m)；7.61-7.64(2H，m)；10.89(1，br s)。

b) 4-Cyclopropylmethoxy-3-fluoro-benzoic acid methyl ester

3-fluoro-4-hydroxy-benzoic acid methyl ester (1.02g, 6.0mmol, 1.0eq) was dissolved in acetone (14mL), NaI (0.45g, 3.0mmol, 0.5eq) and K were added₂CO₃(1.66g, 12.0mmol, 2.0eq) and the mixture was stirred at room temperature for 20 min. (bromomethyl) cyclopropane (0.53mL, 5.4mmol, 0.9eq) was added and the mixture was refluxed for 2 days. The solvent was concentrated under reduced pressure, NaOH 10% was added, extracted with DCM and dried.

0.91g of the title product (69% yield) was obtained and used in the next step without further purification.

C₁₂H₁₃FO₃

¹H-NMR(DMSO-d₆)：0.34-0.37(2H，m)；0.57-0.62(2H，m)；1.22-1.26(1H，m)；3.82(3H，s)；3.99(2H，d，J＝6.8Hz)；7.26(1H，t，J＝8.4Hz)；7.67-7.77(2H，m)。

c)3- (4-Cyclopropylmethoxy-3-fluoro-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole starting from 4-cyclopropylmethoxy-3-fluoro-benzoic acid methyl ester (scheme A1 bis). 0.84g of the title compound was extracted from water and dried over sodium sulfate (88% yield) and used directly in the next step.

C₁₃H₁₂FNO₂

d)5- (4-cyclopropylmethoxy-3-fluoro-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a silica gel column with gradient eluent: 100% Ethyl acetate to EtOAc-MeOH 90: 10 gave the title compound (576mg, 65% yield).

C₁₃H₁₄FN₃O

Mass (calculated value) [247](ii) a (found value) [ M + H [)⁺]＝248。

LC Rt 2.19min, 99% (10 min method)

¹H-NMR(CD3OD)：0.33-0.38(2H，m)；0.59-0.65(2H，m)；1.22-1.31(1H，m)；2.90-3.92(2H，m)；7.02-7.20(2H，m)；7.34-7.40(2H，m)。

e) N- [5- (4-cyclopropylmethoxy-3-fluoro-phenyl) -2H-pyrazol-3-yl ] -4-pyrrolidin-1-yl-butyramide

The product was prepared according to the general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amide, starting from 5- (4-cyclopropylmethoxy-3-fluoro-phenyl) -2H-pyrazol-3-ylamine (123.5mg, 0.5mmol, 1.0eq) using the amino acid route. Purification by preparative HPLC afforded 130mg of the title compound as formate (67% yield).

C₂₁H₂₇N₄O₂F

Mass (calculated) [ 386)](ii) a (found value) [ M + H [)⁺]＝387。

LC Rt 2.01min, 100% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 0.32-0.36(2H, m); 0.56-0.61(2H, m); 1.21-1.28(1H, m); 1.73-1.84(5H, m); 2.36(2H，t，J＝7.2Hz)；2.67-2.77(6H，m)；3.92(3H，d，J＝7.2Hz)；6.79(1H，s)；7.18(1H，t，J＝8.8Hz)；7.45-7.47(1H，m)；7.55-7.59(1H，m)；8.19(1H，s)；10.49(1H，s)

Example 24

N- [4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-yl ] -4-pyrrolidin-1-yl-butyramide

a) N- [4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-yl ] -acetamide

Acetylguanidine (2.6g, 25.7mmol, 3.0eq) was dissolved in anhydrous DMF (40mL), 2-bromo-1- (4-difluoromethoxy-phenyl) -ethanone (2.3g, 8.5mmol, 1.0eq) was added, and the mixture was stirred at room temperature for 4 days. DMF was evaporated, the residue was washed with water, filtered and dried. The crude product was crystallized from methanol to yield 1.2g of the title compound (yield: 53%).

C₁₂H₁₁F₂N₃O₂

¹H-NMR(DMSO-d₆)：3.40(3H，br s)；7.10-7.47(4H，m)；7.82(2H，d，J＝8.4Hz)；11.32(1H，s)；11.73(1H，br s)。

b)4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-ylamine

N- [4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-yl ] -acetamide (1.2g, 4.5mmol, 1.0eq) was dissolved in water (30mL) and methanol (30mL), 30 drops of sulfuric acid were added. The reaction was refluxed for 2 days, then the mixture was evaporated to dryness, the residue was diluted with water, adjusted to pH 8 with NaOH 2N, the product was extracted with DCM and concentrated under reduced pressure to give 1.0g of the title compound (yield: 99%).

C₁₀H₉F₂N₃O

¹H-NMR(DMSO-d₆)：5.59(2H，br s)；6.98-7.35(4H，m)；7.60-7.62(2H，m)。

c) N- [4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-yl ] -4-pyrrolidin-1-yl-butyramide

To a suspension of 4-pyrrolidin-1-yl-butyric acid (386mg, 2.0mmol, 4.0eq) in 1, 2-dichloroethane (3mL) was added N, N' -carbonyldiimidazole (300mg, 1.8mmol, 3.7eq) and N, N-diisopropylethylamine (87 μ L, 0.5mmol, 1.0eq) and the mixture was stirred at room temperature for 1 hour until the amino acid was fully activated and the suspension was completely dissolved.

4- (4-difluoromethoxy-phenyl) -1H-imidazol-2-ylamine (112.5mg, 0.5mmol, 1.0eq) was added and the reaction stirred at room temperature for 1 day and then at 50 ℃ for 2 days (LC-MS monitoring showed the reaction was slow and not complete).

The solvent was evaporated under reduced pressure, and the crude product was purified by preparative HPLC to give 80mg (yield: 44%) of the title compound as a white solid.

C₁₈H₂₂N₄O₂F₂

Mass (calculated) [364](ii) a (found value) [ M + H [)⁺]＝365，[M/2]＝183。

LC Rt 1.18min, 100% (10 min method)

¹H-NMR(DMSO-d₆)：1.74-1.84(6H，m)；2.38(2H，t，J＝7.6Hz)；2.70-2.79(6H，m)；6.99-7.37(4H，m)；7.71(2H，d，J＝8.8Hz)；8.23(1H，br s)

Example 25

N- [5- (5-chloro-2-methoxy-phenyl) -2H-pyrazol-3-yl ] -4-cis-2, 6-dimethyl-piperidin-1-yl) -butyramide

a)4- (2, 6-dimethyl-piperidin-1-yl) -butyric acid ethyl ester

To a solution of cis-2, 6-dimethylpiperidine (6.9mL, 51.3mmol, 2.5eq) in toluene (25mL) was added ethyl 4-bromobutyrate (2.9mL, 20.5mmol, 1eq) and the reaction mixture was refluxed for 2 days. The mixture was cooled to room temperature and the white solid was filtered off and washed with diethyl ether. The crude product was diluted with HCl 1N (8mL, 1eq) then washed with EtOAc, treated with NaOH 1N (16mL, 2eq) and extracted with ethyl acetate to give the title compound (1.51g, yield 32%) which was used in the next step without further purification.

