OA18860A - Pyrazolo[1,5-A)pyrazin-4-yl derivatives as Jak-inhibitors - Google Patents

Pyrazolo[1,5-A)pyrazin-4-yl derivatives as Jak-inhibitors Download PDF

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OA18860A
OA18860A OA1201800314 OA18860A OA 18860 A OA18860 A OA 18860A OA 1201800314 OA1201800314 OA 1201800314 OA 18860 A OA18860 A OA 18860A
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alkyl
compound
pyrazol
hydroxy
pharmaceutically acceptable
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OA1201800314
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Matthew Frank Brown
Matthew Merrill Hayward
Stephen Wayne Wright
Andrew Fensome
Alpay Dermenci
Brian Stephen Gerstenberger
Dafydd Rhys Owen
Li Huang XING
Xiaojing Yang
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Pfizer Inc.
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Publication of OA18860A publication Critical patent/OA18860A/en

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Abstract

A compound compound having the structure (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate of said compound or pharmaceutically acceptable salt, wherein A, A' and A" are independently O, C=O, C-R' or N-R", where R' and R" may independently be H, amino, -NR7COR6, COR6 , -CONR7R8, C1-C6 alkyl, or hydroxy(C1-C6 alkyl), and R" may be present or absent, and is present where the rules of valency permit, and where not more than one of A, A' and A" is O or C=0; R0 and R are independently H, Br, CI, F, or C1-C6 alkyl; R1 is H, C1-C6 alkyl, or hydroxy(C1-C6 alkyl); R2 is selected from the group consisting of H, C1-C6 alkyl, C1-C6 alkoxy, hydroxy(C1-C6 alkyl), phenyl(C1-C6 alkyl), formyl, heteroaryl, heterocyclic, -COR6, -OCOR6, COOR6, -NR7COR6, -CONR7R8, and -(CH2)n-W, where W is cyano, hydroxy, C3-C8 cycloalkyl, SO2NR7R8, and -SO2-R9, where R9 is C1-C6 alkyl, C3-C8 cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or C1-C6 alkyl; X is C-R3 or N, where R3 may be H or C1-C6 alkyl; R4 and R5 are independently H, amino, C1C6 alkyl, or hydroxy(C1-C6 alkyl); R6, R7 and R8 are each independently H, C1-C6 alkyl, C1-C4 alkoxy(C1-C6 alkyl), or C3-C8 cycloalkyl, said C1C6 alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and R8 together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or C1-C6 alkyl; and, n is 0, 1, 2 or 3. Also provided are methods of treatment as Janus Kinase inhibitors and pharmaceutical compositions containing the compounds of the invention and combinations thereof with other therapeutic agents.

Description

PYRAZOLO[1,5-A]PYRAZIN-4-YL DERIVATIVES AS JAK-INHIBITORS
FIELD OF THE INVENTION
The présent invention provides pharmaceutically active pyrazolo[1,5-a]pyrazin~4-yl TYK2 ligands and analogues. Such compounds are useful for inhibiting Janus Kinases (JAKs). This invention also is directed to compositions comprising methods for making such compounds, and methods fortreating and preventing conditions mediated by JAK.
BACKGROUND OF THE INVENTION
Protein kinases are families of enzymes that catalyze the phosphorylation of spécifie residues in proteins, broadly classified into tyrosine and serine/threonine kinases. Inappropriate kinase activity, arising from mutation, over-expression, or inappropriate régulation, dys-regulation or de-regulation, as well as over- or under-production of growth factors or cytokines has been implicated in many diseases, including but not limited to cancer, cardiovascular diseases, allergies, asthma and other respiratory diseases, autoimmune diseases, inflammatory diseases, bone diseases, metabolic disorders, and neurological and neurodegenerative disorders such as Alzheimer's disease. Inappropriate kinase activity triggers a variety of biological cellular responses reiating to cell growth, cell différentiation, cell function, survival, apoptosis, and cell mobility implicated in the aforementioned and related diseases.
Thus, protein kinases hâve emerged as an important class of enzymes as targets for therapeutic intervention. In particular, the JAK family of cellular protein tyrosine kinases (JAK1, JAK2, JAK3, and Tyk2) play a central rôle in cytokine signaling (Kisseleva et al., Gene, 2002, 285, 1; Yamaoka et al. Genome Biology 2004, 5, 253)). Upon binding to their receptors, cytokines activate JAK which then phosphorylate the cytokine receptor, thereby creating docking sites for signaling molécules, notably, members of the signal transducer and activator of transcription (STAT) family that ultimately lead to gene expression. Numerous cytokines are known to activate the JAK family. These cytokines include, the interferon (IFN) family (IFN-alpha, IFN-beta, IFN-omega, Limitin, IFN-gamma, IL-10, IL-19, IL-20. IL-22), the gp130 family (IL-6, IL-11, OSM, LIF, CNTF, NNT-1/EJSF-3, G-CSF, ΟΤΙ, Leptin, IL-12, IL-23), gamma C family (IL-2, IL-7, TSLP, !L-9, IL-15, IL-21, IL-4, IL-13), IL-3 family (IL-3, IL-5, GM-CSF), single chain family (EPO, GH, PRL, TPO), receptor tyrosine kinases (EGF, PDGF, CSF-1, HGF), and G-protein coupled receptors (AT1).
There remains a need for new compounds that effectively and selectively inhibit spécifie JAK enzymes: TYK2 in particular. TYK2 is a JAK kinase family member, and is important in the signaling of the type l interferons (including IFNalpha, INFbeta), IL-6, IL-10, 1L-12 and IL-23 (Liang, Y. et al., Expert Opinion on Therapeutic Targets, 18, 5, 571-580 (2014)). As such, TYK2 signais with other members of the JAK kinase family in the following combinations: TYK2/JAK1, TYK2/JAK2, TYK2/JAK1/JAK2. TYK2 has been shown to be important in the différentiation and function of multiple cell types important in inflammatory disease and autoimmune disease including natural killer celle, B cells, and T helper cell types. Aberrant TYK2 expression is associated with multiple autoimmune or inflammatory conditions. Modulation of immune activity through inhibition of TYK2 kinase activity can prove useful in the treatment of various immune disorders (O'Shea JJ, Plenge R, Immunity, 36, 54250 (2012); Murray, P.J., J. tmmunol., 178, 2623-2629 (2007); Kisseleva, T., et al., Gene, 285, 1-24 (2002)) while avoiding JAK2 dépendent erythropoietin (EPO) and thrombopoietin (TPO) signaling (Neubauer H., et al„ Cell, 93(3), 397-409 (1998); Parganas E„ et al., Cell, 93(3), 385-95 (1998)).
SUMMARY OF THE INVENTION
The présent invention provides a compound of formula I having the structure:
(l) or a pharmaceutically acceptable sait thereof, or a pharmaceuticaliy acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: A, A’ and A are independently O, C=O, CR' or N-R, where R' and R may independently be H, amino, -NR?CORe, COR(l) * * * * 6, -CONRîRb, Ci-Cb alkyl, or hydroxy(Ci-Ce alkyl), and R” may be présent or absent, and is présent where the rules of valency permit, and where not more than one of A, A' and A is O or C=O; Ro and R are independently H, Br, Cl, F, or Ci-Ce alkyl; Ri is H, Ci-Cb alkyl, or hydroxy(Ci-Ce alkyl); R2 is selected from the group consisting of H, Ci-Cb alkyl, Ci-C6 alkoxy, hydroxy(Ci-Ce alkyl), phenyl(Ci-Ce alkyl), forrnyl, heteroaryl, heterocyclic, -CORe, -OCORe, -COORe, -NR?CORb, -CONRtRs, and -(CH2)n-W, where W is cyano, hydroxy, Ca-Ce cycloalkyl, -SO2NR7R8, and -SO2-R9, where Rg is Ci-Cb alkyl, Cs-Cb cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Cb alkyl; X is C-R3 or N, where R3 may be H or Ci-Cb alkyl; Ri and Rs are independently H, amino, Ci-Ce alkyl, or hydroxy(Ci-Ce alkyl); Rb, R7 and Ra are each independently H, Ci-C6 alkyl, Ct-C4 alkoxy(Ct-Ce alkyl), or C3-C0 cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Rb together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ca alkyl; and, n is 0, 1, 2 or 3.
In other aspects, the présent invention also provides:
pharmaceutical compositions which comprise a pharmaceutically acceptable carrier and a compound of formula I;
methods for treating conditions or disorders including inflammation, autoimmune disease, systemic lupus erythematous, lupus nephritis, discoid lupus, cutaneous lupus, central nervous System iupus, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, asthma, allergie asthma, Type I diabètes, polymyositis, dermatomyositis, type I interferonopathies including Aicardi-Goutières syndrome and other mendelian diseases of overexpression of type I interferon, multiple sclerosis, primary progressive multiple sclerosis, relapsing remitting multiple sclerosis, primary biliary cirrhosis also known as primary biliary cholangitis, primary sclerosing cholangitis, autoimmune hepatitis, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, psoriasis, dermatomyositis, scleroderma, atopie dermatitis, vitiligo, alopecia areata, spondylopathy, ankylosing spondylitis, Alzheimer’s disease, neuro-inflammation myositis, vasculitis, pemphigus, Crohn’s disease, lupus, nephritis, psoriasis, multiple sclerosis, major dépréssion disorder, allergy, asthma, Sjogren's disease, dry eye syndrome, transplant rejection, cancer, inflammatory bowel disease, septic shock, cardiopulmonary dysfunction, vitiligo, alopecia, acute respiratory disease, ankylosing spondylitis, autoimmune hepatitis, primary sclerosing cholangitis, primary biliary cirrhosis, Alzheimer’s disease, or cachexia by administering to a subject in need a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable sait thereof;
Methods for treating conditions or disorders including atopie dermatitis, eczema, psoriasis, scleroderma, lupus, pruritus, fatigue, other pruritic conditions, allergie reactions including allergie dermatitis in mammal, horse allergie diseases including bite hypersensitivity, summer eczema, sweet itch in horses, heaves, inflammatory airway disease, récurrent airway obstruction, airway hyperresponsiveness, and chronic obstruction pulmonary disease by administering to a mammal in need a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable sait thereof; and, methods for the préparation of compounds of the présent invention.
The présent invention will be further understood from the following description given by way of example only. The présent invention is directed to a class of pyrazolo[1,5-a]pyrazin-4-yl dérivatives. In particular, the présent invention is directed to pyrazolo[1,5-a]pyrazin-4-yl compounds useful as inhibitors of JAKs, and particularly TYK2. While the présent invention is not so limited, an appréciation of various aspects of the invention will be gained through the following discussion and the examples.
The terrn “alkyl, alone or in combination, means an acyclic, saturated hydrocarbon group of the formula Cnhbn+i which may be linear or branched. Examples of such groups include methyl, ethyl, n-propyl, isopropyl, π-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl and hexyl. Unless otherwise specified, an alkyl group comprises from 1 to 6 carbon atoms. The carbon atom content of alkyl and various other hydrocarbon-containing moieties is indicated by a prefix designating a lower and upper number of carbon atoms in the moiety, that is, the prefix Ci-Cj indicates a moiety of the integer i to the integer j carbon atoms, inclusive. Thus, for example, Ci-Ce alkyl refers to alkyl of one to six carbon atoms, inclusive.
The terrn “hydroxy,” as used herein, means an OH group. The term “heterocyclic refers to a saturated or partially saturated {i.e., non aromatic) heterocycle which contains three to ten ring atoms where one or more, preferably, one, two or three ring atoms, are heteratom(s) selected from N, O and S, the remaining being carbon, and which may be attached via a ring nitrogen atom or a ring carbon atom. Equally, when substituted, the substituent may be located on a ring nitrogen atom (if the substituent is joined through a carbon atom) or a ring carbon atom (in ail cases). Spécifie examples include oxiranyl, aziridinyl, oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, 1,4-dioxanyl, morpholinyi, piperazinyl, azepanyl, oxepanyl, oxazepanyl and diazepinyt.
The term aryl refers to an aromatic monocyclic or bicyclic hydrocarbon containing six to ten ring carbon atoms which may be attached via one of the ring carbon atoms. Equally, when substituted, the substituent may be located on a ring carbon atom. Spécifie examples include, but are not limited to, phenyl, tolyl, xylyl, trimethylphenyl, and naphthyl. Examples of aryl substituents include, but are not limited to, alkyl, hydroxyl, halo, nitrile, alkoxy, trifluoromethyl, carboxamido, SOîMe, benzyl, and substituted benzyl.
The term “heteroaryl refers to a monovalent aromatic monocyclic or bicyclic heterocycle of five to ten ring atoms where one or more, preferably, one, two or three ring atoms, are heteratom(s) selected from N, O, and S, the remainîng being carbon, and which may be attached via a ring carbon atom or a ring nitrogen atom with an appropriate valency. Equally, when substituted, the substituent may be located on a ring carbon atom or a ring nitrogen atom with an appropriate valency. Spécifie examples include, but are not limited to, thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl and pyrazinyl. The term cycloalkyl means a monocyclic, saturated hydrocarbon group of the formula Cnhkn-i. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Unless otherwise specified, a cycloalkyl group comprises from 3 to 8 carbon atoms.
The terms “halo” and “halogen refer to fluoride (F), chloride (Cl), bromide (Br) or iodide (I).
The term mammal refers to human, livestock or companion animais.
The term companion animal” or “companion animais refers to animais kept as pets or household animal. Examples of companion animais include dogs, cats, and rodents including hamsters, guinea pigs, gerbils and the like, rabbits, ferrets and birds.
The term livestock refers to animais reared or raised in an agricultural setting to make Products such as food or fiber, or for its labor. In some embodiments, livestock are suitable for consumption by mammals, for example humans. Examples of livestock animais include cattle, goats, horses, pigs, sheep, including lambs, and rabbits, as well as birds, such as chickens, ducks and turkeys.
The term “treating or treatment” means an alleviation of symptoms associated with a disease, disorder or condition, or hait of further progression or worsening of those symptoms. Depending on the disease and condition of the patient, the term “treatment as used herein may include one or more of curative, palliative and prophylactic treatment. Treatment can also include administering a pharmaceutical formulation of the présent invention in combination with other thérapies.
The term therapeutically-effective indicates the capability of an agent to prevent, or improve the severity of, the disorder,. The phrase therapeutically-effective is to be understood to be équivalent to the phrase effective for the treatment, prévention, or amelioration, and both are intended to qualify the amount of an agent - which will achieve the goal of improvement in the severity of cancer, cardiovascular disease, or pain and inflammation and the frequency of incidence over treatment of each agent by itself.
“Pharmaceutically acceptable means suitable for use in mammals, companion animais or livestock animais.
If substituents are described as being “independently selected” from a group, each substituent is selected independent of the other. Each substituent therefore may be identical to or different from the other substituent(s).
DETAILÈD DESCRIPTION OF THE INVENTION
The présent invention is related to novel compounds which are TYK2 modulators useful for the treatment of diseases and conditions associated with dysrégulation of TYK2. The présent invention further provides pharmaceutical compositions comprising such JAK enzyme modulators as well as methods of treating and/or preventing such diseases and conditions. Accordingly, the présent invention provides a compound of formula I as represented above having the structure (I):
The invention also provides a compound having the structure (la):
or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: A, A' and A are independently O, C=O( C-R' or N-R, where R’ and R may independently be H, amino, -NRrCORa, COR6, -CONRtRs, Ci-Ca alkyl, or hydroxy(Ct-Ca alkyl)-, and R” may be présent or absent, and is présent where the rules of valency permit, and where not more than one of A, A’ and A is O or C=O; Ro and R are independently H, Br, Cl, F, or Ci-Ce alkyl; Ri is H, Ci-Ca alkyl, or hydroxy(Ci-Ca alkyl)-; R2 is selected from the group consisting of H, Ci-Ca alkyl, Ci-Ce alkoxy-, hydroxy(Ci-Ce alkyl)-, phenyl(Ci-Ca alkyl)-, formyl, heteroaryl, heterocyclic, -CORb, -OCORe, -COORa, -NRrCORe, -CONRtRb, and -(CH2)n-W, where W is cyano, hydroxy, Ca-Ca cycloalkyl, -SO2NR7R8, and -SO2-R9, where Rs is Ci-Ca alkyl, Ca-Ca cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ce alkyl; R3 may be H or Ci-Cb alkyl; R« and Rs are independently H, amino, Ci-C6 alkyl, or hydroxyfCT-Ca alkyl); Ra, R7 and Ra are each independently H, Ci-Ca alkyl, C1-C4 alkoxy(Ci-Ce alkyl), or Ca-Ca cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ca alkyl; and, n is 0, 1,2 or 3.
The invention further provides a compound having the structure (Ib):
or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: R is H, -CORs, -CONRtRb, Ci-Ca alkyl-, or hydroxy(Ci-Ca alkyl)-; Ro and R are independently H, Br, Cl, F, or Ci-Ce alkyl; Ri is H, Ci-Ca alkyl, or hydroxy(Ci-Ca alkyl); R2 is selected from the group consisting of H, Ci-Ca alkyl, Ci-Ca alkoxy, hydroxy(Ci-Ca alkyl), phenyl(Ci-Ca alkyl), formyl, heteroaryl, heterocyclic, -CORa, -OCORe, -COORa, NRrCORe, -CONRrRa, and -(CFhJn-W, where W is cyano, hydroxy, Ca-Ca cycloalkyl, -SO2NR7R8, and -SO2-R9, where R9 is Ci-Ca alkyl, Ca-Ca cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ca alkyl; R3 may be H or Ci-Ca alkyl; Ra is H, amino, Ct-Ca alkyl, or hydroxy(Ci-Ca alkyl); Ra, R? and Ra are each independently H, Ci-Ca alkyl, C1-C4 alkoxy(Ci-Ce alkyl), or Ca-Ca cycloalkyl, said Ci-Ca alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Re together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ce alkyl; and, n is 0, 1, 2 or 3.
The invention also provides a compound having the structure (le):
or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: R” is H, -CORe, -CONRyRa, Ci-Ce alkyl, or hydroxy(Ci-Cs alkyl); R2 is selected from the group consisting of H, Ci-Ce alkyl-, Ci-Cs alkoxy-, hydroxy(Ci-Ce alkyl)-, phenyl(Ci-C6 alkyl)-, formyl, heteroaryl, heterocyclic, -CORe, -OCORs, -COORs, -NR7COR6, -CONR7R8, and -(CH2)n-W, where W is cyano, hydroxy, C3-Ce cycloalkyl, -SO2NR7R8, and -SO2-R9, where Rg is Ci-C6 alkyl, C3-Cs cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-C6 alkyl; R3 is H or Ci-C6 alkyl; Rs is H, amino, Ci-Ce alkyl-, or hydroxy(Ci-C6 alkyl)-; Re, R7 and Re are each are each independently H, Ci-Ce alkyl-, C1-C4 alkoxy(Ci-Ce alkyl)-, or C3-Cb cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ca alkyl; and, n is 1, 2 or 3. In a particular embodiment, the invention provides said compound wherein R” is Ci-Ce alkyl and Rs is H.
The invention also provides a compound having the structure (Id);
(Id) or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: R is H, -CORg, -CONRtRs, Ct-Cg alkyl, or hydroxy(Ci-C6 alkyl); Rz is selected from the group consisting of H, Ci-Ce alkyl, Ci-Cg alkoxy, hydroxy(Ci-Cg alkyl), phenyl(Ci-Ce alkyl), formyl, heteroaryl, heterocyclic, -CORg, -OCORg, -COORg, NRrCORe, -CONR7R0, and -(CHzJrrW, where W is cyano, hydroxy, C3-C8 cycloalkyl, -SOzNRzRg, and -SO2-R9, where Rg is Ci-Cg alkyl, C3-C8 cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Cg alkyl; Rs is H, amino, Ct-Cg alkyl-, or hydroxy(Ci-Cg alkyl)-; Rg, R7 and Rg are each independently H, Ct-Cg alkyl, C1-C4 alkoxy(Ci-C6 alkyl), or Ca-Cg cycloalkyl, said Ci-Cg alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Rg together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Cb alkyl; and, n is 1, 2 or 3. In a particular embodiment, the invention provides said compound wherein R is Ci-Cg alkyl and Rg is H.
The invention also provides a compound having the structure (le):
(le) or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: R2 is selected from the group consisting of H, Ct-Cg alkyl-, Ct-Cg alkoxy-, hydroxy(Ct-Cg alkyl)-, phenyl(Ci-Cg alkyl)-, formyl, heteroaryl, heterocyclic, -COR6, -OCOR6, -COOR6, -CONR7R8, and -(CH2)n-W, where W is cyano, hydroxy, C3Ce cycloalkyl, -SOzNR7R8, and -SO2-R', where R' is Ct-Cg alkyl, Cs-Ca cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ct-Cg alkyl; Rg, R7 and Rg are each are each independently H, Ct-Cg alkyl-, C1-C4 alkoxy(Ct-Cg alkyl)- or C?,-Ca cycloalkyl said Ct-Cg alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ce alkyl; and, π is 1, 2 or 3. In a particular embodiment, the invention provides said compound wherein R2 is -(CH2)n-W, where W is cyano and π is 1,2 or 3.
The invention also provides a compound having the structure (If):
<lf) or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein: R is H, -CORb, -CONRïRs, Ct-Ce alkyl-, or hydroxy(Ci-C6 alkyl)-; Rs is H, amino, Ci-Ce alkyl-, or hydroxy(Ci-Ce alkyl)-; Re, R? and Re are each are each independently H, Ci-Cs alkyl-, C1-C4 alkoxy(Ct-C6 alkyl)-, or Ca-Ca cycloalkyl, said C1-C6 alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Cj-Ca alkyl; and, n is 0, 1. 2 or 3. In a particuîar embodiment, the invention provides said compound wherein R” is Ci-Ce alkyl. In another particuîar embodiment, the invention provides said compound wherein R is methyl.
In certain preferred embodiments, the invention provides a compound selected from the group consisting of:
(1r,3r)-3-(4-(6-(3-amino-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)-3-(cyanomethyl)cyctobutane-1-carbonitrile;
2,2-(3-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)azetidine1,3-diyl) diacetonitrrle;
2-((1s,3r)-1-(4-(6-(5-(hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol1-yl)-3-methoxycyclobutyl)acetonitrile;
5-(4-(1-((1 s, 3r)-1-(cyanomethyl)-3-methoxycyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin6-yl)-1H-pyrazole-3-carboxamide;
(1s,3s)-3-(cyanomethyl)-3-(4-(6-(5-(hydroxymethyl)isoxazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)1 H-pyrazol-1 -yl)cyclobutane-1 -carbonitrile;
(1 r,3r)-3-(cyanomethyl)-3-(4-(6-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 Hpyrazol-1 -yl)cyclobutane-1 -carbonitrile;
(1s,3s)-3-(cyanomethyl)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-yl)cyclobutane-1-carbonitrile;
(1r,3r)-3-(cyanomethyl)-3-(4-(3-methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile;
2-((1 r,3s)-1-(4-(6-(3-amino-1H-pyrazol-5-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -yl)-3 methoxycyclobutyl)acetonitrile;
(1 r,3r)-3-(cyanomethyl)-3-(4-(6-(1-(hydroxymethyl)-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile; and,
2-(1-ethyl-3-(4-(6-(5-(hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol1-yl)azetidin-3-yl)acetonitrile, or, a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is (1r,3r)-3-(4-(6-(3amino-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is 2,2'-(3-(4-(6-(1-methyl1H-pyrazol-4-yl)pyrazolo[1.5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidine-1,3-diyl)diacetonitrile, or a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is 2-((1s,3r)-1-(4-(6-(5(hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrite, or a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is 5-(4-(1-((1 s,3r)-1(cyanomethyl)-3-methoxycyciobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6-yl)-1H-pyrazole-3carboxamide, or a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is (1s,3s)-3(cyanomethyl)-3-(4-(6-(5-(hydroxymethyl)isoxazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
In a certain embodiment, the invention provides a compound which is (1r,3r)-3-(cyanomethyl)3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1.5-a]pyrazin-4-yl)-1H-pyrazol-1-yt)cyclobutane-1carbonitrile, or a pharmaceutically acceptable sait thereof.
In another certain embodiment, the invention provides a compound which is (1s,3s)-3(cyanomethyi)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
In yet another embodiment, the invention provides a compound which is (1r,3r)-3(cyanomethyl)-3-(4-(3-methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
In a certain other embodiment, the invention provides a compound which is 2-((1r,3s)-1-(4-(6(3-amino-1H-pyrazol-5-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile, or a pharmaceutically acceptable sait thereof.
In another certain embodiment, the invention provides a compound which is 2-(1 -ethyl-3-(4(6-(5-(hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidin-3yl)acetonitrile, or, a pharmaceutically acceptable sait thereof.
The invention further provides a pharmaceutical or a veterinary composition comprising a compound of formula I and la-f or a pharmaceutically acceptable sait thereof, and a pharmaceutically acceptable carrier.
The invention also provides a method of treating a disease or condition for which a Tyk2 inhibitor is indicated, in a subject in need of such treatment, comprising administering to the subject a
H therapeutically effective amount of a compound of formula I or la-f, or a pharmaceuticaily acceptable sait thereof, or a pharmaceuticaily acceptable solvaté of said compound or sait.
The invention also provides a method for treating or preventing a disorder or condition selected from allergie rhinitis, nasal congestion, rhinorrhea, perennial rhinitis, nasal inflammation, asthma of ail types, chronic obstructive pulmonary disease, chronic or acute bronchoconstriction, chronic bronchitîs, small airways obstruction, emphysema, chronic éosinophilie pneumonia, adult respiratory distress syndrome, exacerbation of airways hyper-reactivity conséquent to other drug therapy, pulmonary vascular disease, pulmonary arterial hypertension, acute lung injury, bronchiectasis, sinusitis, allergie conjunctivitis, idiopathic pulmonary flbrosis or atopie dermatitis, comprising administering to the subject a therapeutically effective amount of a compound of formula I and la-f, or a pharmaceuticaily acceptable sait thereof, or a pharmaceuticaily acceptable solvaté of said compound or sait.
The invention also provides a method of treating primary biliary cirrhosis comprising administering to the subject a therapeutically effective amount of a compound of formula I or la-f, or a pharmaceuticaily acceptable sait thereof, or a pharmaceuticaily acceptable solvaté of said compound or sait.
The invention also provides a method of treating a disease or condition selected from inflammation, inflammation, autoimmune disease, systemic lupus erythematous, lupus nephritis, discoid lupus, cutaneous lupus, central nervous System lupus, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, asthma, allergie asthma, Type I diabètes, polymyositis, dermatomyositis, type I interferonopathies including Aicardi-Goutières syndrome and other mendelian diseases of overexpression of type I interferon, multiple sclerosis, primary progressive multiple sclerosis, relapsing remitting multiple sclerosis, primary biliary cirrhosis also known as primary biliary cholangitis, primary sclerosing cholangitis, autoimmune hepatitis, nonalcoholic fatty liver disease, non-alcoholic steatohepatitis, psoriasis, dermatomyositis, scleroderma, atopie dermatitis, vitiligo, alopecia areata, spondylopathy, ankylosing spondylitis, Alzheimer’s disease, neuro-inflammation comprising administering to the subject a therapeutically effective amount of a compound of formula I or la-f, or a pharmaceuticaily acceptable sait thereof, or a pharmaceuticaily acceptable solvaté of said compound or sait.
The invention also provides a method of treating the symptoms of inflammatory or autoimmune disease, including pruritis and fatigue.
In certain embodiments, the therapeutically effective amount used in accord with the method is from 0.01 mg/kg of body weight/day to 100 mg/kg of body weight/day. In certain other embodiments, the therapeutically effective amount used in accord with the method is wherein the therapeutically effective amount is from 0.1 mg/kg of body weight/day to 10 mg/kg of body weight/day.
Compounds of the invention that hâve the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed isomers. Isomers that differ in the arrangement of their atoms in space are termed stereoisomers. It will be appreciated by those skilled in the art that the compound of formula I can exist as cis- and transachiral diastereomers.
Included within the scope of the described compounds are al! isomers (e.g., cis-, trans-, or dia stéréo mers) of the compounds described herein alone as well as any mixtures. Ail of these forms, including enantiomers, diastereomers, cis, trans, syn, anti, solvatés (including hydrates), tautomers, and mixtures thereof, are included in the described compounds. Stereoisomeric mixtures, e.g., mixtures of diastereomers, can be separated into their corresponding isomers in a known manner by means of suitable séparation methods. Diastereomeric mixtures for example may be separated into their individual diastereomers by means of fractionated crystallization, chromatography, solvent distribution and similar procedures. This séparation may take place either at the level of one of the starting compounds or in a compound of formula I itself. Enantiomers may be separated through the formation of diastereomeric salts, for example by sait formation with an enantiomer-pure chiral acid, or by means of chromatography, for example by HPLC, using chromatographie substrates with chiral ligands. The présent invention includes ail pharmaceutically acceptable isotopically-labelled compounds of formula I wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number which prédominâtes in nature.
Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as 2H and 3H, carbon, such as 11C, 13C and 1«C, chlorine, such as “Cl, fluorine, such as 18F, iodine, such as 123l and 125l, nitrogen, such as 13N and 15N, oxygen, such as 15O, 17O and 1BO, phosphorus, such as 32P, and sulphur, such as “S.
Certain isotopically-labelled compounds of formula 1, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e., 3H, and carbon-14, i.e., t4C, are particularly useful for this purpose in view of their ease of incorporation and ready means of détection.
Substitution with heavier isotopes such as deuterium, i.e., 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds of formula I can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Préparations using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.
In therapeutic use for treating disorders in a mammal, a compound of the présent invention or its pharmaceutical compositions can be administered orally, parenterally, topicaîly, rectally, transmucosally, or intestinally. Parentéral administrations include indirect injections to generate a systemic effect or direct injections to the afflicted area. Topical administrations include the treatment of skin or organs readily accessible by local application, for example, eyes or ears. It aiso includes transdermal detivery to generate a systemic effect. The rectal administration includes the form of suppositories. The preferred routes of administration are oral and parentéral.
I3
Pharmaceutically acceptable salts of the compounds of formula I and la-f include the acid addition and base salts thereof. Suitable acid addition salts are formed from acids which form nontoxic salts. Examples include the acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate, saccharate, stéarate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate salts.
Suitable base salts are formed from bases which form non-toxic salts. Examples include the aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnésium, megiumine, olamine, potassium, sodium, tromethamine and zinc salts.
Hemisalts of acids and bases may also be formed, for example, hemisulfate and hemicalcium salts. For a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Sélection, and Use by Stahl and Wermuth (Wiley-VCH, 2002).
Pharmaceutically acceptable salts of compounds of formula I and la-f may be prepared, respectively, by one or more of three methods: (i) by reacting the compound of formula I and la-f with the desired acid or base; (ii) by removing an acid- or base-labile protecting group from a suitable precursor of the compound of formula I and la-f or by ring-opening a suitable cyclic precursor, for example, a lactone or lactam, using the desired acid or base; or (iii) by converting one sait of the compound of formula I or la-f to another by reaction with an appropriate acid or base or by means of a suitable ion exchange column. Ail three reactions are typically carried out in solution. The resulting sait may precipitate out and be collected by filtration or may be recovered by évaporation of the solvent. The degree of ionization in the resulting sait may vary from completeiy ionized to almost nonion ized.
Pharmaceutical compositions of the présent invention may be manufactured by methods well known in the art, e.g., by means of conventional mixing, dissolving, granulation, dragee-making, levigating, emulsifying, encapsuiating, entrapping, lyophilizing processes or spray drying.
Pharmaceutical compositions for use in accordance with the présent invention may be formulated in conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facititate Processing of the active compound into préparations, which can be used pharmaceutically. Proper formulation is dépendent upon the route of administration chosen. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the instant invention. Such excipients and carriers are described, for example, in Remington's Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991). The formulations of the invention can be designed to be short-acting, fast-releasing, long-acting, and sustained-releasing. Thus, the pharmaceutical formulations can also be formulated for controlled release or for slow release.
Pharmaceutical compositions suitable for use in the présent invention include compositions wherein the active ingrédients are contained in an amount sufficient to achieve the intended purpose,
i.e., control or the treatment of disorders or diseases. More specifically, a therapeutically effective amoLint means an amount of compound effective to prevent, alleviate or ameliorate symptoms/signs of disease or prolong the survival of the subject being treated.
The quantity of active component, which is the compound of this invention, in the pharmaceutical composition and unit dosage form thereof, may be varied or adjusted widely depending upon the manner of administration, the potency of the particular compound and the desired concentration. Détermination of a therapeutically effective amount is well within the capability of those skilled in the art. Generally, the quantity of active component will range between 0.01% to 99% by veight of the composition.
Generally, a therapeutically effective amount of dosage of active component will be in the range of about 0.01 to about 100 mg/kg of body weight/day, preferably about 0.1 to about 10 mg/kg of body weight/day, more preferably about 0.3 to 3 mg/kg of body weight/day, even more preferably about 0.3 to 1.5 mg/kg of body weight/day It is to be understood that the dosages may vary depending upon the requirements of each subject and the severity of the disorders or diseases being treated.
The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrète loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.
Also, it is to be understood that the initial dosage administered may be increased beyond the above upper ievel in order to rapidly achieve the desired plasma concentration. On the other hand, the initial dosage may be smaller than the optimum and the daily dosage may be progressively increased during the course of treatment depending on the particular situation. If desired, the daily dose may also be divided into multiple doses for administration, e.g., two to four times per day..
The présent invention also provides any of the uses, methods or compositions as defined above wherein the compound of formula I or la-f, or a pharmaceuticaîly acceptable sait thereof, or pharmaceutically acceptable solvaté of said compound or sait, is used in combination with another pharmacologically active compound, particularly one of the functionally-defined classes or spécifie compounds listed below. These agents may be administered as part of the same or separate dosage forms, via the same or different routes of administration, and on the same or different administration schedules according to standard pharmaceutical practice known to one skilled in the art.
Suitable agents for use in combination therapy with a compound of formula I or la-f, or a pharmaceutically acceptable sait thereof, or pharmaceutically acceptable solvaté of said compound or sait, sulfasalazine, mesalazine, prednisone, azathioprine, infliximab, adalimumab, belimumab, becertolizumab, natalizumab, vedolizumab, hydrocortisone, budesonide, cyclosporin, tacrolimus, fexofenadine, 6-mercaptopurine, methotrexate, ursodeoxycholic acid, obeticholic acid, antihistamines, rifampin, prednisone , methotrexate, azathioprine, cyclophosphamide, hydroxychloroquine, mofetil, sodium mycophenolate, tacrolimus, leflunomide, chloroquine and quinacrine, thalidomide, rituxan, NSAIDs, solumedrol, depomedrol and dexamethasone.
