# Patent

## Title
Substituted pyrazolo[3,4-b]pyridines as medicaments

## Abstract

The present invention relates to a compound of following formula (I):




or a pharmaceutically acceptable salt or solvate of same, a tautomer of same, or a stereoisomer or mixture of stereoisomers of same in any proportions, such as a mixture of enantiomers, notably a racemic mixture; as well as to the use of same as a drug, notably intended for the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease; to the use of same as a kinase inhibitor; to the pharmaceutical compositions comprising same; and to methods for the preparation of same.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a Continuation of PCT International Application No. PCT/EP2012/051283 filed on Jan. 27, 2012, which claims priority under 35 U.S.C. §119(a) to Patent Application No. 1150651 filed in France on Jan. 27, 2011, all of which are hereby expressly incorporated by reference into the present application.


FIELD OF THE INVENTION
The present invention relates to azaindazole and diazaindazole fused bicyclic derivatives, as well as to the therapeutic use of same, notably in the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease, as well as to methods for synthesizing same.
BACKGROUND OF THE INVENTION
Protein kinases are enzymes that play a key role in cell signal transduction. They are involved in physiological processes such as cell proliferation, mitosis, differentiation, cell invasion and mobility, and apoptosis, for example.
Deregulation of the physiological mechanisms controlled by protein kinases is central to the appearance and development of many pathologies, notably including cancers. It is of particular note that many oncogenes and proto-oncogenes correspond to protein kinases.
Consequently, these enzymes are seen to play an important role during the various stages of tumor development and thus they constitute important pharmaceutical targets for cancer treatments.
Tyrosine kinase receptors (TKRs) form a particular class of protein kinases among which, among others, mention may be made of ALK, EGFR, Her2, PDGFR, Kit, VEGFR, IGFR, FGFR, Trk, Axl, Mer, Met, Ron and Ret. In this subfamily, ALK is regarded as a particularly relevant target because it is genetically modified in certain tumor pathologies and thus acquires an oncogenic nature. More precisely, chromosomal translocations leading to the production of fused protein kinases (ALK-X) which are then constitutively activated cause the development of certain cancers. ALK in oncogenic form is expressed by various tumor pathologies of different histological types. These pathologies are thus ALK-dependent. ALK in oncogenic form exists only in tumor cells and is not expressed by normal cells. For this reason, this protein kinase provides the opportunity to specifically target ALK-dependent tumor tissues while saving healthy tissues from significant toxic effects (Ott G. R. et al., Anticancer Agents Med. Chem., 2010, 10(3), 236-49).
Several cases of chromosomal translocations involving ALK, related to cancer pathologies, have already been documented. For example, the fusion protein NPM-ALK is associated with anaplastic large-cell lymphoma (ALCL) for which an optimal treatment remains to be developed. Similarly, the fusion protein EML4-ALK is associated with tumor development in a subpopulation of patients suffering from non-small cell lung cancer. Mutated forms of ALK have also been observed in neuroblastoma.
c-Src is also a protein kinase whose activation state proved to be negatively correlated with the survival of patients suffering from various forms of cancer, including non-small cell lung cancer (Byers L. A. et al., Clin. Cancer Res. 2009, 15(22), 6852-6861).
For this reason, and because of its involvement in many key mechanisms such as cell cycle progression, adhesion, proliferation, migration and control of apoptosis, this protein is also regarded as a target of interest in oncology.
It has been shown in particular that the inhibition of this target, by both biochemical and pharmacological means, induced effects such as a reduction in cell proliferation, a stopping of the mitotic cycle and a slowing of tumor growth in vivo. In the particular case of non-small cell lung cancer, the inhibition of c-Src by an inhibitor (dasatinib) led to the observation, in vitro, of inhibition of the migration and the invasion of the cells concerned.
Nevertheless, in terms of the control of tumor cell proliferation, it has been proposed that c-Src inhibition alone only induces a partial and/or transitory pharmacological response.
Consequently, there continues to be a need for inhibitors with a composite mode of action that are capable of intervening at several targets, in particular at several targets of the same signaling pathway, proposed as being more effective, with an improved therapeutic index and less likely to give rise to phenomena of compensation, resistance or therapeutic escape.
BRIEF SUMMARY OF THE INVENTION
The compounds of the present invention thus have the property of inhibiting or modulating the enzymatic activity of protein kinases in general and ALK and c-Src in particular. Consequently, said compounds can be used as drug in the treatment of proliferative diseases such as cancer.
Additional indications in inflammation or in affections of the central nervous system may also be pursued.


DETAILED DESCRIPTION OF THE INVENTION
More particularly, the present invention thus has as an object a compound of following general formula (I):




or a pharmaceutically acceptable salt or solvate of same, a tautomer of same, a stereoisomer or a mixture of stereoisomers of same in any proportions, such as a mixture of enantiomers, notably a racemic mixture,

wherein:

    
    
        Y1 and Y4 each represent, independently of each other, a CH group or a nitrogen atom,
        Y2 represents a nitrogen atom or a CH or C—X—Ar group,
        Y3 represents a nitrogen atom or a C—X—Ar or C—W group, on the condition that:
        
            at least one and at most two Y1, Y2, Y3, and Y4 groups represent a nitrogen atom,
            Y2 and Y4 cannot represent a nitrogen atom at the same time,
            when Y2═C—X—Ar, then Y3 represents a nitrogen atom or a C—W group, and
            when Y3═C—X—Ar, then Y2 represents a nitrogen atom or a CH group,
        
        
        Ar represents an aryl or heteroaryl group optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, NR24SO2R25, and R26NR27R28 and/or optionally fused to a heterocycle,
        X represents a divalent group selected from O, S, S(O), S(O)2, NR4, S(NR4), S(O)(NR4), S(O)2(NR4), NR4S, NR4S(O), NR4S(O)2, CH2, CH2S, CH2S(O), CH2S(O)2, SCH2, S(O)CH2, S(O)2CH2, CH2CH2, CH═CH, C≡C, CH2O, OCH2, NR4CH2, and CH2NR4,
        W represents an R5, SR5, OR5 or NR5R6 group,
        U represents a CH2 or NH group, one or more hydrogen atoms which may be replaced by a (C1-C6)alkyl group,
        V represents C(O), C(S) or CH2,
        n represents 0 or 1,
        R1 represents a hydrogen atom, or an OR7 or NR7R8 group,
        R2 represents a hydrogen atom, an optionally substituted heterocycle, NO2, OR9 or NR9R10,
        R3, R4, R11 to R25 and R27 to R28 each represent, independently of each other, a hydrogen atom or a (C1-C6)alkyl group,
        R5 and R6 each represent, independently of each other, a hydrogen atom or a (C1-C6)alkyl, optionally substituted aryl or optionally substituted benzyl group,
        R7, R8, R9 and R10 each represent, independently of each other, a hydrogen atom or an optionally substituted (C1-C6)alkyl or (C3-C12)cycloalkyl group or an optionally substituted heterocycle, and
        R26 represents (C1-C6)alkyl.
    
    


In the preceding definitions, all the combinations of substituents or variables are possible insofar as they lead to stable compounds.
The term “halogen” refers to fluorine, chlorine, bromine or iodine.
The term “(C1-C6)alkyl” refers to saturated linear or branched hydrocarbon chains comprising 1 to 6 carbon atoms. It may be a methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl or hexyl group.
The term “(C1-C6)alkoxy” refers to a (C1-C6)alkyl chain linked to the rest of the molecule via an oxygen atom. As an example, mention may be made of methoxy, ethoxy, propoxy, isopropoxy, butoxy or tert-butoxy groups.
The term “(C1-C6)thioalkoxy” refers to a (C1-C6)alkyl chain linked to the rest of the molecule via a sulfur atom. As an example, mention may be made of thiomethoxy, thioethoxy, thiopropoxy, thioisopropoxy, thiobutoxy or thio-tert-butoxy groups.
The term “(C1-C6)haloalkyl” refers to a (C1-C6)alkyl chain such as defined above wherein one or more hydrogen atoms are replaced by a halogen atom such as defined above. It may be in particular a trifluoromethyl group.
The term “(C1-C6)haloalkoxy” refers to a (C1-C6)alkoxy chain such as defined above wherein one or more hydrogen atoms are replaced by a halogen atom such as defined above. It may be in particular a trifluoromethoxy group.
The term “(C1-C6)halothioalkoxy” refers to a (C1-C6)thioalkoxy chain such as defined above wherein one or more hydrogen atoms are replaced by a halogen atom such as defined above. It may be in particular a trifluorothiomethoxy group.
The term “(C3-C12)cycloalkyl” refers to cyclic hydrocarbon systems comprising from 3 to 12 carbon atoms and comprising one or more rings, in particular fused rings. As an example, mention may be made of an adamantyl or cyclohexyl group.
The term “aryl” refers to an aromatic hydrocarbon group preferably comprising from 6 to 14 carbon atoms and comprising one or more fused rings, such as, for example, a phenyl or naphthyl group. Advantageously, it is a phenyl group.
The term “heteroaryl” refers to a cyclic aromatic group comprising 5 to 7 atoms included in the ring or a bicyclic aromatic group comprising 8 to 11 atoms included in the rings, wherein 1 to 4 of the atoms included in the rings are a heteroatom selected independently from sulfur, nitrogen and oxygen atoms, and wherein the other atoms included in the rings are carbon atoms. Examples of heteroaryl groups include furyl, thienyl, pyridinyl, and benzothienyl groups.
The term “heterocycle” refers either to a stable monocycle containing from 4 to 7 cyclic atoms, or to a stable bicycle containing from 8 to 11 cyclic atoms, which may be either saturated or unsaturated, wherein 1 to 4 of the cyclic atoms are a heteroatom selected independently from sulfur, nitrogen and oxygen atoms, and wherein the other cyclic atoms are carbon atoms. As an example, mention may be made of furan, pyrrole, thiophene, thiazole, isothiazole, oxadiazole, imidazole, oxazole, isoxazole, pyridine, piperidine, pyrazine, piperazine, tetrahydropyran, pyrimidine, quinazoline, quinoline, quinoxaline, benzofuran, benzothiophene, indoline, indolizine, benzothiazole, benzothienyl, benzopyran, benzoxazole, benzo[1,3]dioxole, benzisoxazole, benzimidazole, chromane, chromene, dihydrobenzofuran, dihydrobenzothienyl, dihydroisoxazole, isoquinoline, dihydrobenzo[1,4]dioxane, imidazo[1,2-a]pyridine, furo[2,3-c]pyridine, 2,3-dihydro-1H-indene, [1,3]dioxolo[4,5-c]pyridine, pyrrolo[1,2-c]pyrimidine, pyrrolo[1,2-a]pyrimidine, tetrahydronaphthalene, benzo[b][1,4]oxazin.
In the context of the present invention, “optionally substituted” means that the group in question is optionally substituted by one or more substituents which may be selected in particular from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, NR24SO2R25, and R26NR27R28, wherein R11 to R28 are such as defined above.
In the present invention, “pharmaceutically acceptable” refers to that which is useful in the preparation of a pharmaceutical composition that is generally safe, nontoxic and neither biologically nor otherwise undesirable and that is acceptable for veterinary and human pharmaceutical use.
“Pharmaceutically acceptable salt or solvate” of a compound refers to salts and solvates which are pharmaceutically acceptable, as defined herein, and which has the desired pharmacological activity of the parent compound.
Acceptable salts for the therapeutic use of the compounds of the present invention include the conventional nontoxic salts of the compounds of the invention such as those formed from pharmaceutically acceptable organic or inorganic acids or from pharmaceutically acceptable organic or inorganic bases. As an example, mention may be made of salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid and sulfuric acid, and those derived from organic acids such as acetic acid, trifluoroacetic acid, propionic acid, succinic acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, glutamic acid, benzoic acid, salicylic acid, toluenesulfonic acid, methanesulfonic acid, stearic acid and lactic acid. As an example, mention may be made of salts derived from inorganic bases such as soda, potash or calcium hydroxide and salts derived from organic bases such as lysine or arginine.
These salts may be synthesized from the compounds of the invention containing a basic or acidic part and the corresponding acids or bases according to conventional chemical methods well known to the person skilled in the art.
Acceptable solvates for the therapeutic use of the compounds of the present invention include conventional solvates such as those formed during the last step of the preparation of the compounds of the invention due to the presence of solvents. As an example, mention may be made of solvates due to the presence of water or ethanol.
In the context of the present invention, “stereoisomer” refers to a geometric isomer or an optical isomer.
Geometric isomers result from the different position of substituents on a double bond which can then have a Z or E configuration.
Optical isomers result notably from the different position in space of substituents on a carbon atom comprising four different substituents. This carbon atom thus constitutes a chiral or asymmetrical center. Optical isomers include diastereoisomers and enantiomers. Optical isomers that are mirror images of each other but are non-superimposable are enantiomers. Optical isomers that are not mirror images of each other are diastereoisomers.
In the context of the present invention, “tautomer” refers to a constitutional isomer of the compound obtained by prototropy, i.e., by migration of a hydrogen atom and a change in location of a double bond. The different tautomers of a compound are generally interconvertible and are in equilibrium in solution in proportions which may vary according to the solvent used, the temperature or the pH.
According to a first embodiment, Y4═N.
Y2═C—X—Ar and Y3 preferably represents a C—W group.
In particular:

    Y1═CH or N, and advantageously CH,
    Y2═C—X—Ar,
    Y3═C—W, and
    Y4═N.


According to a second embodiment, Y1 and/or Y4 represent a nitrogen atom.
In this case, Y2 and Y3 preferably do not represent a nitrogen atom.
In particular:

    Y1 and/or Y4═N,
    Y2═CH or C—X—Ar, and
    Y3═C—W or C—X—Ar.