C₁₃H₂₅NO₂

¹H-NMR(CD₃OD)：0.99(6H，d，J＝6.0Hz)；1.07-1.21(6H，m)；1.45-1.58(5H，m)；2.20(2H，t，J＝6.8Hz)；2.30-2.35(2H，m)；2.53-2.57(2H，m)；4.02(2H，q，J＝7.2Hz)。

b)4- (2, 6-dimethyl-piperidin-1-yl) -butyric acid

To a suspension of 4- (2, 6-dimethyl-piperidin-1-yl) -butyric acid ethyl ester (1.5g, 6.7mmol) in water (5mL) and MeOH (1mL) was added NaOH (266mg, 6.7mmol, 1.0eq) and the mixture was refluxed for 22 hours. The reaction was then cooled to room temperature, adjusted to pH 4 with HCl 2N at 0 ℃ and concentrated under reduced pressure. The residue was treated with ethanol and the precipitated sodium chloride was filtered off. Evaporation of the solvent under reduced pressure gave 950mg of the title compound as a white solid (51% yield).

C₁₁H₂₁NO₂

¹H-NMR(CD₃OD)：1.28-1.34(6H，m)；1.46-1.74(5H，m)；1.81-1.91(4H，m)；2.36-2.40(2H，m)；3.20-3.27(3H，m)。

c) N- [5- (5-chloro-2-methoxy-phenyl) -2H-pyrazol-3-yl ] -4- ((cis) -2, 6-dimethyl-piperidin-1-yl) -butyramide

The general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides according to the one-pot method was prepared starting from commercially available 5- (5-chloro-2-methoxy-phenyl) -2H-pyrazol-3-ylamine (111.8mg, 0.5mmol, 1.0eq) and 4- (2, 6-dimethyl-piperidin-1-yl) -butyric acid (149.0mg, 0.8mmol, 1.5 eq).

Purification by preparative HPLC according to the general procedure gave 80mg of the title compound as formate salt (40% yield).

C₂₁H₂₉N₄O₂Cl

Mass (calculated value) [404 ]](ii) a (found value) [ M + H [)⁺]＝405

LC Rt 2.03min, 100% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.12(6H, d, J ═ 6.4 Hz); 1.27-1.32(3H, m); 1.57-1.59(3H, m); 1.68-1.74(2H, m); 2.27-2.31(2H, m); 2.72-2.82(4H, m); 3.87(3H, s); 6.92(1H, s); 7.14(1H, d, J ═ 9.2 Hz); 7.33-7.36(1H, m); 7.70(1H, d, J ═ 2.8 Hz); 8.26(1H, s); 10.48(1H, br s)

Example 26

N- [5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -4- ((S) -2-methyl-pyrrolidin-1-yl) -butyramide

a)4- ((S) -2-methyl-pyrrolidin-1-yl) -butyric acid ethyl ester

(S) -2-methyl-pyrrolidine hydrochloride (0.8g, 6.6mmol, 1.1eq) was dissolved in 2-butanone (20mL) and potassium carbonate (1.7g, 12.6mmol, 2.1eq) was added. Ethyl 4-bromobutyrate (0.86mL, 6.0mmol, 1.0eq) was then added and the reaction mixture was refluxed for 2 days. The mixture was cooled to room temperature and the solid was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give 1.20g of the title compound (yield 99%), which was used in the next step without further purification.

C₁₁H₂₁NO₂

¹H-NMR(DMSO-d₆)：0.95(3H，d，J＝6.0Hz)；1.13-1.17(3H，m)；1.20-1.28(1H，m)；1.59-1.64(4H，m)；1.77-1.86(1H，m)；1.90-2.00(2H，m)；2.10-2.23(1H，m)；2.25-2.31(2H，m)；2.62-2.66(1H，m)；2.96-2.99(1H，m)；3.98-4.03(2H，m)。

b)4- ((S) -2-methyl-pyrrolidin-1-yl) -butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Evaporation under reduced pressure gave 1.1g of the title compound (76% yield) as the hydrochloride salt.

C₉H₁₇NO₂

¹H-NMR(DMSO-d₆Hydrochloride salt): 1.22-1.27(3H, m); 1.62-1.64(1H, m); 2.03-2.09(6H, m); 2.19-2.28(1H, m); 2.47-2.58(1H, m); 2.86-2.92(1H, m); 3.15-3.40(1H, m); 3.69-3.75(2H, m); 7.25(1H, s).

c) N- [5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -4- ((S) -2-methyl-pyrrolidin-1-yl) -butyramide

General procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides according to the one-pot method starting from 5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine (112.5mg, 0.5mmol, 1.0eq) and 4- ((S) -2-methyl-pyrrolidin-1-yl) -butyric acid (155.0mg, 0.8mmol, 1.5 eq). After purification by preparative HPLC 120mg of the title compound were obtained as formate salt (69% yield).

C₁₉H₂₄N₄O₂F₂

Mass (calculated) [ 378)](ii) a (found value) [ M + H [)⁺]＝379

LC Rt 1.64min, 98% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.04(3H, d, J ═ 6.0 Hz); 1.30-1.37(1H, m); 1.65-1.89(5H, m); 2.16-2.26(2H, m); 2.28-2.40(2H, m); 2.80-2.82(1H, m); 3.12-3.17(2H, m); 6.79(1H, s); 7.07-7.44(3H, m); 7.73-7.75(2H, m); 8.18(1H，s)；10.44(1H，br s)

example 27

N- [5- (1H-indol-3-yl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a)3- (1H-indol-3-yl) -3-oxo-propionitrile

In a flask, cyanoacetic acid (5.0g, 58.8mmol, 1.2eq) was dissolved in acetic anhydride (50mL) and heated at 50 ℃. Indole (5.8g, 50.0mmol, 1.0eq) was added and the reaction mixture was heated at 80 ℃ for 5 min. A white precipitate precipitated out of solution, the reaction was cooled to room temperature and then filtered. The resulting solid (620.0mg, 85% yield) was used in the next step without further purification.

C₁₁H₈N₂O

¹H-NMR(DMSO-d₆)：4.48(2H，s)；7.21-7.24(2H，m)；7.48-7.50(1H，m)；8.12-8.14(1H，m)；8.37(1H，d，J＝3.2Hz)；12.17(1H，s)。

b)5- (1H-indol-3-yl) -2H-pyrazol-3-ylamine

To a solution of 3- (1H-indol-3-yl) -3-oxo-propionitrile (6.4g, 34.7mmol, 1.0eq) in anhydrous EtOH (40mL) was added hydrazine monohydrate (5.0mL, 104.1mmol, 3.0eq) and the reaction was heated at reflux for 24H. The reaction mixture was cooled to room temperature, the solid was filtered off and Et₂Washing with O/EtOAc 10/1 gave 3.0g of the title product (74% yield).

C₁₁H₁₀N₄

Mass (calculated value) [198](ii) a (found value) [ M + H [)⁺]＝199。

LC Rt 0.98min, 90% (5 min method)

¹H-NMR(DMSO-d₆)：4.57(2H，bs)；5.70(1H，s)；7.00-7.19(2H，m)；7.33-7.46(1H，m)；7.59(1H，s)；7.69-7.90(1H，bs)；11.11-11.36(1H，bs)；11.37-11.77(1H，bs)。

c) N- [5- (1H-indol-3-yl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

To a suspension of 4-piperidin-1-yl-butyric acid (621.0mg, 3.0mmol, 1.5eq) in 1, 2-dichloroethane (6mL) was added N, N' -carbonyldiimidazole (453.0mg, 2.8mmol, 1.4eq) and the mixture was stirred at room temperature for 1 hour. A solution of 5- (1H-indol-3-yl) -2H-pyrazol-3-ylamine (400.0mg, 2.0mmol, 1.0eq) in 1, 2-dichloroethane (6mL) was added, and the reaction mixture was stirred at room temperature for 2 days and then at 70 ℃ for 1 day to completely transfer the acyl group on the ring nitrogen atom to the exocyclic nitrogen. The reaction was then cooled to room temperature, the mixture was washed with saturated aqueous sodium carbonate solution and then evaporated under reduced pressure. The crude product was purified by preparative HPLC to give 320.0mg (yield: 41%) of the title compound as a formate salt.