Other suitabie agents for use in combination therapy with a compound of formula l or la-f, or a pharmaceutically acceptable sait thereof, or pharmaceutically acceptable solvaté of said compound or sait, include: a 5-lipoxygenase activating protein (FLAP) antagonist; a leukotriene antagonist (LTRA) such as an antagonist of LTB4, LTCî, LTD4, LTE4, CysLTi or CysLTz, e.g., montelukast or zafirlukast; a histamine receptor antagonist, such as a histamine type 1 receptor antagonist or a histamine type 2 receptor antagonist, e.g., loratidine, fexofenadine, desloratidine, levocetirizine, methapyrilene or cetirizine; an a1-adrenoceptor agonist or an a2-adrenoceptor agonist, e.g., phenylephrine, methoxamine, oxymetazoline or methylnorephrine; a muscarinic M3 receptor antagonist, e.g. tiotropium or ipratropium; a dual muscarinic M3 receptor antagononist/p2 agonist; a PDE inhibitor, such as a PDE3 inhibitor, a PDE4 inhibitor or a PDE5 inhibitor, e.g., theophylline, sildenafil, vardenafil, tadalafil, ibudilast, cilomilast or roflumilast; sodium cromoglycate or sodium nedocromil; a cyclooxygenase (COX) inhibitor, such as a non-selective inhibitor (e.g., aspirin or ibuprofen) or a sélective inhibitor (e.g. celecoxib or valdecoxib); a glucocorticosteroid, e.g., fluticasone, mometasone, dexamethasone, prednisolone, budesonide, ciclesonide or beclamethasone; an anti-inflammatory monoclonal antibody, e.g., infliximab, adalimumab, tanezumab, ranibizumab, bevacizumab or mepolizumab; a β2 agonist, e.g., salmeterol, albuterol, salbutamol, fenoterol or formoterol, particularly a long-acting β2 agonist; an intigrin antagonist, e.g., natalizumab; an adhesion molécule inhibitor, such as a VLA-4 antagonist; a kinin B1 or B2 receptor antagonist; an immunosuppressive agent, such as an inhibitor of the IgE pathway (e.g., omalizumab) or cyclosporine; a matrix metalloprotease (MMP) inhibitor, such as an inhibitor of MMP-9 or MMP-12; a tachykinin NKi, NK2 or NK3 receptor antagonist; a protease inhibitor, such as an inhibitor of elastase, chymase or catheopsin G; an adenosine A2a receptor agonist; an adenosine A2b receptor antagonist; a urokinase inhibitor; a dopamine receptor agonist (e.g., ropinirole), particularly a dopamine D2 receptor agonist (e.g., bromocriptine); a modulator of the NFkB pathway, such as an IKK inhibitor; a further modulator of a cytokine signalling pathway such as an inhibitor of JAK kinase, syk kinase, p38 kinase, SPHK-1 kinase, Rho kinase, EGF-R or MK-2; a mucolytic, mucokinetic or anti-tussive agent; an antibiotic; an antiviral agent; a vaccine; a chemokine; an épithélial sodium channel (ENaC) blocker or Epithelial sodium channel (ENaC) inhibitor; a nucléotide receptor agonist, such as a P2Y2 agonist; a thromboxane inhibitor; niacin; a 5-lipoxygenase (5-LO) inhibitor, e.g., Zileuton; an adhesion factor, such as VLAM, ICAM or ELAM; a CRTH2 receptor (DP2) antagonist; a prostaglandin D2 receptor (DPr) antagonist; a haematopoietic prostaglandin D2 synthase (HPGDS) inhibitor; interferon-β; a soluble human TNF receptor. e.g., Etanercept; a HDAC inhibitor; a phosphoinositotide 3-kinase gamma (PI3Ky) inhibitor; a phosphoinositide 3-kinase delta (PI3KÔ) inhibitor; a CXCR-1 or a CXCR-2 receptor antagonist; an [RAK-4 inhibitor; and, a TLR-4 or TLR-9 inhibitor, including the pharmaceutically acceptable salts of the specifically named compounds and the pharmaceutically acceptable solvatés of said specifically named compounds and salts.
Accordingly, the invention provides methods of treating or preventing a disease, condition or disorder associated with JAK in a subject, such as a human or non-human mammal, comprising administering an effective amount of one or more compounds described herein to the subject. Suitabie subjects that can be treated include domestic or wild animais, companion animais, such as dogs, cats, horses and the like; livestock including, cows and other ruminants, pigs, poultry, rabbits and the like; primates, for example monkeys, such as rhésus monkeys and cynomolgus (also known as crab-eating or long-tailed) monkeys, marmosets, tamarins, chimpanzees, macaques and the like; and rodents, such as rats, mice, gerbils, guinea pigs and the like. In one embodiment, the compound 5 is administered in a pharmaceutically acceptable form, optionally in a pharmaceutically acceptable carrier.
Conditions in which sélective targeting of the JAK pathway or modulation of the JAK kinases, particularlyTYK2, are contemplated to be therapeutically useful include, inter alia, arthritis, asthma, autoimmune diseases, cancers or tumors, diabètes, certain eye diseases, disorders or conditions, 10 inflammation, intestinal inflammations, allergies or conditions, neurodegenerative diseases, psoriasis, and transplant rejection. Conditions which can benefit from sélective inhibition of ΤΎΚ2 are discussed in greater detail below.
Accordingly, the compound of formula I or la-f or its pharmaceutically acceptable salts and solvatés, and pharmaceutical compositions thereof, can be used to treat a variety of conditions or 15 diseases such as the following:
Arthritis, including rheumatoid arthritis, juvénile arthritis, and psoriatic arthritis;
Autoimmune or inflammatory diseases or disorders, for example Hashimoto's thyroiditis, autoimmune hemolytic anémia, autoimmune atrophie gastritis of pernicious anémia, autoimmune encephatomyelitis, autoimmune orchitis, Goodpasture's disease, autoimmune thrombocytopenia, 20 sympathetic ophthalmia, myasthenia gravis, Graves' disease, primary biliary cirrhosis, autoimmune hepatitis, primary sclerosing cholangitis, chronic aggressive hepatitis, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis ulcerative colitis and membranous glomerulopathy, systemic lupus erythematosis, rheumatoid arthritis, psoriatic arthritis, Sjogren’s syndrome, Reiter's syndrome, polymyositis, dermatomyositis, type I interferonopathies including Aicardi-Goutières syndrome and 25 other mendelian diseases of overexpression of type I interferon systemic sclerosis, polyarteritis nodosa, multiple sclerosis, relapsing remitting multiple sclerosis, primary progressive multiple sclerosis, secondary progressive multiple sclerosis, and bullous pemphigoid, and additional autoimmune diseases, which can be O-cell (humoral) based or T-cell based, including Cogan's syndrome, ankylosing spondylitis, Wegener's granulomatosis, autoimmune alopecia, Type I or 30 juvénile onset diabètes, or thyroiditis;
Cancers or tumors, including alimentary/gastrointestinal tract cancer, colon cancer, liver cancer, skin cancer including mast cell tumor and squamous cell carcinoma, breast and mammary cancer, ovarian cancer, prostate cancer, lymphoma, leukemia, including acute myelogenous ieukemia and chronic myelogenous leukemia, kidney cancer, lung cancer, muscle cancer, bone cancer, bladder 35 cancer, brain cancer, melanoma including oral and metastatic melanoma, Kaposi's sarcoma, myelomas including multiple myeloma, myeloproliferative disorders, proliférative diabetic retinopathy, or angiogenic-associated disorders including solid tumors;
Diabètes, including Type 1 diabètes or complications from diabètes;
Eye diseases, disorders or conditions including autoimmune diseases of the eye, 40 keratoconjunctivitis, vernal conjunctivitis, uveitis including uveitis associated with Behcet's disease
I7 and lens-induced uveitis, keratitis, herpetic keratitis, conical keratitis, corneal épithélial dystrophy, keratoleukoma, ocular premphigus, Mooren's ulcer, scleritis, Grave‘s ophthalmopathy, Vogt-KoyanagiHarada syndrome, keratoconjunctivitis sicca (dry eye), phlyctenule, iridocyclitis, sarcoidosis, endocrine ophthalmopathy, sympathetic ophthalmitis, allergie conjunctivitis, or ocular neovascularization;
intestinal inflammations, including Crohn's disease, ulcerative colitis, inflammatory bowel disease, celiac diseases, proctitis, éosinophilie gastroenteritis, or mastocytosis;
Neurodegenerative diseases including motor neuron disease, Alzheimer's disease, Parkinson's disease, amyotrophie latéral sclerosis, Huntington's disease, cérébral ischemia, or neurodegenerative disease caused by traumatic injury, strike, glutamate neurotoxicity or hypoxia; ischemic/re perfusion injury in stroke, myocardial ischemica, rénal ischemia, heart attacks, cardiac hypertrophy, atherosclerosis and arteriosclerosis, organ hypoxia, or platelet aggregation;
Skin diseases, conditions or disorders including atopie dermatitis, eczema, psoriasis, scleroderma, pruritus or other pruritic conditions, vitiligo, alopecia;
Allergie reactions including allergie dermatitis in mammal (including horse allergie diseases such as bite hypersensitivity), summer eczema, sweet itch in horses, heaves, inflammatory airway disease, récurrent airway obstruction, airway hyper-responsiveness, or chronic obstruction pulmonary disease;
Asthma and other obstructive airways diseases, including chronic or inveterate asthma, late asthma, bronchitis, bronchial asthma, allergie asthma, intrinsic asthma, extrinsic asthma, or dust asthma;
Transplant rejection, including pancréas islet transplant rejection, bone marrow transplant rejection, graft-versus-host disease, organ and cell transplant rejection such as bone marrow, cartilage, cornea, heart, intervertébral dise, islet, kidney, limb, liver, lung, muscle, myoblast, nerve, pancréas, skin, small intestine, or trachea, or xeno transplantation.
Chemical Synthesis
The skilled person will appreciate that the experimental conditions set forth in the schemes that follow are illustrative of suitable conditions for effecting the transformations shown, and that it may be necessary or désirable to vary the précisé conditions employed for the préparation of compounds of formula (I). It will be further appreciated that it may be necessary or désirable to carry out the transformations in a different order from that described in the schemes, or to modify one or more of the transformations, to provide the desired compound of the invention.
Ail of the dérivatives of formula (I) can be prepared by the procedures described in the general methods presented below or by routine modifications thereof. The présent invention also encompasses any one or more of these processes for preparing the dérivatives of formula (I), in addition to any novel intermediates used therein. The person skilled in the art will appreciate that the following reactions may be heated thermally or under microwave irradiation.
It will be further appreciated that it may be necessary or désirable to carry out the transformations in a different order from that described in the schemes, or to modify one or more of the transformations, to provide the desired compound of the invention.
The routes below, including those mentioned in the Examples and Préparations, illustrate methods of synthesising compounds of formula (I). The skilled person will appreciate that the compounds of the invention, and intemnediates thereto, could be made by methods other than those specifically described herein, for example by adaptation of the methods described herein, for example by methods known in the art. Suitable guides to synthesis, functional group interconversions, use of protecting groups, etc., are for example: Comprehensive Organic Transformations” by RC Larock, VCH Publishers Inc. (1989); Advanced Organic Chemistry by J. March, Wiley interscience (1985); Designing Organic Synthesis” by S Warren, Wiley Interscience (1978); “Organic Synthesis - The Disconnection Approach” by S Warren, Wiley Interscience (1982); “Guidebook to Organic Synthesis by RK Mackie and DM Smith, Longman (1982); “Protective Groups in Organic Synthesis by TW Greene and PGM Wuts, John Wiley and Sons, Inc. (1999); and Protecting Groups by PJ, Kocienski, Georg Thieme Verîag (1994); and any updated versions of said standard works.
In addition, the skilled person will appreciate that it may be necessary or désirable at any stage in the synthesis of compounds of the invention to protect one or more sensitive groups, so as to prevent undesirable side reactions. In particular, it may be necessary or désirable to protect amino or carboxylic acid groups. The protecting groups used in the préparation of the compounds of the invention may be used in conventional manner. See, for example, those described in ’Greene's Protective Groups in Organic Synthesis' by Théodore W Greene and Peter G M Wuts, third édition, (John Wiley and Sons, 1999), in particular chapters 7 (“Protection for the Amino Group) and 5 (“Protection for the Carboxyi Group), incorporated herein by référencé, which also describes methods for the removal of such groups.
In the general synthetic methods below, unless otherwise specified, the substituents are as defined above with référencé to the compounds of formula (I) above.
Where ratios of solvents are given, the ratios are by volume.
The compounds of the invention may be prepared by any method known in the art for the préparation of compounds of analogous structure. In particular, the compounds of the invention can be prepared by the procedures described by référencé to the Schemes that follow, or by the spécifie methods described in the Examples, or by similar processes to either.
The skilled person will appreciate that the experimental conditions set forth in the schemes that follow are illustrative of suitable conditions for effecting the transformations shown, and that it may be necessary or désirable to vary the précisé conditions employed for the préparation of compounds of formula (I).
In addition, the skilled person will appreciate that it may be necessary or désirable at any stage in the synthesis of compounds of the invention to protect one or more sensitive groups, so as to prevent undesirable side reactions. In particular, it may be necessary or désirable to protect amino or carboxylic acid groups. The protecting groups used in the préparation of the compounds of the invention may be used in conventional manner. See, for example, those described in ’Greene’s
Protective Groups in Organic Synthesis' by Theodora W Greene and Peter G M Wuts, third édition, (John Wiley and Sons, 1999), in particular chapters 7 (“Protection for the Amino Group) and 5 (“Protection for the Carboxyl Group), incorporated herein by référencé, which also describes methods for the removal of such groups.
Ail of the derivatives of formula (I) can be prepared by the procedures described in the general methods presented below or by routine modifications thereof. The présent invention also encompasses any one or more of these processes for preparing the derivatives of formula (I), in addition to any novel intermediates used therein. The person skilled in the art wili appreciate that the following reactions may be heated thermally or under microwave irradiation.
It will be further appreciated that it may be necessary or désirable to carry out the transformations in a different order from that described in the schemes, or to modify one or more of the transformations, to provide the desired compound of the invention. The schemes are représentative of methods useful in synthesizing the compounds of the présent invention. They are not to constrain the scope of the invention in any way.
According to a first process, compounds of formula I may be prepared from compounds of formulae (A), (B), (C) and (D), as illustrated by Scheme 1.
Y= Hal (i), B(Pin) (ii)
In scheme 1, compound of the Formula Bi (where Y=Hal) is converted to a compound of Formula Bii (Y=B(0R*)2) by treatment with a suitable boronate such as B2(Pin)2, in the presence of a suitable base, such as K2CO3, and a suitable catalyst, such as Pd(dppf)Cl2 in a suitable solvent, such as dioxane. A skilled person also knows that alternative organometallic coupling strategies can be used involving alternative coupling partners, metals and solvent combinations. A compound of the Formula Bii is prepared and isolated as described above or prepared in situ without isolation in a sequential cross-coupling strategy that is well understood by a skilled person. Thus, a compound of Formula Bii is cross-coupled with a compound of Formula A in the presence of a suitable catalyst, such as Pd(dppf)Cl2, with a suitable base, such as K2CO3 in a suitable solvent such as dioxane at a suitable température from room température to reflux température. The resulting compound of Formula C is cross-coupled with a compound of the Formula D containing a suitable leaving group, such as BusSn or (Pin)2B, with a suitable métal catalyst, such as Pd(PPhs)4, in a suitable solvent, such as MeCN at room or elevated températures.
According to a second process a compound of the Formula I is aiso be prepared by the organometallic cross-coupling reaction of compounds of the Formula F with compounds of Formula B, scheme 2.
Y= Hal (i), B(OR*)2 (ii)
Scheme 2
Compound of the Formula Bi (where Y=Hal) is converted to a compound of Formula Bii (Y=B(OR*)z) by treatment with a suitable boronate such as B(Pin)2, in the presence of a suitable base, such as K2CO3, and a suitable catalyst, such as Pd(dppf)Cl2 in a suitable solvent, such as dioxane. A skilled person also knows that alternative organometallic coupling strategies can be used involving alternative coupling partners, metals and solvent combinations. A compound of the Formula Bii is prepared and isolated as described above or prepared in situ without isolation in a sequential crosscoupling strategy that is well understood by a skilled person. Thus. a compound of Formula Bii is cross-coupled with a compound of Formula F in the presence of a suitable catalyst, such as Pd(dppf)Cl2, with a suitable base, such as K2CO3 in a suitable solvent such as dioxane at a suitable température from room température to reflux température.
According to a third process, compound of Formula I is prepared by the alkylation, acylation, sulfonylation etc., of a compound of Formula G, Scheme 3.
(G)
According to a Fourth process, compound of Formula I is prepared by the Michael addition of a compound of Formula H with a compound of Formula J in the presence of a suitable non-nucleophilic base, such as DBU in a suitable solvent, such as MeCN at a suitable température, scheme 4.
DBU
Scheme 4
Scheme 5 illustrâtes a method for the préparation of compounds of Formula F. A diester of Formula Ei is treated with an alkylating agent of Formula K and a base such as K2CO3 in a suitable solvent such as MeCN. The resulting diester of Formula Eii is then cyclized in the presence NHUOAc in a suitable solvent such as EtOH at elevated température to give bicyciic heterocyclic compound of Formula Eiii. The compound of Formula Eiii is hydrolysed to a compound of Formula Eiv with a suitable base, such as LiOH, in a suitable solvent such as MeOH. The resulting carboxylic acid of Formula Eiv is thermally decarboxylated in a suitable solvent, such as sulfolane at elevated température such as 280°C. The resulting compound of Formula Ev is chlorinated by treatment with a suitable reagent, such as POCI3, in a suitable solvent, such as MeCN, at a suitable température such as reflux to préparé compounds of the Formula F.
(Ei)
R' (Eii)
The following schemes and written descriptions provide general details regarding the préparation of the compounds of the invention. The compounds of the invention may be prepared by any method known in the art for the préparation of compounds of analogous structure. In particuiar, the compounds of the invention can be prepared by the procedures described by référencé to the schemes that follow, or by the spécifie methods described in the examples, or by similar processes to either.
The skiiled person will appreciate that the experimental conditions set forth in the schemes that follow are illustrative of suitable conditions for effecting the transformations shown, and that it may be necessary or désirable to vary the précisé conditions employed for the préparation of compounds of formula (I).
In addition, the skiiled person will appreciate that it may be necessary or désirable at any stage in the synthesis of compounds of the invention to protect one or more sensitive groups, so as to prevent undesirable side reactions. In particuiar, it may be necessary or désirable to protect amino or carboxylic acid groups. The protecting groups used in the préparation of the compounds of the invention may be used in conventional manner. See, for example, those described in Protective Groups in Organic Synthesis by Théodore W. Greene and Peter G. M. Wuts, 3rd édition, (John Wiley and Sons, 1999), in particuiar chapters 7 (Protection for the Amino Group”) and 5 (“Protection for the Carboxyl Group”), incorporated herein by référencé, which also describes methods for the removal of such groups.
Ail of the dérivatives of formula I and ta-f can be prepared by the procedures described in the general methods presented below or by routine modifications thereof. The présent invention also encompasses any one or more of these processes for preparing the dérivatives of formula (I), in addition to any novel intermediates used therein. The person skilled in the art will appreciate that the following reactions may be heated thermaliy or under microwave irradiation.
In executing the synthesis of the compounds of the invention, one skilled in the art will recognize the need to sample and assay reaction mixtures prior to work up in order to monitor the progress of reactions and décidé whether the reaction should be continued or whether it is ready to be worked up to obtain the desired product. Common methods for assaying reaction mixtures incïude thin-layer chromatography (TLC), liquid chromatography/mass spectroscopy (LCMS), and nuclear magnetic résonance (NMR).
One skilled in the art will also recognize that the compounds of the invention may be prepared as mixtures of diastereomers or géométrie isomers (e.g., cis and trans substitution on a cycloalkane ring). These isomers can be separated by standard chromatographie techniques, such as normal phase chromatography on siiica gel, reverse phase préparative high pressure liquid chromatography or supercritical fluid chromatography. One skilled in the art will also recognize that some compounds of the invention are chiral and thus may be prepared as racemic or scalemic mixtures of enantiomers. Several methods are available and are well known to those skilled in the art for the séparation of enantiomers. A preferred method for the routine séparation enantiomers is supercritical fluid chromatography employing a chiral stationary phase.
EXPERIMENTAL SECTION
Except where otherwise noted, reactions were run under an atmosphère of nitrogen. Chromatography on siiica gel was carried out using 250-400 mesh siiica gel using pressurized nitrogen (—10-15 psi) to drive solvent through the column (“flash chromatography). Where indicated, solutions and reaction mixtures were concentrated by rotary évaporation under vacuum.
The nomenclature in this patent is written as described by IUPAC (International Union of Pure and Applied Chemistry and using ChemBioDraw Ultra 13.0, Perkin Elmer to generate names.
The following non-limiting Préparations and Examples illustrate the préparation of compounds and salts of the présent invention. In the Examples and Préparations that are set out below, and in the aforementioned Schemes, the following abbreviations, définitions and analytical procedures may be referred to. Other abbreviations common in the art may also be used. Standard IUPAC nomenclature has been used.
AcOH is acetic acid;
aq. is aqueous;
Boc is tert-butoxycarbonyl;
br is broad;
brine is a saturated solution of sodium chloride in water;
t-Bu is tert-butyl;
n-BuLi is n-butyllithium;
°C is degrees ceicius;
Cbz is carbobenzyloxy;
CDCh is deutero-chloroform;
CDl is 1,T-carbonyldiimidazole;
conc. is concentrated (in référencé to reagents);
CS2CO3 is césium carbonate;
δ is Chemical shift;
d is doublet;
DBU is 1,8-diazabicyclo[5.4.0]undec-7-ene;
DCM is dichloromethane;
DHP is 3,4-dihydro-2H-pyran;
DIPEA is N,N-diisopropylethylamine;
DMAP is 4-dimethylaminopyridine;
DMF is N,N-dimethylformamide;
DMSO is dimethyl sulfoxide;
Et2Û is diethyl ether;
EtOAc is ethyl acetate;
EtOH is éthanol;
(EtO)2P(O)CH2CN is diethyl (cyanomethyl)phosphonate;
g is gram;
GCMS is gas chromatography mass spectrometry
HCl is hydrochloric acid;
HCO2H is formic acid;
HPLC is high performance liquid chromatography;
hrs is hours;
H2SO4 is sulfuric acid;
K2CO3 is potassium carbonate;
KH2PO4 is potassium dihydrogen phosphate
K2HPO4 is potassium monohydrogen phosphate;
K3PO4 is potassium phosphate (tribasic);
KOAc is potassium acetate
L is liter;
LCMS is liquid chromatography mass spectrometry;
LiBr is lithium bromide;
LiOH is lithium hydroxide;
m is multiplet;
M is molar;
MeCN is acetonitrile;
MeOH is methanol;
mg is milligram;
MgSOi is magnésium sulfate;
MHz is megaHertz;
min is minutes;
mL is milliliter;
mmol is millimole;
mol is mole;
MS m/z is mass spectrum ion peak;
MTBE is methyl t-butyl ether
NaBH(0Ac)3 is sodium triacetoxyborohydride;
NasCCh is sodium carbonate;
NaHCOa is sodium hydrogen carbonate;
NaHzPCU is sodium dihydrogen phosphate;
NazHPÛA is sodium monohydrogen phosphate;
Nal is sodium iodide;
NalOi is sodium periodate;
NaOAc is sodium acetate;
NaOCI is sodium hypochlorite;
NaOH is sodium hydroxide;
NHa is ammonia;
NHîCI is ammonium chloride;
NHîOH is ammonium hydroxide;
NH4OAC is ammonium acetate;
NMR is nuclear magnetic résonance;
OsÛ4 is osmium tetroxide;
Pd/C is palladium on carbon;
Pd(dppf)Cl2 is 1,1-bis(diphenylphosphino)ferrocene palladium(ll)dichloride (CAS: 72287-26-4);
Pd(dppf)Cl2.DCM is 1,1-bis(diphenylphosphino)ferrocene palladium(ll)dichloride; complexwith dichloromethane (CAS: 95464-05-4);
Pd(OAc)z is palladium acetate;
Pd(PPh3)4 is tetrakis(triphenylphosphine)palladium;
PMB-CI is (4-methoxy)benzyl chloride;
POCh is phosphorus(V) oxychloride;
ppm is parts per million;
psi is pounds per square inch;
PTSA is para-toluenesulfonic acid
PyHBrsis pyridine hydrobromide perbromide
PyHCI is pyridine hydrochloride q is quartet;
Rt is rétention time;
Rhz(OAc)4 is rhodium (11) acetate dimer;
RuCh hydrate is ruthenium(ll) chloride hydrate;
s is singlet;
SOCI2 is thionyi chloride;
t is triplet;
TBAB is tetrabutylammonium bromide
TEA is triethylamine;
TFA is trifluoroacetic acid;
THF is tetrahydrofuran;
TMSCI is chorotrimethylsilane;
pL is microliter;
pmol is micromole
XPhos Pd G2 is chloro(2-dicyclohexylphosphino-2,,4,,6'-triisopropyl-1,T-biphenyl)[2-(2,-amino-1ITbiphenyl)]palladium(ll); CAS 1310584-14-5.
’H Nuclear magnetic résonance (NMR) spectra were in ail cases consistent with the proposed structures. Characteristic Chemical shifts (δ) are given in parts-per-million downfield from tetramethylsilane using conventional abbreviations for désignation of major peaks: e.g. s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad. The following abbreviations hâve been used for common NMR solvents: CDsCN, deuteroacetonitrile; CDCh, deuterochloroform; DMSO-de, deuterodimethylsulfoxide; and CD3OD, deuteromethanol. Where appropriate, tautomers may be recorded within the NMR data; and some exchangeable protons may not be visible.
Mass spectra were recorded using électron impact ionization (El), electrospray ionisation (ESI) or atmospheric pressure Chemical ionisation (APCI). The observed ions are reported as MS m/z and may be positive ions of the compound [M]+, compound plus a proton [MH]\ or compound plus a sodium ion [MNa]4·. In some cases the only observed ions may be fragment ions reported as [MH(fragment lost)]*. Where relevant, the reported ions are assigned for isotopes of chlorine (35CI and/or 37CI), bromine (79Br and/or 81Br) and tin (120Sn).
Wherein TLC, chromatography, or HPLC has been used to purify compounds, one skilled in the art may choose any appropriate solvent or combination of solvents to purify the desired compound. Chromatographie séparations (excluding HPLC) were carried out using silica gel adsorbent unless otherwise noted.
Ail reactions were carried out using continuous stirring under an atmosphère of nitrogen or argon gas unless otherwise noted. In some cases, reactions were purged with nitrogen or argon gas prior to the start of the reaction. In these cases, the nitrogen or argon gas was bubbled through the liquid phase of the mixture for the approximate specified time. Solvents used were commercial anhydrous grades. Ail starting materials were commercially available products. In some cases, the Chemical Abstracts Service (CAS) identification number is provided to assist with clarity. In some cases, starting materials were prepared according to reported literature procedures as indicated by an asterisk (*). It will be apparent to one skilled in the art that the word “concentrated as used herein generally refers to the practice of évaporation of solvent under reduced pressure, typically accomplished by the use of a rotary evaporator.
GCMS Conditions
Column: 12m x 0.2mm, HP-1 Methyl Siloxane, 0.33 pm film, 1.0 ml/min column flow.
Methods: 7.6 min: Initial Oven Temp 105 ’C; 0.1 min hold; 30 ’C/min ramp to 300 ’C endpoint at 7.6 min; or 7.6 min: Initial Oven Temp 60 ’C; 0.1 min hold; 40 ’C/min ramp to 320 ’C endpoint at 7.6 min; or 5.1 min: Initial Oven Temp 40 °C; 0.1min hold; 30 ’C/min ramp to 150 ’C endpoint at 5.1min.
GC Inlet Parameters: Front Inlet, Split 30:1, He, 8 psi pressure, 250 ’C Injecter, 33.9 ml/min total flow.
MSD Tune: 230 °C Source Temp, 150 ’C Quad Temp, 280 ’C Aux2 Temp
Injection Volume: 1.0 pL
System Components: Agilent 5890 GC Oven with Agilent 5973 Mass Sélective Detector
LCMS Conditions
Acid: Waters Acquity HSS T3, 2.1 mm x 50 mm, C18, 1.7 pm; Column Température 60 ’C
Base: Waters Acquity UPLC BEH, 2.1 mm x 50 mm, C18, 1.8 pm; Column Température 60 ’C
Mobile Phase: A: 0.1% formic acid in water (v/v); Mobile phase B: 0.1% formic acid in acetonitrile (v/v).
Mobile Phase A: 0.1% ammonia in water (v/v); Mobile phase B: 0.1% ammonia in acetonitrile (v/v) Gradient Profiles: 1.5 min Run: Initial conditions: A-95%:B-5%; hold at initial from 0.0-0.1min; Linear Ramp to A-5%:B-95% over 0.1-1.0min; hold at A-5%:B-95% from 1.0-1.1min; return to initial conditions 1.1-1.5min
Purification methods (PM)
The compounds of the Examples were purified according to one of the Purification Methods (PM) referred to below unless otherwise described:
Purification Method A: Préparative HPLC using [Agella venusil ASB C18 150x21.2mmx5pm, from 16% MeCN in water (0.225% formic acid) to 36% MeCN in water (0.225% formic acid)]
Purification Method B: Préparative HPLC using [Phenomenex Gemini C18 250x21 2mmx8um or 150mmx25mmx5pm; from 16-55% MeCN in water (0.1% ammonia) to 36-60% MeCN in water (0.1% ammonia)]
Purification Method C: [YMC -Actus Triart C18 150x30pm, from 24% MeCN in water (0.1 % ammonia) to 44% MeCN in water (0.1 % ammonia)]
Purification Method D: Préparative HPLC using [Phenomenex Gemini C18 250x21.2mmx8pm, from 25% MeCN in water (ammonia pH=10) to 45% MeCN in water (ammonia pH=10)] followed by chiral chromatography using AS 250x25mm LD. 20 μΜ column, with supercritical CO2: EtOH or IPA (0.05% aqueous ammonia) 70:30 at from 50-80 mL/min
Purification Method E: Préparative HPLC using [Phenomenex Gemini C18 250x21 2mmx8pm, from 25% MeCN in water (0.225% ammonia) to 45% MeCN in water (0.225% ammonia) followed by chiral chromatography using AD 250mmx30mmx20pm column with mobile phase A: supercriticai CO2 and mobile phase B MeOH with 0.1% ammonia A:B 50:50 at 180 mL/min
Purification Method F: Silica gel column chromatography eluting with 100% DCM to 12% MeOH with 1% NHîOH.
Purification Method G: Silica gel column chromatography eluting with 97:2:1 DCM:MeOH:NH3 followed by préparative HPLC.
Purification Method H: Préparative HPLC using Column: Waters XBridge C18 19mmx100mm, 5μ; Mobile phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in acetonitrile (v/v); from 5-20% B to 40-100% B at 25 mL/min flow rate.
Purification Method I: Préparative HPLC using Column: Waters Sunfire C18 19mmx100mm, 5μ; Mobile phase A: 0.05% TFA in water (v/v); Mobile phase B: 0.05% TFA in acetonitrile (v/v); from 20% B to 40% B at 6.75 minutes, then to 100% B at 7 minutes at 30 mL/min flow rate.
Spécifie Rotation
Spécifie rotations based on the équation (a] = (100 a)/(l-c) and are reported as unitless numbers where the concentration c is in g/ΙΟΟ mL and the path length I is in decimeters. The units of the spécifie rotation, (degmL)/(gdm), are implicit and are not included with the reported value.
Préparation 1
Ethyl 1-{cyanomethyl)-1H-pyrazole-3-carboxylate
and
Ethyl 1-(cyanomethyl)-1H-pyrazole-5-carboxylate
O
To a suspension of CS2CO3 (2100 g, 6.44 mol) in DMF (12 L) was added ethyl 1 H-pyrazole-3carboxylate (750 g, 5.36 mol), followed by 2-chloroacetonitrile (450 g, 5.96 mol) and the mixture was stirred at about 25 °C for about 16 hrs. The reaction was poured into water (12 L) and extracted with EtOAc (5x5 L). The combined EtOAc extracts were washed with brine (2 x 5 L), dried (NazSOi) and concentrated to afford a residue which was purified by chromatography to afford ethyl 1(cyanomethyl)-1H-pyrazole-3-carboxylate (398 g, 39%) as a yellow oil and ethyl 1-(cyanomethyl)-1H pyrazole-5-carboxylate (680 g). The ethyl 1-(cyanomethyl)-1H-pyrazole-5-carboxylate was dissolved in MTBE (15 L) and washed with brine (3x5 L), dried (NaaSO-i) and concentrated to afford the compound as a yellowoil (489 g, 51%).
’H NMR (400 MHz, CDCh) δ: 7.49 (s, 1 H), 6.82 (s, 1 H). 5.45 (s, 2 H). 4.29 (q, 2 H), 1.29 (t, 3 H).
LCMS m/z = 180.1 [MH]*
Préparation 2
Ethyl 1-(2-amino-2-oxoethyl)-1H-pyrazole-5-carboxylate
O
Two identical reactions were carried out in parallel.
To a solution of ethyl 1-(cyanomethyl)-1H-pyrazole-5-carboxylate (Préparation 1,235.5 g, 1.32 mol) in TFA (1.2 L) was added conc. H2SO4 (377 mL, 7.04 mol) at about 25 °C. The reaction mixture was stirred at about 25 °C for about 16 hrs before being combined with the parallel reaction and concentrated to remove most of the TFA. The residue was poured into ice-water (5 L) and extracted with EtOAc (5 L). The aqueous phase was further extracted with EtOAc (10 x 5 L) and the combined EtOAc extracts were washed with saturated aq. NaHCOs (2x10 L), dried (NazSCh) and concentrated to afford the title compound as a yellow solid (477 g, 92%).
1H NMR (400 MHz, CDCh) δ: 7.60 (s, 1 H), 6.93 (s, 1 H), 5.90 (br. s, 1 H), 5.71 (br. s. 1 H), 5.27 (s, 2 H), 4.35 (q, 2 H), 1.37 (t. 3 H).
LCMS m/z = 198.2 [MH]*
Préparation 3 Pyrazolo[1,5-a]pyrazine-4,6(5H,7H)-dione
O
To a solution of ethyl 1-(2-amino-2-oxoethyî)-1H-pyrazole-5-carboxylate (Préparation 2, 466 g, 2.17 mol) in EtOH (56 L) was added NaOtBu (498 g, 5.20 mol) in THF (4 L) at about 25 °C. A white suspension developed during the addition and the mixture was then heated to about 70 °C for about 16 hrs. The reaction mixture was cooled to about 25 °C and acidified to about pH 6 with 12 M aq. HCl (500 mL), resulting in the formation of a white suspension. The mixture was concentrated to afford the title compound (admixed with sodium chloride) as yellow solid (783 g). This was used without further purification.