Advantageously, X represents a divalent group selected from O, S, S(O), S(O)2, NR4, CH2, CH2S, CH2S(O), CH2S(O)2, NHS(O)2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, CH2CH2, CH═CH, C≡C, CH2O, OCH2, NR4CH2, and CH2NR4.
In particular, X represents a divalent group selected from S, S(O), S(O)2, NR4, CH2, CH2S, CH2S(O), CH2S(O)2, NHS(O)2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, CH2CH2, C≡C, CH2O, OCH2, NR4CH2, and CH2NR4.
More particularly, X may be selected from S, S(O), S(O)2, CH2, CH2S, CH2S(O), CH2S(O)2, NHS(O)2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, CH2CH2, CH═CH, and C≡C.
In particular, X may be selected from S, S(O)2, CH2, SCH2, S(O)2CH2, S(O)2NH, CH2S, CH2S(O)2, NHS(O)2, CH2CH2, and C≡C.
X may notably be selected from S, S(O), S(O)2, NR4, CH2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, CH2CH2, C≡C, OCH2, and NR4CH2; notably from S, S(O)2, CH2, SCH2, S(O)2CH2, S(O)2NH, CH2CH2, and C≡C, wherein the first atom of these groups is bound to atom C of the C—X—Ar chain.
X may be in particular S, S(O)2, SCH2, S(O)2CH2, S(O)2NH, CH2S, CH2S(O)2, or NHS(O)2; and notably S, S(O)2, SCH2, S(O)2CH2, or S(O)2NH, wherein the first atom of these groups is bound to atom C of the C—X—Ar chain.
Advantageously, Ar represents a heteroaryl group, such as pyridine, or an aryl group, such as phenyl, optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, and NR24SO2R25; and/or optionally fused to a heterocycle.
More particularly, Ar may represent an aryl group, such as phenyl, optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, and NR24SO2R25.
Ar may notably represent an aryl group, such as phenyl, optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, and CONR16R17, and in particular from a halogen atom such as fluorine, (C1-C6)alkyl such as methyl, and CONR16R17 such as CONH2.
Ar can also represent a pyridine group.
Ar may notably be selected from the following groups:




notably from the following groups:





in particular, from the following groups:





Ar may advantageously represent the group:




W may advantageously represent an R5, SR5, OR5 or NR5R6 group, and preferably R5, OR5 or NR5R6, with R5 and R6 representing, independently of each other, a hydrogen atom or a (C1-C6)alkyl group.
W may represent in particular H, OMe, Me, OH or NH2, and notably H.
Advantageously, R3 represents a hydrogen atom.
U may represent more particularly a CH2 or NH group.
Advantageously, n may represent 0.
V may represent more particularly a C(O) or C(S) group, and advantageously a C(O) group.
According to a particular embodiment of the invention:

    
    
        R3═H,
        U═CH2 or NH,
        V═C(O) or C(S), and notably C(O), and
        n=0 or 1, and notably 0.
    
    


According to another particular embodiment of the invention:

    
    
        V═C(O) or C(S), and notably C(O), and
        n=0.
    
    


According to still another particular embodiment of the invention:

    
    
        R3═H,
        V═C(O) or C(S), and notably C(O), and
        n=0.
    
    


R1 may represent more particularly a hydrogen atom or an NR7R8 group, with R7 notably representing a hydrogen atom and R8 notably representing an optionally substituted (C3-C12)cycloalkyl group or an optionally substituted heterocycle.
The (C3-C12)cycloalkyl group may be in particular a cyclohexyl. It may be substituted by one or more halogen atoms. It may be in particular the group:




The heterocyclic group may be in particular a tetrahydropyran, notably unsubstituted. It may thus be the following group:




R1 may thus represent more particularly one of the following groups: H,




and notably H and





and advantageously





R2 may represent more particularly an optionally substituted heterocycle (notably substituted by (C1-C6)alkyl or NH2), NO2 or NR9R10, with notably R9═R10═H or else R9 and R10 each represent H or an optionally substituted (C1-C6)alkyl.
R2 may represent in particular an optionally substituted heterocycle, notably substituted by (C1-C6)alkyl or NH2. The heterocycle may be in particular a heterocycle with 5 or 6 members comprising at least one nitrogen atom, and in particular one or two. The heterocycle may thus be selected from piperazine, piperidine and pyrrolidine.
R2 may notably represent one of the following groups:




and notably NH2, NO2,





and in particular





and more particularly





The compounds of the present invention may be selected from the compounds cited in the following table:













14-2






 


14-10






 


14-11






 


15






 


26-4






 


26-8






 


27






 


27-1






 


28






 


29






 


29-a






 


30






 


30-1






 


30-3






 


30-4






 


30-5






 


30-8






 


30-9






 


30-10






 


30-11






 


30-12






 


30-a






 


31






 


32






 


32-1






 


33






 


35






 


26-12






 


30-69






 


27-2






 


27-3






 


27-4






 


30-73






 


14bis






 


30-70






 


30-71






 


30-72






 


27-5






 


30-13






 


30-14






 


30-15






 


30-16






 


30-17






 


30-18






 


30-19






 


30-20






 


30-21






 


30-22






 


30-23






 


30-24






 


30-25






 


30-26






 


30-27






 


30-28






 


30-29






 


30-30






 


30-31






 


30-32






 


30-33






 


30-34






 


30-35






 


30-36






 


30-37






 


30-38






 


30-39






 


30-40






 


30-41






 


30-42






 


30-43






 


30-44






 


30-45






 


30-46






 


30-47






 


30-48






 


30-49






 


30-50






 


30-51






 


30-52






 


30-53






 


30-54






 


30-55






 


30-56






 


30-57






 


30-58






 


30-59






 


30-60






 


30-61






 


30-62






 


30-63






 


30-64






 


30-65






 


30-66






 


30-67






 


30-68






 


31-1













The present invention also has as an object a compound according to the invention of formula (I) such as defined above, to be used as a drug, notably intended for the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease, in particular cancer.
The present invention also relates to the use of a compound of formula (I) such as defined above, for the manufacture of a drug, notably intended for the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease, in particular cancer.
The present invention also relates to a method for the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease, in particular cancer, comprising the administration to a person in need thereof of an effective dose of a compound of formula (I) such as defined above.
The cancer may be more particularly in this case colon cancer, breast cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, glioblastoma, non-small cell lung cancer, neuroblastoma, inflammatory myofibroblastic tumor, diffuse B-cell lymphoma or anaplastic large-cell lymphoma.
The present invention also relates to a compound according to the invention of formula (I) such as defined above, to be used as a drug intended for the treatment of a disease associated with a kinase, and in particular a tyrosine kinase such as the kinases ALK, Abl and/or c-Src, and in particular ALK. The disease may be in particular associated with ALK and at least one other kinase, for example Abl or c-Src, in particular ALK and c-Src.
The present invention also has as an object a compound according to the invention of formula (I) such as defined above, to be used as a kinase inhibitor, and in particular an inhibitor of tyrosine kinases such as ALK, Abl and/or c-Src, and in particular ALK. The compounds according to the invention may notably be used as an inhibitor of ALK and at least one other kinase, for example Abl or c-Src. Preferentially, the compounds according to the invention are inhibitors of ALK and c-Src.
In the context of the present invention, “disease associated with a kinase” or “kinase-associated disease” refers to any diseases, and in particular diseases related to deregulation of cell proliferation, in particular cancers, due to deregulation of the expression or activity of said kinase compared to its normal state of expression or activity. Deregulation of the expression of said kinase may be modification of the sequence expressed or modification of the quantity of protein expressed. These deregulations may lead to changes in cells which may, in particular, result in proliferative disorders including cancers. Preferentially, according to the invention, kinase-associated diseases may be cancers related to deregulation of ALK and/or c-Src activity such as, for example, colon cancer, breast cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, glioblastoma, non-small cell lung cancer, neuroblastoma, inflammatory myofibroblastic tumors, diffuse B-cell lymphoma and anaplastic large-cell lymphoma.
According to the invention, the expression “inhibitor of kinases” or “kinase inhibitor” refers to molecules that are able to interact with the kinase and to reduce its activity. Preferentially, the use of a kinase inhibitor according to the invention makes it possible to suppress the activity of said kinase.
The present invention also relates to a pharmaceutical composition comprising at least one compound of formula (I) such as defined above, and at least one pharmaceutically acceptable excipient.
The pharmaceutical compositions according to the invention may be formulated notably for oral administration or for injection, wherein said compositions are intended for mammals, including humans.
The active ingredient may be administered in unit dosage forms of administration, in mixture with standard pharmaceutical carriers, to animals or to humans. The compounds of the invention as active ingredients may be used in doses ranging between 0.01 mg and 1000 mg per day, given in a single dose once per day or administered in several doses throughout the day, for example twice a day in equal doses. The dose administered per day advantageously is between 5 mg and 500 mg, even more advantageously between 10 mg and 200 mg. It may be necessary to use doses outside these ranges as determined by the person skilled in the art.
The pharmaceutical compositions according to the invention may further comprise at least one other active ingredient, such as an anticancer agent.
The present invention also has as an object a pharmaceutical composition comprising:

    
    
        (i) at least one compound of formula (I) such as defined above, and
        (ii) at least one other active ingredient, such as an anticancer agent, as a combination product for simultaneous, separate or sequential use.
    
    


The present invention also relates to a pharmaceutical composition such as defined above to be used as a drug, notably intended for the treatment of cancer, inflammation and neurodegenerative diseases such as Alzheimer's disease, in particular cancer.
The present invention also has as an object method for the preparation of the compounds of formula (I) according to the invention.
According to a first embodiment, the present invention relates to a method for the preparation of a compound of formula (I) according to the invention wherein V═C(O) or C(S), preferably C(O), and notably U═CH2, comprising the following successive steps:

    (a1) coupling between a compound of following formula (A):






    
    
        wherein Y1, Y2, Y3 and Y4 are such as defined above, and R29 represents a hydrogen atom or an N-protecting group,
        with a compound of following formula (B):
    
    






    
    
        wherein R1, R2, U and n are such as defined above, V═C(O) or C(S), and R30═OH or a leaving group such as Cl,
        to yield a compound of following formula (C):
    
    






    
    
        wherein Y1, Y2, Y3, Y4, R1, R2, R29, U and n are such as defined above and V═C(O) or C(S),
    
    
    (b1) optionally substitution of the nitrogen atom bound to V of the compound of formula (C) obtained in the preceding step with an R3 group other than H and/or deprotection of the nitrogen atom carrying an R29 group representing an N-protecting group to yield a compound of formula (I) with V═C(O) or C(S), and
    (c1) optionally forming of a salt of the compound of formula (I) obtained in the preceding step to yield a pharmaceutically acceptable salt of same.


In the context of the present invention, “N-protecting group” refers to any substituent that protects the NH or NH2 group against undesirable reactions such as the N-protecting groups described in Greene, “Protective Groups in Organic Synthesis” (John Wiley & Sons, New York (1981)) and Harrison et al., “Compendium of Synthetic Organic Methods”, Vols. 1 to 8 (J. Wiley & Sons, 1971 to 1996). N-protecting groups include carbamates, amides, N-alkylated derivatives, amino acetal derivatives, N-benzylated derivatives, imine derivatives, enamine derivatives and N-heteroatom derivatives. In particular, the N-protecting group consists of formyl, acetyl, benzoyl, pivaloyl, phenylsulfonyl, trityl(triphenylmethyl), tert-butyl, benzyl (Bn), t-butyloxycarbonyl (BOC), benzyloxycarbonyl (Cbz), p-methoxybenzyloxycarbonyl, p-nitrobenzyl-oxycarbonyl, trichloroethoxycarbonyl (TROC), allyloxycarbonyl (Alloc), 9-fluorenylmethyloxycarbonyl (Fmoc), trifluoro-acetyl, benzyl carbamates (substituted or not) and the like. It may be in particular a trityl, tert-butyl or BOC group.
In the context of the present invention, “leaving group” refers to a chemical group which may be easily displaced by a nucleophile during a nucleophilic substitution reaction, wherein the nucleophile is more particularly an amine, and notably a primary or secondary amine. Such a leaving group may be more particularly a halogen atom such as a chlorine atom, a mesylate (CH3—S(O2)O—), a triflate (CF3—S(O)2O—) or a tosylate (p-Me-C6H4—S(O)2O—).
Step (a1):
Coupling between (A) and (B) may be carried out by techniques well known to the person skilled in the art.
When R30═OH, the coupling may be carried out under peptide coupling conditions. It may thus be carried out in the presence of a coupling agent such as diisopropylcarbodiimide (DIC), dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), carbonyldiimidazole (CDI), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU) or O-(7-azobenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU); optionally combined with a secondary coupling agent such as N-hydroxysuccinimide (NHS), N-hydroxybenzotriazole (HOBt), 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazole (HOOBt), 1-hydroxy-7-azabenzotriazole (HAt) or N-hydroxysulfosuccinimide(sulfo NHS). Peptide coupling may moreover be carried out in an aprotic solvent such as tetrahydrofuran, dioxane and dichloromethane.
When R30 is a leaving group such as Cl, coupling may be carried out in the presence of a base such as pyridine, triethylamine or diisopropylethylamine (DIPEA). The reaction may be carried out in an aprotic solvent such as tetrahydrofuran, toluene or dichloromethane, or in a base solvent such as pyridine.
The compounds of formula (A) and (B) can be prepared by the methods described in further detail below.
Step (b1):
In the context of the present invention, “deprotection” refers to the process by which a protecting group is eliminated once the selective reaction is completed. Certain protecting groups may be preferred over others due to their convenience or their relative ease of elimination.
The deprotection step may be carried out under conditions well known to the person skilled in the art.
The substitution step may also be carried out by techniques well known to the person skilled in the art. If necessary, functionalities that may be sensitive to the reaction conditions of the substitution step may be protected beforehand and then deprotected once substitution is carried out.
Thus, if a step of deprotection of the nitrogen atom carrying an R29 group representing an N-protecting group and a step of substitution of the nitrogen atom bound to V with an R3 group must be carried out, the order in which these two steps are carried out will depend on the reaction conditions of each of these steps.
Moreover, it may also be necessary to carry out additional steps of functionalization of the molecule by techniques known to the person skilled in the art.
Step (e1):
This step may be carried out in the presence of a pharmaceutically acceptable organic or inorganic acid or a pharmaceutically acceptable organic or inorganic base such as defined above.
According to a second embodiment, the present invention relates to a method for the preparation of a compound of formula (I) according to the invention wherein V═CH2, and notably U═CH2, comprising the following successive steps:

    (a2) reducing amination reaction between a compound of formula (A) such as defined above and an aldehyde of following formula (D):






    
    
        wherein R1, R2, U and n are such as defined above,
        to yield a compound of following formula (E):
    
    






    
    
        wherein Y1, Y2, Y3, Y4, R1, R2, R29, U and n are such as defined above,
    
    
    (b2) optionally deprotection of the nitrogen atom carrying an R29 group representing an N-protecting group and/or substitution of the nitrogen atom bound to V with an R3 group other than H of the compound of formula (E) obtained in the preceding step to yield a compound of formula (I) with V═CH2, and
    (c2) optionally forming of a salt of the compound of formula (I) obtained in the preceding step to yield a pharmaceutically acceptable salt of same.


Step (a2):
This step is carried out in the presence of a reducing agent such as a borohydride and in particular NaBH4, NaBH(OAc)3 or NaBH3CN.
This reaction is more particularly carried out at room temperature, i.e., at a temperature ranging between 15° C. and 40° C., in particular between 20° C. and 30° C.
The reaction may be typically carried out in a solvent such as dichloroethane (DCE), tetrahydrofuran (THF) or acetonitrile, optionally in the presence of water, acetic acid or trifluoroacetic acid.
The compounds of formula (A) and (D) can be prepared by the methods described in further detail below.
Step (b2): see step (b1)
Step (c2): see step (c1)
According to a third embodiment, the present invention relates to a method for the preparation of a compound of formula (I) according to the invention wherein V═C(O) or C(S), n=1 and U═NH, comprising the following successive steps:

    (a3) coupling between a compound of formula (A) such as defined above and a compound of following formula (F):






    
    
        wherein R1 and R2 are such as defined above and Z═O or S,
        to yield a compound of following formula (G):
    
    






    
    
        wherein Y1, Y2, Y3, Y4, R1, R2, R29, and Z are such as defined above,
    
    
    (b3) optionally deprotection of the nitrogen atom carrying an R29 group representing an N-protecting group and/or substitution of the nitrogen atom bound to V with an R3 group other than H of the compound of formula (G) obtained in the preceding step to yield a compound of formula (I) with V═C(O) or C(S), n=1 and U═NH, and
    (c3) optionally forming of a salt of the compound of formula (I) obtained in the preceding step to yield a pharmaceutically acceptable salt of same.