C₂₀H₂₅N₅O

Mass (calculated value) [351](ii) a (found value) [ M + H [)⁺]＝352。

LC Rt ═ 1.42min, 95% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.37-1.39(2H, m); 1.50-1.54(4H, m); 1.72-1.80(2H, m); 2.30-2.34(2H, m); 2.40-2.48(6H, m); 6.78(1H, s); 7.08-7.17(2H, m); 7.43(1H, d, J ═ 7.6 Hz); 7.71(1H, d, J ═ 2.8 Hz); 7.76(1H, d, J ═ 7.6 Hz); 8.19(1H, s); 10.39(1H, s); 11.39(1H, s)

Example 28

N- [5- (4-isopropoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a) 4-Isopropoxy-benzoic acid methyl ester

3.0g 4-isopropoxy-benzoic acid (16.7mmol, 1.0eq) was dissolved in MeOH (20mL), a catalytic amount of sulfuric acid was added and the mixture was heated under reflux for 2 days. The solvent was evaporated and the residue was dissolved in DCM and washed with 10% NaOH. The organic phase was dried and evaporated to give 2.2g of the title product (yield 67%).

C₁₁H₁₄O₃

¹H-NMR(DMSO-d₆)：1.25(6H，d，J＝6.4Hz)；3.77(3H，s)；4.67-4.70(1H，m)；6.96-6.98(2H，m)；7.84-7.87(2H，m)。

b)3- (4-isopropoxy-phenyl) -3-oxo-propionitrile

To a solution of 4-isopropoxy-benzoic acid methyl ester (2.2g, 11.2mmol, 1.0eq) in dry toluene (15mL) was added NaH (50-60% dispersion in mineral oil, 1.1g, 22.4mmol, 2.0eq) under nitrogen. Heating the mixture at 80 deg.C, and adding dry CH dropwise₃CN (2.8mL, 56.0mmol, 5.0 eq). The reaction was heated for 18 hours, then cooled to room temperature and acidified with HCl 2N. The organic layer was recovered to give 2.0g of crude product, which was used in the next step without further purification.

C₁₁H₁₄O₃

c)5- (4-isopropoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazole (scheme a2) starting from 3- (4-isopropoxy-phenyl) -3-oxo-propionitrile. The solvent was removed under reduced pressure and water (10mL) was added to precipitate the title compound as a yellow solid (1.0g, 94% yield), which was used in the next step without further purification.

C₁₂H₁₅N₃O

Mass (calculated value) [217 ]](ii) a (found value) [ M + H [)⁺]＝218。

LC Rt 1.36min, 95% (5 min method)

¹H-NMR(DMSO-d₆)：1.24(6H，d，J＝6.0Hz)；4.57-4.69(3H，br m)；5.64(1H，s)；6.89(2H，d，J＝8.8Hz)；7.51(2H，d，J＝8.8Hz)

d) N- [5- (4-isopropoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

Prepared according to the general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides starting from 5- (4-isopropoxy-phenyl) -2H-pyrazol-3-ylamine (86.0mg, 0.4mmol, 1.0eq) using the amino acid route. The crude product was purified by preparative HPLC to give the title compound (56.0mg, 38% yield) as the formate salt.

C₂₁H₃₀N₄O₂

Mass (calculated value) [370 ]](ii) a (found value) [ M + H [)⁺]＝371，[M+2/2]＝165。

LC Rt 1.91min, 96% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.25(6H, d, J ═ 6 Hz); 1.33-1.41(2H, m); 1.48-1.53(4H, m); 1.71-1.77(2H, m); 2.29(2H, t, J ═ 7.2 Hz); 2.35(2H, t, J ═ 7.2 Hz); 2.42-2.47(4H, m); 4.60-4.66(1H, m); 6.71(1H, s); 6.94(2H, d, J ═ 8.8 Hz); 7.58(2H, d, J ═ 8.8 Hz); 8.17(1H, s); 10.38(1H, s).

Example 29

N- [5- (1-ethyl-1H-indol-3-yl) -2H-pyrazol-3-yl ] -4-pyrrolidin-1-yl-butyramide

a) 1-Ethyl-1H-indole-3-carboxylic acid methyl ester

To a suspension of NaH (50-60% dispersion in mineral oil, 548.0mg, 11.4mmol, 2.0eq) in THF (20mL) was added methyl 1H-indole-3-carboxylate (1.0g, 5.7mmol, 1.0eq), after 20 min ethyl iodide (507.0 μ L, 6.3mmol, 1.1. eq). The reaction was heated at 70 ℃ for 1 hour, then cooled to 0 ℃ and water (10mL) was carefully added. AcOEt was added, the organic phase was collected and concentrated to give the crude product which was purified on a silica gel column (10g) eluting with a gradient of 100% cyclohexane to cyclohexane-EtOAc 80: 20 to give the title product (860mg, 74% yield).

C₁₂H₁₃NO₂

¹H-NMR(DMSO-d₆)：1.36(3H，t，J＝7.2Hz)；3.77(3H，s)；4.26(2H，q，J＝7.2)；7.16-7.27(2H，m)；7.55-7.59(1H，m)；7.97-7.99(1H，m)；8.15(1H，s)。

b)3- (1-ethyl-1H-indol-3-yl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole (scheme A1bis) starting from 1-ethyl-1H-indole-3-carboxylic acid methyl ester (860.0mg, 4.2mmol, 1.0 eq). 820.0mg of the title compound (91% yield) were obtained and used directly in the next step. C₁₃H₁₂N₂O

c)5- (1-ethyl-1H-indol-3-yl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme a2) starting from 3- (1-ethyl-1H-indol-3-yl) -3-oxo-propionitrile (820mg, 3.87mmol, 1.0 eq). The solvent was removed under reduced pressure and the solid residue was washed with ethanol to give the title product (612mg, 70% yield).

C₁₃H₁₄N₄

Mass (calculated value) [226](ii) a (found value) [ M + H [)⁺]＝227。

LC Rt ═ 1.30min, 69% (5 min method)

d) N- [5- (1-ethyl-1H-indol-3-yl) -2H-pyrazol-3-yl ] -4-pyrrolidin-1-yl-butyramide

Prepared according to the general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides starting from 5- (1-ethyl-1H-indol-3-yl) -2H-pyrazol-3-ylamine (99.0mg, 0.5mmol, 1.0eq) and 4-pyrrolidin-1-yl-butyric acid (118mg, 0.75mmol) using the amino acid route. The crude product was purified by preparative HPLC to give the title compound as formate salt (77.0mg, 42% yield).