1H NMR (400 MHz, DMSO-de) δ: 11.82 (s, 1 H), 7.74 (s. 1 H), 6.97 (s. 1 H), 5.19 (s, 2 H).
LCMS m/z= 152.1 [MH]*
Préparation 4 4,6-Dictiloropyrazolo[1,5-a]pyrazine
Cl
Three identical reactions were carried out in parallel.
Pyrazolo[1,5-a]pyrazine-4,6(5H,7H)-dione (Préparation 3, 278 g. 1.14 moi) was added to POCI3 (1.84 kg, 12 mol) at about 25 °C, followed by PyHCI (131 g, 1.14 mol). The reaction mixture was heated at about 120 °C for about 16 hrs. The reactions were cooled to about 25 DC and concentrated to remove most of the POCh. Each residue was diluted with EtOAc (2 L) and the three EtOAc extracts were combined and poured into 1 M aq. NaH2PO« (7.5 L) at about 25 °C and filtered through a pad of Celite®. The filter cake was washed with EtOAc (3 x 2 L) and ail filtrâtes were combined and separated from the aqueous phase. The aqueous phase was extracted with MTBE (10 L). The combined EtOAc and MTBE extracts were washed with brine (2x5 L), dried (Na2SO4) and concentrated. The residue was purified by chromatography and the product was triturated with Petroleum ether (300 mL) and filtered. The filter cake was dried to afford the title compound as a white solid (110 g, 22%).
1H NMR (400 MHz, CDCI3) δ: 8.42 (s. 1 H), 8.06 (s, 1 H), 6.93 (s, 1 H).
LCMS m/z = 189.8 [MH]* (3TCI isotope)
Préparation 5
-(1 -Methy 1-1 H-py razol-4-yl)ethan-1 -one
I
Two identical reactions were carried out in parallel.
To a mixture of 1-methylpyrazole (750 g, 9.16 mol) and acetic anhydride (1.7 kg, 16.67 mol) was added concentrated H2SO4 (75 g, 0.75 mol) at about 20 °C. The reaction mixture was heated at about 150 °C for about 3 hrs. After coolîng, the two mixtures were combined, poured into ice-water
3l (15 L), adjusted to about pH 10 with 20% aq. NaOH and extracted with DCM (4 x 10 L). The combined DCM extracts were dried (NaaSOi) and concentrated to afford the title compound as a brown oil (1240 g, 72%).
1H NMR (400 MHz, CDCh) 5: 7.86 (s, 1 H), 7.84 (s, 1 H), 3.92 (s, 3 H), 2.40 (s, 3 H).
GCMS m/z = 109.0 [M-CH3]+
Préparation 6 2-Bromo-1-(1-methyl-1H-pyrazol-4-yl)ethan-1-one
Two identical reactions were carried out in parallel.
To a solution of 1-(1-methyl-1H-pyrazol-4-yl)ethan-1-one (Préparation 5, 620 g, 5 mol) in DCM (12 L) and éthanol (3 L) was added PyHBra (1.6 kg, 5 mol) at about 15 °C. The mixture was stirred at about 15 ’C for about 18 hrs. The two reaction mixtures were combined, quenched with water (10 L), separated, and the aqueous phase was extracted with DCM (4x10 L). The combined DCM extracts were dried (NajSOi) and concentrated to remove about 69 L of solvent. The residue was diluted with Petroleum ether (5 L), stirred at about 15 °C for about 30 min and the mixture was filtered. The precipitate was dried to afford the title compound as a yellow solid (1.73 kg, 85%).
’H NMR (400 MHz, CDCh) δ: 7.97 (s, 1 H), 7.91 (s, 1 H), 4.17 (s, 2 H), 3.93 (s, 3 H).
LCMS m/z = 203.1 [MH]* (79Br isotope)
Préparation 7
Diethyl 1-(2-(1-methyl-1H-pyrazol-4-yl)-2-oxoethyl)-1H-pyrazole-3,5-dicarboxylate
Two reactions were carried out in parallel.
To a mixture of 2-bromo-1-(1-methyl-1H-pyrazol-4-yl)ethan-1-one (Préparation 6; 500 g, 2.46 mol) and diethyl-1 H-pyrazole-3,5-dicarboxylate (580 g. 2.73 mol) in DMF (8 L) was added CS2CO3 (1050 g, 3.23 mol) at about 20 °C. After about 18 hrs, the two reaction mixtures were combined, diluted with water (10 L) and extracted with DCM (3x10 L). The combined DCM extracts were dried (NasSCh) and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (1.53 kg, 93%).
1H NMR (400 MHz, CDCh) δ: 7.96 (s, 2 H), 7.46 (s, 1 H), 5.86 (s, 2 H), 4.45 (q, 2 H), 4.32 (q, 2 H),
3.99 (s, 3 H), 1.44 (t. 3 H). 1.36 (t, 3 H).
LCMS m/z = 335.0 [MH]+
Préparation 8
Ethyl 4-hydroxy-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxylate
OH
Three identical reactions were carried out in parallel.
To a solution of diethyl 1-(2-(1 -methyl-1H-pyrazol-4-yl)-2-oxoethyl)-1H-pyrazole-3,5-dicarboxylate (Préparation 7; 510 g, 1.52 mol) in éthanol (6 L) was added NH«OAc (352 g, 4.57 mol) at about 20 °C. The mixture was heated in an autoclave at about 130 °C for about 24 hrs. The reaction mixtures were cooled to about 50 °C and were combined and fiîtered. The precipitate was dried to afford the title compound (1090 g, 83%) as an off-white solid.
Ή NMR (400 MHz, DMSO-d6) δ: 11.35 (br. s, 1 H), 8.31 (s, 1 H), 8.20 (s, 1 H), 8.05 (s, 1 H), 7.38 (s, 1 H), 4.34 (q, 2 H), 3.89 (s, 3 H). 1.33 (t, 3 H).
LCMS m/z = 288.0 [MH]
Préparation 9 4-Hydroxy-6-(1-methyi-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxylic acid
OH
Two identical reactions were carried out in parallel.
To a suspension of ethyl 4-hydroxy-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxylate (Préparation 8, 545 g, 1.9 mol) in MeOH (10 L) was added 1 M aq. NaOH (5.75 L) at about 20 °C. After about 30 min, the suspension became a clear solution and stirring was continued at about 20 °C for about 18 hrs. The reaction mixtures were adjusted to about pH 2 with 12 M aq. HCl (650 mL), combined, and concentrated to remove most of the MeOH. The residue was filtered and the precipitate was dried to afford the title compound as an off-white solid (1040 g, 100%).
Ή NMR (400 MHz, DMSO-de) δ: 13.25 (br. s, 1 H), 11.67 (s, 1 H), 8.34 (s, 1 H), 8.16 (s, 1 H), 8.06 (S, 1 H), 7.32 (s, 1 H), 3.88 (s, 3 H).
LCMS m/z = 260.0 [MH]*
Préparation 10
6-(1-Methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-ol
Five identical réactions were carried out in parallel.
4-Hydroxy-6-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxyîic acid (Préparation 9, 85 g, 0.328 mol) was added in portions to pre-heated sulfoiane (800 mL) at about 280 °C. The five reaction mixtures were stirred at about 280 °C for about 2 hrs, cooled to about 25 °C, and stirred for about 18 hrs. The reaction mixtures were combined and the mixture was purified by chromatography eluting with petroleum ether-EtOAc (10:1 to 0:1), followed by DCM-MeOH (10:1) to afford the title compound as a yellow solid (490 g, 75%).
1H NMR (400 MHz, DMSO-de) δ: 11.45 (s, 1 H), 8.28 (s, 1 H), 8.10 (s, 1 H), 8.04 (s, 1 H), 7.88 (s, 1 H), 6.99 (s, 1 H), 3.88 (s, 3 H).
LCMS m/z = 216.0 [MH]’
Préparation 11
4-Chloro-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine
Two identical reactions were carried out in parallel.
To a suspension of 6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-ol (Préparation 10, 307 g, 1.43 mol) in MeCN (7.5 L) was added POCÎ3 (2006 g, 13 mol) at about 25 °C. The mixture was heated at about 85 °C for about 48 hrs. The reaction mixtures were combined and filtered. The precipitate was washed with EtOAc and dried under vacuum. The dried precipitate was purified by chromatography to afford a yellow solid which was dissolved in DCM (15 L) and washed with 1 M aq.
NaHCOa (5 L). The DCM concentrated to remove about 13 L of solvent and the residue was diluted MTBE (2 L) and petroleum ether (2 L). The mixture was fîltered and the precipitate was dried to afford the title compound as a yellow solid (385 g, 58%).
’H NMR (400 MHz, DMSO-ds) δ: 9.22 (s, 1 H), 8.27 (s, 1 H), 8.21 (s, 1 H), 8.04 (s, 1 H), 7.02 (s, 1 H), 3.88 (s, 3 H).
LCMS m/z = 233.8 [MH]+ (3SCI isotope)
Préparation 12 1-(4-Methoxybenzyl)-1H-pyrazole-4-carboxyiic acid —O
Part 1 : Three identical reactions were carried out in paraitel.
To a stirred solution of ethyl 1 H-pyrazole-4-carboxylate (16 g, 110 mmol) in MeCN (160 mL) was added PMB-Cl (85.8 g, 548 mmol) and K2CO3 (23.7 g, 171 mmol) and the mixture was heated under reflux for about 18 hrs. The three batches were cooled, combined and fîltered. The filtrate was concentrated to afford ethyl 1-(4-methoxybenzyl)-1H-pyrazole-4-carboxylate as a yellow oïl that was used without further purification.
Part 2: Three identical reactions were carried out in parallel.
To a stirred solution of crude ethyl 1-(4-methoxybenzyl)-1 H-pyrazole-4-carboxylate (Part 1, 50.0 g, 96 mmol) in THF (150 mL) and MeOH (150 mL) was added a solution of LiOH (10 g, 238 mmol) in water (75 mL). The mixture was heated at about 60 °C for about 18 hrs. The three batches were combined and evaporated to dryness. The residue was diluted with water (800 mL) and MeOH (150 mL) and washed with EtOAc (2 x 500 mL). The EtOAc extracts were discarded and the aqueous solution was acidified to about pH 2 with 6 M aq. HCl and extracted with EtOAc (2 x 800 mL). The combined EtOAc extracts were washed with brine (500 mL), dried (Na2SO4) and concentrated to afford the title compound as a white solid (33.0 g, 83% for the two steps).
’H NMR (400 MHz, DMSO-de) δ: 12.34 (br. s, 1 H), 8.32 (s, 1 H), 7.79 (s, 1 H), 7.23 (m, 2 H), 6.89 (m. 2 H), 5.26 (s, 2 H), 3.72 (s. 3 H).
Préparation 13
-(4-Methoxybenzyl)-1 H-pyrazole-4-carbonyl chloride
A solution of 1-(4-methoxybenzyl)-1H-pyrazole-4-carboxylic acid (Préparation 12, 25.0 g, 110 mmo!) in SOCh (40 mL) was stirred at about 60 °C for about 5 hrs. The solution was concentrated to afford title compound as a brown oil (27.0 g, 100%) which was used without further purification or characterization.
Préparation 14 N-Methoxy-1-(4-methoxybenzyl)-N-methyl-1H-pyrazole-4-carboxamide
To a solution of Ν,Ο-dimethylhydroxylamine hydrochloride (26.3 g, 269 mmol) and TEA (131.0 g, 1.29 mol) in DCM (200 mL) was slowly added a solution of 1-(4-methoxybenzyl)-1H-pyrazole-4-carbonyl chloride (Préparation 13, 27.0 g, 108 mmol) in DCM (50 mL). After the addition was complété, the reaction mixture was stirred at about 20 °C for about 5 hrs. The mixture was diluted with DCM (150 mL) and water (300 mL). The combined DCM extracts were washed with brine, dried (NaaSCU) and concentrated to afford the title compound (20.0 g, 67%) as a brown oil which was used without further purification.
1H NMR (400 MHz, CDCI3) 6: 7.99 (s, 1 H), 7.90 (s. 1 H), 7.22 (d, 2 H), 6.89 (d, 2 H), 5.25 (s, 2 H), 3.81 (s, 3 H), 3.69 (s, 3 H), 3.31 (s, 3 H).
LCMS m/z = 275.0 [MH]+
Préparation 15 1-(1-(4-Methoxybenzyl)-1H-pyrazoi-4-yl)ethan-1 -one —O
Two identical reactions were carried out in parailel.
To a solution of N-methoxy-1-(4-methoxybenzyl)-N-methyl-1H-pyrazole-4-carboxamide (Préparation 14, 10.0 g, 36.3 mmol) in THF (120 mL) was added dropwise 3 M méthylmagnésium bromide in ether (24.2 mL) at about 0 °C. The reaction mixture was warmed to about 25 °C and stirred for about 5 hrs. The reaction was quenched by the addition of saturated aq. NH4CI (100 mL) and extracted with EtOAc (2 x 200 mL). The combined EtOAc extracts were dried (Na2SO4) and concentrated. The concentrated residues from both experiments were combined and purifted by chromatography to afford the title compound (10.0 g, 60%) as a brown solid.
Ή NMR (400 MHz, DMSO-de) δ: 8.47 (s, 1 H), 7.91 (s, 1 H), 7.25 (m, 2 H). 6.90 (m, 2 H), 5.27 (s, 2 H), 3.72 (s, 3 H), 2.34 (s, 3 H).
LCMS m/z = 231.7 [MH]’
Préparation 16 2-Bromo-1 -(1 -(4-methoxybenzyl)-1 H-pyrazoI-4-yl)ethan-1 -one —O
N
Two identical reactions were carried out in parailel.
To a solution of 1-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)ethan-1-one (Préparation 15, 8.0 g, 34.7 mmol) in DCM (96 mL) and EtOH (24 mL) was added PyHBra (13.3 g, 41.7 mmol) at about 20 °C. The reaction mixtures were kept at about 25 °C for about 18 hrs and quenched with water (100 mL) before being combined and extracted with EtOAc (2 x 300 mL). The combined EtOAc extracts were washed with brine, dried (Na2SÛ4) and concentrated. The residue was purified by chromatography to afford a yellow solid. This was triturated with MTBE (100 mL) to afford the title compound (15.0 g, 70%) as a yellow solid. An additional sample (5.0 g, 23%) of slightly impure product was obtained as a yellow solid by concentration of the trituration liquors.
1H NMR (400 MHz, CDCh) δ: 7.99 (s, 1 H), 7.89 (s, 1 H), 7.23 (m, 2 H), 6.92 (m, 2 H), 6.92 (m, 2 H), 5.25 (s, 2 H). 4.15 (s, 2 H), 3.81 (s, 3 H).
LCMS m/z = 333.0 [MNa]* (81 Br isotope)
Préparation 17
Dimethyl 1-(2-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)-2-oxoethyl)-1H-pyrazole-3,5-dicarboxylate
To a mixture of dimethyl 1H-pyrazoie-3,5-dicarboxylate (1 g, 5 mmol) and 2-bromo-1-(1-(4methoxybenzyl)-1H-pyrazol-4-yl)ethan-1-one (Préparation 16, 2.18 g, 7.06 mmol) in DMF (20 mL) was added CS2CO3 (2.3 g, 7.06 mmol) at about 20 °C. After about 2 days, the mixture was concentrated to dryness. The residue was dissolved in DCM and washed once with saturated aq. NHïCI. The DCM was concentrated and the residue was purified by chromatography. The product was stirred in EtOAc (20 mL) at about 20 °C for about 18 hrs. The solid formed was frltered and dried to afford the title compound (1.38 g, 60%).
’H NMR (400 MHz, CDCh) δ: 7.96 (s, 1 H), 7.82 (s, 1 H), 7.42 (s, 1 H), 7.24 (d, 2 H), 6.93 (d, 2 H), 5.80 (s, 2 H), 5.27 (s, 2 H), 3.95 (s, 3 H), 3.84 (s, 3 H), 3.83 (s, 3 H).
LCMS m/z = 413.1 [MH]+
Préparation 18
Methyl 4-hydroxy-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2carboxylate
Ethyl 4-hydroxy-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazîne-2-carboxylate
Six identical reactions were carried out in parallel.
To each of six vials was added dimethyl 1-(2-(1-(4-methoxybenzyi)-1H-pyrazol-4-yl)-2-oxoethyî)-1Hpyrazole-3,5-dicarboxylate (Préparation 17, 300 mg, 0.73 mmol), NFUOAc (336 mg, 4.37 mmol) and EtOH (6 mL). The mixtures were heated under microwave irradiation at about 150 °C for about 2 hrs, then cooled to about 20 °C, stirred for about 1 hrs, and filtered. The combined solids were dried to afford a mixture of both title compounds which was used without further purification in the next step (161 g).
1H NMR (400 MHz, DMSO-ds) δ: 11.65 (br. s„ 1 H), 8.38 (s, 1 H), 8.17 (s, 1 H), 8.07 (s, 1 H), 7.36 (s, 1 H), 7.26 (d, 2 H), 6.93 (d, 2 H), 5.75 (s. 1 H), 5.28 (s, 2 H). 3.86 (s, 3 H), 3.74 (s, 3 H). This is the methyl ester which was the major component.
LCMS m/z= 380.1 [MH]*, 394.1 [MH]*
Préparation 19 4-Hydroxy-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxylic acid
To a solution of the mixture of methyl 4-hydroxy-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazine-2-carboxylate and ethyl 4-hydroxy-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazine-2-carboxy!ate (Préparation 18, 524 mg, about 1.38 mmol) in MeOH (10 mL) was added 1 M aq. NaOH (4.83 mL). The mixture was kept at about 20 °C for about 18 hrs before additional 1 M aq. NaOH (1.38 mL) was added. The mixture was kept at about 20 °C for about 24 additional hours. The MeOH was evaporated and the residue was diluted with water (2 mL) and stirred at about 40 °C until ail solid was dissolved. The solution was acidified with 12 M aq. HCl and stirred at about 0 °C for about 10 min. The resulting precipitate was filtered and the precipitate was washed with water. The solid was dried under vacuum to afford the title compound (487 mg, 96%).
’H NMR (400 MHz, DMSO-ds) ô: 11.65 (s, 1 H), 8.38 (s, 1 H), 8.18 (m, 1 H), 8.10 (s, 1 H), 7.25 - 7.32 (m, 4 H). 6.95 (m, 2 H). 5.28 (s, 2 H), 3.78 (s, 3 H).
LCMS m/z = 366.0 [MH]*
Préparation 20 6-(1-(4-Methoxybenzyl)-1H-pyrazol-4-yl)38yrazole[1,5-a]38yrazole-4-ol
Three réactions were carried out in parallel.
Sample 1:
4-Hydroxy-6-(1-(4-methoxybenzyl)-1 H-pyrazol-4-yl)39yrazole(1,5-a]pyrazine-2-carboxyiic acid (Préparation 19, 40 mg, 0.11 mmol) was heated at about 350 °C for about 10 seconds until the off white solid melted and turned into dark brown liquid.
Sample 2:
4-hydroxy-6-(1 -(4-methoxybenzyl)-1 H-pyrazol-4-yl)39yrazole[1,5-a]pyrazine-2-carboxylic acid (Préparation 19, 120 mg, 0.33 mmol) was heated at about 350 °C for about 15 seconds until the off white solid melted and turned into dark brown liquid.
Sample 3:
4-hydroxy-6-(1-(4-methoxybenzyl)-1 H-pyrazol-4-yi)39yrazole[1,5-a]pyrazine-2-carboxylic acid (Préparation 19, 310 mg, 0.85 mmol) was heated at about 350 °C for about 15 seconds until the off white solid ail melted and turned into dark brown liquid.
Ail three batches were cooled, combined and concentrated twice with toluene to afford the title compound which was used without additional purification in the next step.
1H NMR (400 MHz. DMSO-ds) δ: 11.43 (s, 1 H), 8.37 (s, 1 H), 8.11 (s. 1 H), 8.08 (s, 1 H), 7.88 (d, 1 H), 7.28 (d, 2 H), 6.99 (d, 1 H), 6.94 (d, 2 H), 5.28 (s, 2 H), 3.76 (s, 3 H).
LCMS m/z = 322.1 [MH]+
Préparation 21 4-Chloro-6-(1-(4-methoxybenzyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine
6-(1-(4-Methoxybenzyl)-1H-pyrazol-4-yi)pyrazolo[1,5-a]pyrazin-4-oi (Préparation 20, 390 mg, 1.21 mmol), PyHCI (143 mg, 1.21 mmol) and POCh (10 mL) were heated at about 120 °C for about 18 hrs. The mixture was concentrated and the residue was treated with aq. NaHaPCU solution to maintain about pH 4. The resuiting solution was stirred at about 20 °C for about 10 min and extracted three times with DCM. The combinée! DCM extracts were dried and concentrated to afford the title compound as a brown solid (300 mg, 72%).
Ή NMR (400 MHz, CDCb) δ: 8.74 (s, 1 H), 8.50 (s, 1 H), 8.22 (m, 2 H), 7.45 (m, 2 H), 6.92 - 7.02 (m, 3 H), 3.88 (s, 2 H). 2.15 (s, 3 H).
LCMS m/z = 340.0 [MH]4
Préparation 22 tert-Butyl 4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazole-1-carboxylate
N
N—N // Λ
A solution of 4,6-dichloropyrazolo[1,5-a]pyrazine (Préparation 4, 700 mg, 3.72 mmol), tert-butyl 4(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-carboxylate (1100 mg, 3.72 mmol), 2 M aq. K3PCU (3 mL, 6 mmol) in 1,4-dioxane (10.0 mL) was purged with argon for about 5 min. To this was added bis(tri-t-butylphosphine)palladium(0) (96.1 mg, 0.19 mmol) and the reaction was kept at about 20 °C for about 18 hrs. The solvent was concentrated to afford an amber residue which was taken up in DCM and purified by chromatography to afford the title compound (710 mg, 60%).
1H NMR (400 MHz. CDCb) δ: 8.82 (m, 1 H), 8.44 (m. 3 H). 8.14 (s, 1 H). 1.60 (s, 9 H).
LCMS m/z = 220.1 [MH-BOC]4
Préparation 23 tert-Butyl 4-(6-vinylpyrazolo[1,5-a]pyrazin-4-yl)-1 H-py razole-1 -carboxylate
N-N // 3 'N
A solution of tert-butyl 4-(6-chloropyrazoto[1,5-a]pyrazin-4-yl)-1H-pyrazole-1-carboxylate (Préparation 22, 700 mg, 2.19 mmol) and tributyl(vinyl)stannane (694 mg, 2.19 mmol) in 1,4-dioxane (30 mL) was purged with argon for about 5 min followed by the addition of XPhos Pd G2 (344 mg, 0.44 mmol). The mixture was heated to about 55 ®C for about 18 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound (449 mg, 66%).
4l ’H NMR (400 MHz, CDCh) δ: 8.79 (s, 1 H), 8.46 (s, 1 H), 8.26 (s, 1 H), 8.07 (d, 1 H), 6.95 (dd, 1 H),
6.77 (dd, 1 H), 6.43 (dd, 1 H), 5.52 (dd, 1 H), 1.74 (s, 9 H).
LCMS m/z = 312.3 [MH]*
Préparation 24 tert-Butyl 4-(6-formylpyrazolo[1,5-a]pyrazine-4-yl)-1 H-pyrazole-1 -carboxylate
O
A solution of tert-butyl 4-(6-vinylpyrazolo[1,5-a]41yrazine-4-yl)-1H-pyrazole-1-carboxylate (Préparation 23; 446 mg, 1.43 mmol) and 2,6-lutidine (767 mg, 7.16 mmoi) in 1,4-dioxane (9 mL) and water (3 mL) was cooled to about 0 °C, and NaICh (1530 mg, 7.16 mmol) and 4% aq. OsCU solution (0.54 mL) were added. The mixture was allowed to warm to about 20 °C for about 3 hrs. The solids were removed by filtration and washed with ether. The combined 1,4-dioxane and ether were concentrated and the residue was purified by chromatography to afford the title compound as an off white solid (289 mg, 65%).
‘H NMR (400 MHz, CDCh) δ: 10.20 (s, 1 H), 9.01 (s, 1 H), 8.85 (s, 1 H), 8.50 (s, 1 H), 8.30 (d, 1 H), 7.08-7.12 (m, 1 H). 1.73 (s, 9 H).
LCMS m/z = 314.2 [MH]*
Préparation 25 tert-Butyl €-4-(6-((hydroxyimino)methyl)41yrazine[1,5-a]41yrazine-4-yl)-1H-pyrazole-1carboxylate
Hydroxylamine HCl (112 mg, 1,58 mmol) was added to a mixture of tert-butyl 4-(6-formy!pyrazolo[1,5a]42yrazine-4-yl)-1H-pyrazole-1-carboxylate (Préparation 24, 450 mg, 1.44 mmol), and NazCOa (196 mg, 1.58 mmol) in MeOH (20 mL). The mixture was kept at about 20 °C for about 1.5 hrs. The mixture was concentrated, water (30 mL) was added, and the mixture was stirred for about 5 min before the solid was filtered and dried to yield the title compound as an off white solid (325 mg, 69%). ’H NMR (400 MHz, CDCh) δ: 10.46 (br. s., 1 H), 9.53 (s, 1 H), 8.81 (s, 1 H), 8.46 (s, 1 H), 8.41 (s, 1 H), 8.19 (d, 1 H), 7.03 (d, 1 H), 1.73 (s, 9 H).
LCMS m/z = 329.2 [MH]*
Préparation 26 (3-(4-(1 H-Pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6-yl)isoxazol-5-yl)methanol
Sodium hypochlorite solution (about 12% to 15%, 0.19 mL about 3.0 mmol) was added dropwise to a solution of tert-butyl (E)-4-(6-((hydroxyimino)methyl)pyrazoio[1,5-a]pyrazin-4-yl)-1 H-pyrazole-1 carboxylate (Préparation 25, 200 mg, 0.61 mmol) and propargyl alcohol (171 mg, 3.05 mmol) in DCM (5 mL) at about 0 °C. The mixture was allowed to warm to about 20 °C for about 18 hrs. The resulting solid was filtered to afford the title compound (115 mg, 67%).
’H NMR (400 MHz, DMSO-de) δ: 9.14 (s, 1 H), 8.56 (s, 2 H), 8.31 (d, 1 H), 7.50 (d, 1 H), 7.09 (s, 1 H), 4.57-4.75 (m, 2 H).
LCMS m/z = 283.1 [MH]’
Préparation 27 3-(Cyanomethylene)cyclobutane-1-carbonitrite
A solution of 3-oxocyclobutane-1-carbonitrile* (CAS: 20249-16-5, 14.5 g, 152 mmol) in THF (250 mL) was added to a mixture of (EtO)2P(O)CH2CN (31.1 g, 175 mmol), LiBr (19.9 g, 229 mmol) and TEA (30.9 g, 305 mmol) in THF (300 mL) at about 25 °C. After about 16 hrs, the mixture was filtered and the filtrate was concentrated. The residue was purified by chromatography to afford the title compound as a light yellow oil (16.01 g, 89%).
*See Synthetic Communications 2005, 35, 657 - 662.
Ή NMR (400 MHz, CDaCN) δ: 5.38 (s, 1 H), 3.30 - 3.43 (m, 2 H), 3.16 - 3.30 (m, 3 H).
LCMS m/z = 119.1 [MH]*
Example 1 (1s,3s)-3-(Cyanomethyl)-3-(4-(6-(5-(hydroxymethyl)isoxazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (cis isomer)
DBU (89.0 mg, 0.58 mmol) was added to a solution of (3-(4-(1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6yl)isoxazol-5-yl)methanol (Préparation 26; 55.0 mg, 0.19 mmol) and 3-(cyanomethylene)cyclobutane1-carbonitrile (Préparation 27; 23.0 mg, 0.19 mmol) in MeCN (4 mL). The reaction was purged with nitrogen and stirred at about 20 °C for about 20 hrs. The mixture was partitioned between EtOAc (5 mL) and 1 M aq. NaH2PO4 (5 mL). The EtOAc was separated, dried (Na2SO4), and concentrated. The residue was purified chromatography afford the title compound (5 mg, 6%).
1H NMR (400 MHz, DMSO-de) δ: 9.26 (m, 1 H), 8.92 (m, 1 H), 8.54 (m, 1 H), 8.40 (m, 1 H), 7.52 (m, 1 H), 7.14 (m, 1 H), 5.55 (br s, 1 H), 4.70 (s, 2 H), 3.60 (m, 3 H), 3.42 - 3.45 (m, 2 H), 2.78 - 2.83 (m, 2 H).
LCMS m/z = 401.4 [MH]*
Préparation 28 tert-Butyl 3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1yl)azetidine-1-carboxylate
To a solution of tert-butyl 3-(cyanomethylene)azetidine-1-carboxylate (CAS 1153949-11-1, 7.00 g, 36.1 mmol) in MeCN (100 mL) was added 4-pyrazoleboronic acid pinacol ester (7.71 g, 39.7 mmol) and DBU (2.75 g. 18.0 mmol) at about 25 °C. After about 18 tirs, the mixture was concentrated and the residue was purified column chromatography to afford the title compound as a white solid (11 g, 78%).
’H NMR (400 MHz, CDCh) S: 7.92 (s, 1 H), 7.86 (s, 1 H), 4.40 (m, 2 H), 4.21 (m, 2 H), 3.52 (s, 2 H), 1,44 (s, 9 H), 1.32 (s, 12 H).
LC-MS m/z = 333.0 [MH-CiHb] *
Préparation 29 tert-Butyl 3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine1-carboxylate
To a solution of tert-butyl 3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1Hpyrazol-1-yi)azetidine-1-carboxylate (Préparation 28, 362 mg, 0.93 mmol) and 4,6dichloropyrazolo[1,5-a]pyrazine (Préparation 4; 167 mg, 0.89 mmol) in 1,4-dioxane (5 mL) was added 2 M aq. K3PO4 (1.40 mL) at about 25 °C. The mixture was purged with argon for about 2 min and bis(tri-t-butylphosphine)paladium(0) (94.3 mg, 0.184 mmol) was added. The mixture was stirred at about 20 °C for about 2 hrs. The mixture was diluted with DCM, separated, and the aqueous phase was extracted twice with DCM. The combined DCM extracts were dried (NasSCh) and concentrated. The residue was purified by chromatography to afford the title compound as a white solid (295 mg, 75%).
1H NMR (400 MHz, CDCh) δ: 8.42 (s, 1 H), 8.41 (s, 1 H), 8.31 (s, 1 H), 8.10 (d, 1 H), 7.02 (d, 1 H), 4.54 (d, 2 H), 4.31 (d, 3 H). 3.33 (s, 2 H), 1.49 (s, 9 H).
LCMS m/z = 358.1 [MH-C^Ha]*
Préparation 30 2-(3-(4-(6-Chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-1-(cyclopropylmethyl)azetidin-3yljacetonitrite
Part 1
To a solution of tert-butyl 3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)azetidine-l-carboxylate (Préparation 29, 0.56 g, 1.35 mmol) in DCM (13.5 mL) was added TFA (7 mL) at about 25 °C. After about 4 hrs, the mixture was concentrated to dryness to afford 2-(3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a yellow solid (578 mg, about 100%) which was used without further purification.
Part 2
To a solution of 2-(3-(4-(6-chloropyrazolo[1,5-a]pyrazin~4-yl)-1 H-pyrazol-1 -yl)azetidin-3-yl)acetonitrile (Part 1, 1.35 mmol) and bromomethylcyclopropane (365 mg, 2.71 mmol) in DMF (13.5 mL) was added TEA (548 mg, 5.41 mmol) at about 25 °C. The mixture was heated at about 50 °C for about 14 hrs. The cooled solution was diluted with water (20 mL) and extracted with EtOAc (3 x 20 mL). The combined EtOAc extracts were washed with brine (30 mL), dried (NasSOa) and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (314 mg, 63%). ’H NMR (400 MHz, CDCh) δ: 8.39 (s, 1 H), 8.37 (s, 1 H), 8.27 (s, 1 H), 8.08 (d. 1 H), 7.02 (d, 1 H),
3.78 - 3.84 (m, 2 H), 3.59 (d, 2 H), 3.42 (s, 2 H), 2.45 (d, 2 H), 0.77 - 0.87 (m, 1 H), 0.48 - 0.55 (m, 2 H), 0.14 (q, 2 H).
LCMS m/z = 367.9 [MH]4· (35CI isotope)
Préparation 31
Ethyl 3-(tributyIstanny 1)-1 H-pyrazole-5-carboxylate
Ethynyltributylstannane (50 g, 158 mmol) was added to a solution of ethyldiazoacetate (19.9 g, 175 mmol) in toluene (500 mL) at about 25 °C. The solution was heated for about 16 hrs at about 100 °C, then was concentrated to afford the crude product as a yellowoil (110 g). This was combined with the crude product from an équivalent reaction carried out with ethynyltributylstannane (22 g, 70 mmol) and ethyldiazoacetate (8.76 g, 77 mmol), and the combined residues were purified by column chromatography on alumina to afford the title compound as a yellow oil (42 g, 61%).
’H NMR (400 MHz, CDCh) ΰ: 10.28 (br. s.. 1 H), 6.84 - 6.99 (m, 1 H), 4.41 (q, 2 H), 1.47 -1.67 (m, 6 H), 1.41 (t, 3 H), 1.28 -1.39 (m, 6 H), 1.04 - 1.24 (m, 6 H), 0.90 (t, 9 H).
LCMS m/z 431.2 [MH]+ (’2°Sn isotope).
Préparation 32 (3-(Tributylstannyi)-1 H-pyrazol-5-yl)methanol
Ethyl 3-(tributylstannyl)-1H-pyrazole-5-carboxylate (Préparation 31, 6000 mg, 13.98 mmol) in THF (200 mL) was added to a stirred suspension of lithium aluminum hydride (3108 mg, 83.9 mmol) in THF (200 mL) at about -10 °C. After about 4 hrs, the mixture was quenched with Na2SO« decahydrate at about -10 °C until effervescence ceased. The mixture was filtered and the filter cake was washed with THF (500 mL) and DCM (5 x 500 mL). The combined filtrâtes were concentrated to afford the title compound as a white solid (4460 mg, 82%).
’H NMR (400 MHz, CDCh) 5: 10.35 (br. s., 1 H). 6.28 - 6.43 (m, 1 H). 4.75 (s, 2 H). 1.49 - 1.60 (m, 6 H), 1.29- 1.39 (m, 6 H), 1.08-1.15 (m. 6 H), 0.87-0.93 (m, 9 H).