Step (a3):
This step may be carried out under conditions well known to the person skilled in the art.
A polar or non-polar protic solvent may be more particularly used such as dichloromethane, acetone, acetonitrile, tetrahydrofuran or dioxane.
The compounds of formula (A) and (F) can be prepared by the methods described in further detail below.
Step (b3): see step (b1)
Step (c3): see step (c1)
Once the compound of formula (I) is obtained by any one of the preceding methods, it may be separated from the reaction medium by techniques well known to the person skilled in the art, and notably by evaporation of the solvent, crystallization and filtration, etc.
The compound obtained may be purified if necessary by techniques well known to the person skilled in the art, and notably by high-performance liquid chromatography (HPLC), silica gel chromatography, recrystallization when the compound is crystalline, etc.
Thus, the compounds of formula (I) according to the present invention can be prepared by the various methods summarized in diagrams 1a and 1b below.








Method A:

According to method A, compounds of formula (I) are obtained by the preliminary synthesis of compounds of general formula (V) characterized by a halogenated heterobicyclic ring having an exocyclic primary amine. These compounds are obtained via the synthesis of intermediates of general formula (II) or (III).
Method A1:
Method A1, presented in diagram 2 (iodized compounds) or 3 (brominated compounds) below, describes the general process giving access to compounds of general formula (V) with W defined as in the description of general formula (I), and notably H, (C1-C6)alkyl or aryl, and Rj═H or N-protecting group.




In the context of diagram 2, the optionally substituted 2-chloro-5-iodonicotinonitrile (IIa) is obtained from the corresponding hydroxynicotinonitrile by the successive use of an iodination agent such as N-iodosuccinimide (NIS), or molecular iodine with an inorganic base such as, for example, K2CO3 or Na2CO3, notably in a polar solvent such as hot DMF, followed by treatment with phosphorus oxychloride, pure or diluted in a high boiling-point non-polar solvent, or any other equivalent chlorination agent well known to the person skilled in the art. Reaction temperatures are between −20° C. and 200° C. The compound (IIa) thus obtained is then transformed into optionally substituted 5-iodo-pyrazolo[3,4-b]pyridine-3-amine (Va) by its reaction, preferably under heat, in the presence of a hydrazine optionally carrying an N-protecting group such as trityl, tert-butyl or BOC.
The brominated analogues of general formula (V) as described in diagram 1a may be obtained by the use of the method described in the following references: Witherington et al., Bioorg. Med. Chem. Lett., 2003, 13, 1577-1580 and Lijuan Chen et al., Bioorg. Med. Chem. Lett., 2010, 20, 4273-4278. For reasons of convenience, these molecules were obtained by the use of the reaction sequence presented in following diagram 3.




The optionally functionalized 2-methoxy-nicotinotrile is obtained, for example, by reaction of sodium methanolate in methanol at a temperature between −20° C. and the boiling point of the mixture. Alternatively, this compound may be obtained by methylation of 2-hydroxynicotinonitrile or other methods described above. Bromination of 2-methoxy-nicotinonitrile is typically carried out with dibromine in acetic acid at a temperature varying between 20° C. and 110° C. Formation of the pyrazole is typically carried out by reaction of an excess of hydrazine, functionalized or not, at a temperature varying between 20° C. and 100° C. in the presence of a polar solvent such as water, ethanol, tetrahydrofuran (THF) or any other solvent with comparable properties. Alternatively, the use of hydrazine in a saline or hydrated form, without solvent, is also possible.
Method A2:
Method A2 relates to the synthesis of the functionalized pyrazolopyrazines presented in diagram 4 below with Rj═H or N-protecting group, Hal=halogen and in particular W═H, (C1-C6)alkyl or aryl.




The optionally functionalized 3-amino-6-iodopyrazine-2-carboxamides are typically obtained in two steps from the corresponding methyl 3-aminopyrazine-2-carboxylates by iodination in the presence of N-iodosuccinimide or molecular iodine optionally in the presence of a cofactor such as KIO3, AgCO2CF3, Ag2SO4, AlCl3, CuCl2 or HgO, followed by a conversion reaction of the methyl ester function into carboxamide, notably by the use of ammonia in a polar solvent such as water, methanol or THF at temperatures varying between 0° C. and 100° C. The carboxamide function of the optionally functionalized 3-amino-6-iodopyrazine-2-carboxamide is then converted into nitrile by the use of dehydration agents such as, in particular, CCl4/PPh3, SOCl2, PhSO2Cl, P2O5, TsCl, COCl2, DCC/py (N,N′-dicyclohexylcarbodiimide/pyridine) or (COCl)2 used as the case may be in the presence of an organic base such as pyridine. The preferred method involves the use of phosphorus oxychloride in dimethylformamide (DMF). Deprotection of the dimethylformimidamide function is carried out by treatment with acid such as aqueous hydrochloric acid or any other reagent with equivalent properties. Formation of the pyrazole ring is carried out by a Sandmeyer reaction, well known to the person skilled in the art, followed by a reaction in the presence of a hydrazine, functionalized or not, under conditions as described in the methods above. Alternatively, the diazonium salt, an intermediate of the Sandmeyer reaction, may be reduced by the use, for example, of tin chloride in an acid medium or any other equivalent agent, in order to form a hydrazine function that can undergo intramolecular cyclization under the effect of heat.
Method A3:
Method A3 aims at obtaining derivatives of general formula (V) featuring a variable function in position 6 of the pyrazolopyridine bicycle. It is detailed in diagram 5 below.




Reaction of the cyanothioacetamide with ethyl 3-ethoxyacrilates variously substituted according to methods described notably by Litrivnor et al. in Russ. Chem. Bull., 1999, 48(1), 195-196 and Tsann-Long Su et al. in J. Med. Chem., 1988, 31, 1209-1215 make it possible to yield access, in two steps, to ethyl 5-cyano-6-(methylthio)nicotinates carrying a variable functionality in position 2. These syntheses are typically carried out, for the first step, in an anhydrous polar solvent such as, for example, ethanol at a temperature ranging between 0° C. and 70° C. in the presence of an organic base such as methylmorpholine, triethylamine, DIPEA (N,N-diisopropylethylamine) or DBU (1,8-diazabicyclo[5,4,0]undec-7-ene). The second step of intramolecular cyclization and of alkylation is typically carried out by the heating to a temperature ranging between 20° C. and 100° C. of a solution of the intermediate thioamidate in a polar solvent, for example ethanol in the presence of a suitable alkylating agent such as alkyl halide or dialkyl sulfate.
The 5-cyano-6-(methylthio)nicotinic acids substituted in position 2 are typically obtained by saponification of the corresponding ethyl esters according to methods well known to the person skilled in the art, notably by the use of hot lithium hydroxide. Decarboxylation of these compounds is carried out by heat treatment in a high boiling-point solvent such as diphenylether at a temperature ranging between 150° C. and 250° C.
Halogenation reactions principally aim at obtaining iodinated, brominated or chlorinated derivatives, more particularly iodinated derivatives. The latter are typically obtained by a molecular iodine treatment in the presence of a silver salt such as, for example, Ag2SO4 in a polar solvent such as ethanol at a temperature ranging between 0° C. and 70° C. Alternative methods, notably those based on other salts such as KIO3, AgCO2CF3, AlCl3, CuCl2 or HgO, or other iodination agents such as N-iodosuccinimide, are also considered. The conceivable bromination methods typically rely on agents such as N-bromosuccinimide or dibromine according to methods well known to the person skilled in the art.
In the case in which W═OH (typically resulting from the use of diethyl 2-(ethoxymethylene)malonate), the corresponding compounds are protected by an alkylation reaction. This reaction is notably carried out by the use of methyl iodide or bromomethane, and silver carbonate in dioxane, THF, acetonitrile or acetone, or any other equivalent agent such as dimethylsulfate. The 5-halo-2-(methylthio)nicotinonitriles obtained are subjected to oxidation of their thiomethoxy function, typically by the use of m-CPBA (m-chloroperbenzoic acid), oxone or any other equivalent agent, to lead to the formation of the corresponding sulfoxide. These compounds, which may contain variable quantities of the corresponding sulfone, are engaged in a reaction in the presence of an optionally substituted hydrazine to form the corresponding 5-halogeno-pyrazolo[3,4-b]pyridin-3-amine carrying a variable functionality in position 6.
Method A4:
Method A4 aims at obtaining derivatives of general formula (V) from the compounds of general formula (III) via intermediate formation of compounds of formula (IV). These compounds are typically obtained by the pathway presented in diagram 6. The following references illustrate the method used: Gueiffier et al. Heterocycles, 1999, 51(7), 1661-1667; Gui-Dong Zhu et al. Bioorg. Med. Chem., 2007, 15, 2441-2452.




The compounds of general formula (IIIa), acetylated beforehand by one or another of the methods well known to the person skilled in the art, are subjected to the action of isoamyl nitrite, sodium nitrite or any other equivalent organic or inorganic nitrite, in water or acetic acid, for periods typically varying from 1 to 3 days at temperatures varying between 0° C. and 40° C. The compounds of general formula (IVa) thus obtained are deprotected in acidic conditions, for example by the use of hydrochloric acid, before being subjected to the action of nitration agents such as concentrated nitric acid or potassium nitrate in sulfuric acid at temperatures varying between 0° C. and 25° C.
It should be noted that the direct conversion of compounds of general formula (IIIa) into deprotected compounds (IVb) is possible in general.
The nitropyrazoles thus obtained are typically reduced into aminopyrazoles of general formula (Ve) by the use of SnCl2 in hydrochloric acid. Alternative methods include the use of iron, zinc or tin in acidic conditions and methods of catalytic hydrogenation in the presence of complexes of platinum, nickel or Pd/C under an atmosphere of hydrogen or in the presence of equivalent agents such as cyclohexadiene, cyclohexene, sodium borohydride or hydrazine.
Method B:
According to method B, the compounds of formula (I) are obtained by the preliminary synthesis of compounds of general formula (VI) characterized by a functionalized heterobicyclic ring possessing an exocyclic amine. These compounds are obtained via the synthesis of intermediates of general formula (VI).
Method B1:
Method B1 is represented in diagram 7 below, with W notably representing H, (C1-C6)alkyl, aryl or benzyl.




The 3-nitro-6-thioxo-1,6-dihydropyridin-2-carbonitrile and 3-nitro-6-thioxo-1,6-dihydropyrazine-2-carbonitrile derivatives, optionally functionalized in position 5, are typically obtained from the corresponding 2,6-dichloro-3-nitropyridines or 2,6-dichloro-3-nitropyrazines by the successive reactions of a cyanide salt, such as copper cyanide, in a high boiling-point polar solvent such as N-methylpyrrolidone at temperatures ranging between 100° C. and 200° C.; followed by the reaction of aqueous sodium hydrosulfite in a polar solvent. These compounds are then alkylated, for example by the use of a substituted benzyl bromide, in basic medium, according to methods well known to the person skilled in the art. The preferred protocol includes the use of an aprotic and anhydrous polar solvent such as acetone carrid at its boiling point and an organic base such as pyridine, triethylamine or DIPEA, or an inorganic base such as sodium, potassium or calcium carbonate. Reactions for reducing the nitro function in amine are preferentially carried out by the use of SnCl2 in hydrochloric acid. Alternative methods include the use of iron, zinc or tin in acidic conditions and methods of catalytic hydrogenation in the presence of complexes of platinum, nickel or Pd/C under an atmosphere of hydrogen or in the presence of equivalent agents such as cyclohexadiene, cyclohexene, sodium borohydride or hydrazine.
In certain cases, the product of the reduction reaction, in addition to having a primary amine, has a carboxamide function resulting from hydrolysis of the nitrile function. In this case, isolation of the corresponding 3-aminopicolinonitriles or 3-aminopyrazine-2-carbonitriles may be carried out by dehydration of the carboxamide into nitrile via the use of phosphorus oxychloride in the presence of DMF or any other method well known to the person skilled in the art. Lastly, formation of the aminopyrazole ring is carried out preferentially by the formation of a diazonium, obtained by the successive reaction at low temperature of isoamyl nitrite, sodium nitrite or any other equivalent organic or inorganic nitrite, in water, hydrochloric acid, acetic acid or sulfuric acid, at temperatures varying between 0° C. and 20° C., followed by its reduction into hydrazine and intramolecular cyclization activated by heating of the reaction medium. The reduction reaction is preferentially carried out with tin chloride in acidic conditions but may also be carried out by catalytic hydrogenation or any other method well known to the person skilled in the art. In an alternative to this last step, it is conceivable that the intermediate diazonium undergoes a Sandmeyer reaction during which this functional group is substituted by a halogen atom, such as iodine, by the reaction of an adequate salt, such as NaI. If this option is preferred, formation of the aminopyrazole ring is carried out by the use of a hydrazine, functionalized or not, in a polar solvent such as ethanol at temperatures varying between 25° C. and 150° C.
Method B2:
Alternatively, it is possible to take advantage of an aromatic nucleophilic substitution reaction to functionalize the pyridine or pyrazine ring in position 6. In this case the nucleophiles used are phenols, thiophenols, benzyl alcohols or thiobenzyl alcohols as well as anilines or benzylamines, functionalized or not. The general reaction diagram 8a is presented below, notably with W═H, (C1-C6)alkyl, aryl or benzyl.




In the case in which X═O or S, the 6-chloro-3-nitropicolinonitriles and 6-chloro-3-nitropyrazine-2-carbonitriles, optionally substituted in position 5, are reacted in the presence of the suitable nucleophile, alcohol or thiol, in a polar solvent such as acetonitrile in the presence of an inorganic base such as potassium or sodium carbonate. Solvents such as DMSO (dimethylsulfoxide), DMF (dimethylformamide), acetone, THF (tetrahydrofuran) or pyridine may also be considered. If necessary, these reactions may be catalyzed by the action of copper and may also be carried out without solvent. Typically, the preferred protocol involves temperatures ranging between 20° C. and 150° C.
Alternatively, the use of bases such as pyridine, DIPEA, diisopropylamine, triethylamine, DBU, potassium tert-butylate, NEt3 or NaH is also possible. In the case in which X═N, toluene is a preferred solvent and triethylamine (NEt3) the base of choice.
The following steps, up to the compounds of general formula (VIIb), are identical to those documented in method B1 above.
Method B3:
Method B3, presented in diagram 8b below, is a variant of method B2 characterized by a first step resulting from a catalytic coupling reaction between a benzyl boronate, in acid or ester form, and a 6-chloro-3-nitropicolinonitrile or 6-chloro-3-nitropyrazine-2-carbonitrile derivative. It is also well known to the person skilled in the art that catalytic coupling reactions using alternative catalysts and benzyl derivatives are also possible. Among these, the Stille reaction, based on tin complexes, or those based on organozinc compounds may be considered.