C₂₁H₂₇N₅O

Mass (calculated value) [365 ]](ii) a (found value) [ M + H [)⁺]＝366。

LC Rt 1.83min, 99% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.38(3H, t, J ═ 7.2 Hz); 1.71-1.81(6H, m); 2.34(2H, t J ═ 7.2 Hz); 2.59-2.65(6H, m); 4.23(2H, q, J ═ 7.2 Hz); 6.76(1H, s); 7.11-7.22(2H, m); 7.53(1H, d, J ═ 8.4 Hz); 7.75-7.79(2H, m); 8.19(1H, br s); 10.40(1H, s).

Example 30

N- [5- (4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

a) 4-Cyclopropylmethoxy-benzoic acid methyl ester

4-hydroxy-benzoic acid methyl ester (2.0g, 13.1mmol, 1.2eq) was dissolved in acetone (20mL), NaI (0.97g, 6.5mmol, 0.5eq) and K were added₂CO₃(3.0g, 21.8mmol, 2.0eq) and the mixture was stirred at room temperature for 20 min. (bromomethyl) cyclopropane (1.1mL, 10.3mmol, 1.0eq) was added and the reaction was refluxed for 2 days. The solvent was evaporated under reduced pressure, NaOH 10% was added, the product was extracted with DCM, Na₂SO₄The organic phase was dried and the solvent was evaporated under reduced pressure to give the title product (1.23g, yield 79%) which was used in the next step without further purification.

C₁₂H₁₄O₃

Quality (meter)Calculated value) [206](ii) a (found value) [ M + H [)⁺]＝207。

LC Rt 2.38min, 86% (5 min method)

¹H-NMR(DMSO-d₆)：033-0.34(2H，m)；0.57-0.59(2H，m)；1.21-1.25(1H，m)；3.81(3H，s)；3.89(2H，d，J＝6.8Hz)；7.02(2H，d，J＝8.8Hz)；7.88(2H，d，J＝8.8Hz)。

b)5- (4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine

Prepared according to the general synthetic method (scheme A1bis) starting from 4-cyclopropylmethoxy-benzoic acid methyl ester (1.17g, 5.9mmol, 1.0 eq). The reaction was cooled to room temperature and the solid formed was filtered off and dissolved in water. The solution was acidified to pH 4 and the solid formed was filtered off to give 1.2g of 3- (4-cyclopropylmethoxy-phenyl) -3-oxo-propionitrile, which was used directly in the next step.

5- (4-Cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine is prepared according to the general synthesis of aminopyrazoles (scheme A2). The reaction was concentrated and the residue was precipitated with water to give 500mg of the title compound (37% yield) which was used directly in the next step.

C₁₃H₁₅N₃O

c) N- [5- (4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-yl ] -4-piperidin-1-yl-butyramide

According to a general method for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides, prepared using the amino acid route starting from 5- (4-cyclopropylmethoxy-phenyl) -2H-pyrazol-3-ylamine (152.9mg, 0.7mmol, 1.0eq) and 4-piperidin-1-yl-butyric acid (168mg, 1.0mmol, 1.5 eq). The crude product was purified by preparative HPLC to give 72.0mg of the title compound (28% yield) as the formate salt.

C₂₂H₃₀N₄O₂

Mass (calculated value) [382](ii) a (found value) [ M + H [)⁺]＝383。

LC Rt 1.99min, 100% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 033-0.34(2H, m); 0.55-0.59(2H, m); 1.19-1.25(1H, m); 1.38-1.40(2H, m); 1.49-1.54(4H, m); 1.70-1.77(2H, m); 2.28-2.41(8H, m); 3.84(2H, d, J ═ 6.8 Hz); 6.74(1H, s); 6.97(2H, d, J ═ 8.8 Hz); 7.60(2H, d, J ═ 8.8 Hz); 8.19(1H, s); 10.40(1H, s).

Example 31

4-azepan-1-yl-N- [5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -butyramide

a) 4-Azepan-1-yl-butyric acid ethyl ester

To a solution of azepane (10.2mL, 102.0mmol, 4.0eq) in toluene (30mL) was added ethyl 4-bromobutyrate (3.8mL, 26.0mmol, 1.0eq) and the reaction mixture was refluxed for 10 hours. The mixture was cooled to room temperature and the solid was filtered off and washed with diethyl ether. The filtrate was concentrated under reduced pressure to give the amino ester, which was used in the next step without purification.

C₁₂H₂₃NO₂

b) 4-Azepan-1-yl-butyric acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Evaporation of water under reduced pressure gave 3.8g of the title compound (80% yield) as the hydrochloride salt.

C₁₀H₁₉NO₂

Mass (calculated value) [185 ]](ii) a (found value) [ M + H [)⁺]＝186。

LC Rt 0.26min, 100% (5 min method)

¹H-NMR(DMSO-d₆Hydrochloride salt)：1.53-1.66(4H，m)；1.77-1.91(6H，m)；2.30(2H，t，J＝7.2Hz)；2.98-3.09(4H，m)；3.27-3.30(2H，m)；10.42(1H，brs)。

c) 4-Difluoromethoxybenzoic acid methyl ester

In a two-necked round bottom flask under nitrogen flow, 1.3g of methyl 4-hydroxy-benzoate (8.3mmol, 1.0eq) and 1.5g of sodium difluorochloroacetate (10.0mmol, 1.2eq) were dissolved in DMF (25mL), potassium carbonate (1.4g, 10.0mmol, 1.2eq) was added and the mixture was heated at 125 ℃ for 3.5 hours. The reaction mixture was then diluted with water and extracted with DCM; the organic phase was dried and then evaporated and the crude product was purified over a silica gel column (eluent: cyclohexane/EtOAc 80/20) to give 0.77g of product (yield 46%), which was used directly in the next step.

C₉H₈F₂O₃

d)3- (4-difluoromethoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazole starting from 872.0mg (4.3mmol, 1.0eq) of 4-difluoromethoxy-benzoic acid methyl ester (scheme A1 bis). 818.5mg of the title compound (yield 90%) were obtained, which was used directly in the next step.

C₁₀H₇F₂NO₂

e)5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified on a silica gel column, eluting with a gradient of 100% EtOAc to EtOAc-MeOH 80: 20, to give the title product (826mg, 59% yield).

C₁₀H₉F₂N₃O

Mass (calculated value) [225](ii) a (found value) [ M + H [)⁺]＝226。

LC Rt 1.34min, 100% (5 min method)

¹H-NMR(DMSO-d₆)：4.82(2H，br s)，5.71(1H，s)，7.15(2H，d，J＝8.4Hz)，7.22(1H，t，J＝74.0Hz)，7.67(2H，d，J＝8.8Hz)；11.58(1H，br s)

f) 4-azepan-1-yl-N- [5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -butyramide

Prepared according to the general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides starting from 5- (4-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine (149.0mg, 0.7mmol, 1.0eq) using the amino acid route. Purification by preparative HPLC afforded 90.0mg of the title compound as the formate salt (35% yield).

C₂₀H₂₆F₂N₄O₂

Mass (calculated) [392 ]](ii) a (found value) [ M + H [)⁺]＝393，[M+2/2]＝197。

LC Rt 2.26min, 100% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.51-1.60(8H, m); 1.72-1.76(2H, m); 2.31(2H, t, J ═ 7.6 Hz); 2.56(2H, t, J ═ 7.2 Hz); 2.69(4H, t, J ═ 5.2 Hz); 6.80(1H, s); 7.08-7.45(3H, m); 7.73-7.76(2H, m); 8.21(1H, s); 10.50(1H, br s).