LCMS m/z = 388.9 [MH]*(’20Sn isotope)
Example 2
2-{1-(Cyclopropylmethyl)-3-(4-(6-(5-(hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile
To a solution of 2-(3-(4-(6-chloropyrazoto[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)-1(cyc!opropylmethyl)azetidin-3-yl)acetonitrile (Préparation 30, 204 mg, 0.55 mmol) and 3(tributylstannyl)-1H-pyrazol-5-yl)methanol (Préparation 32, 215 mg, 0.55 mmol) was added XPhos Pd G2 (43.6 mg, 0.055 mmol) in 1,4-dioxane (5.5 mL). The mixture was heated at about 110 °C for about 4 hrs. The mixture was evaporated to dryness and the residue was combined with the residue from an équivalent reaction using 2-(3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-1(cyclopropylmethyl)azetidin-3-yl)acetonitrile (Préparation 30, 110 mg, 0.27 mmol) and 3(tributylstannyl)-1H-pyrazol-5-yl)methanol (Préparation 32, 105 mg, 0.27 mmol). The combined residues were purified by HPLC to afford the titte compound (112 mg, 31%).
1H NMR (400 MHz, CDCh) δ: 8.66 (s, 1 H), 8.64 - 8.69 (m, 1 H), 8.38 (s, 1 H), 8.23 (s, 1 H), 8.06 (d, 1 H), 6.94 (d, 1 H), 6.67 (s, 1 H), 4.80 (s. 2 H), 3.80 (d, 2 H), 3.62 (d, 2 H), 3.41 (s, 2 H), 2.44 (d, 2 H), 0.76 - 0.86 (m, 1 H), 0.46 - 0.53 (m, 2 H), 0.10 - 0.16 (m, 2 H).
LCMS m/z 430.1 [MH]+
Préparation 33
2-(3-(4-(6-Chloropyrazolo[1l5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile
To a solution of tert-butyl 3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H’Pyrazol-1-yl)-3(cyanomethyl)azetidine-l-carboxylate (Préparation 29, 485 mg, 1.17 mmol) in DCM (20 mL) was added TFA (6 mL) at about 0 °C. The mixture was stirred at about 25 °C for about 4 hrs. The solution was concentrated. The residue was adjusted to about pH 9 with conc. NFUOH (about 0.5 mL) and partitioned between water (10 mL) and DCM (30 mL). The aqueous solution was extracted with DCM (3 x 30 mL). The combined DCM extracts were dried (NasSCXt) and concentrated to afford the title compound as a yellow solid (300 mg, 81%).
1H NMR (400 MHz, CDaOD) δ: 8.91 (s, 1 H), 8.72 (s, 1 H), 8.53 (s, 1 H), 8.20 (d, 1 H), 7.40 (d, 1 H), 4.90 (d, 2 H), 4.66 (d, 2 H), 3.72 (s, 2 H).
LCMS m/z 313.9 [MH]+ (35CI isotope)
Préparation 34 2-(3-(4-(6-Chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-1-ethylazetidin-3-yl)acetonitrile
Sodium acetate (314 mg, 3.82 mmol) and acetaldehyde (842 mg, 19.1 mmol) were added to a solution of 2-(3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -yl)azetidin-3-yl)acetonitrile (Préparation 33, 120 mg, 0.38 mmol) in MeOH (6 mL) and the mixture was stirred for about 4 hrs. Then NaBH(OAc)3 (243 mg, 1.15 mmol) was added and the mixture was stirred for about 16 hrs longer at about 25 °C. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a yellow solid (115 mg, 88%).
1H NMR (400 MHz, CDCI3) δ: 8.41 (s, 1 H), 8.39 (s, 1 H), 8.29 (s, 1 H), 8.10 (d, 1 H), 7.04 (d, 1 H),
3.79 (d, 2 H), 3.59 (d, 2 H), 3.43 (s, 2 H), 2.65 (q, 2 H), 1.06 (t, 3 H).
LCMS m/z = 342.1 [MH]+ (35CI isotope)
Example 3 2-(1-Ε8)νΙ·3-(4-(6-(5*(Κν<ΐΓθχνπΐ61ΙΐνΙ)-1Η-ργηζοΙ-3-γΙ)ρνΓ3ΖθΙο[1,5>3]ρνΓ3ζίη-4-/Ι)-1Η-ρνΓ3ΖθΙ-1yi)azetîdin-3-yl)acetonitrile
N-N A
n-nh oh
To a solution of 2-(3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-1-ethylazetidin-3yl)acetonitrile (Préparation 34, 100 mg, 0.29 mmol) in 1,4-dioxane (5 mL) was added 3(tributylstannyl)-1H-pyrazol-5-yl)methanol (Préparation 32, 136 mg, 0.35 mmol) and XPhos Pd G2 (23.0 mg, 0.029 mmol). The mixture was heated at about 110 °C for about 16 hrs. The mixture was concentrated and the residue was purified by chromatography. The product was further purified by HPLC to afford the title compound as a yellow solid (59 mg, 46%).
1H NMR (400 MHz, DMSO-de) δ: 13.21 (s, 0.5 H), 12.93 (s, 0.5 H), 9.23 (s, 0.5 H), 9.07 (s, 1 H), 8.6 8.89 (m, 1 H), 8.69 (s, 0.5 H). 8.47 (s, 0.5 H), 8.26 (s, 1 H), 7.47 - 7.53 (m, 1 H), 6. - 9-6.94 (m, 1 H). 5.35 (s, 0.5 H), 5.35 (s, 0.5 H). 4.50 - 4.57 (m, 2 H), 3.68 - 3.71 (m, 2 H), 3.57 - 3.54 (m, 4 H), 3.17 3.16 (m, 0.5 H), 2.57 - 2.54 (m, 2 H), 0.96 - 0.93 (m, 3 H). This spectrum was consistent with the presence of distinguishable tautomers.
LCMS m/z = 404.3 [MH]-
Préparation 35 2-Cyclobutylideneacetonitrile
A mixture of (EtO)2P(O)CH2CN (4.48 g, 25.2 mmol), LiBr (1.96 g, 22.6 mmol) and TEA (2.28 g, 22.6 mmol) in dry THF (40 mL) was stirred at about 25 °C for about 2 hrs. To this was added a solution of cyclobutanone (1.58 g, 22.6 mmol) in THF (5 mL) at about 25 °C. After about 16 hrs, the mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oit (1.2 g, 57%).
’H NMR (400 MHz, CDCh) δ: 5.11 (quin), 2.93 - 3.05 (m), 2.82 - 2.92 (m), 2.04 - 2.17 (m).
Préparation 36
2-(1-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazoi-1-yl)cyclobutyl)acetonitrile
To a mixture of 2-cyclobutylideneacetonitrile (Préparation 35, 200 mg, 2.15 mmol) and 4pyrazoleboronic acid pinacoi ester (458 mg, 2.36 mmol) in MeCN (15 mL) was added DBU (981 mg, 6.44 mmol). The reaction was stirred at about 25 °C for about 16 hrs and then heated to about 50 °C for about 24 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oil (150 mg, 24%).
1H NMR (400 MHz, CDCI3) δ: m 7.89 (s, 1 H), 7.86 (s, 1 H), 3.09 (s, 2 H), 2.68 - 2.80 (m, 2 H), 2.45 2.55 (m, 2 H), 2.01 -2.10 (m, 2 H), 1.33 (s, 12 H).
LCMS m/z = 287.9 [MH]*
Example 4
2-(1-(4-(6-(1 -Methyi-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-py razol-1 yl)cyciobutyl)acetonitrile
To a mixture of 2-(1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1yl)cyciobutyl)acetonitrile (Préparation 36, 129 mg, 0.45 mmol) and 4-chloro-6-(1-methyl-1H-pyrazol4-yl)pyrazolo[1,5-a]pyrazine (Préparation 11, 100 mg, 0.43 mmol) in 1,4-dioxane (4.3 mL) was added 2 M aq. K3PO4 (0.85 mL) and PdCh(dppf) (15.7 mg, 0.021 mmol). The mixture was purged with nitrogen for about 1 min and stirred at about 80 °C for about 16 hrs. The reaction mixture was combined with an équivalent reaction conducted using 2-( 1-(4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yi)-1H-pyrazol-1-yl)cyclobutyl)acetonitrile (Préparation 36, 20 mg, 0.07 mmol) and 4chloro-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1.5-a]pyrazine (Préparation 11, 19. mg, 0.083 mmol), 2 M aq. K3PO4 (0.14 mL) and PdCb(dppf) (2.5 mg, 0.0035 mmol) in 1,4-dioxane (2 mL). The combined reaction mixtures were concentrated and the residue was purified by chromatography. The compound was further purified by HPLC to afford the title compound as a white solid (22 mg, 12%).
1H NMR (400 MHz, CD3OD) δ: 8.68 (s, 1 H), 8.63 (s, 1 H), 8.39 (s, 1 H), 8.22 (s, 1 H). 8.07 (d, 1 H), 8.06 (s, 1 H), 7.21 (d, 1 H), 3.97 (s, 3 H), 3.94 - 3.99 (m, 1 H), 3.37 (s, 2 H), 3.35 - 3.39 (m, 1 H), 2.84
- 2.95 (m, 2 H), 2.54 (ddd, 2 H), 2.05 - 2.21 (m, 2 H).
LCMS m/z = 358.9 [MH]*
Préparation 37
2-((1 rl3s)-1-(4-Bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (trans isomer)
and
2-((1s,3r)-1-(4-Bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyi)acetonitrile (cis isomer).
DBU (4.25 mL, 28.4 mmol) was added to a solution of 2-(3-methoxycyclobutylidene)acetonitrile (Préparation 90, 3.50 g, 28.4 mmol) and 4-bromopyrazole (4.18 g, 28.4 mmol) in MeCN (80 mL) at about 25 °C. After about 18 hrs, the mixture was poured into NaHïPOi (17.04 g, 142 mmol) in water and the phases were separated. The aqueous layer was extracted twice with EtOAc and the combined EtOAc extracts were concentrated. The excess 4-bromopyrazole was removed by chromatography eluting with DCM:THF (100:0 to 95:5). The material was further purified by chromatography eluting with ethenheptane to afford 2-((1 r,3s)-1-(4-bromo-1 H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile as a white soiid (trans isomer, 2.19 g, 28%)
Ή NMR (400 MHz, CDCh) δ: 7.62 (s, 1 H), 7.55 (s. 1 H), 3.99 (tt, 1 H), 3.30 (s, 3 H), 3.12 (s, 2 H), 2.96 - 3.04 (m, 2 H), 2.44 - 2.51 (m, 2 H).
LCMS m/z = 270.0 [MH]* (79Br isotope) and 2-((1s,3r)-1-(4-bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile as a colorless oil (cis isomer, 5.00 g, 65%).
Ή NMR (400 MHz, CDCh) δ: 7.60 (s, 1 H), 7.53 (s, 1 H), 4.00 (quin, 1 H), 3.29 (s, 3 H), 2.99 (s, 2 H), 2.85 - 2.96 (m, 2 H), 2.56 - 2.67 (m, 2 H).
LCMS m/z = 270.0 [MH]+ (79Br isotope)
Préparation 38 2-((1r,3s)-3-Methoxy-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1yl)cyclobutyl)acetonitrile.
A mixture of 2-((1 r,3s)-1-(4-bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 37, trans isomer, 3399 mg, 12.58 mmol), bis(pinacolato)diboron (3510 mg, 13.8 mmol) and potassium acetate (3700 mg, 37.7 mmol) in 1,4-dioxane (33 mL) was purged with argon for about 5 min, followed by the addition of XPhos Pd G2 (1980 mg, 2.52 mmol) at about 25 °C. The mixture was heated at about 65 °C for about 4 hrs. The mixture was concentrated and the residue was purified by chromatography to afford a solid. To this solid was added EtOAc (10 mL) and heptane (40 mL) and the mixture was stirred at about 25 °C for about 30 min. The solid was filtered and dried under vacuum to afford the title compound as a white solid (1.95 g, 49%).
1H NMR (500 MHz, CDCh) δ: 7.91 (s, 1 H), 7.87 (s, 1 H), 3.98 (tt, 1 H), 3.28 (s, 3 H), 3.17 (s. 2 H). 2.98 - 3.07 (m, 2 H), 2.45 - 2.53 (m, 2 H), 1.31 (s, 12 H).
LCMS m/z = 318.0 [MH]+
Préparation 39
2-((1 r,3s)-1-(4-(6-Chloropyrazolo[1,5-a]py razin-4-yl)-1 H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile
A solution of 2-((1 r,3s)-3-methoxy-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboro!an-2-yl)-1 H-pyrazol-1 yl)cyclobutyl)acetonitrile (Préparation 38, 1950 mg, 6.15 mmol), 4,6-dichloropyrazolo[1,5-a]pyrazine (Préparation 4, 1160 mg, 6.15 mmol) and 2 M aq. K3PO4 (9.22 mL) in 1,4-dioxane (25 mL) was purged with argon for about 5 min followed by the addition of bis(tri-t-butylphosphine)palladium(0) (157 mg, 0.31 mmol) at about 25 °C. After about 2 hrs, the mixture was diluted with EtOAc, the phases were separated and the aqueous phase was extracted twice with DCM. The combined EtOAc and DCM extracts were dried (Na2S0.t) and concentrated to afford a solid that was recrystallized from a warm (about 40 °C) mixture of DCM and heptane to afford the title compound as an off-white solid (1.12 g, 53%). The filtrate was concentrated and purified by chromatography to afford additional title compound (1.01 g, 47%).
1H NMR (500 MHz, CDCb) δ: m 8.39 (d, 1 H), 8.38 (s, 1 H). 8.28 (s, 1 H), 8.09 (d, 1 H), 7.03 (dd, 1 H), 4.04 - 4.10 (m, 1 H), 3.33 (s, 3 H), 3.25 (s, 2 H), 3.09 - 3.17 (m, 2 H), 2.53 - 2.61 (m, 2 H).
LCMS m/z = 343.1 [MH]+ (35Cl isotope)
Préparation 40
3,5-Dibromo-1 -(tetrahydro-2H-pyran-2-yl)-1 H-py razole
Br à
To a solution of 3,5-dibromopyrazole* (CAS: 67460-86-0, 18.0 g, 79.7 mmol) and DHP (30 mL) was added CF3COOH (73 mg, 0.64 mmol). The mixture was heated at about 95 °C for about 12 hrs. The reaction was quenched with NaOH (96 mg, 2.4 mmol) and then concentrated. The resîdue was purified by chromatography to afford the title compound (11.5 g. 46%).
*See: Justus Liebigs Annalen der Chemie 1959, 625, 55 - 65.
1H NMR (400 MHz, CDCb) δ: 6.35 (s, 1 H), 5.42 (d, 1 H), 4.05 (m, 1 H), 3.65 (m, 1 H), 2.38 - 2.48 (m, 1 H), 2.11 (m, 1 H), 1.90 (m, 1 H), 1.62- 1.77 (m, 3 H).
LCMS m/z = 226.7 [MH-THP]+ (79Br, 81 Br isotope)
Préparation 41 3-Bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-carboxylic acid
n-BuLi (2.5 M, 15.8 mL, 39.5 mmol) was added dropwise to a solution of 3,5-dibromo-1-(tetrahydro2H-pyran-2-yl)-1 H-pyrazole (Préparation 40, 9.4 g, 30.0 mmol) in THF (87 mL) at about -78 °C. The mixture was kept at about -78 °C for about 2 hrs. A solution of CO2 (prepared by bubbling CO2 into anhydrous THF (100 mL) for 20 min at about -70 °C and stirring at that température for about 1.5 hrs) was added dropwise while maintaining the internai reaction température below about -65 °C. The mixture was then stirred at about -70 °C for about 1 hr. The mixture was adjusted to about pH 4 with 1 M aq. HCl at about 0 °C and was extracted with EtOAc (3 x 100 mL). The combined EtOAc extracts were washed with brine (2 x 50 mL), dried (NazSCU) and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (5.0 g, 60%).
1H NMR (400 MHz. DMSO-ds) δ: 6.99 (s, 1 H), 6.17 (dd, 1 H), 3.90 (d, 1 H), 3.49 - 3.66 (m, 2 H), 2.12 -2.28 (m, 1 H), 1.91 -2.04 (m, 1 H), 1.83- 1.92 (m, 1 H), 1.58- 1.70 (m, 1 H), 1.45-1.57 (m, 2 H).
Préparation 42 tert-Butyl (3-bromo-1-(tetrahydro-2H-pyran-2-yi)-1H-pyrazol-5-y1)carbamate
Diphenylphosphoryl azide (10 g, 36.4 mmol) was added to a solution of 3-bromo-1-(tetrahydro-2Hpyran-2-yl)-1H-pyrazole-5-carboxylic acid (Préparation 41, 5 g, 18.17 mmol) and DIPEA (6.4 mL, 37.0 mmol) in t-butanol (60.6 mL). The mixture was heated at about 45 °C for about 30 min, and then heated under reflux for about 5 hrs. The mixture was diluted with EtOAc (300 mL) and washed with saturated aq. NaHCOs (3 x 100 mL), brine (2 x 100 mL), dried (NasSO-O and concentrated. The residue was purified by chromatography to afford the title compound as a light yellow oil (3.36 g, 53%).
1H NMR (400 MHz, DMSO-de) δ: 9.64 (br. s., 1 H), 6.26 (s, 1 H), 5.44 (dd, 1 H), 3.84 (d, 1 H), 3.55 3.70 (m, 1 H), 2.08 - 2.20 (m, 1 H), 1.91 - 2.00 (m, 1 H), 1.75 (dd, 1 H), 1.54 - 1.64 (m, 1 H), 1.48 1.53 (m, 2 H), 1.46 (s, 9 H).
LCMS m/z = 367.9 [MNa]+ (79Br isotope)
Préparation 43 3-Bromo-1-(tetrahydro-2H-pyran-2-yl)-5-(diBoc)-amino-1H-pyrazole
DMAP (27 mg, 0.22 mmol) was added to a solution of tert-butyl (3-bromo-1-(tetrahydro-2H-pyran-2yl)-1H-pyrazol-5-yl)carbamate (Préparation 42, 390 mg, 1.13 mmol), di-tert-butyl dicarbonate (492 mg, 2.25 mmol) and TEA (0.47 mL, 3.38 mmol) in DCM (4 mL) at about 20 °C. After about 18 hrs, the mixture was concentrated and the residue was purified by chromatography to afford the title compound (450 mg, 89%) 1H NMR (400 MHz, CDCh) δ: 6.42 (s, 1 H), 5.15 (m, 1 H), 4.02 (m, 1 H), 3.60 (m, 1 H), 2.40 (m, 1 H),
2.15 (m. 1 H), 1.88 (m, 1 H), 1.58-1.76 (m, 3 H), 1.48 (s, 18 H).
LCMS m/z = 467.9 [MNa]‘ (T9Br isotope)
Préparation 44 2-((1r,3s)-1-(4-(6-(5-(DiBoc)-amino-1-(tetrahydro-2H-pyran-2-yl)-pyrazol-3-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1 -yl)-3-methoxycyclobutyl)acetonitrile
A mixture of KOAc (110 mg, 1.06 mmol), bis(pinacolato)diboron (164 mg, 0.64 mmol), 3-bromo-1(tetrahydro-2H-pyran-2-yl)-5-(diBoc)-amino-1H-pyrazole (Préparation 43, 191 mg, 0.43 mmol) and XPhos Pd G2 (55 mg, 0.07 mmol) in 1,4-dioxane (3.5 mL) was was heated to about 65 °C for about 3.5 hrs. The mixture was cooled to about 25 aC and 2-((1 r,3s)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4yl)-1H-pyrazol-1-yl)-3-methoxy-cyclobutyl)acetonitrile (Préparation 39, 68 mg, 0.20 mmol) was added and the mixture was purged with nitrogen before 2 M aq. K3PO4 (0.53 mL, 1.06 mmol) and XPhos Pd G2 (55 mg, 0.07 mmol) were added. The mixture was heated at about 80 °C for about 3 hrs. The reaction was quenched with brine and extracted with EtOAc. The EtOAc extract was concentrated and the residue was purified by chromatography to afford the title compound as a yellow solid (91 mg, 32%).
Ή NMR (400 MHz, CDCh) S: 9.08 (s, 1 H), 8.39 (s, 1 H), 8.33 (s, 1 H), 8.09 (d, 1 H), 6.97 (d, 1 H), 6.90 (s, 1 H), 5.26 (dd, 1 H), 4.02 - 4.11 (m, 2 H), 3.64 (t, 1 H), 3.34 (s, 3 H), 3.25 (s, 2 H), 3.12 - 3.20 (m, 2H), 2.55-2.62 (m, 2 H), 2.17 - 2.25 (m, 1 H), 1.93 (dd, 1 H), 1.58- 1.82 (m, 4 H), 1.45 (s, 18 H).
LCMS m/z = 674.5 [MH]*
Préparation 45
2-((1 r,3s)-1-(4-(6-(5-Amino-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile
CH, O
TFA (2 mL) was added to 2-((1 r,3s)-1-(4-(6-(5-(diBoc)-amino-1-(tetrahydro-2H-pyran-2-yl)-pyrazol-3yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)-acetonitrile (Préparation 44, 91 mg, 0.13 mmol) in anhydrous DCM (1 mL) at about 20 °C. After about 1 hr, the mixture was concentrated. DCM was added, followed by saturated aq. NaHCOa until the solution pH became basic. The phases were separated and the aqueous phase was extracted twice with DCM. The combined DCM extracts were dried (NajSCU), concentrated, and the residue was purified by chromatography to afford the title compound (50 mg, 95%).
1H NMR (400 MHz, CDCh) δ: 8.53 (s, 1 H), 8.40 (s, 1 H), 8.25 (s, 1 H), 8.06 (s, 1 H), 6.96 (br. s., 1 H), 6.01 (br. s., 1 H), 4.33 (br. s., 1 H), 4.05 (m, 1 H), 3.31 (s, 3 H), 3.25 (s, 2 H), 3.11 (dd, 2 H), 2.55 (dd, 2 H).
LCMS m/z = 390.3 [MH]*
Example 5 N-(3-(4-(1 -((1 r,3s)-1 -(Cyanomethyl)-3-methoxycyclobutyl)-1 H-py razol-4-yl)pyrazolo[1,5a]pyrazin-6-yi)-1 H-pyrazol-5-yl)acetamide
CH, Ο
2-((1 r,3s)-1-(4-(6-(5-Amino-1 H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 45, 39 mg, 0.1 mmol) was placed in a reaction tube, which was then evacuated and backfilled three times with nitrogen. To this was added anhydrous DCM (1 mL). The tube was cooled to about 0 °C before N-methylmorpholine (11 mg, 0.11 mmol) and acetyl chloride (8.6 mg, 0.11 mmol) were added. The mixture was allowed to warm to about 25 °C over about 18 hrs. The mixture was concentrated and the residue was dissolved in MeOH (2 mL). K2CO3 (30 mg, 0.22 mmol) was added at about 0 °C. After about 3 hrs, the mixture was filtered through Celite®. The filtrate was concentrated and the residue was purified by chromatography to afford the title compound (30 mg, 63%).
Ή NMR (400 MHz, CD3OD) δ: 8.80 (s, 1 H), 8.75 (s, 1 H), 8.40 (s, 1 H), 8.09 (d, 1 H), 7.23 (d, 1 H), 6.88 (s, 1 H), 4.02 - 4.12 (m, 1 H), 3.81 (s, 3 H), 3.34 (s. 2 H), 3.15 - 3.24 (m. 2 H), 2.48 - 2.59 (m, 2 H). 2.21 (s, 3 H).
LCMS m/z = 432.2 [MH]*
Préparation 46
Ethyl 5-(4-(1-((1 r,3s)-1-(cyanomethyl)-3-methoxycyclobutyl)-1 H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-6-yl)-1H-pyrazole-3-carboxylate
N-N
A solution of 2-((1 r,3s)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile (Préparation 39, 200 mg, 0.58 mmol) and ethyl 3-(tributylstannyl)-1H pyrazole-5-carboxylate (Préparation 31, 300 mg, 0.70 mmol) in 1,4-dioxane (5.8 mL) was purged with nitrogen and XPhos Pd G2 (45.9 mg, 0.058 mmol) added at about 25 °C. The reaction was heated at about 80 °C for about 16 hrs. The cooted mixture was purifted by chromatography to afford the title compound as a yellow solid (220 mg, 84%).
’H NMR (400 MHz, DMSO-cfe) δ: 14.09 (s, 1 H). 9.39 (s, 1 H), 9.10 (s. 1 H), 8.72 (s, 1 H), 8.30 (d, 1 H), 7.59 (d, 1 H), 7.53 (d, 1 H), 4.33 (q, 2 H), 4.01 (dd, 1 H), 3.44 (s, 3 H), 3.22 (s, 2 H). 3.14 - 3.20 (m, 2 H). 2.43 - 2.48 (m, 2 H), 1.34 (t, 3 H).
LCMS m/z = 447.2 [MH]*
Example 6
5-(4-(1-((1 r,3s)-1-(Cyanomethyl)-3-methoxycyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6yl)-N-methyi-1H-pyrazole-3-carboxamide
N-N
A solution of ethyl 5-(4-(1-((1r,3s)-1-(cyanomethyl)-3-methoxycyclobutyl)-1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-6-yl)-1H-pyrazole-3-carboxylate (Préparation 46, 100 mg, 0.22 mmol) in 30% MeNHz in EtOH solution (35 mL) was sealed in a microwave tube at about 20 °C. After about 16 hrs, the mixture was concentrated and the residue was purified by HPLC to afford the title compound 20 as a white solid (62 mg, 58%).
’H NMR (400 MHz, CD3OD) δ: 9.00 (s, 1 H), 8.91 (s, 1 H), 8.57 (s, 1 H), 8.18 (s. 1 H), 7.37 (br. s., 1 H), 7.26 (s. 1 H), 4.61 (s, 1 H), 4.04 - 4.15 (m, 1 H), 3.37 (s, 3 H), 3.33 (s, 2 H), 3.18 - 3.26 (m, 2 H), 2.95 (s, 3 H), 2.56 (dd, 2 H).
LCMS m/z = 432.1 [M+H]*
Préparation 47 tert-Butyl 3-(cyanomethyl)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1 -yl)azetidine-1-carboxylate
To a vial were added 4-chloro-6-(1-methyl-1H-pyrazol-4-yl)pyrazoio[1,5-a]pyrazine (Préparation 11, 150 mg), tert-butyl 3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1yl)azetidine-1-carboxylate (Préparation 28, 374 mg, 0.96 mmol), 2 M aq. Na2COs (0.96 mL) and 1,4dioxane (4 mL). The mixture was purged with argon for about 5 min, followed by the addition of PdCb(dppf) (93.7 mg. 0.13 mmol). The mixture was heated at about 120 °C for about 1 hr under microwave irradiation. The mixture was diluted with EtOAc, washed with water, dried (Na2SO4), and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (233 mg, 79%).
1H NMR (400 MHz, DMSO-de) δ: 9.03 (s, 1 H), 8.95 (s, 1 H), 8.53 (s, 1 H), 8.37 (s, 1 H), 8.20 (d, 1 H),
8.16 (s, 1 H), 7.46 (s, 1 H), 4.54 (d, 2 H), 4.25 (d, 3 H), 3.91 (s, 3 H), 3.67 (s, 2 H), 1.42 (s, 9 H).
LCMS m/z = 460.2 [MH]+
Préparation 48
2-(3-(4-(6-(1-Methyl-1H-py razol-4-yl)pyrazoio[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidin-3yl)acetonitrile
TFA (1 mL) was added to tert-butyl 3-(cyanomethyl)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate (Préparation 47, 233 mg, 0.51 mmol) in DCM (5 mL) at about 25 °C. After about 90 min, the mixture was concentrated. The residue was concentrated twice with toluene, then dried under vacuum. The residue was dissolved in MeOH and passed through a bed of polymer supported carbonate resin. The eluted solution was concentrated to afford the title compound (200 mg, about 100%) which was used without further purification.
’H NMR (400 MHz, DMSO-de) δ: 9.12 (s, 1 H), 9.06 (s, 1 H), 8.58 (s. 1 H), 8.39 (s, 1 H), 8.22 (d, 1 H),
8.17 (s, 1 H), 7.50 (d, 1 H), 4.68 - 4.77 (m, 2 H), 4.35 - 4.40 (m, 2 H), 3.93 (s, 2 H), 3.91 (s, 3 H).
LCMS m/z = 360.5 [MH]*
Example 7 2,2'-(3-(4-(6-(1-Methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)azetidine-1,3diyljdiacetonitrile
DIPEA (132 CL, 0.76 mmol) was added to a solution of 2-(3-(4-(6-(1-methyl-1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acet-onitrile (Préparation 48, 91 mg, 0.25 mmol) in DMF (2 mL). Bromoacetonitrile (36 mg, 0.30 mmol) was added at about 25 ’C. Aller about 18 hrs, the reaction was quenched with conc. NH-tOH and stirred at about 25 °C for about 10 min. The mixture was concentrated and the residue was diluted with DCM. The DCM was washed twice with saturated aq. NH«CI, then twice with saturated aq. NazCCh. The DCM was dried (NaaSOi) and concentrated. The residue was .purified by chromatography to afford the title compound as an off white solid (68 mg, 68%).
1H NMR (400 MHz, CD3OD) δ: 8.77 (s, 1 H), 8.74 (s, 1 H), 8.45 (s, 1 H), 8.25 (s, 1 H), 8.08 - 8.10 (m, 2 H), 7.25 (d, 1 H), 4.01 (d, 2 H), 3.98 (s, 3 H), 3.86 (d, 2 H), 3.78 (s, 2 H), 3.54 (s, 2 H).
LCMS m/z = 399.3 [MH]‘
Example 8 2-(3-(4-(6-(1 -Methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]py razin-4-yl)-1 H-pyrazol-1 -y l)-1 (methylsulfonyl)azetidin-3-yl)acetonÎtrile
6I
To a solution of 2-(3-(4-(6-(1 -methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)azetidin-3-yl)acetonitrile (Préparation 48. 120 mg, 0.27 mmol) and TEA (331 mg, 3.27 mmol) in DCM (20 mL) was added methanesulfonyl chloride (313 mg, 2.73 mmol) at about 0 °C. The mixture was then stirred at about 10 °C for about 1 hr before being concentrated. The residue was purified using HPLC to afford the title compound as a white soiid (39 mg, 33%).
’H NMR (400 MHz, DMSO-ds) δ: 9.05 (s, 1 H), 8.99 (s, 1 H), 8.57 (s, 1 H), 8.37 (s, 1 H), 8.21 (d, 1 H),
8.17 (s, 1 H), 7.47 (d, 1 H), 4.65 (d, 2 H), 4.30 (d, 2 H), 3.91 (s, 3 H), 3.70 (s, 2 H), 3.14 (s, 3 H).
LCMS m/z = 437.9 [MH]'
Example 9
2-(1-(Cyclopropylsulfonyl)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-yl)azetîdin-3-yl)acetonitrîle
To a mixture of 2-(3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazoi-1yl)azetidin-3-yl)acetonitrile (Préparation 48, 100 mg, 0.25 mmol) in DCM (10 mL) were added TEA (153 mg, 1.52 mmol) and cyctopropanesulfonyl chloride (107 mg, 0.76 mmol) at about 0 °C. The mixture was stirred at about 20 °C for about 16 hrs, then concentrated. The residue was purified by HPLC to afford the title compound as a white solid (60 mg, 46%).
’H NMR (400 MHz. DMSO-d6) δ: 9.05 (s. 1 H), 9.00 (s, 1 H), 8.57 (s, 1 H), 8.37 (s, 1 H), 8.21 (d, 1 H), 8.16 (s, 1 H), 7.46 (d, 1 H), 4.69 (d, 2 H), 4.32 (d, 2 H), 3.91 (s, 3 H). 3.70 (s, 2 H), 2.85 (m, 1 H), 0.96 - 1 09 (m, 4 H).
LCMS m/z = 464.0 [MH]4
Example 10
2-(3-(4-(6-(1 -Methy 1-1 H-py razol-4-y l)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-py razol-1 -yl)-1 propionylazetidin-3-yl)acetonitrile
To a solution of 2-(3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)azetidin-3-yl)acetonitrile (Préparation 48, 90 mg, 0.23 mmoi) in DCM (5 mL) were added TEA (69 mg, 0.68 mmol) and propionic anhydride (59 mg, 0.45 mmol). The reaction was stirred at about 20 °C for about 30 min, then concentrated and the residue was purified by HPLC to afford the title compound as a white solid (28 mg, 30%).
1H NMR (400 MHz, DMSO-ds) δ: 9.05 (s, 1 H), 8.97 (s, 1 H), 8.55 (s, 1 H), 8.37 (s, 1 H), 8.21 (d, 1 H). 8.16 (s, 1 H), 7.46 (d. 1 H), 4.80 (d, 1 H), 4.52 (dd, 2 H), 4.23 (d. 1 H), 3.91 (s. 3 H), 3.70 (s, 2 H), 2.16 (q, 2 H), 0.99 (t, 3 H).
LCMS m/z 416.0 [MH]*
Préparation 49 2-(3-(Benzyloxy)cyclobutylidene)acetonitrile
LiBr (0.270 g, 3.12 mmol) was placed under vacuum, then backfilled with nitrogen and THF (28 mL) was added, folfowed by (EtOJîPOCHaCN (0.53 mL, 3.12 mmol) and TEA (0.79 mL, 5.67 mmol). The resulting solution was stirred at about 20 °C for about 45 min, then a solution of 3(benzyloxy)cycîobutanone (500 mg, 2.84 mmol) in dry THF (3 mL) was added. After about 5 hrs, the mixture was poured into EtOAc (100 mL) and the EtOAc was washed three times with saturated aq. NH4CI (3 x 50 mL) and with brine (25 mL). The EtOAc extract was dried (MgSO<) and concentrated. The residue was chromatographed to afford the title compound (480 mg, 85%).
Ή NMR (400 MHz, CDCh) δ: 7.29 - 7.43 (m, 5 H), 5.24 (quin, 1 H), 4.45 - 4.53 (m, 2 H), 4.19 (quin, 1 H), 3.18 - 3.29 (m, 1 H), 3.03 - 3.13 (m, 1 H), 2.86 - 3.00 (m, 2 H).
Préparation 50
2-(3-Hydroxycyclobutyl)acetonitrile
HO
Palladium hydroxide on carbon (20% Pd, wet. 430 mg) was added to a solution of 2-(3(benzyloxy)cyclobutylidene)acetonitrile (Préparation 49, 430 mg, 2.16 mmol) in THF (6.5 mL). The mixture was pressurized under hydrogen (100 psi) in a Steel reactor and stirred at about 20 °C for about 1 hr. Additional palladium hydroxide on carbon (20% Pd, wet, 430 mg) added and the mixture was re-pressurized under hydrogen (100 psi) and stirred for about 1.5 hrs longer. The mixture was diluted with EtOAc, filtered through Celite®, and the filtrate was concentrated to afford the title compound as a colorless oil (240 mg, 100%) as a mixture of cis and trans isomers.
Isomer 1: 1H NMR (400 MHz, CDCh) δ: 4.52 (quin, 1 H), 2.63 - 2.74 (m, 2 H), 2.19 - 2.28 (m, 4 H), 2.07-2.17 (m, 1 H), 1.83 (br. s, 1 H).