An optionally substituted 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane is obtained beforehand, for example from the corresponding benzyl chloride and octamethyl-bi-dioxaborolane in dioxane in the presence of potassium acetate and Pt(dppf)Cl2 (dppf=1,1′-bis(diphenylphosphino)ferrocene). This compound is brought together with a 6-chloro-3-nitropicolinonitrile, a 6-chloro-3-nitropyrazine-2-carbonitrile optionally substituted in position 5 or a 5-chloro-2-nitronicotinonitrile optionally substituted in position 6 and a palladium catalyst such as Pd(dppf)Cl2 or Pd(PPh3)4, an organic base such as triethylamine or an alcoholate, or an inorganic base such as sodium, potassium or cesium carbonate in a solvent such as toluene, benzene, THF or dioxane. The preferred reaction temperatures are between 20° C. and 100° C. The products of these reactions correspond to substituted 6-benzyl-3-nitropicolinonitrile, 6-benzyl-3-nitropyrazine-2-carbonitrile or 5-benzyl-2-nitronicotinonitrile derivatives for which the following transformation steps are reproduced from method B1 above.
Method B4:
Method B4, presented in diagram 9 below, gives access to pyrazolopyridine and pyrazolopyrazines bicycles featuring optionally functionalized aryl sulfonamide functions, with R1═(C1-C6)alkyl and notably W═H, (C1-C6)alkyl, aryl or benzyl.




The ethyl 2-chloro-5-(chlorosulfonyl)nicotinate derivatives required for this reaction sequence may be obtained according to the methods described by Levett P. C. et al., Org. Proc. Res. Dev., 2002, 6(6), 767-772; WO 01/98284 and WO 2008/010964.
The formation of sulfonamides is typically carried out by mixing the 2-chloro-5-(chlorosulfonyl)nicotinate of interest with a primary or secondary aniline, optionally functionalized, in an aprotic solvent such as dichloromethane, THF, acetone or acetonitrile in the presence of an organic base such as triethylamine (NEt3), pyridine or DIPEA. The use of an inorganic base such as sodium or potassium carbonate may also be considered. The optimal reaction temperatures are between 0° C. and 70° C.
The saponification reaction of the product thus obtained, notably by the use of lithium hydroxide in a THF/water mixture, gives access to the corresponding 2-chloro-5-(N-phenylsulfamoyl)nicotinic acids.
The corresponding acid chlorides are prepared by treatment with thionyl chloride in toluene under reflux or by any other dehydrochlorination method well known to the person skilled in the art. The reaction of these intermediates with aqueous ammonia makes it possible to form optionally functionalized 2-chloro-5-(N-phenylsulfamoyl)nicotinamides which are then engaged in a dehydration reaction, notably by the use of POCl3, at a temperature ranging between 75° C. and 150° C. The alternative use of agents such as P2O5 or trifluoroacetic anhydride and pyridine may also be considered.
Lastly, these derivatives of general formula (VIh) are reacted in the presence of a hydrazine, functionalized or not, in a polar solvent such as ethanol at temperatures varying between 25° C. and 150° C. to form the corresponding derivatives of general formula (VIId).
Method B5:
Method B5, presented in diagram 10 below, gives access to pyrazolopyridine bicycles featuring optionally functionalized benzyl ether functions, notably with (C1-C6)alkyl, aryl or benzyl.




The method described below is inspired by the work of J. Baldwin et al., J. Heterocyclic. Chem., 1980, 17(3), 445-448. The 5-hydroxynicotinonitrile derivatives, optionally functionalized in position 6, are alkylated, typically by the use of an optionally functionalized benzyl halide in the presence of a base. The preferred method requires the use of an aprotic polar solvent such as DMF and a base such as NaH. The optimal reaction temperatures are between 20° C. and 100° C. Alternatively, the solvents which may be used include, for example, THF, DMSO, dioxane, acetonitrile, dichloromethane or acetone and bases such as tBuOK, DIPEA, pyridine, triethylamine, DBU or sodium, potassium or cesium carbonate.
Oxidation of the pyridine ring into pyridine-N-oxide is typically carried out by use of m-CPBA in dichloromethane at room temperature. Nevertheless, many alternative methods are conceivable, notably those based on the use of sodium percarbonate in the presence of a rhenium catalyst, sodium perborate in the presence of acetic acid or the urea-hydrogen peroxide complex.
Treatment of these pyridine-N-oxide derivatives with phosphorus oxychloride leads to the formation of the corresponding 2-chloronicotinonitriles (VI).
Their reaction under heat with a hydrazine, functionalized or not, in a polar solvent such as isopropanol or ethanol leads to the formation of the pyrazolopyridine bicycles (VIIe) sought.
Method B6:
Method B6, presented in diagram 10a below, gives access to optionally functionalized pyrazolopyridine and pyrazolopyrazine bicycles featuring with reversed sulfonamide functions, notably with W═H, (C1-C6)alkyl, aryl or benzyl.




The method described below consists in forming a sulfonamide function from an aromatic amine and an arylsulfonyl halide, or any other equivalent reagent, in the presence of a base, which can optionally be introduced as solvent or co-solvent. Alternatively, the arylsulfonyl halide or its equivalent can be generated in situ.
Their reaction under heat with a hydrazine, functionalized or not, in a polar solvent such as isopropanol or ethanol leads to the formation the desired pyrazolopyridine and pyrazolopyrazine bicycles (VIIf).
Method C:
Method C aims at the preparation of compounds of general formula (XI) as described in diagram 1.
Method C1:
Method C1, presented in diagram 11 below, is intended for the preparation of pyrazolopyridines and pyrazolopyrazines functionalized at position 6 with Rn=halogen, mesylate, tosylate or triflate, X═O, S, NH, N—(C1-C_)alkyl, and optionally CH2 for (Xc) and (Xd), and Rj═H or N-protecting group.
This method can also be used to carry out the synthesis of molecules comprising a diatomic X group corresponding notably to an ArX group representing: —ArCH2NH—, —ArCH2N(R4)—, —ArCH2O—, —ArCH2S—, —ArCH2CH2—, —ArCHCH—, or —ArCC—.




The 6-hydroxy-2-(methylthio)nicotinonitriles or 5-hydroxy-3-(methylthio)pyrazine-2-carbonitriles are subjected to a dehydrochlorination reaction, typically in the presence of phosphorus oxychloride, with or without solvent, at temperatures varying between 70° C. and 180° C. If a solvent is used, a high boiling-point non-polar solvent such as toluene or xylene will be preferred. Alternatively, it is possible to activate the 6-hydroxy-2-(methylthio)nicotinonitriles and 5-hydroxy-3-(methylthio)pyrazine-2-carbonitriles by their derivation into sulfonic esters via the formation of the corresponding tosylates, mesylates or triflates. If this option is preferred, the use of tosyl, mesyl or triflyl chlorides in a solvent such as toluene, dichloromethane, THF, acetonitrile, acetone or dioxane in the presence of an organic or inorganic base gives access to these derivatives.
The 6-chloro-2(methylthio)nicotinonitriles and 5-chloro-3-(methylthio)pyrazine-2-carbonitriles respectively obtained, or their sulfonic ester analogues if this option is preferred, are then reacted with a nucleophile such as a phenol, an aniline or a thiophenol in the context of aromatic nucleophilic substitution. In this case, the reaction is carried out in a polar solvent such as DMSO, DMF, acetone, THF or acetonitrile, in the presence of a base such as potassium tert-butylate or NaH. If necessary, these reactions may be catalyzed by the action of copper and may also be carried out without solvent. Typically, the preferred protocol involves temperatures ranging between 20° C. and 150° C.
Alternatively, the use of organic bases such as pyridine, diisopropylamine, triethylamine or DBU, or inorganic bases such as sodium or potassium carbonate is also possible.
Alternatively, the compounds of formula (IXb) may give rise to a catalytic coupling reaction such as a Suzuki reaction. In this case, these compounds are brought together with an optionally substituted 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane already described in preceding method B3, a palladium catalyst such as Pd(dppf)Cl2 or Pd(PPh3)4, an organic base such as triethylamine or an alcoholate, or an inorganic base such as sodium, potassium or cesium carbonate in a solvent such as toluene, benzene, THF or dioxane. The preferred reaction temperatures are between 20° C. and 100° C.
The derivatives obtained by one or another of these methods are then oxidized, typically by the use of m-CPBA or oxone to form the corresponding methyl sulfoxides or methyl sulfones. These compounds, sometimes obtained as mixtures, are used as-is in the aminopyrazole ring formation reaction by use of an optionally substituted hydrazine in a polar solvent such as ethanol at temperatures varying between 25° C. and 150° C.
Alternatively, it is possible to modify the reaction sequence, notably by reversing the synthesis steps.
Method C2:
Method C2, presented in diagram 12 below, is intended for the preparation of pyrazolopyridines and pyrazolopyridazines functionalized at position 6 with X═O, S, NH, N—(C1-C_)alkyl, or CH2 and Rj═H or N-protecting group.




The 6-hydroxy-4-(methylthio)nicotinonitrile or 6-hydroxy-4-(methylthio)pyridazin-3-carbonitrile derivatives are oxidized, typically by the use of m-CPBA or oxone to form the corresponding methyl sulfoxides or methyl sulfones. These compounds, sometimes obtained as mixtures, are used as-is in the aminopyrazole ring formation reaction by use of an optionally substituted hydrazine in a polar solvent such as ethanol at temperatures varying between 25° C. and 150° C.
The pyrazolopyridines and pyrazolopyridazines thus obtained are subjected to a dehydrochlorination reaction, typically in the presence of phosphorus oxychloride, with or without solvent, at temperatures varying between 70° C. and 180° C. If a solvent is used, a high boiling-point non-polar solvent such as toluene or xylene will be preferred. The optionally substituted 6-chloro-pyrazolo[4,3-c]pyridin-3-amine and 6-chloro-pyrazolo[4,3-c]pyridazin-3-amine respectively obtained are then reacted with a nucleophile such as a phenol, an aniline or a thiophenol in the context of aromatic nucleophilic substitution. In this case, the reaction is carried out in a polar solvent such as DMSO, DMF, acetone, THF or acetonitrile, in the presence of a base such as potassium tert-butylate or NaH. If necessary, these reactions may be catalyzed by the action of copper and may also be carried out without solvent. Typically, the preferred protocol involves temperatures ranging between 20° C. and 150° C.
Alternatively, the use of organic bases such as pyridine, diisopropylamine, triethylamine or DBU, or inorganic bases such as sodium or potassium carbonate is also possible.
Alternatively, the compounds of formula (XIVa) may give rise to a catalytic coupling reaction such as a Suzuki reaction. In this case, these compounds are brought together with an optionally substituted 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane described above in preceding method B3, a palladium catalyst such as Pd(dppf)Cl2 or Pd(PPh3)4, an organic base such as triethylamine or an alcoholate, or an inorganic base such as sodium, potassium or cesium carbonate in a solvent such as toluene, benzene, THF or dioxane. The preferred reaction temperatures are between 20° C. and 100° C.
Method C3:
Method C3, presented in diagram 12a below, is a variant of method C1 based on the regioselective functionalization of 2,6-dichloronicotinonitrile either by an anionic nucleophile such as a phenate or a thiophenate, or by an organometallic such as a benzylzinc chloride. In the latter case, the reaction is catalyzed for example with a palladium(II) complex. The transformation of the chloronicotinonitrile thus obtained in the corresponding pyrazolopyridine, in the case where Y1═CH, is carried out as previously described in method A1.




Method D:

These methods have as an object the synthesis of compounds of general formula (I) or (VII) by the use of various catalytic coupling methods.
Method D1:
Method D1, presented in diagram 13 below, makes use of the coupling reaction as described in J.A.C.S., 1984, 106, 158 between an organozinc compound prepared in situ and an aryl bromide catalyzed by palladium complexes.




The optionally substituted 3-amino-diazaindazoles or 3-amino-azaindazoles are brought together with a zinc benzyl chloride, optionally substituted, in an aprotic polar solvent such as THF or dioxane, in the presence of a catalytic quantity of a palladium complex such as (dppf)2PdCl2.CH2Cl2. The coupling reaction is carried out at temperatures ranging between 25° C. and 100° C.
Method D2:
Method D2, presented in diagram 14 below, makes use of the coupling reaction as described by Gueiffier A. et al., Tetrahedron, 2006, 62, 6042-6049, between a thiol, in particular a thiophenol or a benzylthiol, and an aryl iodide catalyzed by copper complexes.




This reaction is typically carried out in a high boiling-point polar solvent such as 2-propanol in the presence of a catalytic quantity of polyethylene glycol, a metal salt such as copper iodide (CuI) and an excess of an inorganic base such as potassium carbonate, calcium carbonate or sodium carbonate. The reaction temperatures typically vary between 50° C. and 100° C.
Method D3:
Method D3, presented in diagram 15 below, makes use of the coupling reaction as described by Sonogashira, K. et al. in Tetrahedron Lett., 1975, 16, 4467-4470 between an acetylene derivative and an aryl halide catalyzed by copper and palladium complexes.




Such a reaction is typically carried out by the reaction under an inert atmosphere of a heteroaryl halide with a stoichiometric quantity of an optionally substituted ethynylbenzene in the presence of a catalytic quantity of a palladium complex, for example PdCl2(PPh3)2 or Pd(PPh3)4, a catalytic quantity of a copper salt, for example CuI, and an organic base such as triethylamine or DIPEA, or an inorganic base such as potassium or cesium carbonate. The protocol generally involves reaction temperatures ranging between 20° C. and 45° C. in solvents including DMF, THF, dioxane or diethyl ether.
Method E:
The protocols of method E aim at functionalizing the exocyclic amine of aminopyrazole rings by their reaction with an intermediate featuring an electrophile function, optionally generated in situ, such as acid chloride, an isocyanate, a isothiocyanate or an aldehyde.
Method E1:
Method E1, presented in diagram 16 below, aims at the transformation of the primary exocyclic amine function of aminopyrazole compounds into an amide function.




These compounds are synthesized via the corresponding 3-aminopyrazole by the addition of adequate acid chloride prepared beforehand by the use of oxalyl chloride and a catalytic quantity of DMF in a solvent such as tetrahydrofuran. These acid chlorides may be obtained by the use of alternative methods, such as those based on the use of thionyl chloride or phosphorus oxychloride, well known to the person skilled in the art. The condensation of acid chlorides on aminopyrazoles is typically carried out in an aprotic solvent such as tetrahydrofuran, toluene or dichloromethane in the presence of a base such as DIPEA, pyridine or triethylamine.
Alternatively, the use of a base as a solvent, in particular pyridine, is a possibility.
Alternatively, this type of reaction may be conducted in a biphasic system according to the well-known Schotten-Baumann method.
Alternatively, formation of the amide bond may be carried out from the corresponding 3-aminopyrazole and the acid of interest by the use of peptide coupling conditions using reagents such as HOBt (hydroxybenzotriazole), TBTU (O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate), HATU (2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate), EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) or carbonyldiimidazole at a temperature ranging between −20° C. and 100° C. in an aprotic solvent such as tetrahydrofuran, dioxane, dichloromethane or any solvent with similar properties.
Method E2:
Derivatives characterized by the presence of a secondary amine in position 3 of the aminopyrazole ring are synthesized by a reducing amination reaction according to diagram 17 below.