Example 32

Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid (5-o-tolyl-2H-pyrazol-3-yl) -amide

a) Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid ethyl ester

Ethyl 2-formyl-1-cyclopropanecarboxylate (3.0g, 21.1mmol, 1.2eq) and piperidine (1.5g, 17.6mmol, 1.0eq) were dissolved in DCM (45mL) under nitrogen, and after 2 hours at room temperature, the mixture was cooled to 0 ℃ and sodium triacetoxyborohydride (5.6g, 26.4mmol, 1.5eq) was added dropwise. The mixture was stirred at room temperature for 2.5 hours, and then the organic phase was washed with aqueous NaOH solution and water to obtain 3.3g of the title compound (yield 89%).

C₁₂H₂₁NO₂

¹H-NMR(CDCl₃)：0.70-0.75(1H，m)；1.20-1.38(4H，m)；1.39-1.43(3H，m)；1.53-1.61(5H，m)；2.22-2.27(1H，m)；2.34-2.43(5H，m)；4.08-4.17(2H，m)。

b) Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid

The product was prepared according to the general synthesis of omega-amino acids (scheme C2). Water was evaporated under reduced pressure and triturated with ether to give 1.3g of the title compound (33% yield) as the hydrochloride salt.

C₁₀H₁₇NO₂

Mass (calculated value) [ 183)](ii) a (found value) [ M + H [)⁺]＝184。

LC Rt 0.19min (5 min method)

¹H-NMR(DMSO-d₆Hydrochloride salt): 0.96-1.01(1H, m), 1.06-1.11(1H, m), 1.27-1.41(1H, m), 1.62-1.85(7H, m), 2.82-3.06(4H, m), 3.36-3.37(2H, m), 10.88(1H, bs), 12.38(1H, bs)

c) Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid (5-o-tolyl-2H-pyrazol-3-yl) -amide

Prepared according to the general procedure for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides starting from commercially available 5-o-tolyl-2H-pyrazol-3-ylamine (152.0mg, 0.9mmol, 1.0eq) using the amino acid route. The crude product was purified by preparative HPLC and silica gel column with 100% CH gradient eluent₃CN to CH₃CN/2N NH₃A solution in MeOH 80: 20 gave the title compound (18mg, 6% yield).

C₂₀H₂₆N₄O

Mass (calculated) [ 338)](ii) a (found value) [ M + H [)⁺]＝339，[M+2/2]＝170。

LC Rt 1.71min, 100% (10 min method)

¹H-NMR(DMSO-d₆)：0.62(1H，br s)；0.94-0.97(1H，m)；1.27-1.37(3H，m)；1.44-1.49(4H，m)；1.65-1.68(1H，m)；2.08-2.13(1H，m)；2.30-2.35(8H，m)；6.62(1H，s)；7.24-7.27(3H，m)；7.38(1H，d，J＝6.0Hz)；10.64(1H，s)；12.45(1H，s)。

Example 33

Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid [5- (2-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -amide

a) 2-Difluoromethoxybenzoic acid methyl ester

2.0g 2-difluoromethoxy-benzoic acid (10.6mmol, 1.0eq) was dissolved in MeOH (15mL), a catalytic amount of sulfuric acid was added and the mixture was heated under reflux overnight. The solvent was then evaporated, the residue dissolved in DCM and saturated NaHCO₃And (6) washing. The organic phase was dried and evaporated to yield 1.9g of the title product (yield 87%).

C₉H₈F₂O₃

¹H-NMR(DMSO-d₆)：3.82(3H，s)；6.99-7.40(2H，m)；7.31(1H，d，J＝8.4Hz)；7.63-7.67(1H，m)；7.82-7.84(1H，m)。

b)3- (2-difluoromethoxy-phenyl) -3-oxo-propionitrile

The product was prepared according to the general procedure for the synthesis of aminopyrazoles starting from 1.5g (7.4mmol, 1.0eq) of 2-difluoromethoxy-benzoic acid methyl ester (scheme A1 bis). The crude product was used directly in the next step.

C₁₀H₇F₂NO₂

c)5- (2-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine

The product was prepared according to the general procedure for the synthesis of aminopyrazoles (scheme A2). The crude product was purified over a silica gel column with gradient eluent from 100% EtOAc to EtOAc-MeOH 90: 10. The title product was obtained (1.3g, 76% yield).

C₁₀H₉F₂N₃O

¹H-NMR(DMSO-d₆)：4.82(2H，bs)，5.79(1H，s)，7.00-7.37(4H，m)，7.79(1H，d)，11.74(1H，bs)

d) Trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid [5- (2-difluoromethoxy-phenyl) -2H-pyrazol-3-yl ] -amide

According to a general method for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides, prepared using the amino acid route starting from trans (±) -2-piperidin-1-ylmethyl-cyclopropanecarboxylic acid (99.1mg, 0.6mmol, 1.3eq) and 5- (2-difluoromethoxy-phenyl) -2H-pyrazol-3-ylamine (125.7mg, 0.4mmol, 1.0 eq). The crude product was purified by column chromatography over silica gel eluting with a gradient of 100% DCM to DCM-NH₃A solution in MeOH 2N 80: 20 gave the title compound (39.9mg, 23% yield).

C₂₀H₂₄F₂N₄O₂

Mass (calculated value) [390](ii) a (found value) [ M + H [)⁺]＝391。

LC Rt 1.68min, 100% (10 min method)

¹H-NMR(DMSO-d₆)：0.62-0.65(1H，m)；0.96-1.00(1H，m)；1.21-1.69(7H，br m)；2.13(1H，br s)；2.30-2.49(3H，m)；3.29-3.31(3H，m)；6.91-7.42(5H，m)；7.72(1H，d，J＝7.2Hz)；10.67(1H，s)；12.68(1H，s)

Example 34

N- [5- (4-chloro-phenyl) -2H-pyrazol-3-yl ] -2-methyl-4-pyrrolidin-1-yl-butyramide

According to a general method for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides, prepared using the amino acid route starting from 5- (4-chloro-phenyl) -2H-pyrazol-3-yl-amine (58.0mg, 0.3mmol, 1.0eq) and 2-methyl-4-pyrrolidin-1-yl-butyric acid (77.0mg, 0.45mmol, 1.5 eq). The crude product was purified by preparative HPLC to give 21.1mg of the title compound as formate salt (18% yield).

C₁₈H₂₃ClN₄O

Mass (calculated value) [346 ]](ii) a (found value) [ M + H [)⁺]＝347，[M+2/2]＝174。

LC Rt 1.84min, 100% (10 min method)

¹H-NMR(DMSO-d₆HCOOH salts): 1.07(3H, d, J ═ 6.8 Hz); 1.47-1.52(1H, m); 1.64-1.67(4H, m); 1.74-1.79(1H, m); 2.38-2.58(4H, m); 3.79(3H, s); 6.87-6.90(1H, m); 7.25-7.27(2H, m); 7.33(1H, t, J ═ 8.4 Hz); 10.42(1H, br s)

Example 35

5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide

a) 5-amino-3- (4-methoxy-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

A solution of di-tert-butyl dicarbonate (605.0mg, 2.8mmol, 1.0eq) in DCM (3mL) was added to a vigorously stirred mixture of 5-amino-3- (4-methoxy-phenyl) -pyrazole (500.0mg, 2.7mmol, 1.0eq), DCM (20mL) and KOH4.5M aqueous solution (4.7mL, 21.1mmol, 8eq) and stirred at room temperature for 20 minutes. The organic layer was collected and washed with water/brine 1/1 solution. Evaporation of the solvent gave a crude product, which was purified by silica gel column eluting with DCM to give the title compound (720mg, 94% yield).