Isomer 2: 1H NMR (400 MHz, CDCh) δ: 4.14-4.25 (m, 1 H), 2.55 - 2.64 (m, 2 H), 2.48 (dd, 4 H), 1.83 (br. s, 1 H), 1.67- 1.79 (m, 1 H).
Préparation 51 2-(3-Oxocyclobutyl)acetonitrile
To a solution of 2-(3-hydroxycyclobutyl)acetonitrile (Préparation 50, 50 mg, 0.45 mmol) in dry THF (1.8 mL) was added Dess-Martin periodinane (CAS: 87413-09-0, 216 mg, 0.49 mmol). The vial was sealed under ambient atmosphère and stirred at about 50 °C for about 2 hrs. The mixture was diluted with EtzO (10 mL) folfowed by saturated aq. NaHCOs (4 mL). Sodium thiosulfate pentahydrate (954 mg, 3.82 mmol) was added and the mixture was stirred vigorously for about 10 min until ali solids had dissolved. The phases were separated and the aqueous phase extracted once more with Et20. The combined EhO extracts were dried (MgSO<) and concentrated. The residue was purified by chromatography to afford the title compound as a colorless oil (27 mg, 55%).
1H NMR (400 MHz, CDCh) δ: 3.23 - 3.37 (m, 2 H), 2.89 - 3.01 (m, 2 H), 2.75 - 2.89 (m, 1 H), 2.69 (d, 2 H).
13C NMR (101 MHz, CDCh) δ: 203.6, 117.6, 52.3, 23.1, 20.6.
Préparation 52
2-(3-(Cyanomethyl)cyclobutylidene)acetonitrile
The title compound was prepared (75mg, 89%) using the method of Préparation 35 using 2-(3oxocyclobutyljacetonitrite (Préparation 51, 70 mg, 0.64 mmol), (EtOJîPOCHîCN (213 mg, 1.15 mmol), LiBr (100 mg, 1.15 mmol) and TEA (0.27 mL, 1.92 mmol). The title compound was used directly in the next reaction (Example 11 and 12).
’H NMR (400 MHz, CDCh) δ: 5.25 (m, 1 H), 3.10 - 3.30 (m, 2 H), 2.80 (m, 3 H), 2.60 (m, 2 H).
GCMS m/z = 132 [M]*
Préparation 53
6-(1 -Methyl-1 H-pyrazol-4-yl)-4-(1 H-pyrazol-4-yl)py razolo[1,5-a]pyrazine
N-NH // Δ
Pd(dppf)Cb (5.01 g, 6.85 mmol) was added a mixture of 4-chloro-6-(1-methyl-1 H-pyrazol-4yl)pyrazolo[1,5-a]pyrazine (Préparation 11, 8 g, 34 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-1H-pyrazoïe-1-carboxy!ate (12.1 g, 41.1 mmol) and 2 M aq. K2CO3 (34.2 mL) in 1,4-dioxane (30 mL) and toluene (30 mL) at about 10 °C while a stream of nitrogen was bubbled through the solution. The mixture was stirred at about 100 °C for about 16 hrs, then kept at about 10 °C for about 48 hrs. The mixture was filtered and the filtrate was concentrated. The residue was purified by chromatography to afford the title compound as a grey oil (6.3 g, 69%).
Ή NMR (400 MHz, DMSO-de) δ: 8.97 (s, 1 H), 8.72 (s, 1 H), 8.40 (s, 1 H), 8.35 (s, 1 H), 8.15 (d. 1 H), 8.12 (s, 1 H), 7.36 (d, 1 H), 3.91 (s, 3 H).
LCMS m/z = 265.8 (MH]*
Example 11
2,2'-(( 1 s,3s)-1 -(4-(6-(1 -Methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 yl)cyclobutane-1,3-diyl)diacetonitrile (cis isomer)
and
Example 12 2,2'-(( 1 r,3r)-1 -(4-(6-(1 -Methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-py razol-1 yl)cyclobutane-1,3-diyl)diacetonitrile (trans isomer)
DBU (34 mg, 0.22 mmol) was added to a siurry of 2-(3-(cyanomethyl)cyclobutylidene)acetonitrile (Préparation 52, 32 mg, 0.24 mmol) and 6-(1-methyl-1 H-pyrazol-4-yl)-4-(1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazine (Préparation 53, 53 mg, 0.20 mmol) in MeCN (4.8 mL). The mixture was heated at about 50 °C for about 16 hours, then concentrated. The residue was purified by chromatography to afford a mixture of the title compounds as 1:1 mixture of cis/trans isomers (80 mg, 83%). The isomers were separated by HPLC to afford 2,2'-((1s,3s)-1-(4-(6-(1-methyl-1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1,3-diyl)diacetonitrile as a solid (cis isomer, 27 mg, 36%).
Ή NMR (500 MHz, 9:1 CDCI3-CD3OD) δ: 8.45 (s, 1 H), 8.35 (s, 1 H), 8.25 (s, 1 H), 7.99 (d, 1 H), 7.95 (s, 1 H), 7.92 (S, 1 H), 6.93 (s, 1 H), 3.95 (s, 3 H), 3.72 (s, 2 H), 3.18 (s, 2 H), 2.81 - 2.91 (m, 2 H), 2.72 - 2.81 (m, 1 H), 2.72 - 2.81 (m, 1 H). 2.64 - 2.72 (m. 2 H), 2.61 (d, 2 H).
LCMS m/z= 399.1 [MH] + and 2,2-((1 r,3r)-1 -(4-(6-(1 -methyl-1 H-pyrazo!-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)cyclobutane-1,3-diyl)diacetonitrile as a solid (trans isomer, 14 mg, 18%).
Ή NMR (500 MHz, CDCh) δ: 8.49 (s. 1 H), 8.46 (s, 1 H), 8.32 (s, 1 H), 8.05 (d, 1 H), 7.96 (m, 2 H), 6.95 (d, 1 H), 4.00 (s, 3 H), 3.75 (br. s., 1 H), 3.15 - 3.25 (m, 2 H), 3.11 (s, 2 H), 2.88 (tt. 1 H). 2.64 (d, 2 H), 2.48-2.59 (m, 2 H).
LCMS m/z = 399.1 [MH]
Préparation 54
2-(3-( Benzyloxy)cyclobutylidene)acetonitrile
To a solution of (EtO)2P(O)CHïCN (24.1 g, 136 mmol) in dry THF (500 mL) was added LiBr (11.8 g, 136 mmol) and TEA (18.4 g, 182 mmol) and the resulting mixture was stirred at about 20 °C for about 1 hr. To this was added 3-(benzyloxy)cyclobutan-1-one (16.00 g, 90.8 mmol). The mixture was stirred at about 20 °C for about 20 hrs. The mixture was filtered and the filtrate was concentrated. The residue was purified by chromatography to afford the title compound as a pale yellow liquid (18 g, 99%).
1H NMR (400 MHz, CDCh) δ: 7.29 - 7.40 (m, 5 H), 5.24 (dt, 1 H), 4.45 - 4.53 (m, 2 H), 4.19 (quin, 1 H), 3.19 - 3.29 (m, 1 H), 3.03 - 3.13 (m, 1 H), 2.86 - 3.00 (m, 2 H).
LCMS m/z = 200.1 [MH]
Préparation 55 2-(3-(Benzyloxy)-1-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1I5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutyl)acetonitrile (mixture of cis and trans isomers)
DBU (1.448 g, 9.4 mmol) was added to a solution of 6-(1-methyl-1H-pyrazoi-4-yl)-4-(1H-pyrazoi-4yl)pyrazolo[1,5-a]pyrazine (Préparation 53, 313 mg, 1.18 mmol) and 2-(318860 (benzytoxy)cyclobutyiidene)acetonitrile (Préparation 54, 470 mg, 2.36 mmol) in MeCN (6.38 mL). The mixture was stirred at about 20 °C for about 18 hrs, then it was poured into aq. NaHîPCh solution to maintain a pH of about 5. The mixture was extracted three times with DCM. The combined DCM extracts were concentrated and the residue was purified by chromatography to afford the title 5 compound (474 mg, 86%).
1H NMR (400 MHz, CD3OD) δ: 8.67 - 8.72 (m, 2 H), 8.63 (s, 1 H), 8.39 (s, 1 H), 8.22 (s, 1 H), 8.04 8.10 (m, 2 H), 7.25 - 7.42 (m, 5 H), 4.53 (s, 2 H), 4.24 - 4.34 (m, 1 H), 3.97 (s, 3 H), 3.29 (s, 2 H). 2.92 - 3.02 (m, 2 H), 2.77 - 2.88 (m, 2 H).
LCMS m/z = 465.3 [MH]*
Example 13
2-((1 s,3r)-3-Hydroxy-1 -(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-y l)-1 H-pyrazol-1 yl)cyclobutyl)acetonitrile (cis isomer)
N-N
and
Example 14 PF-06877900
2-((1 r,3s)-3-Hydroxy-1-(4-(6-(1-methyl-1 H-pyrazoi-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 yl)cyclobutyl)acetonitrile (trans isomer)
Sodium iodide was dried with a heating gun at full heat for 5 min then cooled to about 25 °C under nitrogen prior to use. The dried Nal (1.21 g, 8.1 mmol) was added at about 20 °C to a stirred solution of (2-(3-(benzyloxy)-1-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutyl)acetonitrile (Préparation 55, 377 mg, 0.81 mmol) in MeCN (12 mL), followed byTMSCI (1.05 mL, 8.1 mmol). The mixture was stirred at about 50 °C for about 18 hrs. An additional portion of both Nal and TMSC! were added and the mixture was maintained at about 50 °C for about 8 hrs additional. The cooled mixture was poured into ice cold saturated aq. NaHCOa containing sodium thiosulfate pentahydrate (10.1 g, 40.6 mmol) and extracted three times with EtOAc. The combined EtOAc extracts were concentrated to afford a mixture of the two title compounds. This mixture was purified by chromatography to afford 2-((1s,3r)-3-hydroxy-1-(4-(6-(1-methyl-1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutyl)acetonitrile (trans isomer, 80 mg, 26%)
Ή NMR (400 MHz, CDsOD) δ: 8.72 (s, 1 H), 8.71 (s, 1 H), 8.42 (s, 1 H), 8.24 (s, 1 H), 8.05 - 8.10 (m, 2 H), 7.24 (d, 1 H), 4.34 - 4.44 (m, 1 H), 3.98 (s, 3 H), 3.35 (s, 2 H), 3.22 - 3.28 (m, 2 H), 2.49 (dd, 2 H).
LCMS m/z = 375.4 [MH]* and 2-((1 r,3s)-3-hydroxy-1-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutyl)acetonitrile (cis isomer, 180 mg, 59%).
1H NMR (400 MHz, CDaOD) δ: 8.67 (s, 1 H), 8.62 (s, 1 H), 8.38 (s, 1 H), 8.21 (s, 1 H), 8.07 (d, 1 H), 8.05 (s, 1 H), 8.04 - 8.06 (m, 1 H), 7.21 (d, 1 H), 4.41 (quin, 1 H), 3.97 (s, 3 H), 3.28 (s, 2 H), 2.95 3.04 (m, 2 H), 2.68 - 2.78 (m, 2 H).
LCMS m/z = 375.4 [MH]*
Example 15
2-((1 r,3s)-3-Methoxy-1-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutyl)acetonitrile (trans isomer)
and
Example 16
2-((1 s,3r)-3-Methoxy-1 -(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1yl)cyciobutyl)acetonitrile (cis isomer)
A solution of a mixture of 2-((1 s,3r)-3-hydroxy-1-(4-(6-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutyl)acetonitrile and 2-((1r,3s)-3-Hydroxy-1-(4-(6-(1-methyl-1Hpyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazoî-1-yl)cyclobutyl)acetonitrile (Examples 13 and 14, mixture of cis and trans isomers, 48 mg, 0.13 mmol) in THF (0.2 mL) was sequentially treated with tetrabutylammonium bromide (TBAB, 126 mg, 0.38 mmol), 1 M aq. NaOH (1.02 mL) and dimethyl sulfate (6 CL). The reaction vial was sealed and stirred vigorously at about 20 °C for about 1 hr. Additional dimethyl sulfate (14 CL) and TBAB (10 mg) were added and the reaction was kept at about 20 °C for about 2 hrs. The mixture was extracted three times with EtOAc and the combined EtOAc extracts were dried (NasSCU) and concentrated. The residue was purified by chromatography to afford a mixture of cis and trans isomers. Further purification by HPLC afforded 2-((1r,3s)-3-methoxy1-(4-(6-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)cyclobutyl)acetonitrile as a solid (trans isomer, 10 mg, 20%) ’H NMR (400 MHz, CD3CN) Ô: 8.61 (d, 1 H), 8.55 (s, 1 H), 8.38 (s, 1 H), 8.12 (s, 1 H), 8.05 (d, 1 H), 7.99 (s, 1 H), 7.13 (d, 1 H), 3.99 - 4.08 (m, 1 H), 3.93 (s, 3 H), 3.27 (s, 5 H), 3.13 - 3.21 (m, 2 H), 2.45 -2.53 (m,2H).
LCMS m/z = 389.1 [MH]’ and 2-((1 s,3r)-3-methoxy-1-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 yl)cyclobutyl)acetonitrile as a solid (cis isomer, 17 mg, 33%).
1H NMR (400 MHz, CDsCN) δ: 8.60 (d, 1 H), 8.49 (s, 1 H), 8.36 (s, 1 H), 8.12 (s, 1 H), 8.04 (d, 1 H), 7.98 (s, 1 H), 7.12 (dd, 1 H), 4.03 (quin, 1 H), 3.92 (s, 3 H), 3.26 (s, 3 H), 3.19 (s. 2 H), 2.89 - 2.97 (m, 2 H), 2.65 -2.74 (m, 2 H).
LCMS m/z= 389.1 [MH]*
Préparation 56
Benzyl (R)-2-methyl-3-oxoazetidine-1 -carboxylate
Part 1
A solution of N,4-dimethyl-N-nitrosobenzenesulfonamide (Diazald®, 21.25 g, 99.19 mmol) in Et2O (150 mL) was added dropwise to a solution of KOH (6 g, 106.9 mmol) and 2-(2-ethoxyethoxy)ethanol (30 mL) and water (10 mL) at about 70 °C. The mixture was heated at about 70 °C and an ethereal solution of diazomethane was collected as a yellow liquid by distillation (100 mL, estimated to contain 66 mmol of diazomethane) using a dry-ice/acetone condenser and used immediately in Part 2.
Part 2
Ethyl chloroformate (2.430 g, 22.4 mmol) was added drop wise to a solution of Cbz-D-alanine (5.00 g, 22.4 mmol) and TEA (2.27 mg, 22.4 mmol) in THF (50 mL) at about -15 °C. The reaction mixture was warmed to about 0 °C and the diazomethane solution from Part 1 (100 mL, about 66 mmol) was added dropwise and stirred at about 20 °C for about 16 hrs. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (2 x 30 mL). The combined EtOAc extracts were dried (NazSOi) and concentrated. The residue was purified by chromatography to afford the diazo-ketone intermediate (4.5 g, 81%) as a white solid. The material was used in Part 3.
Part 3
To a solution of the diazo-ketone intermediate of Part 2 (4.50 g, 18.2 mmol) in DCM (450 mL) were added TEA (18 mg, 0.18 mmol) and Rh2(OAc)4 (40 mg, 0.091 mmol) at about 0 °C. The mixture was stirred for about 16 hrs at about 25 'C. The mixture was quenched with water (50 mL) and the DCM phase was separated and concentrated. The residue was purified by chromatography on afford the title compound as a white solid (1.20 g, 30%).
1H NMR (400 MHz, CDCb) δ: 7.34 - 7.38 (m. 5 H), 5.18 (m, 2 H), 5.03 (m, 1 H), 4.65 - 4.81 (m, 2 H), 1.49 (d, 3 H).
GCMS m/z = 219 [M]*
Préparation 57
Benzyl {R)-3-(cyanomethy1ene)-2-methyiazetidine-1-carboxylate
7I
To a mixture of LiBr (166 mg, 1.92 mmol) and TEA (277 mg, 2.74 mmol) in THF (10 mL) was added (EtO)ïP(O)CH2CN (339 mg, 1.92 mmol) at about 25 °C. After about 2.5 hrs, benzyl (R)-2-methyl-3oxoazetidine-1-carboxylate (Préparation 56, 300 mg, 1.37 mmol) in THF (2 mL) was added at about 25 °C and the mixture was stirred for about 16 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as mixture of E/Z olefin isomers as colorless oil (290 mg, 87%).
1H NMR (400 MHz, CDCh) δ: 7.33 - 7.40 (m, 5 H), 5.36 (m, 1 H), 5.10 - 5.17 (m, 2 H), 4.97 (m, 1 H), 4.63-4.77 (m, 2 H), 1.50-1.67 (m. 3 H).
Préparation 58
Benzyl (2R)-3-(cyanomethyl)-2-methyl-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin4-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate
To a solution of 6-(1-methyl-1H-pyrazol-4-yl)-4-(1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine (Préparation 53, 268 mg, 1.01 mmol) in MeCN (15 mL) were added benzyl (R)-3-(cyanomethylene)-2methylazetidine-1-carboxylate (Préparation 57, 294 mg, 1.21 mmol) and DBU (77 mg, 0.51 mmol) at about 15 °C. The mixture was stirred at about 25 °C for about 7 hrs before being diluted with EtOAc (50 mL) and washed with 1 M aq. citric acid (15 mL) followed by brine (15 mL). The EtOAc extract was concentrated and the residue was purified by chromatography to afford the title compound as a yellow gum (500 mg, 97%).
Ή NMR (400 MHz, CDCh) S: 8.49 (s, 1 H), 8.33 (s, 1 H), 8.32 (s, 1 H), 8.05 (d, 1 H), 7.96 (s, 1 H), 7.95 (s, 1 H), 7.33 - 7.40 (m, 5 H), 6.93 (d, 2 H), 5.15 (s, 2 H), 4.87 (m. 1 H), 4.61 (d, 1 H), 4.26 (d. 1 H), 4.01 (s, 3 H), 3.27 (s, 2 H), 1.69 (d, 3 H).
LCMS m/z = 530.1 [M+Na]*
Préparation 59 2-((2R)-2-M ethyl-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 yl)azetidin-3-yl)acetonitrile
To a mixture of Nal (2.36 g, 15.8 mmol) in MeCN (16 mL) was added TMSCI (2 mL, 15.8 mmol) at about 0 °C. The mixture was stirred at about 15 °C for about 4 hrs. A solution of benzyl (2R)-3(cyanomethyl)-2-methyl-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)azetidine-1-carboxylate (Préparation 58, 400 mg, 0.79 mmol) in MeCN (4 mL) was added at about 0 °C. Stirring was continued at about 15 °C for about 3 hrs. The mixture was cooled to about 10 °C and quenched by the addition of TEA (2 mL). The mixture was concentrated. The residue was dissotved in EtOAc (20 mL) and MeOH (2 mL) and the solids présent were removed by filtration. The filtrate was concentrated and the residue was purified by TLC to afford the title compound as a yellow solid (200 mg, 68%).
’H NMR (400 MHz, DMSO-de) 5: 9.06 (s, 1 H), 9.03 (s, 1 H), 8.59 (s, 1 H), 8.39 (s, 1 H), 8.22 (d, 1 H),
8.16 (s, 1 H), 7.47 (s, 1 H), 5.09 (m, 1 H), 4.74 (d, 1 H), 4.24 (d, 1 H). 3.91 (s, 3 H), 2.99 (s, 2 H), 1.65 (d, 3 H).
LCMS m/z = 374.0 [MH]*
Example 17 2-((2R,3S)-2-Methyl-3-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)py razolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 y 1)-1-(2,2,2-trifluoroethyl)azetidin-3-yl)acetonitrile
To a solution of 2-((2R)-2-methyl-3-(4-(6-(1-methyl-1H-pyrazo!-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-y!)azetidin-3-yl)acetonitrile (Préparation 59, 200 mg, 0.43 mmol) in DMF (5 mL) were added 2,2,2-trifluoroethyl trifiuoromethanesulfonate (298 mg, 1.29 mmol) and DIPEA (332 mg, 2.57 mmol). The mixture was stirred at about 10 °C for about 36 hrs before being difuted with EtOAc (30 mL) and washed with brine (15 mL). The EtOAc extract was concentrated the residue was purified by HPLC to afford the title compound as a mixture of diastereomers (80 mg, 41%) as a white solid. Further HPLC purification afforded the title compound as a white solid (41.8 mg, 21%, 94.7% ee).
Ή NMR (400 MHz, CD3CN) δ: 8.59 (s, 1 H), 8.45 (s, 1 H), 8.36 (s, 1 H), 8.10 (s, 1 H), 8.04 (d, 1 H), 7.98 (s, 1 H), 7.09 - 7.13 (m, 1 H), 3.92 - 3.96 (m, 1 H). 3.91 (s, 3 H), 3.84 - 3.89 (m, 1 H), 3.77 - 3.84 (m, 1 H), 3.38 (s, 2 H), 3.28 - 3.43 (m, 1 H), 3.15 (dq, 1 H), 1.36 (d, 3 H).
LCMS m/z = 456.2 [MH]*
Example 18 2-((1 r,3r)-1-(4-(6-(1 -Methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)-3-(1 Hpyrazol-5-yl)cyclobutyl)acetonitrîle
To a solution of LiEJr (70 mg, 0.81 mmol), TEA (205 pL, 1.47 mmol) in THF (10 mL), was added (EtO)2P(O)CHzCN (143 mg, 0.81 mmol). The mixture was stirred at about 20 °C for about 30 min. 3(1 H-Pyrazol-5-yl)cyclobutan-1-one (Préparation 99, 100 mg, 0.74 mmol) was added and mixture was kept at about 20 °C for about 18 hrs. About 30% of the reaction solution was taken out and 6-(1methyl-1H-pyrazol-4-yl)-4-(1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine (Préparation 53, 58 mg, 0.22 mmol) and DBU (110 pL, 0.73 mmol) in MeCN (5 mL) were added. Stirring at about 20 °C was maintained for about 20 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound (6 mg, 6%).
Ή NMR (500 MHz, CD3OD) δ: 8.85 (s, 1 H), 8.73 (s, 1 H), 8.46 (s, 1 H), 8.26 (s, 1 H), 8.10 (s, 2 H), 7.60 (br. s., 1 H), 7.28 (d, 1 H), 6.32 (d, 1 H), 3.95 - 4.04 (m. 3 H), 3.69 - 3.78 (m, 1 H), 3.33 - 3.43 (m, 2 H), 2.77-2.89 (m, 2 H).
LCMS m/z = 425.4 [MH]*
Example 19 (1s,3s)-3-(Cyanomethyl)-3-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 Hpyrazol-1-yl)cyclobutane-1-carbonitrile
To a solution of (1s,3s)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1Hpyrazol-1-yl)cycîobutane-1-carbonitrile (Préparation 91, 539 mg, 1.72 mmol) and 4-chloro-6-(1methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine (Préparation 11, 350 mg, 1.5 mmol) in 1,4-dioxane (7.5 mL) was added 2 M aq. K3PO4 (2.25 mL) at about 25 °C. The mixture was placed under nitrogen, then XPhos Pd G2 (11.8 mg, 0.0150 mmol) was added. The mixture was heated at about 40 °C for about 6 h, then it was heated to about 80 °C to dissolve the precipitated product. The aqueous layer was removed while maintaining the température at about 80 °C, then the 1,4-dioxane phase was added to EtOH (70 mL, previously preheated to about 50 °C). The mixture was stirred for about 10 min at about 50 °C before being removed from heat. Stirring at about 25 °C was continued for about 18 h. The solid was fîltered and washed with EtOH (2 x 25 mL) and dried under vacuum to afford the title compound as a white solid (483 mg, 84%).
mp 217-220 °C ’H NMR (400 MHz, DMSO-de) δ: 9.02 (s, 1H), 8.87 (s, 1H), 8.51 (s, 1H), 8.37 (s, 1H), 8.20 (s, 1H),
8.16 (s, 1H), 7.45 (s, 1H), 3.92 (s, 3H), 3.65 - 3.59 (m, 1H), 3.57 (s, 2H), 3.26-3.16 (m, 2H), 2.88 (m, 2H).
13C NMR (101 MHz, DMSO-de) δ: 145.21, 142.50, 140.09, 137.15, 133.80, 131.39, 129.54, 129.24, 121.94, 121.03, 120.33, 117.69, 114.55, 99.94, 59.62, 39.28, 36.90, 27.42, 14.64.
LCMS m/z = 384.2 [MH]+
Example 20 (1 r,3r)-3-(Cyanomethyl)-3-(4-(6-(1-methyl-1 H-py razol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-yi)cyclobiJtane-1-carbonitrile
To a solution of (1r,3r)-3-(cyanomethyl)-3-(4-(4,4,5.5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol1-yl)cyclobutane-1-carbonitrile (Préparation 91, 3.38 g, 10.8 mmol) and 4-chloro-6-(1-methyl-1Hpyrazol-4-yl)pyrazolo[1,5-a]pyrazine (Préparation 11, 2.20 g, 9.4 mmol) in 1,4-dioxane (47.1 mL) was added 2 M aq. K3PO4 (14.1 mL). Nitrogen was bubbled through the mixture for about 5 min at about 25 °C, then XPhos Pd G2 (37.0 mg, 0.047 mmol) was added. The mixture was heated at about 40 °C for about 18 h, then it was heated to about 80 °C to dissolve the precipitated product. The aqueous layer was removed while maintaining the température at about 80 °C, then the 1,4-dioxane phase was added to EtOH (471 mL, previously preheated to about 50 °C). The mixture was stirred for about 10 min at about 50 °C before being removed from heat. Stirring at about 25 °C was continued for about 6 h. The solid was filtered and washed with EtOH (2 x 25 mL), water ( 2 x 50 mL), and EtOH (2 x 25 mL). The precipitate was dried under vacuum to afford the title compound as a white solid (3.61 g, 74%).
The title compound (500 mg, 1.30 mmol) was heated in 1,4-dioxane (6.5 mL) at about 80 °C until ail of the material was dissolved. 1,2-Bis(diphenylphosphino)ethane (7.8 mg, 0.019 mmol) was added and heating at about 80 °C was continued for about 4 hrs, then the 1,4-dioxane phase was added to EtOH (58.7 mL, previousiy preheated to about 50 °C). An additional 6.5 mL of preheated EtOH was used to rinse the reaction vessel. The mixture was removed from heat. Stirring at about 25 °C was continued for about 18 h. The solid was filtered and washed with EtOH (2x5 mL), water (5 mL), then EtOH (3x5 mL). The precipitate was dried under vacuum to afford the title compound as a white solid (450 mg, 90%).
Melting point 213 - 215 °C 1H NMR (400 MHz, DMSO-de) δ: 9.02 (s, 1 H) 8.92 (s, 1 H) 8.52 (s, 1 H) 8.37 (s, 1 H) 8.19 (s, 1 H)
8.16 (s, 1 H) 7.45 (s, 1 H) 3.91 (s, 3 H) 3.55 - 3.65 (m, 1 H) 3.52 (s, 2 H) 3.22 - 3.38 (m, 2 H) 2.85 3.00 (m, 2 H).
13C NMR (126 MHz, DMSO-de) δ: 145.17, 142.46, 140.20, 137.13, 133.78, 131.38, 129.82, 129.51, 122.28, 121.13, 120.33, 117.30, 114.54, 99.91,61.51, 39.26, 36.26, 29.65, 16.05.
LCMS m/z = 384.1 [MH]4.
Préparation 60 (1r,3r)-3-(Cyanomethyl)-3-(3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol1 -yl)cyclobutane-1 -carbonitrile (trans isomer)
and (1 s,3s)-3-(Cyanomethyl)-3-(3-methyl-4-(4,4,5,546^3171611^1-1,3,2-01(^36010130-2^1)-11-1^^2011-yi)cyclobutane-1-carbonitrile (cis isomer)
To a solution of 3-methyl-4-(4.4l5l5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (600 mg, 2.88 mmol) and 3-(cyanomethylene)cyclobutane-1-carbonitrile (Préparation 27, 341 mg, 2.88 mmol) in MeCN (28.8 mL) was added DBU (439 mg, 2.88 mmol) at about 20 °C. After about 18 hrs at about 20 °C, the mixture was poured into EtOAc and 10% aq. K2HPO4. The EtOAc was separated and the aqueous phase was extracted twice more with EtOAc. The combined EtOAc extracts were washed with brine, dried (Na2SÛ4) and concentrated. The residue was purified by column chromatography to afford (1 r,3r)-3-(cyanomethyl)-3-(3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yi)-1 H-pyrazol1-yl)cyclobutane-1-carbonitrile (trans isomer, 325 mg, 35%) ’H NMR (400 MHz, CDCh) δ: 7.79 (s, 1 H), 3.17 - 3.28 (m, 3 H), 3.16 (s, 2 H), 2.81 - 2.89 (m, 2 H), 2.39 (s, 3 H), 1.32 (s, 12 H).
LCMS m/z = 327.2 [MH]* and (1 s,3s)-3-(cyanomethyl)-3-(3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1 yl)cyclobutane-1-carbonitrile (cis isomer, 171 mg, 18%).
’H NMR (400 MHz, CDCh) δ: 7.75 (s, 1 H), 3.18 - 3.28 (m, 1 H), 3.08 - 3.18 (m, 2 H), 3.05 (s, 2 H), 2.93 - 3.02 (m, 2 H), 2.39 (s, 3 H), 1.32 (s. 12 H).
LCMS m/z = 327.2 [MH]*
Example 21 ( 1 r, 3 r)-3-(Cy anomethy l)-3-(3-methy 1-4-(6-(1 -methy 1-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile
To a solution of (1r,3r)-3-(cyanomethyl)-3-(3-methyl-4-(4,4.5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (Préparation 60, trans isomer, 209 mg, 0.64 mmol) in 1,4dioxane (4.3 mL) was added 4-chloro-6-(1-methyl-1H-pyrazol-4-yi)pyrazolo[1,5-a]pyrazine (Préparation 11, 150 mg, 0.64 mmol) and 2 M aq. KaPCU (0.96 mL). The mixture was purged with nitrogen for about 5 min, then XPhos Pd G2 (101 mg, 0.13 mmol) was added. The mixture was heated at about 40 °C for about 2 hrs, then concentrated. The residue was dissolved in EtOAc and the EtOAc was washed with water. The aqueous phase was extracted twice more with EtOAc, then once with DCM. The combined EtOAc and DCM extracts were dried (Na2SOi), concentrated, and the residue was purified by chromatography to afford a solid which was recrystallized from MeCN to afford the title compound (120 mg, 47%).
1H NMR (400 MHz, DMSO-de) δ: 9.00 (s, 1 H), 8.74 (s, 1 H), 8.26 (s, 1 H), 8.18 (s, 1 H), 8.08 (s, 1 H), 7.32 (s, 1 H), 3.91 (s, 3 H), 3.55 (quin, 1 H), 3.48 (s, 2 H), 3.22 - 3.30 (m, 2 H), 2.84 - 2.93 (m, 2 H), 2.63 (s, 3 H).
LCMS m/z = 398.3 [MH]*
Préparation 61 4-Bromo-1 -methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-1H-pyrazole
Q
Το a solution of 4-bromo-1-methyl-1H-pyrazole-3-methanol (720 mg, 3.77 mmol) in THF (30 mL) was added DHP (951 mg, 11.3 mmol) and PTSA (14 mg, 0.075 mmol). The solution was stirred at about 50 DC for about 16 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oil (1.0 g, 84%).
1H NMR (400 MHz. CDCh) δ: 7.37 (s, 1 H), 4.81 (t, 1 H), 4.74 (d, 1 H), 4.46 (d, 1 H), 3.96 - 4.03 (m, 1 H), 3.88 (s, 3 H), 3.56 - 3.63 (m, 1 H), 1.80- 1.93 (m. 1 H), 1.59- 1.78 (m, 3 H), 1.48- 1.57 (m, 2 H). LCMS m/z = 190.7 [MH-THP]‘
Préparation 62 (1s,3s)-3-(Cyanomethyl)-3-(4-(6-(1-methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-1H-pyrazol4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yi)cyclobutane-1-carbonitrile
Part 1
To a solution of 4-bromo-1-methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-1H-pyrazoie (Préparation 61, 200 mg, 0.73 mmol) in 1,4-dioxane (10 mL) were added KOAc (313 mg, 3.19 mmol), and bis(pinacolato)diboron (405 mg, 1.59 mmol). The mixture was purged with nitrogen for about 5 min before the addition of Pd(dppf)Ch (78 mg, 0.11 mmol). The mixture was heated at about 90 °C for about 18 hrs. The mixture was concentrated to afford an impure sample of 1-methyl-3(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole as a black oil (234 mg), which was used in Part 2 below without further purification.
Part 2
A mixture of (1r,3r)-3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 75, 85 mg, 0.25 mmol), 1-methyl-3(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-4-(4,4,5.5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Part 1, 81 mg, 0.25 mmol), and 2 M aq. K3PO4 (1.0 mL) in 1,4-dioxane (3.0 mL) was purged with argon for about 2 min, after which XPhos Pd G2 (39 mg, 0.050 mmol) was added. The reaction mixture was heated at about 45 °C for about 45 min. The 1,4-dioxane phase was separated from the aqueous phase, which was extracted further with EtOAc (5 mL). The combined 1,4-dioxane and EtOAc extracts were dried (NazSO.*) and concentrated. The residue which was purified by chromatography to afford the title compound (50 mg, 40%).
’H NMR (400 MHz, CDCh) δ: 8.92 (s, 1 H), 8.45 (s, 1 H), 8.33 (s. 1 H), 8.07 - 8.10 (m, 1 H), 8.00 (s, 1 H), 6.96 (d, 1 H), 5.02 (d. 1 H), 4.87 - 4.91 (m, 1 H), 4.70 (m. 1 H), 4.59 (d, 1 H), 4.51 (d, 1 H), 3.99 (s, 3 H), 3.51 - 3.62 (m, 2 H), 3.34-3.44 (m, 1 H). 3.29 (s, 2 H), 2.99 (m. 2 H), 1.80 - 1.96 (m, 3 H), 1.681.80 (m, 3 H).
LCMS m/z = 498.3 [MH]*
Example 22 (1r,3r)-3-(Cyanomethyl)-3-(4-(6-(3-(hydroxymethyl)-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yi)cyclobutane-1-carbonitrile
PTSA (10 mg, 0.052 mmol) was added to solution of (1s,3s)-3-(cyanomethyl)-3-(4-(6-(1-methyl-3(((tetrahydro-2H-pyran-2-yl)oxy)methyI)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1yl)cyclobutane-1-carbonitrile (Préparation 62, 50 mg, 0.10 mmol) in MeOH (10 mL). The reaction mixture was kept at about 20 °C for about 18 hrs. The precipitated solid was filtered to afford the title compound (20 mg, 48%).