Reducing amination reactions are typically carried out by mixing adequate stoichiometric quantities of aminopyrazole and aldehyde in a solvent such as DCE (dichloroethane), THF or acetonitrile, optionally in the presence of a quantity of water, TFA (trifluoroacetic acid) or acetic acid, by adding successive fractions of a reducing agent such as NaBH4, NaBH(OAc)3 or NaBH3CN. These reactions are typically carried out at room temperature.
Method E3:
Derivatives carrying a 3-ureido or 3-thioureido function are obtained by the reaction, presented in diagram 18 below, of an aminopyrazole with an isocyanate or isothiocyanate obtained according to methods well known to the person skilled in the art.




In a typical reaction, the reaction mixture is prepared in a polar or non-polar aprotic solvent such as dichloromethane, acetone, DMF, DMA, acetonitrile, THF or dioxane carried at temperatures varying between 20° C. and the boiling point of the chosen solvent. If necessary, recourse to a weakly nucleophilic organic or inorganic base may prove to be necessary. In this case, sodium hydride is a possible option.
Method F: Post-Synthetic Deprotections and Modifications
Method F1: Deprotections
The trifluoroacetate protecting groups are removed by the action of an organic base such as triethylamine or pyridine in a polar solvent such as methanol, ethanol or THF at the reflux temperatures of the solvents used.
The tert-butyl or trityl protecting groups carried by the pyrazole rings are displaced by the action of a strong acid, typically TFA, in a non-polar solvent such as dichloromethane or DCE.
Method F2: Alkyne Reductions




Reactions for reducing diaryl alkynes into diaryl alkanes are typically carried out by catalytic hydrogenation, under hydrogen pressure, in the presence of catalysts such as PtO2, Pt, Pd/C, Ni or Rh. Alternatively, the use of DIBAL-H (diisobutylaluminum hydride) in the presence or the absence of a catalyst such as Cp2TiCl2 is conceivable.
Method F3: Oxidation of Sulfides into Sulfones and Sulfoxides




Oxidation reactions of sulfides into sulfoxides are typically carried out via the use of oxone in a mixture of polar solvents such as THF/MeOH or DMF/water. The optimal reaction temperatures are typically between 25° C. and 50° C.
Many alternative methods are available, and some give the possibility of producing semi-oxidized derivatives, namely sulfoxides. Such alternative methods include the use of m-CPBA, KMnO4/MnO2 in dichloromethane, H2O2 (30%) in a biphasic medium and the presence of a phase transfer catalyst or a catalyst in the form of a urea complex (UHP).
The combined use of H2O2 and metal complexes such as Sc(OTf)3 promotes partial oxidation derivatives.
Other known methods include, for example, the use of CAN/NaBrO3 (CAN=ceric ammonium nitrate).
The examples which follow illustrate the invention without limiting its scope in any way.
EXAMPLES
The following abbreviations are used:

    DMSO Dimethylsulfoxide
    EI Electron impact
    ES Electrospray
    LCMS Liquid chromatography-mass spectrometry
    mg milligram
    mL milliliter
    NMR Nuclear magnetic resonance


I. Synthesis of the Compounds According to the Invention
Examples of Method A1
Example 1
5-iodo-1H-pyrazolo[3,4-b]pyridine-3-amine




Example 1a
2-hydroxy-5-iodonicotinonitrile
9 g (0.5 eq) of N-iodosuccinimide at room temperature is added to a solution of 10 g (83 mmol) of 2-hydroxynicotinonitrile in 150 ml of anhydrous dimethylformamide. The reaction mixture is stirred at 60° C. After 30 minutes of stirring, 9 g (0.5 eq) of N-iodosuccinimide is added and then the reaction mixture is stirred at 60° C. for 5 hours. The solvent is evaporated and the precipitate formed is filtered, rinsed with water and with diethyl ether and then dried under vacuum to yield 18.5 g (90%) of 2-hydroxy-5-iodonicotinonitrile in the form of a beige powder.
LCMS (EI, m/z): (M+1) 246.93
1H NMR: δH ppm (400 MHz, DMSO): 12.79 (1H, s, OH), 8.36 (1H, d, CHarom), 8.04 (1H, d, CHarom).
Example 1b
2-chloro-5-iodonicotinonitrile
30.7 ml (329 mmol) of phosphorus oxychloride at 0° C. and 6 drops of sulfuric acid are added to 9 g (6.6 mmol) of 2-hydroxy-5-iodonicotinonitrile. The reaction mixture is heated at 110° C. for 5 hours and then at room temperature overnight. The reaction mixture is poured in a beaker containing ice and a little water, and a precipitate is formed. The mixture is allowed to gradually return to room temperature and then is filtered and rinsed with water. The solid is dried to yield 6.8 g (70%) of 2-chloro-5-iodonicotinonitrile.
LCMS (EI, m/z): (M+1) 265.45
1H NMR: δH ppm (400 MHz, DMSO): 9.61 (1H, d, CHarom), 9.14 (1H, d, CHarom).
Example 1
5-iodo-1H-pyrazolo[3,4-b]pyridine-3-amine
Hydrazine (3.86 ml, 79 mmol) is added at room temperature to 7 g (26.5 mmol) of a solution of 2-chloro-5-iodonicotinonitrile in 25 ml of propan-2-ol. The reaction mixture is heated at 85° C. for 7 hours and then at room temperature overnight. The suspended solid is filtered, rinsed with isopropanol and then with ether and dried in an oven at 50° C. to give 6 g (87%) of 5-iodo-1H-pyrazolo[3,4-b]pyridine-3-amine.
LCMS (EI, m/z): (M+1) 260.95
1H NMR: δH ppm (400 MHz, DMSO): 12.12 (1H, bs, NH), 8.51 (1H, d, CHarom), 8.45 (1H, d, CHarom), 5.64 (2H, bs, NH2).
The following compounds were obtained according to the same method.


















 












Ex.** 
W
Rj
Compound name
Yield
Mass MH+





1-2
H
t-butyl
1-tert-butyl-5-iodo-1H-pyrazolo[3,4-b]pyridin-3-amine
68%
317.05


1-3
Me
H
5-iodo-6-methyl-1H-pyrazolo[3,4-b]pyridin-3-amine
93%
275.02





**1H NMR, DMSO-d6,


Ex. 1-2: 8.55 (1H, bs, CHarom), 8.42 (1H, bs, CHarom), 6.33 (1H, bs, CHarom), 1.57 (9H, s, CH).;


1-3: 11.92 (1H, s, NH), 8.55 (1H, s, CHarom), 5.59 (2H, bs, NH2), 2.66 (3H, s, CH3).






Example 2
5-bromo-1H-pyrazolo[3,4-b]pyridine-3-amine




Example 2a
2-methoxy-nicotinonitrile
4.98 g (217 mmol) of sodium is added to 80 ml of anhydrous methanol. The reaction medium is stirred at room temperature for 10 minutes and then 10 g (72.2 mmol) of 2-chloronicotinonitrile is added at 0° C. The reaction medium is stirred at 25° C. for 16 hours. The reaction is hydrolyzed by slowly adding water at 0° C. After returning to room temperature, the precipitate obtained is filtered, rinsed with water and then dried at 50° C. to yield 7.85 g (81%) of 2-methoxy-nicotinonitrile in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 135.04
1H NMR: δH ppm (400 MHz, DMSO): 8.46-8.48 (1H, dd, CHarom), 8.25-8.27 (1H, dd, CHarom), 7.17-7.20 (1H, dd, CHarom), 3.99 (3H, s, CH3).
Example 2b
5-bromo-2-methoxy-nicotinonitrile
12.23 g (149 mmol) of sodium acetate and then 7.66 ml (149 mmol) of bromine at 0° C. are added to 10 g (74.6 mmol) of a solution of 2-methoxy-nicotinonitrile in 29 ml of acetic acid. The reaction mixture is heated at 70° C. overnight. After returning to room temperature, the reaction medium is added to an ice bath and the precipitate obtained is filtered, rinsed with water and then dried at 50° C. to yield 11.6 g (73%) of 5-bromo-2-methoxy-nicotinonitrile in the form of a white solid.
LCMS (EI, m/z): (M+1) 214.95
1H NMR: δH ppm (400 MHz, DMSO): 8.61 (1H, d, CHarom), 8.60 (1H, d, CHarom), 3.98 (3H, s, CH3)
Example 2
5-bromo-1H-pyrazolo[3,4-b]pyridine-3-amine
35 ml (23.47 mmol) of hydrazine is added at room temperature to 5 g (23.47 mmol) of 5-bromo-2-methoxynicotinonitrile. The reaction medium is carried at 100° C. for 3 hours. After returning to room temperature, the precipitate obtained is filtered, rinsed with water and then dried at 50° C. to yield 3.6 g (72%) of 5-bromo-1H-pyrazolo[3,4-b]pyridine-3-amine in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 214.05
1H NMR: δH ppm (400 MHz, DMSO): 12.18 (1H, s, NH), 8.38 (1H, d, CHarom), 8.37 (1H, d, CHarom), 5.66 (2H, s, NH).
Examples of Method A2
Example 3
5-iodo-1H-pyrazolo[3,4-b]pyrazine-3-amine




Example 3a
Methyl 3-amino-6-iodopyrazine-2-carboxylate
1.5 equivalents of N-iodosuccinimide are added at room temperature to 5 g (32.7 mmol) of a methyl 3-aminopyrazine-2-carboxylate solution in 25 ml of dimethylformamide. The reaction medium is heated at 65° C. for 1 hour, added together with 0.5 equivalents of N-iodosuccinimide and maintained at 65° C. for 24 hours. After returning to room temperature, the solvent is evaporated and then the product is extracted several times with dichloromethane. The organic phases are combined, washed with 10% sodium bisulfite solution, dried on magnesium sulfate and concentrated to yield 8 g (88%) of methyl 3-amino-6-iodopyrazine-2-carboxylate in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 280
1H NMR: δH ppm (400 MHz, DMSO): 8.50 (1H, s, CHarom), 7.50 (2H, bs, NH2), 3.20 (3H, s, CH3).
Example 3b
3-amino-6-iodopyrazine-2-carboxamide
30 ml of ammonia in water is added under magnetic stirring to 15 g (53.8 mmol) of a solution of methyl 3-amino-6-iodopyrazine-2-carboxylate in 150 ml of methanol. The reaction medium is stirred at 25° C. for 48 hours. After evaporation of the solvents, the precipitate obtained is filtered, rinsed with water and then dried at 50° C. to yield 12.50 g of 3-amino-6-iodopyrazine-2-carboxamide (88%) in the form of a beige solid.
LCMS (EI, m/z): (M+1) 265.02
1H NMR: δH ppm (400 MHz, DMSO): 8.35 (1H, s, CHarom), 7.85 (1H, bs, NH), 7.60 (3H, bs, NH), 3.25 (3H, s, CH3)
Example 3c
N′-(3-cyano-5-iodopyrazine-2-yl)-N,N-dimethylformimidamide
13.59 ml (146 mmol) of phosphorus oxychloride is added drop by drop at 0° C. to 11 g (41.7 mmol) of a solution of 3-amino-6-iodopyrazine-2-carboxamide in 80 ml of dimethylformamide. The reaction mixture is stirred at room temperature overnight and then poured in a beaker containing ice and a little water. The pH is adjusted to 8 with 1 N soda solution; a precipitate is formed. The mixture is allowed to gradually return to room temperature and then the solid formed is filtered, rinsed with water and dried at 50° C. to yield 10.50 g of N′-(3-cyano-5-iodopyrazine-2-yl)-N,N-dimethyl formimidamide (84%) in the form of a beige solid.
LCMS (EI, m/z): (M+1) 302.07
1H NMR: δH ppm (400 MHz, DMSO): 8.69 (1H, s, CHarom), 8.67 (1H, s, CHethyl), 3.20 (3H, s, CH3), 3.11 (3H, s, CH3).
Example 3d
3-amino-6-iodopyrazine-2-carbonitrile
77 ml (77 mmol) of 1 M hydrochloric acid solution is added to 7.7 g (25.6 mmol) of N′-(3-cyano-5-iodopyrazin-2-yl)-N,N-dimethylformimidamide. The reaction medium is heated at 50° C. for 4 hours and then stirred at room temperature overnight. The precipitate formed is filtered, rinsed with water and dried at 50° C. to yield 6 g (95%) of 3-amino-6-iodopyrazine-2-carbonitrile in the form of a beige solid.
LCMS (EI, m/z): (M+1) 247.0
1H NMR: δH ppm (400 MHz, DMSO): 8.49 (1H, S, CHarom), 7.53 (2H, bs, NH2).
Example 3e
3-chloro-6-iodopyrazine-2-carbonitrile
64.3 ml of hydrochloric acid is added at −5° C. to 7.7 g (31.3 mmol) of 3-amino-6-iodopyrazine-2-carbonitrile. At this temperature, a sodium nitrite solution (4.32 g, 62.6 mmol) dissolved in 9 ml of water is added to the reaction mixture and is stirred for 4 hours at −50° C. and then at room temperature overnight. Another equivalent of sodium nitrite is added to the reaction mixture and the precipitate formed is filtered, rinsed with water and dried at 50° C. to yield 3.65 g (44%) of 3-chloro-6-iodopyrazine-2-carbonitrile in the form of a beige solid.
LCMS (EI, m/z): (M+1) 266.49
1H NMR: δH ppm (400 MHz, DMSO): 9.13 (1H, s, CHarom)
Example 3
5-iodo-1H-pyrazolo[3,4-b]pyrazine-3-amine
0.74 ml (9.8 mmol) of hydrazine is added to 2.6 g (9.80 mmol) of a solution of 3-chloro-6-iodopyrazine-2-carbonitrile in 15 ml of butanol. The reaction mixture is heated at 110° C. for 5 hours and then left at room temperature overnight. The suspended solid is filtered, rinsed with butanol and then dried in an oven at 50° C. to yield 2.2 g (86%) of 5-iodo-1H-pyrazolo[3,4-b]pyrazine-3-amine in the form of a brown solid.
LCMS (EI, m/z): (M+1) 262.02
1H NMR: δH ppm (400 MHz, DMSO): 12.59 (1H, bs, NH), 8.60 (1H, d, CHarom), 5.83 (2H, bs, NH2).
Examples of Method A3
Example 4
5-iodo-6-methoxy-1H-pyrazolo[3,4-b]pyridin-3-amine