C₁₅H₁₉N₃O₃

Mass (calculated) [ 289)](ii) a (found value) [ M + H [)⁺]＝290

LC Rt 1.43min, 100% (3 min method)

¹H-NMR(DMSO-d₆)：1.58(9H，s)；3.78(3H，s)；5.69(1H，s)；6.36(2H，s)；6.96(2H，br d，J＝8.8Hz)；7.68(2H，br d，J＝8.8Hz)。

b)2- (3-bromo-propyl) -2-methyl-malonic acid dimethyl ester

A dispersion of 60% NaH in mineral oil (1.63g, 40.8mmol, 1.3eq) was washed three times with hexane and then dried. After addition of dry THF (30mL), the suspension was cooled to 0 ℃. Dimethyl methylmalonate (4.7g, 32.3mmol, 1.0eq) was carefully added slowly and gas evolution was observed. The mixture was stirred for 15 minutes, then 1, 3-dibromopropane (24g, 119.0mmol, 3.7eq) was added in one portion. The mixture was warmed to room temperature and stirred for a further 16 hours. NaOH 1.0M solution was added and the crude product was extracted with ethyl acetate, the organic phase collected, dried and the resulting oil purified over a silica gel column eluting with cyclohexane followed by EtOAc to give the title product (6.6g, 76% yield).

C₉H₁₅BrO₄

¹H-NMR(DMSO-d₆)：1.32(3H，s)；1.67-1.72(2H，m)；1.861-1.90(2H，m)；3.51(2H，t，J＝6.4Hz)；3.64(6H，s)。

c) 5-bromo-2-methyl-pentanoic acid

HBr aq 48% (10mL, 88.4mmol) was added to dimethyl 2- (3-bromo-propyl) -2-methyl-malonate (1.80g, 6.74mmol) at room temperature, and the mixture was stirred at 120 ℃ for 24 hours. After cooling to room temperature, NaOH solution was added to adjust to pH 3 and the product was extracted with a mixture of DCM: MeOH 95: 5. The crude product obtained (0.81g, 62% yield) was used in the next step without purification.

C₆H₁₁BrO₂

¹H-NMR(DMSO-d₆)：1.05(3H，d，J＝7.2Hz)；1.41-1.50(1H，m)；1.61-1.70(2H，m)；1.75-1.83(2H，m)；2.31-2.40(1H，m)；3.52(2H，dd，J＝6.8Hz，6.4Hz)。

d)5- (5-bromo-2-methyl-pentanoylamino) -3- (4-methoxy-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

Oxalyl chloride (250.0 μ L, 3.0mmol, 1.5eq) was added slowly to a solution of 5-bromo-2-methyl-pentanoic acid (390.0mg, 2.0mmol, 1.0eq) in DCM (1mL) at room temperature and the mixture was stirred under nitrogen atmosphere for 2 h. The solvent and excess oxalyl chloride were evaporated, the residue was dissolved in DCM (1mL) and added dropwise to a solution of tert-butyl 5-amino-3- (4-methoxy-phenyl) -pyrazole-1-carboxylate (656.0mg, 2.3mmol, 1.15eq) and triethylamine (0.28mL, 2.0mmol, 1.0eq) in DCM (1 mL). The mixture was stirred at room temperature for 48 hours, then a saturated aqueous sodium bicarbonate solution was added, and the organic layer was collected and dried. The crude product was purified by silica gel column eluting with cyclohexane-DCM 10: 0 to 1: 1 to give the title compound (237.0mg, 25% yield).

C₂₁H₂₈BrN₃O₄

Mass (calculated) [466](ii) a (found value) [ M + H [)⁺]＝467

LC Rt 1.83min, 92% (3 min method)

¹H-NMR(DMSO-d₆)：1.14(3H，d，J＝6.8Hz)；1.62(9H，s)；1.72-1.86(4H，m)；2.63-2.70(1H，m)；3.55(2H，dd，J＝6.8Hz，6.4Hz)；3.78(3H，s)；7.01(2H，br d，J＝8.8Hz)；7.07(1H，s)；7.79(2H，br d，J＝8.8Hz)；10.09(1H，s)。

e)5- [5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoylamino ] -3- (4-methoxy-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

Tert-butyl 5- (5-bromo-2-methyl-pentanoylamino) -3- (4-methoxy-phenyl) -pyrazole-1-carboxylate (280.0mg, 0.6mmol, 1.0eq) was dissolved in DCM (3 mL). Triethylamine (80. mu.L, 0.6mmol, 1.0eq) and 1- [1, 4] -diazepan-1-yl-ethanone (158. mu.L, 170.0mg, 1.2mmol, 2.0eq) were added and the mixture was stirred at room temperature for 24 h and then at 50 ℃ for 16 h. Saturated aqueous sodium bicarbonate was added, the organic layer was separated and the solvent was evaporated to give the crude product. Purification on a silica gel column eluting with DCM, DCM: MeOH 99: 1 to 96: 4 gave the title compound (181.3mg, 54% yield).

C₂₈H₄₁N₅O₅

Mass (calculated value) [527 ]](ii) a (found value) [ M + H [)⁺]＝528

LC Rt 1.63min, 100% (5 min method).

¹H-NMR(DMSO-d₆)：1.13(3H，d，J＝6.4Hz)；1.33-1.50(4H，m)；1.62(9H，s)；1.65-1.81(2H，m)；1.96(3H，s)；2.34-2.44(1H，m)；2.52-2.67(3H，m)；2.98-3.13(3H，m)；3.40-3.46(4H，m)；3.80(3H，s)；7.01(2H，br d，J＝8.8Hz)；7.06(1H，s)；7.79(2H，br d，J＝8.8Hz)；10.07(1H，s)。

f)5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoic acid [5- (4-methoxy-phenyl) -2H-pyrazol-3-yl ] -amide

Tert-butyl 5- [5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoylamino ] -3- (4-methoxy-phenyl) -pyrazole-1-carboxylate (181.0mg, 0.34mmol, 1.0eq) was dissolved in DCM (3mL) and 4.0M HCl in dioxane (0.16mL, 0.64mmol, 1.9eq) was added at room temperature. After 3 hours, an additional 1.9eq HCl was added and the mixture was stirred for an additional 3 hours. Saturated aqueous sodium bicarbonate was added, the organic layer was separated and dried. Evaporation of the solvent gave the title product (120 mg; yield 82%).