1H NMR (400 MHz, DMSO-de) δ: 9.00 (s, 1 H), 8.91 (s, 1 H), 8.48 (s, 1 H), 8.35 (s, 1 H), 8.22 (d, 1 H), 7.47 (d, 1 H), 5.41 (br. s., 1 H), 4.66 (s, 2 H), 3.88 (s. 3 H), 3.54 - 3.62 (m, 1 H), 3.52 (s, 2 H), 3.28 3.34 (m, 2 H), 2.90 - 2.98 (m, 2 H).
LCMS m/z = 414.4 [MH]*
Préparation 63 (1r,3r)-3-(Cyanomethyl)-3-(4-(6-(1-methyl-3-nitro-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitriie
Part 1
To a solution of 4-bromo-1-methyl-3-nitro-1H-pyrazole (200 mg, 0.97 mmol) in 1,4-dioxane (10 mL) were added KOAc (285 mg, 2.90 mmol), and bis(pinacolato)diboron (368 mg, 1.45 mmol). The mixture was purged with nitrogen for about 5 min, after which Pd(dppf)Cï2 (70.8 mg, 0.097 mmol) was 10 added. The mixture was heated at about 90 °C for about 18 h. The mixture was concentrated to afford impure 1-methyl-3-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole as a black oil, which was used without further purification in Part 2.
Part 2
A mixture of (1r,3r)-3-(4-(6-chloropyrazo[o[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 75, 130 mg, 0.38 mmol), 1-methyl-3-nitro-4(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Part 1. 97 mg, 0.38 mmol), and 2 M aq. K3PO4 (2.0 mL) in 1,4-dioxane (6.0 mL) was purged with argon for about 2 min, aller which XPhos Pd G2 (60.6 mg, 0.077 mmol) was added. The mixture was heated at about 45 °C for about 45 min. The 20 1,4-dioxane phase was separated from the aqueous phase, which was extracted further with EtOAc (10 mL). The combined 1,4-dioxane and EtOAc extracts were dried (Na2SO4) and concentrated. The residue was purified by chromatography to afford the title compound (120 mg, 72%).
1H NMR (400 MHz, DMSO-ds) δ: 9.10 (s, 1 H), 8.90 (s, 1 H), 8.56 (s, 1 H), 8.43 (s, 1 H), 8.32 (d, 1 H), 7.56 (br. s., 1 H), 4.05 (s. 3 H), 3.54 - 3.60 (m, 1 H), 3.51 (s, 2 H), 3.18 - 3.29 (m, 2 H), 2.88 - 2.98 (m, 25 2 H).
LCMS m/Z = 429.4 [MH]*
Example 23 (1r,3r)-3-(4-(6-(3-Amino-1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1 -yl)-330 (cyanomethyl)cyclobutane-l -carbonitrile
SI
To a solution of (1r,3r)-3-(cyanomethyl)-3-(4-(6-(1-methyl-3-nitro-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazo!-1-yl)cyclobutane-1-carbonitrile (Préparation 63, 120 mg, 0.28 mmol) in EtOH (3 mL) and water (0.5 mL) were added NHîCI (90 mg, 1.68 mmol) and iron powder (50 mg, 0.90 mmol). The reaction was stirred at about 60 °C for about 18 hrs, after which additional portions of iron pwder and NH4CI were added. The mixture was heated at about 100 °C for about 3 hrs. The mixture was concentrated and hot EtOH (20 mL) was added. The undissolved solids were removed by filtration. The filtrate was concentrated and the residue was purified by chromatography to afford the title compound as a white solid (10 mg, 9%).
’H NMR (400 MHz, CD3OD) δ: 8.74 (s, 1 H), 8.67 (s, 1 H), 8.40 (s, 1 H), 8.08 (d, 1 H), 7.94 (s, 1 H), 7.24 (d, 1 H), 3.77 (s, 3 H), 3.47 - 3.55 (m, 1 H), 3.43 (s, 2 H), 3.34 - 3.41 (m, 2 H), 2.98 (dd, 2 H). LCMS m/z = 399.4 [MH]'
Préparation 64 3,5-Dibromo-1-methyl-1H-pyrazole
Br
A solution of 3,5-dibromopyrazole (6.5 g, 28.7 mmol) in THF (30 mL) was added dropwise to an icecooled suspension of NaH (2.88 g, 60% in minerai oil, 71.9 mmol) in THF (45 mL). The mixture was stirred at about 0 °C for about 1 hrs. lodomethane (5.37 mL, 86.3 mmol) was added and the mixture was stirred at about 0 °C for about 3 hrs, then stirring was continued at about 15 °C for about 2 hrs. The mixture was poured into saturated aq. NH-tCI (20 mL) and extracted with EtOAc (40 mL). The EtOAc extract was washed with brine (20 mL), dried (NaaSO^), and concentrated. The residue was purified by chromatography to afford the title compound as a colorless oil (5.5 g, 79%).
’H NMR (400 MHz, CDCfa) δ: 6.31 (s, 1 H), 3.87 (s, 3 H).
LCMS m/z = 240.6 [MH]' (79Br, 8’Br isotope)
Préparation 65 3-Bromo-1-methyl-1H-pyrazole-5-carboxylic acid
OH
Br
N-N
A solution of 3,5-dibromo-1-methyl-1H-pyrazole (Préparation 64, 3.0 g, 12.5 mmol) in THF (30 mL) was treated dropwise with n-BuLi (2.5 M, 6.25 mL, 15.6 mmol) at about -70 °C. After about 30 min at this température, a solution of CO2 in THF (30 mL) was added dropwise while maintaining the internai température below about -65 “C. The mixture was stirred at this température for about 1 hrs. The reaction mixture was then poured into 1 M aq. HCI (50 mL) and the mixture was partially concentrated to remove most of the THF. The aqueous layer was extracted with DCM (50 mL). The DCM extract was dried (NaaSCM and concentrated to afford the title compound as a yellow solid (2.1 g, 81%). ’HNMR (400 MHz, DMSO-de) S: 13.70 (br s, 1H), 6.95 (s, 1H), 4.04 (s, 3H).
LCMS m/z = 206.9 [MH]’ (B1Br isotope)
Préparation 66 tert-Butyl (3-bromo-1-methyl-1 H-pyrazol-5-yl)carbamate
Diphenylphosphoryl azide (18.8 g, 68.5 mmol) was added to a solution of 5-bromo-2-methyl-2Hpyrazole-3-carboxylic acid (Préparation 65, 7.02 g, 34.2 mmol) and DIPEA (11.9 mL, 68.5 mmol) in tbutanol (114 mL). The mixture was stirred at about 45 °C for about 30 min, then heated under reflux for about 5 hrs. The cooled mixture was diluted with EtOAc (60 mL) and washed with saturated aq. NaHCOa (2 x 30 mL) and brine (20 mL). The EtOAc extract was concentrated and the residue was purified by chromatography to afford the title compound as a yellow solid (4.80 g, 51%).
Ή NMR (400 MHz, DMSO-de) δ: 6.28 (br s, 1H), 6.18 (s, 1H), 3.73 (s, 3H), 1.50 (s, 9H).
LCMS m/z = 221.7 [MH-CoHe]* (81 Br isotope)
Préparation 67 3-Bromo-1 -methyl-5-(diBoc)-amino-1 H-pyrazole
Di-fert-butyî bicarbonate (1.58 g, 7.24 mmol) was added to a solution of tert-butyl (3-bromo-1-methyl1H-pyrazol-5-yl)carbamate (Préparation 66, 2.0 g, 7.24 mmol), TEA (4.04 mL, 29.0 mmol), and DMAP (177 mg, 1.45 mmol) in DCM (40 mL). The mixture was stirred at about 20 °C for about 18 hrs. Water (25 mL) was added and the mixture was extracted with DCM (2 x 30 mL). The combined DCM extracts were concentrated and the residue was purified chromatography to afford the title compound (1.85 g, 67)%.
1H NMR (400 MHz. CDCh) δ: 6.06 - 6.20 (m, 1 H), 3.64 (s, 3 H), 1.44 (s, 18 H).
LCMS m/z = 378.2 [MH]+ (81 Br isotope)
Example 24
2-((1 r,3s)-1 -(4-(6-(5-Amino-1 -methyl-1 H-pyrazol-3-yl)pyrazolo[1,5-a]py razin-4-y l)-1 H-pyrazol-1yl)-3-methoxycyclobutyl)acetonitrile
Part 1
A mixture of 3-bromo-1-methyl-5-(diBoc)-amino-1H-pyrazole (Préparation 67, 1200 mg, 3.19 mmol), KOAc (988 mg, 9.57 mmol) and bis(pinacolato)diboron (1210 mg, 4.78 mmol) in 1,4-dioxane (15 mL) was purged with argon for about 5 minutes before XPhos Pd G2 (502 mg, 0.64 mmol) was added. The reaction mixture was heated at about 65 °C for about 3.5 hrs, then 2-((1 s,3r)-1-(4-(6chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclo-butyl)acetonitrile (Préparation 100, 1090 mg, 3.19 mmol), 2 M aq. K3PO4 (4.78 mL) and XPhos Pd G2 (502 mg, 0.64 mmol) were added. The mixture was purged with argon again, then heated at about 80 °C for about 1 hrs. EtOAc was added and the phases were separated. The aqueous phase was extracted twice more with EtOAc and the combined EtOAc extracts were concentrated. The residue was purified by chromatography to afford a mixture of mono- and di-BOC intermediates (710 mg, 36%) which was used in Part 2.
Part 2
TFA (6 mL, 80 mmol) added to a solution of the mono- and di-BOC intermediates of Part 1 (710 mg, 1.18 mmol) in DCM (6 mL) at about 20 °C. Afler about 1 hrs, the mixture was concentrated. DCM was added, followed by sufficient saturated aq. NaHCOa to render the solution pH basic. The phases were separated and the aqueous phase was extracted twice more with DCM. The combined DCM extracts were dried (Na2SO4) and concentrated. An équivalent reaction using the compound of Part 1 (220 mg, 0.36 mmol) and TFA (2 mL, 30 mmol) in DCM (2 mL) was combined with the material from the reaction above and the combined samples were purified by chromatography to afford the title compound as a clear gum (470 mg, 99%).
’H NMR (400 MHz, DMSO-de) δ: 8.81 (s, 1 H), 8.73 (s, 1 H). 8.40 (s, 1 H), 8.21 (d, 1 H), 7.44 (d, 1 H), 5.33 (br. s, 2 H), 3.94 - 4.02 (m, 1 H), 3.62 (s, 3 H), 3.44 (s, 2 H), 3.21 (s, 3 H), 3.14 - 3.20 (m, 2 H), 2.39-2.45 (m,2H).
LCMS m/z = 404.5 [MH]*
Préparation 68
Diethyl 4-bromo-1 H-pyrazole-3,5-dicarboxylate
A mixture of 1H-pyrazole-3,5-dicarboxylic acid diethyl ester (4.0 g, 18.85 mmol) and Nbromosuccinimide (4.03 g, 22.6 mmol) in a mixture of conc. nitric acid and glacial acetic acid (12.0 mL, 5:95 v/v) was heated by microwave irradiation at about 120 °C for about 20 minutes. A total of total of 13.0 g (61.26 mmol) of 1H-pyrazole-3,5-dicarboxylic acid diethyl ester starting material was processed in this manner in parallel batches. The resulting brown crude reaction mixtures were combined, poured into water (260 mL), and treated with sufficient NaHCOa to render the solution pH basic. The mixture was extracted with EtOAc (3 x 300 mL). The combined EtOAc extracts were dried (NazSO4) and concentrated to afford the title compound as an off-white solid (17.0 g, 95%).
’H NMR (400 MHz, CDCh) δ: 8.79 (br. s., 1 H), 4.44 (q, 4 H), 1.42 (t, 6 H).
LCMS m/z = 290.7 [MH]* (79Br isotope)
Préparation 69
Diethyl 4-bromo-1-(2-(1-methyl·1H-pyrazol-4-yl)-2-oxoethyl)-1H-pyrazo[e-3,5-dicaΓboxylate
A solution of 4-bromo-1H-pyrazole-3,5-dicarboxylic acid diethyl ester (Préparation 68, 9.66 g, 33.18 mmol) and 2-bromo-1-(1-methyl-1H-pyrazol-4-yl)-ethanone (Préparation 6, 6.0 g, 29.55 mmol) in MeCN (140 mL) was treated with K2CO3 (5.31 g, 38.40 mmol) and the reaction mixture was stirred at about 25 °C for about 16 hrs. Water (100 mL) was added and the mixture was extracted with EtOAc (3 x 100 mL). The combined EtOAc extracts were dried (NaaSO^) and concentrated. The residue was purified by chromatography to afford the title compound as a white solid (10.0 g, 82%).
’HNMR (400 MHz. CDCI3) δ: 7.95 (s, 1 H), 7.92 (s. 1 H), 5.82 (s, 2 H), 4.43 (q. 2 H), 4.36 (q, 2 H), 3.96 (s, 3 H), 1.45 (t, 3 H), 1.35 (t, 3 H).
LCMS m/z = 436.8 [MNa]’’ (®1Br isotope)
Préparation 70
Diethyl 4-methyl-1-(2-(1 -methyl-1 H-pyrazoI-4-yl)-2-oxoethyl)-1H-pyrazole-3,5-dicarboxylate
A mixture of K2CO3 (11.0 g, 79.9 mmol) and 4-bromo-1 H-[2-(1-methyl-1 H-pyrazo!-4-yl)-2-oxo-ethyl]1H-pyrazole-3,5-dicarboxylic acid diethyl ester (Préparation 69, 11.0 g, 26.62 mmol) in DMF (133.0 mL) was purged with nitrogen at about 25 °C, after which Pd(dppf)Clj (1950 mg, 2.66 mmol) and trimethylboroxine (10.0 g, 79.9 mmol) were added and the solution was heated at about 110 °C for about 5 hrs. The cooled réaction was diluted with EtOAc (100 mL) and the resulting mixture was washed with brine (2 x 100 mL), dried (NasSOi), and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (5.45 g, 50%).
1H NMR (400 MHz, CDCh) δ: 7.92 (s, 1 H), 7.90 (s, 1 H), 5.78 (s, 2 H), 4.41 (q, 2 H), 4.28 (q, 2 H), 3.95 (s, 3 H), 2.58 (s, 3 H), 1.39 (t, 3 H), 1.31 (t, 3 H).
LCMS m/z = 348.9 [MH]’
Préparation 71
Ethyl 4-hydroxy-3-methyl-6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazine-2-carboxyiate
4-Methyl-[2-(1 -methyl-1 H-pyrazol-4-yl)-2-oxo-ethyl]-1 H-pyrazole-3,5-dicarboxylic acid diethyl ester (Préparation 70, 5.35 g, 15.36 mmol) was mixed with dry EtOH (15 mL) and concentrated. The resulting solid was dissolved in EtOH (40 mL) and NH«OAc (3.55 g, 46.1 mmol) was added. The reaction mixture was heated at about 130 °C in an autoclave for about 8 hrs. After cooling, the mixture was filtered and the precipitate was dried to afford the title compound as an off-white solid (3.40 g, 73%).
1H NMR (400 MHz, DMSO-de) δ: 11.45 (br. s., 1 H), 8.27 (s, 1 H), 8.08 (s, 1 H), 8.02 (s, 1 H), 4.31 (m, 2 H), 3.87 (s, 3 H), 2.61 (s, 3 H), 1.24 - 1.40 (m, 3 H).
LCMS m/z = 301.8 [MH]*
Préparation 72
4-Hydroxy-3-methyl-6-(1-methyl-1H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazine-2-carboxy{ic acid
OH
Lithium hydroxide monohydrate (1.42 g, 33.9 mmol) was added to a suspension of 4-hydroxy-3methyl-6-(1-methyl-1H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazine-2-carboxylic acid ethyl ester (Préparation 71, 3.40 g, 11.3 mmol) in THF (50 mL), MeOH (50 mL) and water (25 mL). The mixture was stirred at about 60 °C for about 4 hrs. The cooled mixture was concentrated and water (50 mL) was added. The pH of the mixture was adjusted to about 2 by the addition of 12 M aq. HCl. The resulting precipitate was filtered and washed with water (50 mL), then dried under vacuum to afford the title compound as a solid.
1HNMR (400 MHz. DMSO-ds) δ: 12.90 - 13.20 (br. s, 1 H), 11.40 (s, 1 H), 8.30 (s, 1 H), 8.02 (m, 2 H), 3.90 (S, 3 H), 2.60 (s, 3 H).
LCMS m/z = 301.8 [MH]*, 323.8 [MNa]*
Préparation 73
3-Methyl-6-(1-methyl-1 H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazin-4-ol
OH
4-Hydroxy-3-methyl-6-( 1 -methyl-1 H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazine-2-carboxylic acid (Préparation 72, 2.58 g, 9.44 mmol) was added in portions to preheated sulfolane (18.9 mL) at about 280 °C. Once the addition was complété, the mixture was stirred for an additional 1 hrs at about 280 °C. The cooled mixture was purified directly by chromatography on silica gel (eluting with petroleum ether:EtOAc (100:0 to 50:50), then DCMiMeOH (91:9)) to afford the title compound as a yellow solid (1.50 g, 69%).
’HNMR (400 MHz, DMSO-de) δ: 11.20 (s, 1 H), 8.38 (s, 1 H), 8.02 (s, 1 H), 7.96 (s, 1 H), 7.70 (s, 1 H), 3.85 (s, 3 H), 2.40 (s, 3 H).
Préparation 74
4-Chloro-3-methyl-6-(1 -methyl-1 H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazine
Cl
To a suspension of 3-methyl-6-(1-methyl-1H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazin-4-ol (Préparation 73, 1.40 g, 6.11 mmol) in MeCN (60.0 mL) was added POCb (4.68 g, 30.5 mmol). The reaction mixture was heated at about 80 °C for about 16 hrs. After cooling, the mixture was poured into water (200 mL) at about 25 °C. The mixture was adjusted to about pH 9 by the addition of saturated aq. NaHCCb (200 mL), then extracted with EtOAc (5 x 100 mL). The combined EtOAc extracts were dried (Na2SO4) and concentrated. The residue was purified by chromatography, then further purified by HPLC to afford the title compound as a white solid (163 mg, 11%).
’H NMR (400 MHz, CDCb) δ: 8.37 (s, 1 H), 7.89 (s, 1 H), 7.85 (s, 1 H), 7.80 (s. 1 H), 3.96 (s, 3 H), 2.57 (s. 3 H). LCMS m/z = 247.7 [MH]* (35CI isotope)
Example 25 (1r,3r)-3-(Cyanomethyl)-3-(4-(3-methyl-6-(1-methyl-1 H-py razol-4-yl)pyrazo!o[1,5-a]pyrazin-4-yl)1H-pyrazol-1 -yt)cyclobutane-1-carbonitrile
To a solution of 4-chloro-3-methyl-6-(1-methyl-1H-pyrazol-4-yl)-pyrazolo[1,5-a]pyrazine (Préparation 74, 98 mg, 0.40 mmol) and (1r,3r)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (Préparation 91, 247 mg, 0.79 mmol) in 1,4-dioxane (9 mL) was added 2 M aq. K3PO4 (0.59 mL) at about 25 °C and the mixture was purged with nitrogen for about 2 min before XPhos Pd G2 (62.3 mg, 0.079 mmol) was added. The mixture was purged with nitrogen for about 3 min, then heated at about 80 °C for about 16 hrs. The mixture was fiitered and concentrated. The residue was purified by chromatography, then further purified by HPLC to afford the title compound as a light yellow solid (56 mg, 36%).
1H NMR (400 MHz, DMSO-de): δ: 8.98 (s, 1 H), 8.94 (s, 1 H), 8.60 (s, 1 H), 8.27 (s, 1 H), 8.22 (s, 1 H), 8.10 (s. 1 H), 7.98 (s, 1 H). 3.90 (s, 3 H), 3.52 - 3.57 (m, 3 H), 3.19 - 3.26 (m, 2 H), 2.91 - 2.97 (m, 2 H), 3.06 (s, 3 H).
LCMS m/z = 398.0 [MH]*
Préparation 75 (1 r,3r)-3-(4-(6-Chloropyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -yl)-3-(cyanomethy l)cyclobutane1-carbonitrile
A mixture of 4,6-dichloropyrazolo[1,5-a]pyrazine (Préparation 4, 350 mg, 1.86 mmol), (1r,3r)-3(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)cyclobutane-1carbonitrile, (Préparation 91, trans isomer, 581 mg, 1.86 mmol), and 2 M aq. K3PO4 (2.79 mL) in 1,4dioxane (10 mL) was purged with argon for about 5 min, after which bis(tri-t18860 butylphosphine)palladium(O) (48.0 mg, 0.093 mmol) was added. The mixture was kept at about 25 °C for about 2 hrs then filtered. The precipitate was washed with EtïO and dried. The filtrate was concentrated, triturated with EbO, filtered, washed with EbO and dried. The two précipitâtes were combined to afford the title compound as a white solid (605 mg, 96%).
Ή NMR (400 MHz, DMSO-de) 6: 9.05 (d, 1 H), 8.93 (s, 1 H), 8.46 (s, 1 H), 8.32 (d, 1 H), 7.61 (d, 1 H), 3.55 - 3.62 (m, 1 H), 3.54 (s, 2 H), 3.24 - 3.32 (m, 2 H), 2.89 - 3.00 (m, 2 H).
LCMS m/z = 338.2 [MH]*
Example 26 (1r,3r)-3-(Cyanomethyl)-3-(4-(6-(5-methyl-1 H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-yl)cyclobutane-1-carbonitrile
Part 1
A mixture of (1r,3r)-3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yi)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 75, 85 mg, 0.25 mmol), 3-methyl-1-(tetrahydro2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Préparation 93, 73 mg, 0.25 mmol), and 2 M aq. K3PO4 (1.0 mL) in 1,4-dioxane (3.0 mL) was purged with argon for about 2 min, after which XPhos Pd G2 (39.6 mg, 0.050 mmol) was added. The mixture was heated at about 45 °C for about 45 min before being cooled and concentrated. The residue was purified by chromatography to afford (1 r,3r)-3-(cyanomethyl)-3-(4-(6-(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1 Hpyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane’1-carbonitrile (26 mg, 22%), which was used directly in Part 2.
LCMS m/z = 384.3 [MH-THP]*
Part 2
TFA (1 mL) was added to a solution of (1r,3r)-3-(cyanomethyl)-3-(4-(6-(5-methyl-1-(tetrahydro-2Hpyran-2-yl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazof-1-yl)cyclobutane-1-carbonitrile (26 mg, 0.055 mmol) in DCM (2 mL). The reaction mixture was heated to about 50 °C for about 1 hrs. The mixture was concentrated and the residue was concentrated twice with toluene (5 mL each time). The residue was then purified by chromatography to afford a solid, which was further triturated with ether to afford the title compound (8 mg, 38%).
Ή NMR (400 MHz, CD3CN) δ: 8,87 (s, 1 H), 8.66 (s, 1 H), 8.48 (s, 1 H), 8.15 (d, 1 H), 7.23 (s, 1 H), 6.73 (s, 1 H), 3.46 (dd, 1 H). 3.36 (s, 2 H), 3.32 - 3.41 (m, 2 H), 2.98 (dd, 2 H), 2.37 (s, 3 H).
LCMS m/z = 384.4 [MH]4
Example 27 2-((1st3r)-1-(4-(6-(5-(Hydroxymethyl)-1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yi)-1H-pyrazol-1yl)-3-methoxycyclobutyl)acetonitrile
A solution of 2-((1s,3r)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile (Préparation 82, 100 mg, 0.29 mmol), and (3-(tributylstannyl)-1Hpyrazol-5-yl)methanol (Préparation 32, 113 mg, 0.29 mmol) in 1,4-dioxane (2 mL) was purged with argon for 5 min, followed by the addition of XPhos Pd G2 (45.9 mg, 0.058 mmol). The mixture was heated at about 80 °C for about 18 hrs. The resulting precipitate was filtered, washed with 1,4dioxane and EtîO, and dried to afford the title compound as a white solid (62 mg, 53%).
1H NMR (400 MHz. CDaCN) δ: 8.86 (s, 1 H), 8.58 (s, 1 H), 8.44 (s, 1 H), 8.13 (d, 1 H), 7.21 (d, 1 H), 6.85 (s, 1 H), 4.65 (d, 2 H), 4.05 (quin, 1 H), 3.27 (s, 3 H), 3.21 (s, 2 H), 2.90 - 2.99 (m, 2 H), 2.67 2.76 (m, 2 H).
LCMS m/z = 405.3 [MH]4
Example 28
2-((1 r,3s)-1-(4-(6-(5-(Hydroxymethyl)-1 H-pyrazoi-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1yl)-3-methoxycyclobutyl)acetonitrile
A solution of 2-((1 r,3s)-1 -(4-(6-chtoropyrazolo[1,5-a]pyrazin-4-y[)-1 H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile (Préparation 39; 1000 mg, 2.92 mmol) and (3-(tributylstannyl)-1 Hpyrazol-5-yl)methanol (Préparation 32, 1130 mg, 2.92 mmol) in 1,4-dioxane (2 mL) was purged with argon for 5 min, followed by the addition of XPhos Pd G2 (45.9 mg, 0.058 mmol). The mixture was heated at about 80 °C for about 18 hrs. The precipitate was fîitered and then purified by chromatography to afford an off white solid. The solid was heated in sufficient boiling EtOH until it was entirely dissolved, and then stirred for about 18 hrs at about 20 °C. The precipitate was fîitered and dried under vacuum to afford the title compound as a crystalline solid (507 mg, 43%).
’H NMR (400 MHz, CD3CN) δ: 11.48 (br. s.), 8.89 (s), 8.65 (s). 8.46 (s), 8.15 (d), 7.23 (s), 6.87 (s), 4.67 (d). 4.08 (quin), 3.31 (s), 3.30 (s), 3.15 - 3.25 (m), 2.53 (dd).
LCMS m/z = 405.3 [MH]'
Préparation 76
Ethyl 5-(tributylstannyl)isoxazole-3-carboxylate
TEA (0.69 mL, 4.95 mmol) was added dropwise to an ice-cold solution of ethyl 2-chloro-2(hydroxyimino)acetate (500 mg, 3.30 mmol) and ethynyltributylstannane (1040 mg, 3.30 mmol) in EUO (10 mL). The reaction mixture was warmed to about 20 °C and kept for about 18 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oil (810 mg, 57%).
’H NMR (400 MHz, CDCh) δ; 6.80 (s, 1 H), 6.77 - 6.84 (m, 1 H), 4.45 (q, 2 H), 1.51 - 1.63 (m, 6 H), 1.43 (t, 3 H), 1.34 (dq, 6 H), 1.15-1.26 (m, 6 H), 0.90 (t, 9 H).
LCMS m/z = 454.2 [MNa]' (12QSn isotope)
Préparation 77 Ethyî 5-(4-(1-((1 r,3r)-3-cyano-1-(cyanomethyl)cyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-6-yl)isoxazole-3-carboxylate
A solution of (1r,3r)-3-(4-(6-chioropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 75, 110 mg, 0.33 mmol) and ethy! 5(tributylstannyl)isoxazole-3-carboxylate (Préparation 76, 140 mg, 0.33 mmol) in 1,4-dioxane (3 mL) was purged with argon for about 5 min, followed by the addition of XPhos Pd G2 (51.2 mg, 0.065 mmol). The mixture was heated at about 100 °C for about 5 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a light yellow solid (66 mg, 45%).
1H NMR (400 MHz, DMSO-ds) B: 9.44 (s, 1 H), 9.02 (s, 1 H), 8.60 (s, 1 H), 8.42 (d, 1 H), 7.65 (d, 1 H), 7.62 (s, 1 H), 4.44 (q, 2 H), 3.54 - 3.63 (m, 1 H), 3.53 (s. 2 H), 3.26 - 3.36 (m, 2 H), 2.90 - 2.98 (m, 2 H), 1.37 (t, 3 H).
LCMS m/z = 443.3 [MH]*
Example 29
5-(4-(1-((1 r,3r)-3-Cyano-1-(cyanomethyl)cyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6yl)isoxazole-3-carboxamide
A solution of ethyl 5-(4-(1 -((1r,3r)-3-cyano-1-(cyanomethyl)cyclobutyi)-1H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-6-yl)isoxazole-3-carboxylate (Préparation 77, 54 mg, 0.12 mmol) in methanol (3 mL) was treated with a 7 M solution of ammonia gas in methanol (2 mL, 14 mmol). The reaction vessel was tightly sealed and the mixture was heated at about 95 °C for about 18 hrs. After cooling, the precipitate was filtered, washed with EtOAc followed by EtîO, and dried to afford the title compound as a white solid (45 mg, 89%).
Ή NMR (400 MHz, DMSO-de) δ: 9.36 (s, 1 H), 9.01 (s, 1 H), 8.58 (s, 1 H), 8.41 (d, 1 H), 8.21 (br. s, 1 H), 7.91 (br. s, 1 H), 7.64 (d, 1 H), 7.50 (s, 1 H). 3.55 - 3.62 (m, 1 H), 3.53 (s, 2 H), 3.26 - 3.35 (m, 2 H), 2.90 - 2.98 (m, 2 H).
LCMSm/z = 414.3 [MH]*
Préparation 78
2-((1r,3s)-1-(4-(6-(5-(DiBoc)-amîno-1-(tetrahydro-2H-pyran-2-yl)-pyrazoi-3-yl)pyrazoto[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitnle
A mixture of 3-bromo-1-(tetrahydro-2H-pyran-2-yl)-5-(diBoc)-amino-1H-pyrazole (Préparation 43, 233 mg, 0.52 mmol), KOAc (162 mg, 1.57 mmol), and bis(pinacolato)diboron (199 mg, 0.78 mmol) in 1,4dioxane (4 mL) was purged with argon for about 5 min, after which XPhos Pd G2 (82.1 mg, 0.10 mmol) was added. The mixture was heated at about 65 °C for about 3.5 hrs. After cooling to about 20 °C, 2-((1 r,3s)-1 -(4-(6-ch!oropyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)-3-methoxycyclo-butyl)acetonitrile (Préparation 39, 125 mg, 0.36 mmol), 2 M aq. K3PO4 (0.783 mL) and XPhos Pd G2 (82.1 mg, 0.10 mmol) were added. The mixture was again purged with argon, then heated at about 80 °C for about 1 hrs. After cooling, the mixture was diluted with EtOAc and the phases were separated. The aqueous phase was extracted twice with EtOAc and the combined EtOAc extracts were concentrated. The residue was purified by chromatography to afford the title compound as a clear oil (220 mg, 62%).
1H NMR (400 MHz, CDCh) δ: 9.08 (s, 1 H), 8.39 (s, 1 H), 8.33 (s, 1 H), 8.09 (d, 1 H). 6.97 (d, 1 H), 6.90 (s, 1 H). 5.26 (dd, 1 H), 4.03-4.11 (m, 2 H), 3.64 (t, 1 H), 3.34 (s, 3 H), 3.25 (s, 2 H), 3.11 -3.19 (m, 2 H), 2.55-2.63 (m, 2 H), 2.21 (m, 1 H). 1.93 (dd, 1 H), 1.59 -1.84 (m, 4 H), 1.45 (s, 18 H).
LCMS m/z = 674.5 [MH]*
Example 30 2-((1r,3s)-1-(4-(6-(3-Amino-1H-pyrazol-5-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile
O
TFA (3 mL) was added to a solution of 2-((1 r,3s)-1-(4-(6-(5-(diBoc)-amino-1-(tetrahydro-2H-pyran-2yl)-pyrazo!-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yi)-3-methoxycyclobutyl)acetonitrile (Préparation 78, 220 mg, 0.33 mmol) in DCM (2 mL) at about 20 °C. After about 1 hrs at this température, the mixture was concentrated and the residue was dissoived in DCM and saturated aq. NaHCCh, ensuring that the pH was basic. The DCM was separated and the aqueous phase was extracted twice with DCM. The combined DCM extracts were dried (NazSO4), concentrated, and the residue was purified by chromatography followed by HPLC to afford the title compound as a white solid (17 mg, 13%).
1H NMR (400 MHz, CD3OD) δ: 8.89 (s, 1 H), 8.84 (s, 1 H), 8.54 (s, 1 H), 8.17 - 8.18 (m, 1 H), 7.34 (s, 1 H), 6.22 (s, 1 H), 4.09 - 4.12 (m, 1 H), 3.39 (s, 3 H), 3.22 - 3.27 (m, 2 H), 2.56 - 2.60 (m, 2 H).
LCMS m/z = 390.3 [MH]*
Préparation 79 (4-Bromo-l -methyl-1 H-pyrazol-3-yl)methanol
Sodium borohydride (3.4 g, 90 mmol) was added to a solution of ethyl 4-bromo-1-methyl-1 H-pyrazole-
3-carboxylate (4.2 g, 18 mmol) in anhydrous EtOH (150 mL) at about 5 °C. The mixture was then heated at about 50 °C for about 16 h, after which EtOAc (100 mL) and water (100 mL) were added. The EtOAc was separated, dried (NaîSOi), and concentrated. The residue was purified by chromatography to afford the title compound as a white solid (1.05 g, 31%).
1H NMR (400 MHz, DMSO-de) δ: 7.84 (s, 1 H), 5.01 (td, 1 H), 4.33 (d, 2 H), 3.77 (s, 3 H).
LCMS m/z = 192.7 [MH]+ (81Br isotope)
Préparation 80
4-Bromo-1 -methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)-1 H-pyrazole
To a solution of (4-bromo-1-methyl-1H-pyrazol-3-yl)methanol (Préparation 79, 1.00 mg, 5.23 mmol) in THF (30 mL) were added DHP (1.32 g, 15.7 mmol) and PTSA (19.9 mg, 0.10 mmol). The solution was heated at about 50 °C for about 16 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oil (1.02 g, 71%).
’H NMR (400 MHz, CDCh) δ: 7.36 (s, 1 H), 4.80 (t, 1 H), 4.73 (d, 1 H). 4.45 (d, 1 H), 3.95 - 4.03 (m, 1 H), 3.87 (s, 3 H), 3.53 - 3.64 (m, 1 H), 1.78- 1.93 (m, 1 H), 1.60- 1.77 (m, 3 H), 1.45- 1.60 (m, 2 H).
LCMS m/z = 174.6 [MH-THP]‘
Préparation 81 1-Methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyi)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)-1 H-pyrazole
To a solution of 4-bromo-1-methyl-3-(((tetrahydro-2H-pyran-2-yl)oxy)methyi)-1 H-pyrazole (Préparation 80, 330 mg, 1.20 mmol) in DMF (12 mL) were added bis(pinacolato)diboron (457 mg,
1.8 mmol) and KOAc (353 mg, 3.6 mmol). The mixture was purged with nitrogen for about 2 min, after which Pd(dppf)Cb (87.8 mg, 0.12 mmol) was added. The mixture was heated at about 90 °C for about 16 hrs. The cooled mixture was filtered through a pad of Celite® and the filter was washed with methanol (15 mL). The filtrate was concentrated to afford the impure title compound as dark gum (1.17 g), which was used in Example 31 next step without further purification.