Example 4a
ethyl 5-cyano-2-hydroxy-6-(methylthio)nicotinate
Ethyl 5-cyano-2-hydroxy-6-(methylthio)nicotinate is obtained by following the procedure of Ya. Yu. Yakunin et al., Russian Chemical Bulletin, 1999, 48(1), 195-6 with a total yield of 34%.
LCMS (EI, m/z): (M−1) 237.22
1H NMR: δH ppm (400 MHz, DMSO): 12.72 (1H, bs, OH), 8.40 (1H, s, CHarom), 4.29 (2H, q, CH2), 2.64 (3H, s, CH3), 1.30 (3H, t, CH3).
Example 4b
5-cyano-2-hydroxy-6-(methylthio)nicotinic acid
4.16 g (2 eq) of lithium hydroxide monohydrate is added at room temperature to a solution of 11.8 g (49.5 mmol) of ethyl 5-cyano-2-hydroxy-6-(methylthio)nicotinate in 100 ml of ethanol and 100 ml of water. The reaction mixture is stirred at 60° C. for 2 hours. The ethanol is evaporated and 1 N aqueous soda is added. The aqueous phase is washed with ethyl acetate and then re-acidified by adding 1 N aqueous hydrogen chloride (pH=1). The precipitate formed is filtered, rinsed with water and with diethyl ether and then dried under vacuum to yield 9.9 g (95%) of 5-eyano-2-hydroxy-6-(methylthio)nicotinic acid in the form of a brown powder.
LCMS (EI, m/z): (M−1) 209.09
1H NMR: δH ppm (400 MHz, DMSO): 8.32 (1H, s, CHarom), 2.61 (3H, s, CH3).
Example 4c
6-hydroxy-2-(methylthio)nicotinonitrile
A solution of 6 g (28.5 mmol) of 5-cyano-2-hydroxy-6-(methylthio)nicotinic acid in 35 ml of diphenyl ether is stirred at 250° C. for 4 hours. After returning to room temperature, 100 ml of cyclohexane is added and the reaction medium is triturated for 30 minutes. The solid formed is filtered, rinsed thoroughly with cyclohexane and then dried under vacuum to yield 2.87 g (60%) of 6-hydroxy-2-(methylthio)nicotinonitrile in the form of a brown powder.
LCMS (EI, m/z): (M+1) 167.12
1H NMR: δH ppm (400 MHz, DMSO): 12.16 (1H, bs, OH), 7.92 (1H, d, CHarom), 6.46 (1H, d, CHarom), 2.59 (3H, s, CH3).
Example 4d
6-hydroxy-5-iodo-2-(methylthio)nicotinonitrile
6 g (1.6 eq) of silver sulfate and 4.58 g (1.5 eq) of iodine are added successively to a solution of 2 g (12 mmol) of 6-hydroxy-2-(methylthio)nicotinonitrile in 200 ml of ethanol. The reaction medium is stirred at room temperature for 2 hours. The solid is filtered and the residue rinsed thoroughly with methanol. The filtrate is evaporated and then taken up in ethyl acetate. The organic phase is washed with water three times, dried on magnesium sulfate and evaporated to yield 3.18 g (90%) of 6-hydroxy-5-iodo-2-(methylthio)nicotinonitrile in the form of a yellow powder.
LCMS (EL m/z): (M+1) 292.93
1H NMR: δH ppm (400 MHz, DMSO): 12.96 (1H, bs, OH), 8.38 (1H, s, CHarom), 2.62 (3H, s, CH3).
Example 4e
5-iodo-6-methoxy-2-(methylthio)nicotinonitrile
905 μl (2 eq) of methyl iodide and 2.1 g (1.05 eq) of silver carbonate are added successively to a solution of 2.12 g (7.26 mmol) of 6-hydroxy-5-iodo-2-(methylthio)nicotinonitrile in 20 ml of 1,4-dioxane. The reaction medium is stirred at 60° C. for 5 hours. The solid is filtered and the residue rinsed thoroughly with methanol. The filtrate is evaporated and the residue purified by silica column chromatography (4:6 dichloromethane/cyclohexane as eluent) to yield 1.52 g (68%) of 5-iodo-6-methoxy-2-(methylthio)nicotinonitrile in the form of a white powder.
LCMS (EI, m/z): (M+1) 306.95
1H NMR: δH ppm (400 MHz, DMSO): 8.50 (1H, s, CHarom), 4.04 (3H, s, CH3), 2.63 (3H, s, CH3).
Example 4f
5-iodo-6-methoxy-2-(methylsulfinyl)nicotinonitrile
1.42 g (1.1 eq) of 70% 3-chloroperbenzoic acid is added to a solution of 1.6 g (5.23 mmol) of 5-iodo-6-methoxy-2-(methylthio)nicotinonitrile in 20 ml of dichloromethane. The reaction medium is stirred at room temperature for 1 hour. Ethyl acetate is added and the organic phase is washed with saturated sodium bicarbonate solution, dried on magnesium sulfate, filtered and evaporated to yield 1.63 g (97%) of 5-iodo-6-methoxy-2-(methylsulfinyl)nicotinonitrile in the form of a white powder which may also contain 5-iodo-6-methoxy-2-(methylsulfonyl)nicotinonitrile in small proportions (<20%). If necessary, the mixture is used as-is in the following steps.
LCMS (EI, m/z): (M+1) 322.95
1H NMR: δH ppm (400 MHz, DMSO): 8.86 (1H, s, CHarom), 4.05 (3H, s, CH3), 2.95 (3H, s, CH3).
Example 4
5-iodo-6-methoxy-1H-pyrazolo[3,4-b]pyridin-3-amine
294 μl (1.2 eq) of hydrazine monohydrate is added to a solution of 1.63 g (5.05 mmol) of 5-iodo-6-methoxy-2-(methylsulfinyl)nicotinonitrile in 30 ml of 2-propanol. The reaction medium is stirred at 80° C. for 9 hours. After returning to room temperature, the solid formed is filtered and rinsed with 2-propanol to yield 1.14 g (78%) of 5-iodo-6-methoxy-1H-pyrazolo[3,4-b]pyridin-3-amine in the form of a white powder.
LCMS (EI, m/z): (M+1) 291.00
1H NMR: δH ppm (400 MHz, DMSO): 11.87 (1H, s, NH), 8.49 (1H, s, CHarom), 5.49 (2H, bs, NH2), 3.90 (3H, s, CH3).
Example 5
5-iodo-1H-pyrazolo[3,4-b]pyridine-3,6-diamine




Example 5a
4-methylmorpholinium (2,4)-ethyl-5-amino-2,4-dicyano-5-mercaptopenta-2,4-dienoate
4-methylmorpholinium (2,4)-ethyl-5-amino-2,4-dicyano-5-mercaptopenta-2,4-dienoate is prepared according to the procedure described by V. D. Dyachenko et al., Chemistry of Heterocyclic Compounds, 2005, 41(4), 503-10 with a yield of 50%.
1H NMR: δH ppm (400 MHz, DMSO): 9.60 (1H, bs, NH), 8.66 (1H, s, CH), 8.33 (1H, bs, NH), 7.43 (1H, bs, NH), 4.08 (2H, q, CH2), 3.82-4.02 (2H, m, CH2), 3.55-3.78 (2H, m, CH2), 3.24-3.42 (2H, m, CH2), 3.98-3.17 (2H, m, CH2), 2.81 (3H, s, CH3), 1.19 (3H, t, CH3).
Example 5b
Ethyl 2-amino-5-cyano-6-(methylthio)nicotinate
2.73 ml (1 eq) of methyl iodide is added to a solution of 14.2 g (43.8 mmol) of 4-methylmorpholinium (2,4)-ethyl-5-amino-2,4-dicyano-5-mercaptopenta-2,4-dienoate in 78 ml of N,N-dimethylformamide. The reaction mixture is stirred at room temperature for 1 hour and then at 75° C. for 20 hours. After returning to room temperature, water is added and the solid formed is filtered and dried under vacuum to yield 10.31 g (100%) of ethyl 2-amino-5-cyano-6-(methylthio)nicotinate in the form of a beige powder.
LCMS (EI, m/z): (M+1) 238.20
1H NMR: δH ppm (400 MHz, DMSO): 8.25 (1H, s, CHarom), 8.19 (1H, bs, NH), 7.99 (1H, bs, NH), 4.27 (2H, q, CH2), 2.58 (3H, s, CH3), 1.31 (3H, t, CH3).
Example 5c
2-amino-5-cyano-6-(methylthio)nicotinic acid
3.08 g (2 eq) of lithium hydroxide monohydrate is added at room temperature to a solution of 8.7 g (36.7 mmol) of ethyl 2-amino-5-cyano-6-(methylthio)nicotinate in 87 ml of ethanol and 87 ml of water. The reaction mixture is stirred at 60° C. for 2 hours. The ethanol is evaporated and 1 N aqueous soda is added. The aqueous phase is washed with ethyl acetate and then re-acidified by adding 1 N aqueous hydrogen chloride (pH=1). The precipitate formed is filtered, rinsed with water and with diethyl ether and then dried under vacuum to yield 7.67 g (quantitative) of 2-amino-5-cyano-6-(methylthio)nicotinic acid in the form of a brown powder.
LCMS (EI, m/z): (M+1) 210.16
1H NMR: δH ppm (400 MHz, DMSO): 13.28 (1H, bs, CO2H), 8.21 (1H, s, CHarom), 8.13 (2H, bs, NH2), 2.57 (3H, s, CH3).
Example 5d
6-amino-2-(methylthio)nicotinonitrile
A solution of 3 g (14.3 mmol) of 2-amino-5-cyano-6-(methylthio)nicotinic acid in 30 ml of diphenyl ether is stirred at 255° C. for 60 hours. After returning to room temperature, 60 ml of cyclohexane is added and the reaction medium is triturated for 30 minutes. The solid formed is filtered and then rinsed thoroughly with cyclohexane. The solid is redissolved in ethyl acetate and then the organic phase is washed with water, dried on magnesium sulfate, filtered and then evaporated to yield 1.32 g (55%) of 6-amino-2-(methylthio)nicotinonitrile in the form of a brown powder.
LCMS (EI, m/z): (M+1) 166.13
1H NMR: δH ppm (400 MHz, DMSO): 7.58 (1H, d, CHarom), 7.12 (2H, bs, NH2), 6.20 (1H, d, CHarom), 2.51 (3H, s, CH3).
Example 5e
6-amino-5-iodo-2-(methylthio)nicotinonitrile
3.75 g (1.5 eq) of silver sulfate and 2.85 g (1.4 eq) of iodine are added successively to a solution of 1.32 g (8.02 mmol) of 6-amino-2-(methylthio)nicotinonitrile in 65 ml of ethanol. The reaction medium is stirred at room temperature for 3 hours. The solid is filtered and the residue rinsed thoroughly with methanol. The filtrate is evaporated and redissolved in ethyl acetate. The organic phase is washed with water three times, dried on magnesium sulfate and evaporated to yield 1.89 g (81%) of 6-amino-5-iodo-2-(methylthio)nicotinonitrile in the form of a brown powder.
LCMS (EI, m/z): (M+1) 291.99
1H NMR: δH ppm (400 MHz, DMSO): 8.13 (1H, s, CHarom), 7.19 (1H, broad flat singlet, NH2), 2.51 (3H, s, CH3).
Example 5f
6-amino-5-iodo-2-(methylsulfinyl)nicotinonitrile
1.77 g (1.1 eq) of 70% 3-chloroperbenzoic acid is added to a solution of 1.89 g (6.51 mmol) of 6-amino-5-iodo-2-(methylthio)nicotinonitrile in 60 ml of dichloromethane. The reaction medium is stirred at room temperature for 1 hour. Ethyl acetate is added and the organic phase is washed with saturated sodium bicarbonate solution, dried on magnesium sulfate, filtered and evaporated to yield 1.5 g (75%) of 6-amino-5-iodo-2-(methylsulfinyl)nicotinonitrile in the form of a white powder which may also contain 6-amino-5-iodo-2-(methylsulfonyl)nicotinonitrile in small proportions (<20%). If necessary, the mixture is used as-is in the following steps.
LCMS (EI, m/z): (M+1) 307.98
1H NMR: δH ppm (400 MHz, DMSO): 8.45 (1H, s, CHarom), 7.70 (2H, broad flat singlet, NH2), 2.84 (3H, s, CH3).
Example 5
5-iodo-1H-pyrazolo[3,4-b]pyridine-3,6-diamine
275 μl (2 eq) of hydrazine monohydrate is added to a solution of 872 mg (2.84 mmol) of 6-amino-5-iodo-2-(methylsulfinyl)nicotinonitrile in 11 ml of 2-propanol. The reaction medium is stirred at 80° C. for 3 hours. Water is added and the product is extracted with ethyl acetate. The organic phase is dried on magnesium sulfate, filtered and evaporated. The residue is triturated in a minimum of diisopropyl ether. The solid is filtered to yield 523 mg (67%) of 5-iodo-1H-pyrazolo[3,4-b]pyridin-3,6-diamine in the form of a brown powder.
LCMS (EI, m/z): (M+1) 276.00
1H NMR: δH ppm (400 MHz, DMSO): 11.23 (1H, s, NH), 8.26 (1H, s, CHarom), 6.11 (2H, bs, NH2), 5.25 (2H, bs, NH2).
Examples of Method B1
Example 6
5-(3,5-difluorobenzylthio)-1H-pyrazolo[4,3-b]pyridin-3-amine