C₂₃H₃₃N₅O₃

Mass (calculated) [427 ]](ii) a (found value) [ M + H [)⁺]＝428。

LC Rt 1.58min, 100% (10 min method)

¹H-NMR(DMSO-d₆)：1.05(3H，d，J＝6.4Hz)；1.26-1.40(3H，m)；1.50-1.57(1H，m)；1.62-1.68(1H，m)；1.70-1.76(1H，m)；1.96(3H，s)；2.36-2.42(2H，m)；2.53-2.50(2H，m)；2.59-2.62(1H，m)；3.31-3.34(2H，m)；3.37-3.47(4H，m)；3.78(3H，s)；6.80(1H，s)；7.00(2H，br d，J＝8.8Hz)；7.63(2H，br d，J＝8.8Hz)；10.30(1H，s)；12.6(1H，s)。

Example 36

5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoic acid [5- (4-chloro-phenyl) -2H-pyrazol-3-yl ] -amide

a) 5-amino-3- (4-chloro-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

To a solution of 5-amino-3- (4-chloro-phenyl) -pyrazole (2.8g, 14.5mmol, 1.0eq) in DCM (30mL) was added potassium hydroxide (27mL of a 4.5M solution) followed by di-tert-butyl dicarbonate (3.5g, 16.0mmol, 1.1 eq). The mixture was stirred at room temperature until LC-MS showed complete conversion. The organic layer was collected after extraction of the aqueous layer and evaporated to dryness under reduced pressure. The solid was washed with methanol to give 3.6g of a white solid (yield 85%).

C₁₄H₁₆ClN₃O₂

¹H-NMR(DMSO-d₆)：1.68(9H，br s)；5.34(2H，br s)；7.25-7.27(1H，m)；7.35(2H，d，J＝8.4Hz)；7.74(2H，d，J＝8.4Hz)。

b)5- (5-bromo-2-methyl-pentanoylamino) -3- (4-chloro-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester

To a solution of 5-bromo-2-methyl-pentanoic acid (1.79g, 9.2mmol, 1eq) in anhydrous DCM (8mL) was added oxalyl chloride (1.0mL, 12.0mmol, 1.3eq) dropwise and the mixture was stirred at room temperature for 16 h. After evaporation of the solvent and excess oxalyl chloride, the residue was dissolved in anhydrous DCM (8mL) and a solution of tert-butyl 5-amino-3- (4-chloro-phenyl) -pyrazole-1-carboxylate (2.7g, 9.2mmol, 1.0eq) and triethylamine (1.7mL, 12mmol, 1.3eq) was added dropwise at 0 ℃. The mixture was warmed to room temperature and stirred at room temperature for 24 hours, then 0.5eq of activated 5-bromo-2-methyl-pentanoic acid was added. HCl1M was added and the crude product was extracted with DCM and purified over a silica gel column (eluent DCM) to give 3.3g (97% yield) of the title product.

C₂₀H₂₅BrClN₃O₃

Mass (calculated value) [370 ]](ii) a (found value) [ M + H [)⁺]＝370/372。

LC Rt 2.33, 95% (5 min method)

c)5- (4-acetyl- [1, 4] diazepan-1-yl) -2-methyl-pentanoic acid [5- (4-chloro-phenyl) -2H-pyrazol-3-yl ] -amide

1- [1, 4] is reacted]Diazepan-1-yl-ethanone (1.4mL, 10.8mmol, 1.2eq) was added to a solution of 5- (5-bromo-2-methyl-pentanoylamino) -3- (4-chloro-phenyl) -pyrazole-1-carboxylic acid tert-butyl ester (3.3g, 9.0mmol, 1.0eq) and triethylamine (1.25mL, 9.0mmol, 1.0eq) in 2-butanone (15mL), and the mixture was refluxed with stirring for 48 hours. After removal of the solvent, DCM (5mL) and TFA (3mL) were added and stirred at room temperature for 3 hours. DCM and TFA were evaporated under reduced pressure and the crude product was treated with saturated sodium carbonate solution and extracted with ethyl acetate. The crude product was purified on a silica gel column, 100% DCM to DCM-NH₃Solution in MeOH 2N 92: 8) gradient. 1.7g (yield 44%) of the title product are obtained.

C₂₂H30ClN5O₂

Mass (calculated value) [431](ii) a (found value) [ M + H [)⁺]＝432。

LC Rt 1.80min, 90% (10 min method)

¹H-NMR(CDCl₃)：1.14-1.21(3H，d，J＝6.58Hz)；1.36-1.53(1H，m)；1.53-2.0(6H，m)；2.1(3H，s)；2.48-3.07(6H，m)；3.39-3.77(4H，m)；6.93(1H，s)；7.49(2H，d，J＝8.0Hz)；7.71(2H，d，J＝8.0Hz)；10.40(1H，s)；12.87(1H，s)。

Example 37

4-pyrrolidin-1-yl-pentanoic acid [5- (4-chloro-phenyl) -2H-pyrazol-3-yl ] -amide

a) 4-Pyrrolidin-1-yl-pentanoic acid methyl ester

Pyrrolidine (3mL, 36mmol, 1.2eq) was dissolved in DCM (50mL) and methyl levulinate (4mL, 30mmol, 1.0eq) was added. The solution was stirred at room temperature for 1 hour, then Na (OAc) was added₃BH (7.6g, 36.0mmol, 1.2 eq). The mixture was stirred at room temperature for 16 h, then brine was added and the crude product was extracted with DCM to give 2.0g of the title compound after drying (34% yield).

C₁₀H₁₉NO₂

¹H-NMR(CDCl₃)：1.04(3H，d，J＝6.4Hz)；1.67-1.90(6H，m)；2.26-2.43(3H，m)；2.51-2.54(4H，m)；3.64(3H，s)。

b) 4-pyrrolidin-1-yl-pentanoic acid

To a suspension of 4-pyrrolidin-1-yl-pentanoic acid methyl ester (2.0g, 10.0mmol) in water (20mL) was added NaOH (0.8g, 20.0mmol, 2.0eq) and the mixture was refluxed for 10 hours. The reaction was then cooled to room temperature, adjusted to pH 3 with HCl 37%, and concentrated under reduced pressure. The residue was treated with ethanol, the precipitated sodium chloride was filtered off and the solvent was evaporated under reduced pressure to give 1.7g of the title compound as a white solid (99% yield).

C₉H₁₇NO₂

¹H-NMR(DMSO-d₆)：1.22(3H，d，J＝6.4Hz)；1.64-1.74(1H，m)；1.81-1.96(4H，m)；1.97-2.07(1H，m)；2.23-2.30(1H，m)；2.36-2.44(1H，m)；2.97-3.02(2H，m)；3.20-3.26(1H，m)；3.35-3.46(2H，m)；10.80(1H，s)

c) 4-pyrrolidin-1-yl-pentanoic acid [5- (4-chloro-phenyl) -2H-pyrazol-3-yl ] -amide

According to a general method for the synthesis of omega-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl) -amides, prepared using the amino acid route starting from 5- (4-chloro-phenyl) -2H-pyrazol-3-ylamine (97.0mg, 0.5mmol, 1.0eq) and 4-pyrrolidin-1-yl-pentanoic acid (128.0mg, 0.7mmol, 1.5 eq). The reaction was stirred at room temperature for 16 hours and then at 50 ℃ for 8 hours to complete the formation of the acylated isomer of the exocyclic nitrogen atom. The crude product was purified by preparative HPLC to give 150.3mg of the title compound as formate salt (87% yield).