’H NMR (400 MHz, CDCIa) δ: 7.61 (s. 1 H), 4.75 - 4.77 (m, 1 H), 4.50 - 4.57 (m, 2 H), 4.03 - 4.10 (m, 1 H), 3.91 - 3.98 (m, 1 H), 3.89 (s, 3 H). 1.81 - 1.90 (m, 1 H), 1.61 - 1.76 (m, 3 H), 1.49- 1.56 (m, 2 H), 1.30 (s, 12 H).
LCMS m/z = 239.1 [MH-THP]*
Example 31
2-((1 r,3s)-1-(4-(6-(3-(Hydroxymethyl)-1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yt)-1Hpy razol-1 -y l)-3-methoxycyclobutyl)aceton itrile
Part 1
A mixture of 2-((1 r,3s)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 39, 80 mg, 0.23 mmol), 1-methyl-3-(((tetrahydro-2H-pyran-2yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Préparation 81, 376 mg, 1.17 mmol), and 2 M aq. K3PO4 (0.35 mL) in 1,4-dioxane (2.5 mL) was treated with XPhos Pd G2 (18.4 mg, 0.023 mmol) and the mixture was purged with nitrogen. The mixture was heated at about 60 °C for about 20 hrs. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a yellow gum. This sample was combined with the product from an équivalent reaction conducted using 2-((1 r,3s)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 39, 120 mg, 0.35 mmol), 1-methyl-3-(((tetrahydro2H-pyran-2-yl)oxy)methyt)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Préparation 81, 564 mg, 1.75 mmol), 2 M aq. K3PO4 (0.525 mL), and XPhos Pd G2 (13.8 mg, 0.023 mmol) in 1,4dioxane (3.5 mL) to afford 2-((1r,3s)-3-methoxy-1-(4-(6-(1-methyl-3-(((tetrahydro-2H-pyran-2yl)oxy)methyl)-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yi)-1H-pyrazol-1-yl)cyclobutyl)acetonitrile in a total yield of 150 mg (51%), which was used in Part 2 without further purification.
Part 2
TFA (1 mL) was added to an ice-cooled solution of 2-((1r,3s)-3-methoxy-1-(4-(6-(1-methyl-3(((tetrahydro-2H-pyran-2-yi)oxy)methyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 yl)cyclobutyl)acetonitrile (Part 1, 150 mg, 0.30 mmol) in DCM (3 mL). The mixture was stirred with cooling in ice for about 2 hrs, then concentrated. The residue was purified by HPLC to afford the title compound as a light yellow solid (53 mg, 38%).
1H NMR (400 MHz, CD3OD) Ô: 8.87 (s, 1 H), 8.69 (s, 1 H), 8.40 (s, 1 H), 8.21 (s, 1 H), 8.13 (d, 1 H), 7.28 (d, 1 H), 4.83 (s, 2 H), 4.05 - 4.14 (m, 1 H), 3.96 (s, 3 H), 3.37 (s, 2 H), 3.34 (s, 3 H), 3.18 - 3.27 (m, 2H), 2.53-2.62 (m, 2 H).
LCMS m/z = 419.1 (MH]+
Préparation 82 2-((1s,3r)-1-(4-(6-Chloropyrazolo[1,5-a]pyrazin-4-yI)-1 H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile
Part 1
To a solution of 2-((1s,3r)-1-(4-bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 37, cis isomer, 400 mg, 1.48 mmol) in toluene (9 mL) were added bis(pinacolato)diboron (564 mg, 2.22 mmol) and KOAc (436 mg, 4.44 mmol). The mixture was purged with nitrogen for about 3 min, after which Pd(dppf)Cl2 (108 mg, 0.15 mmol) was added. The mixture was again purged with nitrogen for about 3 min, then heated at about 110 °C for about 16 hrs. The cooled mixture was concentrated and the residue was purified by chromatography to afford impure 2-((1 s,3r)-3-methoxy-1-(4-(4,4,5,5tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1 -yl)cyclobutyl)acetonitri!e as light-yellow oil (500 mg), which was used in Part 2.
Part 2
A solution of 2-((1 s,3r)-3-methoxy-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1yl)cyclobutyl)acetonitrile (Part 1, 470 mg, 1.48 mmol) and 4,6-dichloropyrazolo[1,5-a]pyrazine (Préparation 4. 279 mg, 1,48 mmol), and 2 M aq. KzCCh (2.22 mL) in 1,4-dioxane (10 mL) was purged with nitrogen for about 2 min, after which Pd(dppf)Cl2 (108 mg, 0.15 mmol) was added. The mixture was again purged with nitrogen for about 3 min, then heated at about 90 °C for about 16 hrs.
The cooled mixture was concentrated and the residue was purified by chromatography to afford the title compound as a yellow solid (130 mg, 26%).
Ή NMR (400 MHz, CDCh) δ: 8.40 (s, 1 H), 8.35 (s, 1 H), 8.27 (s, 1 H), 8.08 (d, 1 H), 7.00 (s, 1 H), 4.05 (quin, 1 H). 3.30 (s, 3 H), 3.10 (s, 2 H), 3.00 (m, 2 H), 2.75 (m, 2 H).
LCMS m/z = 343.2 [MH]* (3SCI isotope)
Préparation 83
Ethyl 5-(4-(1-((1 s,3r)-1-(cyanomethyl)-3-methoxycyclobuty!)-1H-pyrazol~4-yl)pyrazolo[1,5a]pyrazîn-6-yl)-1H-pyrazole-3-carboxylate
A solution of 2-((1s,3r)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile (Préparation 82, 120 mg, 0.35 mmol) and ethyl 5-(tributylstannyl)-1Hpyrazole-3-carboxylate (Préparation 31, 150 mg, 0.35 mmol) in 1,4-dioxane (5 mL) was treated with XPhos Pd G2 (13.8 mg, 0.017 mmol) and the mixture was purged with nitrogen for about 2 min. The mixture was heated at about 110 °C for about 2 hrs. The cooled mixture was concentrated and the residue was purified by chromatography to afford the title compound as a yellow solid (180 mg).
’H NMR (400 MHz, CDCh) ô: 9.05 (br. s., 1 H), 8.39 (s, 1 H), 8.32 (s, 1 H), 8.14 (s, 1 H), 7.65 (s, 1 H), 7.02 (s, 1 H), 6.88 (s, 1 H), 4.44 - 4.50 (m, 2 H), 4.08 (t, 1 H), 3.80 (s, 3 H), 3.13 (s, 2 H), 3.00 - 3.08 (m, 2 H), 2.75 - 2.83 (m, 2 H), 1.46 (t, 3 H).
LCMS m/z = 447.1 [MH]*
Example 32
5-(4-(1-((1s,3r)-1-(Cyanomethyl)-3-methoxycyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6yl)-1 H-pyrazole-3-carboxamide
A solution of ethyl 5-(4-(1-((1 s,3r)-1-(cyanomethyl)-3-methoxycyclobutyl)-1 H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-6-yl)-1H-pyrazole-3-carboxylate (Préparation 83, 100 mg, 0.22 mmol) in MeOH (2 mL) was treated with a 4 M solution of ammonia gas in methanol (1 mL). The reaction vessel was tightly sealed and the mixture was heated at about 60 °C for about 16 hrs. The mixture was concentrated and the residue was purified by HPLC to afford the title compound (23 mg, 25%). 1H NMR (400 MHz, DMSO-de) 5: 13.79 (br. s., 1 H), 9.31 (br. s., 1 H), 9.00 (br. s., 1 H), 8.67 (br. s., 1 H), 8.28 (d, 1 H), 7.56 (br. s., 2 H), 7.39 (br. s., 1 H), 7.25 - 7.35 (m, 1 H), 4.08 (quin, 1 H), 3.44 (s, 2 H), 3.21 (s, 3 H), 2.79 - 2.90 (m, 2 H), 2.63 - 2.73 (m, 3 H).
LCMS m/z = 440.1 [MNa]*
Préparation 84
2-(Tetrahydro-2H-pyran-2-yl)-2H-1,2,3-triazole
To a mixture of 1H-1,2,3-triazole (15 g, 220 mmol) in DCM (724 mL) were added DHP (21.9 g, 261 mmol) and PTSA (0.374 g, 2.17 mmol). The mixture was kept at about 25 °C for about 18 hrs, after which NaOH (96 mg, 2.39 mmol) was added. The mixture was stirred at about 25 °C for about 1 hrs then filtered. The filtrate was concentrated and the residue was purified by chromatography to afford the title compound as a colorless oil (18 g, 54%).
’H NMR (400 MHz, CDCh) δ: 7.68 (s, 2 H), 5.74 (dd, 1 H), 4.00 - 4.08 (m, 1 H), 3.69 - 3.81 (m, 1 H), 2.36 - 2.51 (m, 1 H), 2.02 - 2.20 (m, 2 H), 1.61 -1.81 (m, 3 H).
Préparation 85 2-(Tetrahydro-2H-pyran-2-yl)-4-(4,4,515-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-1,2,3-triazole ιοο
Το a solution of 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (24 g, 129 mmol) in pentane (200 mL) were added 4,4'-di-tert-butyl-2,2’-bipyridine (0.315 g, 1.18 mmol) and (1,5cyclooctadiene)(methoxy)iridium(l) dimer (234 mg, 0.35 mmol). The solution rapidly became a red colon and gas évolution was observed. After about 15 min, 2-(tetrahydro-2H-pyran-2-yl)-2H-1,2,3triazole (Préparation 84, 18 g, 117 mmol) was added. The mixture was stirred at about 25 °C for about 6 h. The mixture was concentrated and the residue was purified by chromatography to afford the title compound as a colorless solid (26 g, 79%).
1H NMR (400 MHz, CDCh) δ: 7.99 (s, 1 H), 5.81 (dd, 1 H), 4.07 (d, 1 H), 3.67 - 3.78 (m, 1 H), 2.41 2.55 (m, 1 H), 2.02-2.16 (m, 2 H), 1.60 - 1.80 (m, 3 H), 1.37 (s, 12 H).
LCMS m/z = 113.9 [MH-C2H2N]*
Préparation 86 (1r,3r)-3-(Cyanomethyl)-3-(4-(6-(2-(tetrahydro-2H-pyran-2-yl)-2H-1,2,3-triazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile
A mixture of (1r,3r)-3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 75, 300 mg, 0.89 mmol), 2-(tetrahydro-2Hpyran-2-yl)-4-(4,4,5,5-tetramethyl-1r3,2-dioxaborolan-2-yl)-2H-1,2,3-triazole (Préparation 85, 248 mg, 0.89 mmol), and 2 M aq. K3PO4 (1 mL) in 1,4-dioxane (4 mL) was purged with argon for about 5 min, after which XPhos Pd G2 (140 mg, 0.18 mmol) was added. The mixture was heated at about 50 °C for about 1 hrs, then cooled and diluted with EtOAc. The phases were separated and the aqueous phase was extracted twice with DCM. The combined EtOAc and DCM extracts were dried (Na2SÛ4) and concentrated. The residue was purified by chromatography to afford the title compound as a clear gum (384 mg, 95%).
ΙΟΙ
Ή NMR (400 MHz, CDCb) δ: 9.02 (s, 1 Η), 8.41 (s, 1 Η), 8.35 (s. 1 Η), 8.28 (s, 1 Η), 8.13 (d, 1 Η), 7.00 (d, 1 Η), 5.81 (dd, 1 Η), 4.06 - 4.12 (m, 1 Η), 3.77 - 3.87 (m, 1 Η), 3.35 - 3.47 (m, 3 Η), 3.30 (s, 2 Η). 2.95 - 3.04 (m, 2 Η), 2.45 - 2.59 (m, 1 Η), 2.12 - 2.24 (m, 2 Η), 1.69 -1.87 (m, 3 Η).
LCMS m/z = 455.3 [ΜΗ]*
Préparation 87 (1 r,3r)-3-(4-(6-(2H-1,2,3-T riazol-4-yl)py razolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -y I )-3(cyanomethyl)cyclobutane-l -carbonitrile
A suspension of (1 r,3r)-3-(cyanomethyl)-3-(4-(6-(2-(tetrahydro-2H-pyran-2-yl)-2H-1,2,3-triazol-4yi)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -yl)cyclobutane-1-carbonitrile (Préparation 86, 384 mg, 0.84 mmol) in MeOH (5 mL) was treated with PTSA (16.1 mg, 0.084 mmol). The mixture was heated at about 60 °C for about 3.5 h. The solids dissolved to form a homogeneous solution, after which a white solid precipitated. Heating was continued for about 1 hrs further. The mixture was cooled to about 0 °C for about 30 min and filtered. The precipitate was washed with MeOH and dried under vacuum to afford the title compound as a white solid (204 mg, 65%).
’H NMR (400 MHz, DMSO-de) δ: 9.05 (s. 1 H), 8.97 (s, 1 H), 8.72 (br. s., 1 H), 8.57 (s, 1 H), 8.46 (s, 1 H), 8.30 (s, 1 H), 7.56 (s. 1 H), 3.55 - 3.62 (m, 1 H). 3.53 (s, 2 H), 3.22 - 3.30 (m, 2 H), 2.89 - 3.00 (m, 2 H).
LCMS m/z =371.3 [MH]*
Example 33 (1 r,3r)-3-(Cyanomethyl)-3-(4-(6-(1-methy!-1 H-1,2,3-triazol-4-yl)pyrazolo[1,5-a]py razin-4-yl)-1 Hpy razol-1 -yl)cyclobutane-1 -carbonitrile
I02
To a mixture of (1r,3r)-3-(4-(6-(2H-1,2,3-triazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3(cyanomethyl)cyclobutane-l-carbonitrile (Préparation 87, 100 mg, 0.27 mmol) and K2CO3 (75 mg, 0.54 mmol) in DMF (1 mL) was added iodomethane (0.034 mL, 0.540 mmol). The mixture was stirred for about 2 hrs at about 25 °C. The mixture was filtered and the filter cake was washed twice with DCM. The filtrate was concentrated and the residue was purified by chromatography followed by HPLC to afford the title compound as a white solid (40 mg, 39%).
’H NMR (400 MHz, CDCh) δ: 9.19 (s, 1 H), 8.51 (s, 1 H), 8.33 (s, 1 H), 8.30 (s, 1 H), 8.15 (d, 1 H). 7.03 (s, 1 H), 4.22 (s, 3 H), 3.35 - 3.46 (m, 3 H), 3.30 (s, 2 H), 2.99 (d, 2 H).
LCMS m/z = 385.4 [MH]*
Example 34 (1s,3s)-3-(Cyanomethyl)-1 -methyl-3-(4-(6-(1 -methyl-1 H-pyrazoI-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile
Example 35 (1r,3r)-3-(Cyanomethyl)-1-methyl-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile
103
N—
DBU (327 mg. 2.15 mmol) was added to a solution of 6-(1-methyl-1H-pyrazol-4-yl)-4-(1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazine (Préparation 53, 190 mg, 0.72 mmol) and 3-(cyanomethylene)-15 methylcyclobutane-1-carbonitrile (Préparation 89, 142 mg, 1.07 mmol) in MeCN (15 mL) and the mixture was stirred at about 50 °C for about 4 hrs. The mixture was concentrated and the residue was purified by chromatography to afford a residue which was further purified by TLC to afford a mixture of the two title compounds (200 mg, 70%) as a brown soiid. The mixture of isomers was separated by HPLC to afford (1 r,3r)-3-(cyanomethyl)-1-methyl-3-(4-(6-(1 -methyl-1 H-pyrazol-410 yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile as a light pink solid (trans isomer, 24 mg 8%)
Ή NMR (400 MHz, CD3CN) δ: 8.36 (s, 1 H), 8.28 (s, 1 H), 8.26 - 8.31 (m, 1 H), 8.14 (s, 1 H), 7.86 (s, 1 H), 7.79 (d, 1 H), 7.73 (s, 1 H), 6.87 (d. 1 H), 3.66 (s, 3 H), 3.10 (s, 2 H), 2.80 - 2.92 (m, 4 H). 1.25 (s, 3 H).
LCMS m/z = 398.0 [MH]+ and 420.0 [MNa]’ and (1 s,3s)-3-(cyanomethyl)-1-methyl-3-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile (cis isomer, 24 mg, 8%).
Ή NMR (400 MHz, CD3CN) δ: 8.68 (s, 1 H), 8.63 (s, 1 H), 8.44 (s, 1 H), 8.14 (s, 1 H), 8.05 (d, 1 H), 8.00 (s, 1 H), 7.15 (d, 1 H), 3.92 (s, 3 H), 3.47 - 3.55 (m, 2 H), 3.19 (s, 2 H), 2.74 - 2.82 (m. 2 H), 1.63 20 (s, 3 H).
LCMS m/z = 398.0 [MH]’ and 419.9 [MNa]’
Préparation 88
1-Methyl-3-oxocyclobutane-1 -carbon itrile
Part 1
To a solution of diisopropylamine (1.30 g, 12.9 mmol) in THF (30 mL) was added 2.5 M n-BuLi (5.15 mL) at about 0 °C. The solution was stirred for about 30 min at about 0 °C before being cooled to about -78 °C. 3-Methylenecyclobutane-1-carbonitrile (1.00 g, 10.74 mmol) was added and the solution
104 was stirred for about 1 h at about -78 °C. lodomethane (1.98 g, 14.0 mmol) was added to the solution at about -78 °C, then the mixture was allowed to warm to about 20 °C and was kept at this température for about 0.5 hrs. Saturated aq. NH4CI (30 mL) was added and the mixture was extracted with EtOAc (3 x 30 mL). The combined EtOAc extracts were concentrated to afford 1-methyl-3methylenecyclobutane-1-carbonitrile as a pale yellowoil (1.1 g, 96%).
’H NMR (400 MHz, CDCb) δ: 4.90 - 4.98 (m, 2 H), 3.23 - 3.35 (m, 2 H), 2.64 - 2.75 (m, 2 H), 1.55 (s, 3 H).
Part 2
To a mixture of 1-methyl-3-methylenecyclobutane-1-carbonitrile (1.10 g, 10.3 mmol) and RuCb hydrate (50.9 mg, 0.23 mmol) in a mixture of DCM (20 mL), MeCN (20 mL) and water (40 mL) was added NalÛ4 (8.78 g, 41.1 mmol) in small portions at about 5 °C. The mixture was then stirred at about 25 °C for about 17 hrs. The aqueous phase was separated and extracted with DCM (2 x 50 mL). The DCM extracts were combined with the DCM - MeCN phase and dried (Na2SC>4), then filtered through about 10 g of silica gel. The silica gel was washed with DCM (50 mL). The filtrate was concentrated to afford the title compound as a brown oil (0.80 g, 71%).
1H NMR (400 MHz. CDCb) δ: 3.71 (m, 2 H), 3.14 (m, 2 H), 1.71 (s, 3 H).
Préparation 89
3-(Cyanomethylene)-1-methylcyclobutane-1-carbonitrile
A mixture of 1-methyl-3-oxocyclobutane-1-carbonitrile (Préparation 88, 0.80 g, 7.0 mmol), (EtO)2P(O)CH2CN (1.43 g, 8.06 mmol), LiBr (0.955 g, 11.0 mmol) and TEA (1.48 g, 14.7 mmol) in THF (20 mL) was stirred at about 25 °C for about 16 hrs. Water (30 mL) was added and the mixture was extracted with EtOAc (3 x 50 mL). The combined EtOAc extracts were dried (ΝθςΒΟλ) and concentrated. The residue was purified by chromatography to afford the title compound as a colorless oil (0.65 g, 70%).
1H NMR (400 MHz. CDCb) δ: 5.34 (m, 1 H), 3.47 (m, 2 H), 2.98 (m, 2 H), 1.60 (s. 3 H).
GCMS m/z = 131 [M-H]4
Préparation 90
2-(3-Methoxycyclobutylidene)acetonitrile zN
I05
Part 1
Twelve Identical reactions were carried out in parallel as follows:
For each reaction, a 100 mL sealed tube was charged with methoxyethene (37 g, 637 mmol), DIPEA (9.88 g, 76.4 mmol), and acetyi chloride (5 g, 60 mmol) at about -30 °C. The mixture was then heated at about 70 °C for about 5 hrs. The twelve reactions were combined and washed with 1 M aq. HCl (2 x 100 mL), saturated aq. NaHCOa (2 x 100 mL), dried (NazSCh) and concentrated to afford a crude specimen of 3-methoxycyclobutan-1-one as a black oil (27 g, 37%). This material was about 50% pure as judged by 1H NMR and was used without further purification in Part 2.
Part 2
Two identical reactions were carried out in parallel.
To a mixture of LiBr (13.4 g, 154 mmol) and TEA (39 mL, 280 mmol) in THF (200 mL) was added (EtO)2P(O)CH2CN (26 g, 147 mmol) at about 0 eC. Stirring was continued at about 25 °C for about 2 hrs. To this mixture was added a solution of the crude 3-methoxycyclobutan-1-one prepared in Part 1 (13 g, about 70 mmol) in THF (40 mL) at about 0 °C. The mixture was then stirred at about 25 °C for about 16 hrs. The two reaction mixtures were combined and concentrated. The residue was purified by chromatography to afford the title compound as a light yellow oil (9.2 g, 56%).
1H NMR (400 MHz, CDCh) δ: 5.25 (m, 1 H), 4.05 (m, 1 H), 3.30 (s, 3 H). 3.25 (m,1 H), 3.10 (m, 1 H), 2.85 (m, 2H).
LCMS m/z = 124.08 [MH]*
Préparation 91 (1 r,3r)-3-(Cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1 yl)cyclobutane-1-carbonitrile (trans isomer)
(1 s, 3s)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-l, 3,2-dioxaborolan-2-yl)-1 H-pyrazol-1 yl)cyclobutane-1 -carbonitrile (cis isomer)
106
To a solution of 4-(4,4,5,5,-tetramethyl-1,3.2-dioxaborolan-2-yl)-1H-pyrazole (1.3 kg, 6.67 mol) in MeCN (43 L) were added 3-cyanomethylene)cyclobutane-1-carbonitrile (Préparation 27, 953 g, 8 mol) and DBU (3.06 kg, 20.1 mol) at about 20 °C. Stirring was continued at about 20 °C for about 16 hrs. The mixture was poured into 1 M aq. KHîPO4 (10 L) and extracted with EtOAc (5x5 L). The combined EtOAc extracts were concentrated and the residue was purified by chromatography to afford (1 r,3r)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyt-1,3.2-dioxaborolan-2-yl)-1 H-pyrazol-1yl)cyclobutane-1-carbonitrile as a white solid (trans isomer, 610 g, 30%) melting point 137 - 140 °C ’H NMR (400 MHz, CDCh) δ: 7.90 (s, 1 H), 7.88 (s, 1 H), 3.21 - 3.28 (m, 3 H), 3.19 (s, 2 H), 2.86 2.94 (m, 2 H), 1.33 (s, 12 H).
’3C NMR (101 MHz. CD3OD) δ: 147.54, 136.49, 122.49, 117.11, 84.96, 61.93, 37.52, 30.47, 25.28, 25.18, 17.21.
LCMS m/z = 313.1 [MH]* and (is,3s)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1yl)cyclobutane-1-carbonitrile as an off-white solid (cis isomer, 250 g, 12%).
melting point 95 - 98 °C ’H NMR (400 MHz, CDCI3) δ: 7.86 (s, 2 H), 3.24 - 3.31 (m, 1 H), 3.13 - 3.21 (m, 2 H), 3.07 (s, 2 H), 2.96 - 3.04 (m, 2 H), 1.34 (s, 12 H).
’3C NMR (101 MHz, CD3OD) δ: 147.43, 135.94, 121.94, 117.37, 84.98, 75.96, 60.10, 37.95, 28.42, 25.29, 16.16.
LCMS m/z = 313.1 [MH]'
Préparation 92 5-Methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole
107
3-Methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole
A mixture of 3-methy!pyrazole (1.00 g, 12.18 mmol), DHP (1.54 g, 18.3 mmol) and TFA (0.007 mL, 0.089 mmol) was heated at about 85 °C for about 4 hrs. The mixture was cooled to about 20 °C and NaH (60% in oil, 20 mg, 0.5 mmol) was added. Stirring at about 20 °C was continued for about 18 hrs. The mixture was concentrated the residue was purified by chromatography to afford 5-methyl-1(tetrahydro-2H-pyran-2-yl)-1H-pyrazole as an oil (400 mg, 20%) ’H NMR (400 MHz, CDCh) δ: 7.45 (d, 1 H), 6.05 (d, 1 H), 5.24 - 5.30 (m, 1 H), 4.00 - 4.08 (m, 1 H), 3.61 - 3.70 (m, 1 H), 2.41 -2.55 (m, 1 H), 2.35 (s, 3 H), 2.08 - 2.18 (m, 1 H), 1.93-2.02 (m, 1 H), 1.65 -1.77 (m, 2 H), 1.55-1.64 (m, 1 H).
GCMS m/Z= 166.1 [M]+ and 3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole as an oil (352 mg, 15%).
Ή NMR (400 MHz, CDCb) δ: 7.48 (d, 1 H), 6.08 (d, 1 H), 5.29 (dd, 1 H), 4.08 (dt, 1 H). 3.64 - 3.74 (m, 1 H), 2.30 (s, 3 H), 2.07-2.18 (m, 1 H), 1.98-2.07 (m, 2 H), 1.64- 1.77 (m, 3 H).
GCMS m/z= 166.1 [M]+
Préparation 93 3-Methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5l5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1Hpyrazole
A 2.5 M solution of n-BuLi in hexane (0.44 mL, 1.10 mmol) was added to a solution of 3-methyl-1(tetrahydro-2H-pyran-2-yl)-1H-pyrazole (Préparation 92, 200 mg, 1.10 mmol) in THF (2 mL) at about -70 °C. The mixture was stirred at this température for about 10 min before 2-isopropoxy-4,4,5,5tetramethyl-1,3,2-dioxaborolane (214 mg, 1.15 mmol) was added. The mixture was kept at about -70 °C for about 1 hrs longer, then warmed to about 20 °C and concentrated to afford the title compound admixed with about 65% unreacted 3-methyM-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole. This mixture was used without further purification.
108 1H NMR (400 MHz, CDCh) δ: 6.51 (s, 1 H), 5.76 (dd. 1 H), 4.01 - 4.12 (m, 1 H), 3.59 - 3.74 (m. 1 H), 2.37-2.51 (m, 2 H), 2.29 (s, 3 H), 1.97-2.18 (m, 1 H), 1.62- 1.76 (m, 2 H), 1.47- 1.62 (m, 1 H), 1.33 (s, 12 H).
GCMS m/z = 292.2 [M]“
Préparation 94
3-(Benzyloxy)-N-methoxy-N-methylcycIobutanecarboxamide
To a solution of 3-(benzyloxy)cyclobutanecarboxylic acid (324 g, 1.57 mol) in DCM (1.5 L) was added CDI (280 g, 1.73 mol) in portions. The mixture was heated at reflux for about 2 hrs, after which N,Odimethylhydroxylamine hydrochlorïde (183.7 g, 1.88 mol) and TEA (261 mL, 1.88 mol) were added. The mixture was heated at reflux for about 3 hrs further, then stirred at about 25 °C for about 16 hrs. The DCM was washed with saturated aq. K2CO3. dried (K2CO3), and concentrated. The residue was purified by chromatography to afford the title compound (298 g, 76%).
Ή NMR (400 MHz, DMSO-de) δ: 7.24-7.38 (m, 5 H), 4.37 (s, 2 H), 4.06 - 4.15 (m, 1 H). 3.61 (s, 3 H), 3.09 (s, 3 H), 2.96 (s, 1 H), 2.30 - 2.40 (m, 2 H), 2.09 - 2.22 (m, 1 H), 1.95 - 2.07 (m, 1 H).
Préparation 95
1-[3-(Benzyloxy)cyclobutyl]ethanone
O
To a solution of 3-(benzyloxy)-N-methoxy-N-methylcyclobutanecarboxamide (Préparation 94, 298 g, 1.2 mol) in THF (1.5 L) was added dropwise a solution of méthylmagnésium bromide (1.3 mol) at about -20 °C. After the addition of méthylmagnésium bromide was completed, the cooling bath was removed and the mixture was allowed to warm to about 25 °C. Then 0.5 M aq. HCl (2.5 L) was added, and the mixture was extracted with Et2Û (1 L + 500 mL). The combined EtsO extracts were washed with water, dried (Na2SC>4), and concentrated. The residue was purified by chromatography to afford the title compound (194 g, 79%).
109 ’H NMR (400 MHz, DMSO-ds) δ: 7.24 - 7.38 (m, 5 H), 4.35 (s, 2 H), 3.90 - 3.99 (m, 0.5 H), 3.16 - 3.24 (m, 0.5 H), 2.75-2.87 (m, 1 H), 2.31 -2.40 (m, 2 H), 2.06 - 2.15 (m, 1 H), 2.04 (s, 1.5 H), 1.99 (s, 1.5 H), 1.87- 1.97 (m, 1 H).
Préparation 96
1-[3-(Benzyloxy)cyclobutyl]-3-(dimethylamino)prop-2-en-1-one
To a solution of 1-[3-(benzyloxy)cyclobutyl]ethanone (Préparation 95,194 g, 0.95 mol) in DMF (500 mL) was added dimethylformamide dimethyl acetal (285 g, 2.4 mol). The mixture was heated at about 110 °C for about 12 hrs. The mixture was concentrated to afford the title compound (258 g), which was used without purification or further characterization.
Préparation 97 5-[3-(Benzyloxy)cyclobutyl]-1H-pyrazole
To a solution of 1-[3-(benzyloxy)cyclobutyl]-3-(dimethylamino)prop-2-en-1-one (Préparation 96, 110 g, 0.42 mol) in MeOH (500 mL) was added hydrazine hydrate (30 g, 0.6 mol). The mixture was heated at reflux for about 12 hrs, then concentrated. The residue was purified by chromatography to afford the title compound (79 g, 82%), which was used without purification or further characterization.
Préparation 98
3-(1 H-Pyrazol-5-yl)cyclobutanol \\ / n-nh
I ΙΟ
Το a solution of 5-[3-(benzyloxy)cyclobutyl]-1H-pyrazole (Préparation 97, 79 g, 0.35 mol) in methanol (800 mL) in a 2 L hydrogénation flask was added palladium on carbon (10% Pd, 16 g). The mixture was pressurïzed under hydrogen (30 psi) and the mixture was shaken at about 60°C for about 12 hrs. Additional palladium on carbon (10% Pd, 16 g) was added, and the hydrogénation was continued for about 10 hrs. The mixture was filtered and the filtrate was concentrated. The residue was purified by chromatography to afford the title compound (46 g, 96%), which was used without purification or further characterization.
Préparation 99 3-(1H-pyrazol-5-yl)cyclobutane-1-one
N-NH
To a solution of oxalyl chloride (7.8 mL, 0.09 mol) in DCM (50 mL) at about -78 °C was added dropwise a solution of DMSO (12.7 mL, 0.18 mol) in DCM (50 mL). The mixture was stirred for about 30 min, then 3-(1H-pyrazoi-5-yl)cyclobutanol (Préparation 98, 13.7 g, 0.09 mol) was added dropwise at this température. The resulting mixture was kept for about 30 min, after which TEA (25 mL, 0.18 mol) was added dropwise. The cooling bath was removed and the mixture was allowed to warm to about 25 °C and was kept at that température for about 3 hrs. Then an aqueous solution of K2CO3 (100 mL) was added. The DCM was separated, and the aqueous phase was extracted with DCM. The combined DCM extracts were washed with water, dried (NazSOA), and concentrated. The residue was purified by chromatography to afford the title compound (7.4 g, 55%) as a white solid.
Ή NMR (400 MHz, DMSO-d6) δ: 12.56 (s, 1 H), 7.61 (s, 1 H), 6.21 (d, 1 H), 3.46 - 3.68 (m, 1 H), 3.37 - 3.46 (m. 2 H), 3.15 - 3.25 (m, 2 H).
LCMS m/z = 137.1 [MH]+
Préparation 100
2-((1 r,3s)-1-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3methoxycyclobutyl)acetonitrile (trans isomer)
111
Part 1
To a solution of 2-((1 r,3s)-1-(4-bromo-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile (Préparation 37, trans isomer, 3399 mg, 12.58 mmol) in 1,4-dioxane (33 mL) were added bis(pinacolato)diboron (3510 mg, 13.8 mmol) and KOAc (3700 mg, 37.7 mmol). The mixture was purged with argon for about 5 min, after which XPhos Pd G2 (1980 mg, 2.52 mmol) was added. The mixture was heated at about 65 °C for about 4 hrs. The cooled mixture was concentrated and the residue was purified by chromatography. The product was stirred with EtOAc (10 mL) at about 25 °C, then heptane (40 mL) was added and crystallization was allowed to occur for about 30 min. The precipitate was filtered and dried to afford 2-((1 r,3s)-3-methoxy-1 -(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1yl)cycfobutyl)acetonitrile as a white solid (1950 mg, 49%).
1H NMR (400 MHz, CDCh) δ: 7.91 (s, 1 H), 7.87 (s, 1 H). 3.98 (tt, 1 H), 3.29 (s, 3 H), 3.17 (s, 2 H), 2.97 - 3.07 (m, 2 H), 2.44 - 2.53 (m, 2 H), 1.33 (s, 12 H).
LCMS m/z = 318.0 [MH]*
Part 2
A solution of 2-((1r,3s)-3-methoxy-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazol-1yl)cyclobutyl)acetonitrile (Part 1. 1950 mg, 6.15 mmol) and 4,6-dichloropyrazolo[1,5-a]pyrazine (Préparation 4, 1160 mg, 6.15 mmol), and 2 M aq. K3PO4 (9.22 mL) in 1,4-dioxane (25 mL) was purged with argon for about 5 min, after which bis(tri-t-butylphosphine)palladium(0) (157 mg, 0.31 mmol) was added. The mixture was stirred at about 25 °C for about 2 hrs. The cooled mixture was diluted with EtOAc and the phases were separated. The aqueous phase was extracted twice with DCM. The combined EtOAc and DCM extracts were dried (NazSOi) and concentrated. The residue was dissolved in DCM (20 mL) and heated at about 40 °C until ail was dissolved, then heptane (10 mL) was added and crystallization was allowed to occur for about 30 min. The precipitate was filtered and dried to afford the title compound as an off white solid (1120 mg, 53%). The filtrate was concentrated and the residue was purified by chromatography to afford additional title compound (640 mg, 30%).
Ή NMR (500 MHz, CDCh) δ: 8.39 (d, 1 H), 8.38 (s, 1 H), 8.28 (s, 1 H), 8.08 (d, 1 H), 7.03 (dd, 1 H), 4.03 - 4.10 (m, 1 H), 3.34 (s, 3 H), 3.25 (s, 2 H), 3.08 - 3.16 (m, 2 H), 2.53 - 2.60 (m. 2 H).