Example 6a
6-chloro-3-nitropicolinonitrile
2,6-Dichloro-3-nitropyridine (5.18 mmol, 1 g) is mixed with 5 ml of N-methyl-2-pyrrolidinone in a microwave reactor. The reaction mixture is heated at 180° C. for 15 minutes (6 bars). The crude reaction product is dissolved in ethyl acetate, filtered and washed several times using an aqueous phase. The organic phase is collected, dried on magnesium sulfate and dry concentrated. The crude product thus obtained is purified by silica gel chromatography (heptane/AcOEt) to yield, after concentration, 0.62 g (65%) of a brown oil.
1H NMR: δH ppm (400 MHz, DMSO): 8.81 (1H, d, CHarom), 8.18 (1H, d, CHarom).
Example 6b
3-nitro-6-thioxo-1,6-dihydropyridine-2-carbonitrile
One equivalent of NaSH:H2O is added to a solution of 6-chloro-3-nitropicolinonitrile (5.45 mmol, 1 g) in 20 ml of EtOH. The color turns orange. The reaction medium is stirred at room temperature for 30 minutes. The crude reaction product is then concentrated, redissolved in ethyl acetate and extracted several times using an acidic aqueous phase (1 N HCl) and then a neutral phase. The organic phase is concentrated and the crude reaction product recrystallized in acetone to yield 0.64 g (79%) of yellow crystals.
1H NMR: δH ppm (400 MHz, DMSO): 8.71 (1H, d, CHarom), 8.27 (1H, d, CHarom).
Example 6c
6-(3,5-difluorobenzylthio)-3-nitropicolinonitrile
A mixture of 3-nitro-6-thioxo-1,6-dihydropyridin-2-carbonitrile (4.42 mmol, 1.34 g), 3,5-diflurobenzylbenzylbromide (8.83 mmol, 1.828 g), and K2CO3 (11.04 mmol, 1.525 g) in 5 ml of acetone is heated at 70° C. for 10 hours and then evaporated under reduced pressure. The residue is purified by silica gel chromatography (AcOEt/heptane) to yield 1.33 g (98%) of the expected product.
LCMS (ES−): m/z 306 (M−H+).
1H NMR: δH ppm (400 MHz, DMSO): 8.53 (1H, d, CHarom), 7.91 (1H, d, CHarom), 7.21 (2H, m), 7.17 (1H, m), 4.55 (2H, CH2).
Example 6d
3-amino-6-(3,5-difluorobenzylthio)picolinamide
A mixture of 6-(3,5-difluorobenzylthio)-3-nitropicolinonitrile (0.05 g, 0.163 mmol) and PtO2 (0.739 mg, 3.25 μmol) in 10 ml of MeOH is placed under stirring at atmospheric pressure of hydrogen for 2 hours. The catalyst is filtered, the solution is concentrated and the residue thus obtained is purified by silica gel chromatography (AcOEt/heptane) to yield, after concentration, 0.04 g (83%) of white crystals.
LCMS (ES+) m/z: 296 (MH+).
1H NMR: δH ppm (400 MHz, DMSO): 7.84 (1H, broad s, NH), 7.40 (1H, broad s, NH), 7.14 (1H, d, CHarom), 7.08 (4H, m, CHarom), 6.80 (2H, broad s, NH2), 4.43 (2H, s, CH2).
Example 6e
3-amino-6-(3,5-difluorobenzylthio)picolinonitrile
A mixture of 3-amino-6-(3,5-difluorobenzylthio)picolinoamide (2.37 mmol, 0.7 g) and P2Cl5 (9.48 mmol, 1.346 g), 20 ml of toluene and 1 ml of ionic solvent (1-butyl-3-methylimidazolium tetrafluoroborate) are placed in a microwave reactor and then heated at 140° C. for 30 minutes. The crude reaction product is then concentrated under reduced pressure and the orange crystals thus obtained are redissolved in ethyl acetate and washed using saturated aqueous NaHCO3 solution. The organic phase is dried on magnesium sulfate and then concentrated to yield 0.7 g of a brown oil. This crude reaction product is purified by silica gel chromatography (AcOEt/heptane+0.1% of NEt3) to yield, after concentration, 0.15 g (23%) of orange crystals.
1H NMR: δH ppm (400 MHz, DMSO): 7.73 (1H, d, CHarom), 7.25 (2H, m, CHarom), 7.18 (1H, m), 6.85 (1H, d), 5.43 (2H, CH2).
Example 6
5-(3,5-difluorobenzylthio)-1H-pyrazolo[4,3-b]pyridin-3-amine
A solution cooled to 0° C. of NaNO2 in 3 ml of water is added drop by drop to a solution at 0° C. of 3-amino-6-(3,5-difluorobenzylthio)picolinonitrile (1.587 mmol, 0.44 g) in 15 ml of 6 N HCl solution. After 15 minutes, a solution cooled to 0° C. of SnCl2.2H2O diluted in 4 ml of 12 N HCl is added drop by drop. The reaction medium is then stirred at 25° C. for 1 hour. The solution is extracted with ethyl acetate and then washed using saturated NaHCO3 solution and then saturated NaCl solution. The organic phase is collected, dried on magnesium sulfate and then concentrated under reduced pressure. The residue is purified by silica gel chromatography (AcOEt/heptane) to yield, after concentration of the organic phases, 0.07 g (15%) of black crystals.
1H NMR: δH ppm (400 MHz, DMSO): 11.64 (1H, s, NH), 7.63 (1H, d, CHarom), 7.21 (2H, m, CHarom), 7.13 (1H, d, CHarom), 7.04 (1H, m, CHarom), 5.38 (2H, s, NH2), 4.51 (2H, s, CH2).
The following compounds are obtained by a similar method:
















 














Ex.**
ArX
W
Y4
Rj
Compound names
Yield
Mass MH+





6-2




H
CH
H
5-(2,5-difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
5% 4 steps
293.0


 


6-3




H
CH
H
5-(2,5-dichlorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
3% 4 steps
324.9





**1H NMR: δH ppm (400 MHz, DMSO):


6-2: 11.65 (1H, s, NH), 7.64 (1H, dd, CHarom, J = 8.8 Hz), 7.42-7.51 (1H, m, CHarom), 7.20-7.25 (1H, m, CHarom), 7.14 (1H, dd, CHarom, J = 8.8 Hz), 7.01-7.11 (1H, m, CHarom), 5.37-5.41 (2H, m, NH2), 4.49 (2H, s).


6-3: 11.65 (1H, s, NH), 7.83 (1H, m, CHarom), 7.61 (1H, dd, CHarom, J = 8.8 Hz), 7.50 (1H, m, CHarom), 7.28-7.32 (1H, m, CHarom), 7.10 (1H, dd, CHarom, J = 8.8 Hz), 7.01-7.11 (1H, m, CHarom), 5.42 (2H, s, NH2), 4.47 (2H, s).






Examples of Method B2
Example 7
5-(3,5-dichlorophenylthio)-1H-pyrazolo[4,3-b]pyridin-3-amine




Example 7a
6-(3,5-dichlorophenylthio)-3-nitropicolinonitrile
A mixture of 6-chloro-3-nitropicolinonitrile (3.70 g, 0.02 mol), 3,5-dichlorobenzenethiol (3.60 g, 0.02 mol) and K2CO3 (5.6 g, 0.04 mol) in 100 ml of acetonitrile is carried at 70° C. for 16 hours. The crude reaction product is diluted in an ethyl acetate fraction and washed using an aqueous phase. The organic phase is dried with sodium sulfate and the residue is purified by silica gel chromatography (AcOEt/petroleum ether) to yield 5.4 g (80%) of a yellow solid.
Example 7b
3-amino-6-(3,5-dichlorophenylthio)picolinonitrile
10 ml of concentrated HCl is added to a solution of 6-(3,5-dichlorophenylthio)-3-nitropicolinonitrile (3.4 g, 0.01 mol) in 50 ml of methanol under stirring. The reaction medium is refluxed, added together with 1.68 g (0.03 mol) of iron and stirred for 10 minutes. After returning to room temperature, the reaction mixture is added together with 100 ml of ethyl acetate and 50 ml of water. The pH is adjusted to 10 using 30% soda solution and the organic phase is extracted and then dried on anhydrous sodium sulfate before being concentrated. The residue is purified by silica gel chromatography (ethyl acetate/petroleum ether) to yield, after concentration of the fractions, 2.82 g (91%) of a yellow solid.
LCMS (m/e): 296(M+H+). %.
Example 7
5-(3,5-dichlorophenylthio)-1H-pyrazolo[4,3-b]pyridin-3-amine
A solution of 350 mg of NaNO2 (5.07 mmol) in water (2 ml) is added to a stirring solution of 1.5 g of 3-amino-6-(3,5-dichlorophenylthio)picolinonitrile (5.07 mmol) in 100 ml of 50% sulfuric acid at 0° C. The mixture is stirred for 20 minutes at 0-5° C. A solution of 2.9 g of SnCl2.2H2O (12.7 mmol, 2.5 eq) in hydrochloric acid (12 N solution, 10 ml) is then added and the solution is stirred for 1 hour at room temperature. The solid formed is filtered and then washed twice with 20 ml of water. The solid is suspended in 100 ml and the pH is adjusted to 10 by adding 30% soda solution. The organic phase is separated and then dried on anhydrous sodium sulfate before being concentrated under vacuum. A light yellow solid is obtained after recrystallization in ethyl acetate (470 mg, 34%).
LCMS m/z 311 (M+H+).
1H NMR: δH ppm (400 MHz, DMSO): 11.91 (1H, bs, NH), 7.79 (1H, d, CHarom), 7.55 (1H, s, CHarom), 7.36 (2H, s, CHarom), 7.33 (1H, m, CHarom), 5.42 (2H, s, NH2).
The following compounds are obtained by a similar method:
















 














Ex.**
ArX
Y4
W
Rj
Compound names
Yield
Mass MH+





7-1




CH
H
H
5-(3,5-difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3-amine
28%
277


 


7-2




CH
H
H
5-(3,5-difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
33% 3 steps
278.9


 


7-3




CH
H
H
5-(2,4-difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
24% 3 steps
279.0


 


7-4




CH
H
H
5-(2,4-dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
24% 3 steps
311.0


 


7-5




CH
H
H
5-(2-(trifluoromethyl)phenylthio)-1H- pyrazolo[4,3-b]pyridin-3-amine
17% 3 steps
311.0


 


7-6




N
H
H
5-(3,5-difluorophenylthio)-1H- pyrazolo[3,4-b]pyrazin-3-amine
6% 7 steps
279.9


 


7-7




N
H
H
5-(2,4-dichlorophenylthio)-1H- pyrazolo[3,4-b]pyrazin-3-amine
4% 7 steps
311.9


 


7-8




N
H
H
5-(2-(trifluoromethyl)phenylthio)-1H- pyrazolo[3,4-b]pyrazin-3-amine
6% 7 steps
311.9


 


7-9




CH
H
H
5-(3,5-difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3-amine
6% 3 steps
277.0


 


7-10




CH
H
H
5-(2,5-difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3-amine
3% 3 steps
277.0


 


7-11




CH
H
H
5-(2,5-dichlorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3-amine
32% 3 steps
309.0


 


7-12




CH
H
H
5-(5-chloro-2-(trifluoromethyl)benzyloxy)- 1H-pyrazolo[4,3-b]pyridin-3-amine
8% 3 steps
343.1


 


7-13




CH
H
H
5-(pyridin-3-ylmethoxy)-1H-pyrazolo[4,3- b]pyridin-3-amine
6% 3 steps
342.1





**1H NMR: δH ppm (400 MHz, DMSO):


7-1: 11.61 (1H, s large, NH), 7,73 (1H, d, CHarom), 7.24 (2H; m, CHarom), 7.18 (1H, m, CHarom), 6.86 (1H, d, CHarom).


7-2: 11.95 (1H, sl, NH), 7.78 (1H, d, CHarom, J = 11.6 Hz), 7.33 (1H, d, CHarom, J = 11.6 Hz), 7.19 (1H, t, CHarom), 7.04 (2H, 2d, CHarom, J = 8.8 Hz), 5.51 (2H, s, NH2).


7-3: 11.80 (1H, sl, NH), 7.70 (1H, d, CHarom, J = 8.8 Hz), 7.60 (1H, t, CHarom), 7.49 (1H, q, CHarom), 7.27-7.33 (1H, m, CHarom), 7.11 (1H, d, CHarom, J = 8.8 Hz), 5.41 (2H, s, NH2).


7-4: 11.93 (1H, sl, NH), 7.80 (1H, d, CHarom, J = 11.6Hz), 7.62 (1H, d, CHarom, J = 11.6 Hz), 7.40 (1H, dd, CHarom, J = 11.2 Hz), 7.29 (1H, d, CHarom, J = 11.6 Hz), 7.1 (1H, s, CHarom), 5.51 (2H, s, NH2).


7-5: 11.86 (1H, sl, NH), 7.87 (1H, d, CHarom, J = 9.6 Hz), 7.73 (1H, d, CHarom, J = 11.6), 7.50-7.68 (2H, m, CHarom), 7.44 (1H, d, CHarom, J = 10.4Hz), 7.11 (1H, d, CHarom, J = 11.6Hz), 5.46 (2H, s, NH2).


7-6: 12.66 (1H, sl, NH), 8.52 (1H, s, CHarom), 7.12-7.20 (1H, m, CHarom), 7.02-7.10 (2H, m, CHarom), 5.90 (2H, s, NH2).


7-7: 12.70 (1H, s, NH), 8.52 (1H, s, CHarom), 7.60 (1H, d, CHarom, J = 8.8 Hz), 7.38 (1H, dd, CHarom, J = 8.4 Hz), 7.12 (1H, s, CHarom), 5.92 (2H, s, NH2).


7-8: 12.66 (1H, s, NH), 8.39 (1H, s, CHarom), 7.84 (1H, d, CHarom, J = 7.6 Hz), 7.58 (1H, t, CHarom), 7.50 (1H, t, CHarom), 7.34 (1H, d, CHarom, J = 7.6 Hz), 5.87 (2H, s, NH2).


7-9: 11.57 (1H, s, NH), 7.74 (1H, d, Charom, J = 9 Hz), 7.25 (3H, m, CHarom), 6.88 (1H, d, Charom, J = 9 Hz), 5.44 (2H, s), 5,08 (2H, s).


7-10: 11.58 (1H, s, NH), 7.73 (1H, d, CHarom, J = 12.0 Hz), 7.48-7.58 (1H, m, CHarom), 7.21-7.37 (2H, m, CHarom), 6.85 (1H, d, CHarom, J = 12.0 Hz), 5.44 (2H, s, CH), 5.10 (2H, sl, NH2).


7-11: 11.60 (1H, sl, NH), 7.70-7.77 (2H, m, CHarom), 7.57 (1H, d, CHarom, J = 11.2 Hz), 7.40-7.50 (1H, m, CHarom), 6.89 (1H, d, CHarom, J = 12.0 Hz), 5.48 (2H, s, CH), 5.06 (2H, sl, NH2).


7-12: 11.60 (1H, sl, NH), 7.91 (1H, s, CHarom), 7.83 (1H, d, CHarom, J = 11.2 Hz), 7.75 (1H, d, CHarom, J = 12.0 Hz), 7.66 (1H, d, CHarom, J = 9.6 Hz), 6.88 (1H, d, CHarom, J = 12.0 Hz), 5.58 (2H, s, CH), 5.01 (2H, sl, NH2).


7-13: 11.56 (1H, sl, NH), 8.77 (1H, s, CHarom), 8.55 (1H, s, CHarom), 7.96 (1H, d, CHarom, J = 10.4 Hz), 7.72 (1H, d, CHarom, J = 12.0 Hz), 7.42 (1H, dd, CHarom, J = 10.0 Hz), 6.83 (1H, d, CHarom, J = 11.6 Hz), 5.45 (2H, s, CH), 5.15 (2H, sl, NH2).