C₁₈H₂₃ClN₄O

Mass (calculated value) [346 ]](ii) a (found value) [ M + H [)⁺]＝347。

LC Rt 1.69min, 100% (10 min method)

¹H-NMR(DMSO-d₆: formate salt): 1.11(3H, d, J ═ 6.4 Hz); 1.63-1.80(5H, m); 1.90-1.99(1H, s); 2.29-2.42(2H, m); 2.80-2.86(5H, m); 6.82(1H, s); 7.46-7.49(2H, m); 7.70-7.73(2H, m); 8.19(1H, s); 10.55(1H, br s)

TABLE 3 examples 38 to 372

A portion of the compounds synthesized according to the method described in the last column of the table and discussed in detail in the synthetic methods of examples 1-37 are given in table 3. When the compound is in the form of the HCl salt, the salt is formed by dissolving the free base in methanol and adding 1eq 1M HCl in ether followed by evaporation of the solvent. When the compound is in the form of a HCOOH (formate) salt, the compound is purified by preparative HPLC.

Biological activity

Cloning of alpha 7 nicotinic acetylcholine receptors to generate recombinant cell lines stably expressing alpha 7nAChR

Full-length cDNAs encoding the α 7 nicotinic acetylcholine receptors were cloned from rat brain cDNA libraries according to standard molecular biology techniques. Rat Rat GH4C1 cells were then transfected with Rat receptor cDNA and assayed for expression of functional α 7 nicotinic receptor by measuring changes in intracellular calcium concentration following cloning using the FLIPR assay. Cells showing the strongest calcium-mediated fluorescent signal upon stimulation with agonist (nicotine) were further cloned and then stained with texas red-labeled α -bungarotoxin (BgTX) to analyze the level and uniformity of α 7 nicotinic acetylcholine receptor expression using confocal microscopy. Three cell lines were then expanded, and the pharmacological profile of one of the cell lines was studied (see table 4 below) and then used to screen compounds.

TABLE 4 study of pharmacological characteristics of stably expressed α 7nAChR in GH4C1 cells Using a functional FLIPR assay

Compound (I)	EC₅₀[μM]
		Acetylcholine	3.05±0.08(n＝4)
Choline	24.22±8.30(n＝2)
		Cytisine	1.21±0.13(n＝5)
DMPP	0.98±0.47(n＝6)
		Epibatidine	0.012±0.002(n＝7)
Nicotine	1.03±0.26(n＝22)

Establishment of functional FLIPR assay for Primary screening

A stable functional FLIPR assay (Z' ═ 0.68) was established using a stable GH4C1 recombinant cell line for screening α 7 nicotinic acetylcholine receptors. The FLIPR system may use Ca²⁺Real-time measurement of Ca in living cells with sensitive fluorescent dyes (e.g., Fluo4)²⁺The concentration changes. The instrument can screen agonists and antagonists of the alpha 7nAChR channel stably expressed in GH4C1 cells.

Cell culture

Stably transfected rat α 7-nAChR GH4C1 cells (as described above) were used. These cells have poor adsorptivity and are therefore usedpoly-D-lysine pretreated flasks and plates. Cells at 150cm²Growing in T-type culture flask, adding 30ml of culture medium, and culturing at 37 deg.C and 5% CO₂Culturing under the condition.

Data analysis

EC was calculated using the IDBS XLFit4.1 software package according to the Sigmoidal concentration response (variable slope) equation below₅₀And IC₅₀：

Y ═ lowest value + ((highest-lowest value)/(1 + ((EC)₅₀/X) ^ slope Hill slope)

Method verification

Functional FLIPR assays were validated with the α 7nAChR agonists nicotine, cytisine, DMPP, Epibatidine, choline, and acetylcholine. Establishing a concentration response curve, EC, between 0.001 and 30. mu.M₅₀The values are shown in Table 2, and the activity sequences of the resulting agonists are consistent with literature reports (Quik et al, 1997, mol. Pharmacol., 51, 499-506).

The assay was further validated with a specific α 7nAChR antagonist MLA (methyl taurine) at 1 μ M to 0.01nM in competition with 10 μ M nicotine. IC calculated from nine independent experiments₅₀The value was 1.31. + -. 0.43 nM.

Establishment of functional FLIPR assay for Selective screening

A functional FLIPR assay was set up to determine the selectivity of compounds for the α 1 (muscle) and α 3 (ganglion) nACh receptors and the structurally related 5-HT3 receptor. The membrane potential sensitive dye-based assay was used to determine the α 1 receptor activity naturally expressed by TE 671 cell line derived from rhabdomyosarcoma, whereas α 3 selectivity was determined by calcium monitoring assay in the native SH-SY5Y cell line. Recombinant cell lines expressing the human 5-HT3A receptor in HEK 293 cells were constructed and selectivity for the 5-HT3 receptor was determined according to the calcium monitoring FLIPR assay.

Screening compounds

Each compound was tested in a recombinant GH4C1 cell line stably expressing α 7nAChR using the functional FLIPR primary screening method described above. Concentration response curves were established to further validate the hits identified. The potency of the compounds of examples 1-372 was 10 nM-10. mu.M, with the majority of the titers ranging from 100 nM-5. mu.M, as determined by the functional FLIPR screening assay.

The compounds were also found to be selective for the α 1 nAChR, α 3 nAChR and the 5HT3 receptor.

Claims

1. Compounds of formula (I) and salts, tautomers, diastereomers and racemic mixtures thereof:

wherein

T is propane-1, 3-diyl, optionally substituted by (C1-C3) alkyl, halogen;

z is CH₂；

Q is

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl, alkoxy, acyl;

j is 0;

r is a 5 to 10 membered aromatic or heteroaromatic ring;

m is 0, 1, 2 or 3;

y is halogen; a hydroxyl group; a mercapto group; a cyano group; a nitro group; an amino group; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl, di-or trihaloalkoxy, alkoxy or alkylcarbonyl; (C3-C6) cycloalkyl- (C1-C6) alkoxy; (C3-C6) cycloalkyl- (C1-C6) alkyl; a linear, branched or cyclic (C1-C6) alkylcarbonylamino group; mono-or di-linear, branched or cyclic (C1-C6) alkylaminocarbonyl; a carbamoyl group; a linear, branched or cyclic (C1-C6) alkylsulfonylamino group; a linear, branched or cyclic (C1-C6) alkylsulfonyl group; mono-or di-linear, branched or cyclic (C1-C6) alkylsulfamoyl; linear, branched or cyclic (C1-C6) alkoxy- (C1-C6) alkyl, wherein when m is greater than 1, Y are independent of each other; or when m ═ 2, two Y substituents may form a ring together with the atom of the R group to which they are attached.

2. The compound of claim 1, wherein

T is propane-1, 3-diyl;

z is CH₂；

q and q' are independently of each other 1 or 2;

p is 0 or 1;

r' is selected from linear, branched or cyclic (C1-C6) alkyl;

q is

j is 0;

r is phenyl, pyridyl, naphthyl;

m is 1 or 2;

y is halogen; a hydroxyl group; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl, di-or trihaloalkoxy, alkoxy; (C3-C6) cycloalkyl- (C1-C6) alkoxy, wherein Y are independent of each other when m is greater than 1.

3. The compound of claim 2, wherein Q-R is

4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier or excipient.

5. Use of a compound according to any one of claims 1-3 for the preparation of a medicament for the treatment of neurological disorders, neurodegenerative disorders, psychiatric disorders, cognitive disorders, immune disorders, inflammatory disorders, metabolic disorders, addiction, nociceptive disorders, and sexual dysfunction.

6. The use according to claim 5 for the treatment of senile dementia, attention deficit disorder, Alzheimer's disease and schizophrenia.