LCMS m/z = 343.3 [MH]* (35CI isotope)
Préparation 101
Ethyl 5-methyl-1-(2-(1-methyl-1H-pyrazol-4-yl)-2-oxoethyl)-1H-pyrazole-3-carboxylate
112 0
O
Λ,Ν-ν Ο-^χ 'fT
I
Ethyl 3-methyl-1H-pyrazole-5-carboxylate (92 mg, 0.6 mmol), 2-bromo-1-(1-methyl-1 H-pyrazol-4yl)ethan-1-one (Préparation 6, 134 mg, 0.66mmol) and K2CO3 (104 mg, 0.75 mmol).were combined in MeCN ( 2 mL) and the suspension was stirred at about 40 °C for about 16 hrs. The solids were filtered and the filtrate was concentrated. The residue was purified by chromatography to afford the title compound as a yellow oil (125 mg, 5.5%).
’H NMR (400 MHz, CDCb) δ: 7.86 (s. 2 H), 6.73 (s, 1 H). 5.65 (s. 2 H), 4.26 (q, 2 H), 3.95 (s, 3 H), 2.32 (s, 3 H), 1.31 (t, 3 H).
LCMS m/z = 277.1 [MH]+
Préparation 102
2-Methyl-6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin~4-ol
OH
To a solution of ethyl 5-methyl-1-(2-(1-methyl-1H-pyrazoî-4-yl)-2-oxoethyl)-1H-pyrazole-3-carboxylate (Préparation 101, 125 mg, 0.45 mmol) in EtOH (5 mL) was added NH^OAc (105 mg, 1.06 mmol). The mixture was heated under microwave irradiation at about 105 °C for about 4 hrs. The mixture was concentrated and the residue was dissolved in EtOH and concentrated again to afford the title compound (110 mg, 85%).
’H NMR (400 MHz, DMSO-de) Ô: 8.25 (s, 1 H), 8.01 (s, 1 H), 7.97 (s, 1 H), 6.76 (s, 1 H), 3.87 (s, 3 H), 2.34 (s. 3 H).
LCMS m/z = 230.0 (MH]+
Préparation 103
4-Chloro-2-methyl-6-(1-methyl-1 H-pyrazol-4-yl)pyrazoto[1,5-a]pyrazine
113
2-Methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-ol (Préparation 102) was suspended in POCh and heated at about 120 °C for about 6 hrs. The solution was concentrated afford an impure sample of the title compound which was used in the next step without further purification. LCMS m/z = 248.0 [MH]* (35CI isotope)
Example 36 (1r,3r)-3-(Cyanomethyl)-3-(4-(2-methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (trans isomer)
A mixture of crude 4-chloro-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazoio[1,5-a]pyrazine (Préparation 103, 100 mg, 0.40 mmol), (1r,3r)-3-(cyanomethyl)-3-(4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (Préparation 91, 132 mg, 0.42 mmol), Pd(dppf)CI2 DCM (16.5 mg, 0.02 mmol), and K2CO3 (167 mg, 1.21 mmol) were combined in a mixture of 1,4-dioxane (2.5 mL) and water (0.5 mL). The mixture was purged with nitrogen for about 5 min, then heated at about 90 °C for about 3 hrs. The mixture was concentrated and the residue was purified by chromatography and HPLC to afford the title compound (5.6 mg, 3% over two steps).
Ή NMR (400 MHz, CDCh) 5: 8.37 (s, 1 H), 8.33 (s, 1 H), 8.30 (s, 1 H), 7.92 (s, 1 H), 7.90 (s, 1 H), 6.68 (s, 1 H), 3.99 (s, 3 H), 3.32 - 3.42 (m, 3 H), 3.27 (s, 2 H), 2.93 - 3.02 (m. 2 H), 2.56 (s, 3 H). LCMS m/z = 398.0 [MH]*
Préparation 104
3-((4-Methoxybenzyl)oxy)-1-methyl-1H-pyrazole
OMe
Il4
To a 100 mL round bottom flask were added 1-methyl-1H-pyrazol-3-ol (1.40 g, 14.3 mmol), DMF (30 mL), and K2CO3 (3.94 g, 28.5 mmol). Lastly 4-methoxybenzyl chloride (2.32 mL, 17.1 mmol) was added to the mixture. The mixture was heated at about 60 °C for about 8 hours. The mixture was then diluted with water (80 mL) and extracted with EtOAc (60 mL x 3). The combined EtOAc extracts were washed with brine (60 mL x 2), dried (Na2SO«) and concentrated. The residue was purified by chromatography to afford the title compound as a colorless oil (2.6 g, 83%).
1H NMR (400 MHz. CDCI3) δ: 7.38 (d, 2 H), 7.13 (d, 1 H), 6.91 (d. 2 H), 5.64 (d, 1 H), 5.11 (s, 2 H), 3.82 (s, 3 H), 3.76 (s, 3 H).
LCMS m/z = 218.9 [MH]*
Préparation 105
4-lodo-3-((4-methoxybenzyl)oxy)-1-methyl-1H-pyrazole
N
To a 100 mL round bottom flask were added 3-((4-methoxybenzyl)oxy)-1-methyl-1H-pyrazole (Préparation 104, 1.0 g, 4.58 mmol) and MeCN (20 mL), after which ceric ammonium nitrate (1.51 g, 2.75 mmol) and iodine (698 mg, 2.75 mmol) were added to the mixture. The brown mixture was stirred at about 20 °C for about 1 h. The mixture was quenched with 5% aqueous sodium bisulfite (50 mL) and extracted with EtOAc (40 mL x 3). The combined EtOAc extracts were dried (NasSOi) and concentrated. The residue was purified by chromatography to afford the title compound as a green oil (800 mg, 51%).
1H NMR (400 MHz, CDCI3) δ: 7.40 (d, 2 H). 7.19 (s, 1 H), 6.91 (d. 2 H). 5.18 (s, 2 H), 3.82 (s, 3 H), 3.77 (S, 3 H).
Préparation 106
3-((4-Methoxybenzyl)oxy)-1 -methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazole
OMe
H5
To a 100 mL round bottom flask were added 4-iodo-3-((4-methoxybenzyl)oxy)-1 -methyl-1 H-pyrazole (Préparation 105, 800 mg, 2.32 mmol) and THF (16 mL). followed by the dropwise addition of isopropylmagnesium chloride (1.3 M in THF, 2.15 mL, 2.79 mmol) to the mixture at about -10 °C. The mixture was stirred at a température between about -18 °C and 10 °C for about 45 min. 2-lsopropoxy4,4,5,5-tetramethyl-1,3,2-dioxaborolane (649 mg, 3.49 mmol) was added to the mixture at about -10 °C and the mixture was aliowed to warm to about 15 °C for about 1.5 h. Additional isopropylmagnesium chloride (1.3 M in THF, 0.72 mL, 0.93 mmol) and 2-isopropoxy-4,4,5,5tetramethyl-1,3,2-dioxaborolane (216 mg, 1.16 mmol) were added to the mixture at about -15 °C. The mixture was aliowed to warm to about 15 °C for about 1 h. The mixture was diluted with EtOAc (40 mL) and washed with saturated aqueous NH4CI (30 mL), brine (30 mL), dried (NaïSO^), and concentrated. The residue was purified by chromatography to afford the title compound as a white solid (600 mg, 75%).
’H NMR (400 MHz, CDCh) δ: 7.40 - 7.45 (m, 3 H), 6.88 (d, 2 H), 5.24 (s, 2 H), 3.81 (s, 3 H), 3.73 (s, 3 H), 1.31 (s, 12 H).
LCMS m/z = 345.1 [MH]*
Préparation 107 (1r,3r)-3-(Cyanomethyl)-3-(4-(6-(3-((4-methoxybenzyl)oxy)-1-methyl-1H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyc1obutane-1-carbonitrile
To a 25 ml round bottom flask were added (1r,3r)-3-(4-(6-chloropyrazolo[1,5-a]pyrazin-4-yl)-1Hpyrazol-1-yl)-3-(cyanomethyl)cyclobutane-1-carbonitrile (Préparation 75, 300 mg. 0.88 mmol), 3-((4methoxybenzyl)oxy)-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (Préparation 106, 367 mg, 1.07 mmol), dioxane (16 mL), XPhos Pd G2 (140 mg, 0.178 mmol) and 2 M aq. K3PO4 (3.55 mL, 7.11 mmol). The mixture was placed under nitrogen, then heated at about 60 °C for about 4 h. The mixture was diluted with water (70 mL) and extracted with EtOAc (50 mL x 3).
The combined EtOAc extracts were dried (Na:SO4) and concentrated. The residue was purified by chromatography to afford the title compound as a yellow solid (450 mg, 98%).
ne ’H NMR (400 MHz, DMSO-dg) δ: 8.92 (s, 1 H), 8.57 (s, 1 H), 8.52 (s, 1 H), 8.31 (s, 1 H), 8.16 (d, 1 H), 7.50 (d, 2 H), 7.45 (s, 1 H), 7.00 (d, 2 H), 5.29 (s, 2 H), 3.95 (s, 3 H), 3.82 (s, 3 H), 3.55 - 3.59 (m, 1 H), 3.52 (s, 2 H), 3.25 - 3.32 (m, 2 H), 2.94 (dd, 2 H).
LCMS m/z = 542.1 [MNa]+
Example 37 ( 1r,3r)-3-(Cyanomethy 1)-3-(4-(6-(1-methy l-3-oxo-2,3-d ihyd ro-1 H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (trans isomer)
(1r,3r)-3-(Cyanomethyl)-3-(4-(6-(3-((4-methoxybenzyl)oxy)-1-methyl-1H-pyrazol-4-yl)pyrazolo(1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (Préparation 107, 450 mg, 0.86 mmol) and TFA (13 mL) were stirred at about 10 °C for about 4 h. The mixture was concentrated and the residue was diluted with DCM (40 mL) and MeOH (40 mL) and neutralized with solid NaHCO3. The mixture was filtered. The filtrate was concentrated and the residue was purified by HPLC to afford the title compound as a yellow solid (54 mg, 16%).
1H NMR (400 MHz, DMSO-de) δ: 10.76 (br. s., 1 H), 8.92 (s, 1 H), 8.71 (s, 1 H), 8.51 (s, 1 H), 8.18 (d, 1 H), 8.16 (s, 1 H), 7.45 (d, 1 H), 3.73 (s, 3 H), 3.55 - 3.62 (m, 1 H), 3.53 (s, 2 H), 3.25 - 3.33 (m, 2 H), 2.90-2.99 (m, 2 H).
LCMS m/z = 400.1 [MH]+
Bioloqical Evaluation
Compounds of the invention were evaluated by in vitro methods to détermine their respective ability to inhibit the JAK kinases (TYK2, JAK1, JAK2, JAK3).
Assay Format
The human JAK inhibitory activity was determined by using a microfluidic assay to monitor phosphorylation of a synthetic peptide by the recombinant human kinase domain of each of the four members of the JAK family, JAK1, JAK2, JAK3 and TYK2. Reaction mixtures contained 1 μΜ of a fluorescently labeled synthetic peptide, a concentration less than the apparent Km. and 1 mM ATP.
117
Each assay condition was optimized for enzyme concentration and room température incubation time to obtain a conversion rate of 20% to 30% phosphorylated peptide product. Reactions were terminated by the addition of stop buffer containing EDTA. Utilizing the LabChip 3000 mobility shift technology (Caliper Life Science), each assay reaction was sampled to détermine the level of phosphorylation. This technology is separation-based, allowing direct détection of fluorescently labeled substrates and products. Séparations are controlled by a combination of vacuum pressure and electric field strength optimized for each peptide substrate.
Assay Protocol
JAK Caliper Enzyme Assay at 1mM ATP
Compounds were added to a 384-well plate. Reaction mixtures contained 10 mM HEPES, pH 7.4, 10 mM MgCh, 0.01% BSA, 0.0005% Tween 20, 1 mM ATP and 1 μΜ peptide substrate. The JAK1 and TYK2 assays contained 1 μΜ of the IRStide peptide (5FAM-KKSRGDYMTMQID) and the JAK2 and JAK3 assays contained 1 μΜ of the JAKtide peptide (FITC-KGGEEEEYFELVKK). The assays were initiated by the addition of 20 nM JAK1, 1 nM JAK2, 1 nM JAK or 1 nM TYK2 enzyme and were incubated at room température for three hours for JAK1, 60 minutes for JAK2, 75 minutes for JAK3 or 135 minutes for TYK2. Enzyme concentrations and incubation times were optimized for each new enzyme preps and were modified slightly over time to ensure 20% to 30% phosphorylation. The assays were stopped with 15 uL of 180 mM HEPES, pH 7.4, 20 mM EDTA, and 0.2% Coating Reagent 3. The assay plates were placed on a Caliper Life Science LC3000 instrument, and each well was sampled using appropriate séparation conditions to measure the unphosphorylated and phosphorylated peptide.
Data Analysis
The data was cottected using the HTS Well Analyzer software from Caliper Life Sciences. The data output for data analysis is the percent product converted calculated on peak height (Equation 1). Equation 1 : % product converted = 100‘((product)/(product + substrate))
The percent effect at each compound concentration was calculated based on the positive and négative control well contained within each assay plate (Equation 2). The positive control wells contained a saturating concentration of a control compound that produced a level of phosphorylation comparable to background (i.e., completely inhibited JAK1, JAK2, JAK3 or TYK2). The négative control wells contained DMSO aione (at the same concentration as the compound wells) that was used to set the baseline activity in the assay (i.e., uninhibited JAK1, JAK2, JAK3 or TYK2).
Equation 2: % effect = 100*((sample well - négative control)/(positive control-negative control))
The percent effect was plotted against the compound concentration compound. An unconstrained sigmoid curve was fitted using a 4 parameter logistic model and the compound concentration required for 50% inhibition (ICso) was determined (Equation 3).
Equation 3: y = ((max - m/n)/(1 + ((x/ICso)As))) + min
Where max is the maximum asymptote (complété inhibition), min is the minimum asymptote (no inhibition) and s is the slope factor. IC50 values are reported in nM for each compound:
us
Table I. JAK Caliper Data
Example No. TYK2 IC50 (nM) JAK1 IC50 (nM) JAK2 IC50 (nM) JAK3 IC50 (nM)
1 18 291 40 >9788
2 62 1057 299 >10000
3 64 2292 487 >10000
4 55 1338 141 >10000
5 35 1917 472 >10000
6 21 5720 500 >10000
7 7 250 37 6682
8 11 265 42 >9282
9 8 185 49 >10000
10 580 5709 1601 >10000
11 8 273 38 >6170
12 24 764 159 >10000
13 149 3228 487 >10000
14 81 1061 367 >10000
15 22 1495 228 >9121
16 16 664 99 10000
17 35 1728 205 10000
18 47 1079 206 10000
19 6 21 8 1051
H9
20 16 383 74 >10000
21 34 1288 109 >10000
22 30 2544 127 >10000
23 9 431 26 9410
24 25 3550 432 >10000
25 23 319 99 >10000
26 21 713 158 >10000
27 24 737 171 >10000
28 27 1362 249 >10000
29 352 3932 3041 >10000
30 7 174 71 >10000
31 38 >9857 324 >10000
32 17 2254 339 >10000
33 52 8717 444 >10000
34 11 96 19 3263
35 136 1915 268 >10000
36 1605 3521 755 >10000
37 32 3489 166 >10000
Selected compounds were assessed for their ability to inhibit IL-12 signaling in a human whole blood flow cytometry assay. IL-12 signais through TYK2 and JAK2.
Human whole blood IL-12 induced STAT4 phosphorylation Assay
Test articles were prepared as 30 mM stocks in DMSO. An 11-pofnt 2.5 dilution sériés was created in DMSO with a top concentration of 10 mM. Further dilution was done by adding 4 pL of the
120 above test article solutions into 96 μΐ_ of PBS with a top concentration of 400 μΜ. Human whole blood was collected from heaîthy donors via vein puncture into Vacutainer collection tubes containing sodium heparin (Catalog No. 366480; Becton Dickinson, Franklin Lakes, NJ). Blood was warmed to 37OC prior to use. Human whole blood was aliquoted (90 mL/well) in 96-well, deep-well, V-bottom plates and treated with compounds at 11 different concentrations (0.2% DMSO final) at 37°C for 60 minutes. This was followed by a challenge with IL-12 (5 mL/well; final, 5 ng/mL) for 15 minutes. Samples were treated with warm 1X Lyse/Fix buffer (700 mL/well) to terminale activation and further incubated at 37°C for 20 minutes to lyse red blood cells. Plates were centrifuged at 300 x g for 5 minutes, supernatant was aspirated, and cells were washed with 800 mL per well of staining buffer (PBS containing 0.5% fêtai bovine sérum and 0.01% sodium azide). The washed cell pelîets were resuspended with 350 mL/well of pre-chilled 90% methanol, and incubated at 4°C for 30 minutes. Plates were centrifuged at 300 x g for 5 minutes, supernatant containing 90% methanol was aspirated. and cells were washed with 800 mL /well of staining buffer. Cell pellets were resuspended in staining buffer containing anti-pSTAT4-AlexaFluor647 (1 to 150 dilution, 150 mL/well), and incubated at room température in the dark overnight.
Samples were transferred to 96-well U-bottom plates and flow cytométrie analysis was performed on a FACSCalibur or LSRFortessa equipped with a HTS plate loader (BD Biosciences). The lymphocyte population was gated for histogram analysis of pSTAT4. Background fluorescence was defined using unstimulated cells and a gâte was placed at the foot of the peak to include ~0.5% gated population. The histogram statistical analysis was performed using CelIQuestÔ Pro version 5.2.1 (BD Biosciences) or FACSDiva version 6.2 (BD Biosciences) software. Relative fluorescence unit (RFU), which measures the level of phospho STAT4, was calculated by multiplying the percent positive population and its mean fluorescence. Data from 11 compound concentrations (singlicate at each concentration) was normalized as a percentage of control based on the formula:
% of Control = 100 x (A - B)/(C - B) where A is the RFU from wells containing compound and IL-12, B is the RFU from welts without IL-12 and compound (minimum fluorescence) and C is the RFU from wells containing only IL-12 (maximum fluorescence). Inhibition curves and IC» values were determined using the Prism version 5 software (GraphPad, La Jolla, CA).
121
Table II. Human whole blood IL-12 Data.
Example Number HWB IL-12 ICso (nM)
1 162
3 203
7 41
19 28
20 50
23 34
25 59
27 86
30 46
32 130

Claims (15)

    CLAIMS What is claimed is:
  1. (1 r,3r)-3-(Cyanomethyl )-3-(4-(6-( 1-methyl-3-oxo-2,3-dihydro-1 H-pyrazol-4-yl)pyrazolo[1,5a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile (trans isomer);
    (1r,3r)-3-(cyanomethyi)-3-(4-(6-(1-(hydroxymethyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile; and, or, a pharmaceutically acceptable sait thereof.
    (1 r,3r)-3-(cyanomethyl)-3-(4-(6-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-y[)-1 Hpyrazol-1-yl)cyclobutane-1-carbonitrile;
    (1s,3s)-3-(cyanomethyl)-3-(4-(6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 Hpyrazol-1-yl)cyclobutane-1-carbonitrile;
    (1r,3r)-3-(cyanomethyl)-3-(4-(3-methyl-6-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile;
    1. A compound having the structure (I):
    or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    A, A' and A” are independently O, C=O, C-R' or N-R, where R' and R may independently be H, amino, -NRrCORe, COR6, -CONRrRe, Ci-Ce alkyl, or hydroxy(Ci-C6 alkyl)-, and R” may be présent or absent, and is présent where the rules of valency permit, and where not more than one of A, A’ and A is O or C=O;
    Ro and R are independently H, Br, Cl, F, or Ci-Cs alkyl;
    Ri is H, Ci-Ce alkyl, or hydroxy(Ci-Cg alkyl)-;
    R2 is seiected from the group consisting of H, Ci-Ce alkyl-, Ci-Ce alkoxy-, hydroxy(Ci-Ce alkyl)-, phenyl(Ci-C6 alkyl)-, formyl, heteroaryl, heterocyclic, -CORe, -OCORs, -COORe, -NR7COR6, CONRrRs, and -(CH2)n-W, where W is cyano, hydroxy, C3-CB cycloalkyl, -SOïNRrRa, and -SO2-R9, where Rs is C1Ce alkyl, Ca-Ce cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ce alkyl;
    X is C-R3 or N, where R3 may be H or Ci-C6 alkyl;
    R4 and Rs are independently H, amino, Ci-Ce alkyl, or hydroxy(Ci-C6 alkyl)-;
    R6, R? and Re are each independently H, Ci-Cb alkyl, C1-C4 alkoxy(Ci-C6 alkyl) or C3-C8 cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Rs together with the atom bonded
    123 thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ce alkyl;
    and, n is 0, 1, 2 or 3.
  2. 2-(1-ethyl-3-(4-(6-(5-(hydroxymethyl)-1H-pyrazo!-3-yl)pyrazolo[1,5-ajpyrazin-4-yl)-1H-pyrazol1 -yl)azetidin-3-yl)acetonitrile;
    2-((1 r,3s)-1-(4-(6-(3-amino-1H-pyrazol-5-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)-3methoxycyclobutyl )acetonitrile ;
    2-((1 s,3r)-1-(4-(6-(5-(hydroxymethyl)-1 H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol1-yl)-3-methoxycyclobutyl)acetonitrile;
    2,2'-(3-(4-(6-( 1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1 -yl)azetidine1,3-diyl)diacetonitrile;
    2. The compound of claim 1 having the structure (la):
    or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    A, A and A ’ are independently O, C=O, C-R' or N-R”, where R' and R may independently be H, amino, -NR7COR6, CORe, -CONRtRb, Ci-Cg alkyl, or hydroxy(Ct-Ce alkyl), and R” may be présent or absent, and is présent where the rules of valency permit, and where not more than one of A, A' and A is O or C=O;
    Ro and R are independently H, Br, Cl, F, or Ci-Ce alkyl;
    Ri is H, Ci-Cg alkyl, or hydroxy(Ci-Ce alkyl);
    R2 is selected from the group consisting of H, Ci-Ce alkyl-, Ci-C6 alkoxy-, hydroxy(Ci-Ce alkyl)-, phenyl(Ci-Ce alkyl)-, formyl, heteroaryl, heterocyclic, -CORe, -OCORe, -COORe, -NR7CORg, -CONR7R8, and -(CHz)n-W, where W is cyano, hydroxy, C3-Cb cycloalkyl. -SO2NR7Ra, and -SO2-R9, where Rg is CtCe alkyl, C3-C8 cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl. heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ce alkyl;
    R3 may be H or Ci-Ce alkyl;
    Ri and Re are independently H, amino, Ci-C6 alkyl, or hydroxy(Ci-Ce alkyl)-;
    124
    Re, R? and Ra are each independently H, Ci-Ca alkyl, C1-C4 alkoxy(Ci-Cs alkyl)-, or Ca-Ca cycloalkyl, said Ci-Ca alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ce alkyl;
    and, n is 0, 1, 2 or 3.
  3. 3. The compound of claim 1 having the structure (Ib):
    or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    R is H, -CORe, -CONRzRa, Ci-Ce alkyl, or hydroxy(Ci-Ce alkyl)-;
    Ro and R are independently H, Br, Cl, F, or Ci-Ce alkyl;
    Ri is H. Ci-Cs alkyl, or hydroxy(Ci-C6 alkyl);
    R2 is selected from the group consisting of H, Ct-Ce alkyl, Ci-Ca alkoxy, hydroxy(Ci-Cealkyl), phenyl(CiCe alkyl), formyl, heteroaryl, heterocyclic, -COR6, -OCORe, -COOR6, -NRzCORe, -CONRzRe, and (CH2)n-W, where W is cyano, hydroxy, Ca-Ca cycloalkyl, -SOzNRzRe, and -SO2-R9, where Rg is Ci-Ce alkyl, C3-Ce cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ce alkyl;
    R3 may be H or Ci-Ce alkyl;
    Rs is independently H, amino, Ci-Ce alkyl, or hydroxy(Ci-CB alkyl);
    125
    Re, Rz and Ra are each independently H, Ci-Ca alkyl, C1-C4 alkoxy(Ci-Ca alkyl), or Ca-Ce cycloalkyl, said Ci-Ca alkyl is optionally substituted by halo, CN or hydroxy; or, Rz and Ra together with the atom bonded thereto fomn a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ca alkyl;
    and, n is 0, 1, 2 or 3.
  4. 4. The compound of claim 3 wherein R is Ci-Ca alkyl and Rs is H.
  5. 5-(4-(1-((1 s, 3r)-1-(cyanomethyl)-3-methoxycyclobutyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin6-yl)-1H-pyrazoie-3-carboxamide;
    (1s,3s)-3-(cyanomethyl)-3-(4-(6-(5-(hydroxymethyl)isoxazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)1H-pyrazol-1-yl)cyclobutane-1-carbonitriie;
    5. The compound of claim 1 having the structure (le):
    NC R2
    Z'' R3 N—N
    or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    R is H, -CORa, -CONRzRe, Ct-Cs alkyl-, or hydroxy(Ci-Ca alkyl)-;
    R2 is selected from the group consisting of H, Ci-C6 alkyl, Ci-Ce alkoxy-, hydroxy(Ci-Ca alkyl)-, phenyl(Ci-C6 alkyl-), formyl, heteroaryl, heterocyclic, -CORa, -OCOR6, -COORa, -NRzCORa, -CONRzRe, and -(CHz)n-W, where W is cyano, hydroxy, C3-C8 cycloalkyl, -SO2NRzRe, and -SO2-R9, where R9 is C1Ce alkyl, Ca-Ca cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ca alkyl;
    R3 is H, Ci-Ce alkyl, amino, cyano, or Ci-Ca alkoxy-;
    Rs is H, amino, Ci-Ce alkyl, or hydroxy(Ci-Ce alkyl)-;
    Re, Rz and R8 are each are each independently H, Ci-C6 alkyl, C1-C4 alkoxy(Ci-C6 alkyl)-, or C3-C8 cycloalkyl, said Ct-Ca alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Ra together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Ca alkyl;
    126 and, n is 0, 1, 2 or 3.
  6. 6. The compound of daim 5 wherein R” is Ci-Ce alkyl and Rs is H.
  7. 7. The compound of claim 1 having the structure (Id):
    (Id) or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    R” is H, -CORe, -CONRrRe, Ci-Ce alkyl, or hydroxy(Ci-Ce alkyl)-;
    R2 is selected from the group consisting of H, Ci-Ce alkyl, Ci-Ce alkoxy-, hydroxy(Ci-Ce alkyl)-, phenyl(Ci-Ce alkyl)-, formyl, heteroaryl, heterocyclic, -CORe, -OCOR6, -COOR6, -CONRtRb, and (CH2)n-W. where W is cyano, hydroxy, C3-C8 cycloalkyl-, -SOsNRyRa, and -SOZ-R9, where Rg is Ci-C6 alkyl, C3-Ca cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Ce alkyl;
    Rs is H, amino, Ci-Cs alkyl-, or hydroxy(Ci-Ce alkyl)-;
    Re, Rr and Ra are each are each independently H, Ci-C6 alkyl, C1-C4 alkoxy(Ci-C6 alkyl), or C3-CB cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R7 and Re together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-Cs alkyl;
    and, n is 0, 1, 2 or 3.
  8. 8. The compound of claim 7 wherein R is Ci-Ce alkyl and R5 is H.
    127
  9. 9. The compound of claim 1 having the structure (le):
    or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    R2 is selected from the group consisting of H, Ci-Ce alkyl, Ci-CB alkoxy-, hydroxy(Ci-CB alkyl)-, phenyl(Ci-CB alkyl)-, formyl, heteroaryl, heterocyclic -COR6, -OCOR6, -COOR6, -CONR7RB, and -(CH2)nW, where W is cyano, hydroxy, C3-Ca cycloalkyl, -SO2NR7R8, and -SO2-R9, where R9 is Ct-C6 alkyl, C3Ce cycloalkyl, heteroaryl, or heterocyclic; wherein each of said alkyl, cycloalkyl, heterocyclic, or heteroaryl may be unsubstituted or substituted by halo, cyano, hydroxy, or Ci-Cs alkyl;
    Re, R? and Re are each are each independently H, Ct-Ce alkyl, C1-Q1 alkoxy(Ci-C6 alkyl), or C3-Ce cycloalkyl, said Ci-Ce alkyl is optionally substituted by halo, CN or hydroxy; or, R? and Rb together with the atom bonded thereto form a 5- or 6-membered ring, said ring being optionally substituted by halo, hydroxy, CN, or Ci-CB alkyl;
    and, n is 0, 1, 2 or 3.
  10. 10. The compound of claim 9 wherein R2 is -(CH2)n-W, where W is cyano and n is 1, 2 or 3.
  11. 11. The compound of claim 1 having the structure (If):
    128 (If) or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or pharmaceutically acceptable sait, wherein:
    R is H, Ci-Ce alkyl, or hydroxy(Ci-Ce alkyl)-; and,
    Rs is H, amino, Ci-Ce alkyl, or hydroxy(Ci-C6 alkyl)-.
  12. 12. The compound of claim 11 wherein R is Ci-Ce alkyl and Rs is H.
  13. 13. The compound of claim 1 selected from the group consisting of:
    (1r,3r)-3-(4-(6-(3-amino-1-methyl-1H-pyrazol-4-y[)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1yl)-3-(cyanomethyl)cyclobutane-1-carbonitrile;
  14. 14. The compound of claim 1 wherein the compound is (1r,3r)-3-(4-(6-(3-amino-1-methyl-1 Hpyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-(cyanomethyl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
    129
    15. The compound of claim 1 wherein the compound is 2,2'-(3-(4-(6-(1 -methyl-1 H-pyrazol-4yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)azetidine-1,3-diyl)diacetonitrile, or a pharmaceutically acceptable sait thereof.
    16. The compound of claim 1 wherein the compound is 2-((1 s,3r)-1 -(4-(6-(5-(hydroxymethy[)IH-pyrazol-S-yQpyrazolotl.S-alpyrazin-A-ylJ-IH-pyrazol-l-yQ-S-methoxycyclobutyOacetonitrile, or a pharmaceutically acceptable sait thereof.
    17. The compound of claim 1 wherein the compound is 5-(4-(1-((1 s,3r)-1-(cyanomethyl)-3methoxycyclobutyl)-1 H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-6-yl)-1 H-pyrazole-3-carboxamide, or a pharmaceutically acceptable sait thereof.
    18. The compound of claim 1 wherein the compound is (1s,3s)-3-(cyanomethyl)-3-(4-(6-(5(hydroxymethyl)isoxazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1-carbonitrile, or, a pharmaceutically acceptable sait thereof.
    19. The compound of claim 1 wherein the compound is (1r,3r)-3-(cyanomethyl)-3-(4-(6-(1methyl-1H-pyrazol-4-yl)pyrazoIo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
    20. The compound of claim 1 wherein the compound is (1s,3s)-3-(cyanomethyl)-3-(4-(6-(1methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1 H-pyrazol-1-yl)cyclobutane-1-carbonitrile, or a pharmaceutically acceptable sait thereof.
    21. The compound of claim 1 wherein the compound is (1r,3r)-3-(cyanomethyl)-3-(4-(3methyl-6-(1-methyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)cyclobutane-1carbonitrile, or a pharmaceutically acceptable sait thereof.
    22. The compound of claim 1 wherein the compound is 2-((1 r,3s)-1-(4-(6-(3-amino-1H-pyrazol5-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazol-1-yl)-3-methoxycyclobutyl)acetonitrile, or a pharmaceutically acceptable sait thereof.
    23. The compound of claim 1 wherein the compound is 2-(1-ethyl-3-(4-(6-(5-(hydroxymethyl)1H-pyrazol-3-yl)pyrazolo[1,5-a]pyrazin-4-yl)-1H-pyrazo!-1-yl)azetidin-3-yl)acetonitrile, or, a pharmaceutically acceptable sait thereof.
    24. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait, and a pharmaceutically acceptable excipient.
    130
    25. A compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait for use in a melhod of treating a disease or condition for which a Tyk2 inhibitor is indicated.
    26. A compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait for use in a method of treating an inflammatory or autoimmune condition.
    27. A compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait for use in a method of treating a disease or condition selected from inflammation, autoimmune disease, systemic lupus erythematous, lupus nephritis, discoid lupus, cutaneous lupus, central nervous System lupus, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, asthma, allergie asthma, Type I diabètes, polymyositis, dermatomyositis, type I interferonopathies including Aicardi-Goutières syndrome and other mendelian diseases of overexpression of type I interferon, multiple sclerosis, primary progressive multiple sclerosis, relapsing remitting multiple sclerosis, primary biliary cirrhosis also known as primary biliary cholangitis, primary sclerosing cholangitis, autoimmune hepatitis, nonalcoholic fatty liver disease, non-alcoholic steatohepatitis, psoriasis, dermatomyositis, scleroderma, atopie dermatitis, vitiligo, alopecia areata, spondylopathy, ankylosing spondylitis, Alzheimer's disease, and neuro-inflammation.
    28. A compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait for use in a method of treating a symptom of an autoimmune or inflammatory disease.
    29. The compound for use of claim 28, wherein the symptom is selected from the group consisting of pruritis and fatigue.
    30. Use of a compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait in the manufacture of a médicament for treating a disease or condition for which a Tyk2 inhibitor is indicated.
    31. Use of a compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait in the manufacture of a médicament for treating an inflammatory or autoimmune condition.
    32. Use of a compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait in the manufacture of a médicament for treating a disease or condition selected from inflammation, autoimmune disease, systemic lupus
    131 erythematous, lupus nephritis, discoid lupus, cutaneous lupus, central nervous System lupus, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, asthma, allergie asthma, Type I diabètes, polymyositis, dermatomyositis, type I interferonopathies including Aicardi-Goutières syndrome and other mendelian diseases of overexpression of type I 5 interferon, multiple sclerosis, primary progressive multiple sclerosis, relapsing remitting multiple sclerosis, primary biliary cirrhosis also known as primary bitiary cholangitis, primary sclerosing cholangitis, autoimmune hepatitis, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, psoriasis, dermatomyositis, scleroderma, atopie dermatitis, vitiligo, alopecia areata, spondylopathy, ankylosing spondylitis, AJzheimer's disease, and neuro-inflammation.
    33. Use of a compound of claim 1, or a pharmaceutically acceptable sait thereof, or a pharmaceutically acceptable solvaté of said compound or sait in the manufacture of a médicament for treating a symptom of an autoimmune or inflammatory disease.
  15. 15 34· The use of c|airri 33, wherein the symptom is selected from the group consisting of pruritis and fatigue.
OA1201800314 2016-02-24 2017-02-10 Pyrazolo[1,5-A)pyrazin-4-yl derivatives as Jak-inhibitors OA18860A (en)

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