Example 8
N5-(3,5-difluorophenyl)-1H-pyrazolo[4,3-b]pyridine-3,5-diamine




Example 8a
6-(3,5-difluorophenylamino)-3-nitropicolinonitrile
A mixture of 6.5 g of 6-chloro-3-nitropicolinonitrile (0.065 mol) and 6.2 g of 3,5-difluoroaniline (0.048 mol) in 100 ml of toluene is heated at 70° C. for 5 hours. The crude reaction product is diluted in an ethyl acetate fraction and washed using saturated NaCl solution. The organic phase is dried with sodium sulfate and the residue purified by silica gel chromatography (AcOEt/petroleum ether) to yield 3.9 g (33%) of a yellow solid.
Example 8b
3-amino-6-(3,5-difluorophenylamino)picolinonitrile
10 ml of concentrated HCl is added to a solution of 6-(3,5-dichlorophenylthio)-3-nitropicolinonitrile (3.9 g, 0.0141 mol) in 150 ml of ethanol under stirring. The reaction medium is refluxed, added together with 2.4 g of iron (0.0423 mol) and stirred at 80° C. for 1 hour. After returning to 0° C. the pH is adjusted to 8 using 1 N soda solution and the reaction medium is filtered on Celite. The reaction mixture is added together with 100 ml of ethyl acetate and 50 ml of methanol. The organic phase is extracted and the aqueous phase is extracted several times by ethyl acetate fractions. The organic phases are combined and then dried on anhydrous sodium sulfate before being concentrated to yield, after concentration, 2.3 g (66%) of a brown solid.
Example 8
5-(3,5-difluorophenylamino)-1H-pyrazolo[4,3-b]pyridin-3-amine
A solution of 713 mg of NaNO2 (10.3 mmol) in water (5 ml) is added, drop by drop, to a stirring solution of 2.3 g of 3-amino-6-(3,5-difluorophenylamino)picolinonitrile (9.4 mmol) in 100 ml of 6 N hydrochloric acid at 0° C. The mixture is stirred for 20 minutes at 0-5° C. A solution of 5.3 g of SnCl2.2H2O (23.5 mmol, 2.5 eq) in hydrochloric acid (12 N solution, 30 ml) is then added drop by drop and the solution is stirred for 1 hour at room temperature. The reaction medium is then cooled at 0° C. and basified to pH 8 using 30% soda solution. The mixture is extracted with ethyl acetate and washed using saturated NaCl solution and the organic phase is dried on anhydrous sodium sulfate before being concentrated under vacuum. The residue is purified by silica column chromatography (AcOEt). A light yellow solid is obtained (530 mg, 22%).
LCMS: m/z 262 (M+H+).
1H NMR: δH ppm (400 MHz, DMSO): 11.47 (s, 1H), 9.45 (s, 1H), 7.65 (m, 3H), 6.87 (d, 1H, J=7.8 Hz), 6.60 (m, 1H), 5.09 (s, 2H).
The following compounds are obtained by a similar method:
















 














Ex.**
ArX
Y4
W
Rj
Compound names
Yield
Mass MH+





8-1




CH
H
H
N-(2,5-difluorophenyl)-1H- pyrazolo[4,3-b]pyridine-3,5-diamine
4% 4 steps
262.0


 


8-2




CH
H
H
N-(2,5-dichlorophenyl)-1H- pyrazolo[4,3-b]pyridine-3,5-diamine
9% 4 steps
294.0





**1H NMR: δH ppm (400 MHz, DMSO):


8-1: 11.46 (1H, s, NH), 8.75-8.82 (2H, m, CHarom), 7.65 (1H, dd, CHarom, J = 9.2 Hz), 7.19-7.31 (2H, m, CHarom), 6.67-6.63 (1H, sl, CHarom), 5.06 (2H, s, NH2).


8-2: 11.58 (1H, sl, NH), 8.65 (1H, s, CHarom), 8.35 (1H, s, CHarom), 7.69 (1H, d, CHarom, J = 12.0 Hz), 7.45 (1H, d, CHarom, J = 11.6 Hz), 7.24 (1H, d, CHarom, J = 12.0 Hz), 6.96 (1H, dd, CHarom, J = 11.2 Hz), 5.03 (2H, sl, NH2).






Example of Method B3
Example 9
5-(3,5-difluorobenzyl)-1H-pyrazolo[4,3-b]pyridin-3-amine




This compound can be prepared from the following intermediates, according to method B3.
Example 9a
2-(3,5-difluorobenzyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
Example 9b
6-(3,5-difluorobenzyl)-3-nitropicolinonitrile
Example 9c
3-amino-6-(3,5-difluorobenzyl)picolinonitrile
Example of Method B4
Example 10
3-amino-N-(3,5-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-5-sulfonamide




Example 10a
5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinic acid
2.74 g (9.64 mmol) of ethyl 2-chloro-5-(chlorosulfonyl)nicotinate in solution in 20 ml of anhydrous dichloromethane is added, drop by drop at 0° C., to a mixture of 623 mg (4.82 mmol) of 3,5-difluoroaniline and 1.68 ml (12.05 mmol) of triethylamine diluted in 10 ml of anhydrous dichloromethane. The solution is stirred at room temperature for 3 hours. The solvent is evaporated to yield a light brown solid. The solid is triturated in 20 ml of methanol, filtered and then rinsed with 3 ml of methanol to yield 2.85 g of a white solid.
This solid is redissolved in 25 ml of tetrahydrofuran and is added together with a solution of 0.421 g (10.04 mmol) of lithium monohydrate hydroxide in 10 ml of water. The reaction mixture is left under stirring for 3 hours at 35° C. and then diluted in water, acidified with 1 N hydrochloric acid and extracted with ethyl acetate. The organic phase is collected, dried on sodium sulfate, filtered and concentrated to yield 1.12 g of 5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinic acid in the form of an orange solid (yield=67%).
1H NMR: δH ppm (400 MHz, DMSO): 8.91 (1H, s, CHarom), 8.51 (1H, s, CHarom), 7.02 (1H, dd, CHarom), 6.83 (2H, d, CHarom).
Example 10b
2-chloro-5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinamide
0.288 ml (3.87 mmol) of thionyl chloride and a drop of DMF are added successively to 0.450 g (1.29 mmol) of 2-chloro-5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinic acid in 5 ml of anhydrous toluene. The mixture is placed under stirring, at reflux of toluene, for 2 hours. The acid chloride reaction mixture is then added drop by drop to an iced solution, under stirring, of 4.5 ml of 25% ammonium hydroxide. A release of gas is observed. The reaction medium is left under stirring at room temperature for 30 minutes. The reaction medium is extracted several times with ethyl acetate. The combined organic phases are dried on anhydrous sodium sulfate and then concentrated. 0.315 g of 2-chloro-5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinamide in the form of a light brown solid is obtained (yield=72%).
1H NMR: δH ppm (400 MHz, DMSO): 11.18 (1H, bs, NH), 8.86 (1H, s, CHarom), 8.22 (1H, s, CHarom), 8.21 (1H, bs, NH), 7.98 (1H, bs, NH), 6.96 (1H, dd, CHarom), 6.79 (2H, d, CHarom).
Example 10c
6-chloro-5-cyano-N-(3,5-difluorophenyl)pyridine-3-sulfonamide
3.4 ml (36.2 mmol) of phosphoryl chloride and a drop of concentrated sulfuric acid are added to 0.315 g (0.906 mmol) of 2-chloro-5-(N-(3,5-difluorophenyl)sulfamoyl)nicotinamide. The reaction mixture is stirred for 2 hours at 90° C. and then added drop by drop to ice. The brown solid is filtered, rinsed with water and then dried under vacuum. 0.217 g of 6-chloro-5-cyano-N-(3,5-difluorophenyl)pyridine-3-sulfonamide is obtained in the form of a light brown solid (yield=72%).
1H NMR: δH ppm (400 MHz, DMSO): 11.34 (1H, bs, NH), 9.04 (1H, s, CHarom), 8.92 (1H, s, CHarom), 7.03 (1H, dd, CHarom), 6.85 (2H, d, CHarom).
Example 10
3-amino-N-(3,5-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-5-sulfonamide
0.377 ml (2.63 mmol) of 35% hydrazine is added to 0.217 g (0.658 mmol) of 6-chloro-5-cyano-N-(3,5-difluorophenyl)pyridine-3-sulfonamide diluted in 6 ml of isopropanol. The solution is heated at 75° C. for 2 hours. The solvent is evaporated to yield 0.214 g of 3-amino-N-(3,5-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-5-sulfonamide in the form of a yellow solid (yield=100%).
1H NMR: δH ppm (400 MHz, DMSO): 8.74 (1H, d, CHarom), 8.68 (1H, d, CHarom), 6.88 (1H, dd, CHarom), 6.80 (2H, d, CHarom), 6.04 (2H, bs, NH).
Examples of Method B5
Example 11
5-(3,5-difluorobenzyloxy)-1H-pyrazolo[3,4-b]pyridin-3-amine




This compound can be prepared from the following intermediates, according to method B5.
Example 11a
5-hydroxynicotinonitrile
A mixture of 1 g of 5-methoxynicotinonitrile (7.46 mmol) and 8.62 g of pyridine hydrochloride is heated at 200° C. for 2 hours. The crude reaction product is diluted in a water fraction several times with diethyl ether. The aqueous phase is basified by adding sodium bicarbonate and then extracted again with diethyl ether. The organic phase is dried and then concentrated to yield 850 mg of 5-hydroxynicotinonitrile (95%) in the form of a beige solid.
LCMS: m/z 120.94 (M+H+).
1H NMR: δH ppm (400 MHz, DMSO): 10.79 (s, 1H), 8.46 (s, 1H, CHarom.), 8.42 (s, 1H, CHarom.), 7.60 (s, 1H, CHarom.).
Example 11b
5-(3,5-difluorobenzyloxy)nicotinonitrile
876 mg (2 eq) of sodium hydride is added gradually at 0° C. under nitrogen to a solution of 865 mg of 5-hydroxynicotinonitrile (7.2 mmol) in 15 mL of dimethylacetamide. The mixture is stirred 10 min at 0° C. before adding 2.24 g (1.5 aq) of 3,5-difluorobenzyl bromide. The mixture is placed under stirring for 2.5 additional hours before being diluted in an ethyl acetate fraction and being washed with aqueous fractions. The organic phases are isolated, dried and concentrated. The solid residue obtained is recrystallized in methanol to yield 1.1 g (68% of 5-(3,5-difluorobenzyloxy)nicotinonitrile in the form of a beige powder.
LCMS: m/z 247.11 (M+H+).
1H NMR: δH ppm (400 MHz, DMSO): 8.69 (s, 1H, CH), 8.65 (s, 1H, CH), 8.08 (s, 1H, CH), 7.26 (m, 3H, CH), 5.28 (d, 2H, CH2).
Example 11c
3-cyano-5-(3,5-difluorobenzyloxy)pyridine 1-oxide
224 mg of m-CPBA is added at 0° C. to a solution in acetonitrile of 250 mg of 5-(3,5-difluorobenzyloxy)nicotinonitrile. The reaction medium is stirred for 20 hours while a precipitate is formed progressivelt. This solid is then filtered and washed to yield 200 mg (75%) of 3-cyano-5-(3,5-difluorobenzyloxy)pyridine 1-oxide in the form of a white powder.
LCMS: m/z 263.06 (M+H+).
Example 11d
2-chloro-5-(3,5-difluorobenzyloxy)nicotinonitrile
A mixture of 650 mg of 3-cyano-5-(3,5-difluorobenzyloxy)pyridine 1-oxide in 2.3 mL of POCl3 added with few drops of H2SO4 is heated at 110° C. for 1 h 30. The crude reaction medium is then poured in ice and the precipitate thus formed is isolated by filtration and dried under vacuum to yield 600 mg of a beige solid in the form of a mixture of regioisomers comprising mainly the desired 2-chloro-5-(3,5-difluorobenzyloxy)nicotinonitrile which is used without further purification.
LCMS: m/z 281.02 (M+H+).
Example 11
5-(3,5-difluorobenzyloxy)-1H-pyrazolo[3,4-b]pyridin-3-amine
313 mg of hydrazine hydrate (5 eq) is added to a solution of 1.6 g of 2-chloro-5-(3,5-difluorobenzyloxy)nicotinonitrile (450 μmol) in 10 mL of propan-2-ol. The reaction mixture is heated at 100° C. for 6 hours. After return to room temperature leading to a precipitation, the crude reaction medium is filtered, the solid is removed and the filtrate is dry evaporated. It is then purified by chromatography on a silica column eluted with a gradient of ethyl acetate and methanol, whereas the more polar fraction is isolated, concentrated and suspended again in a small fraction of methanol under stirring. The solid thus obtained is isolated and dried to yield 221 mg of 5-(3,5-difluorobenzyloxy)-1H-pyrazolo[3,4-b]pyridin-3-amine in the form of a beige solid wich is used without further purification.
LCMS: m/z 277.07 (M+H+).
Example of Method B6
Example 11bis
N-(3-amino-1H-pyrazolo[3,4-b]pyridin-5-yl)-3,5-difluorobenzene sulfonamide




Example 11bis-a
N-(6-chloro-5-cyanopyridin-3-yl)-3,5-difluorobenzene-sulfonamide
1.132 g (5.32 mmol) of 3,5-difluorobenzene-1-sulfonyle chloride is added under argon to a solution of 545 mg (3.55 mmol) of 5-amino-2-chloronicotinotrile in 20 mL of an anhydrous 1:1 mixture of THF and pyridine. The reaction medium is heated to 70° C. for 3 hours and let 12 additional hours under stirring at room temperature. The solvent is dry evaporated and the crude reaction product is redissolved in ethyl acetate and washed with several aqueous fractions. The organic phase is dried on magnesium sulfate, filtered, concentrated and then purified by silica gel chromatography to yield 784 mg (67%) of N-(6-chloro-5-cyanopyridin-3-yl)-3,5-difluorobenzene-sulfonamide.
1H NMR: δH ppm (400 MHz, DMSO): 11.39 (1H, s1, NH), 8.34 (1H, m, CHarom), 8.10 (1H, m, CHarom), 7.67 (1H, m, CHarom), 7.59 (2H, m, CHarom).
Example 11bis
N-(3-amino-1H-pyrazolo[3,4-b]pyridin-5-yl)-3,5-difluorobenzene-sulfonamide
1.786 g (35.7 mmol) of hydrazine hydrate is added under argon to a solution of 784 mg (2.38 mmol) of N-(6-chloro-5-cyanopyridin-3-yl)-3,5-difluorobenzene-sulfonamide in 6 mL of ethanol. The solution is heated to 100° C. for 20 hours and then cooled to room temperature. The solvent is evaporated to yield 810 mg of N-(3-amino-1H-pyrazolo[3,4-b]pyridin-5-yl)-3,5-difluorobenzene-sulfonamide (100%) which is used without further purification in the following steps.
LCMS: m/z 326.07 (M+H+).
Example of Method C1
Example 12
N6-(2,4-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-3,6-diamine




This compound can be prepared from the following intermediates, according to method C1.
Example 12-a
5-cyano-6-(methylthio)pyridin-2-yl trifluoromethanesulfonate
15.26 mL (1.2 eq) of potassium 2-methylpropan-2-olate and then 9.03 g (1.2 eq) of 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide are added dropwise to a solution of 3.5 g (21.06 mmol) of 6-hydroxy-2-(methylthio)nicotinonitrile in 180 mL of tetrahydrofurane under nitrogen. The reaction mixture is stirred at room temperature for 2 h 45. Water is added and the product is extracted with ethyl acetate. The organic phase is dried on anhydrous magnesium sulfate, filtered and evaporated to yield an orange solid. The product is purified on a silica gel column (eluent: cyclohexane/dichloromethane 5:5) to yield 5.31 g (85%) of 5-cyano-6-(methylthio)pyridin-2-yl trifluoromethanesulfonate in the form of a yellow solid.
1H NMR: δH ppm (400 MHz, DMSO): 8.57 (1H, d, CH), 7.52 (1H, d, CH), 2.59 (3H, s, CH3).
Example 12-b
6-(2,4-difluorophenylamino)-2-(methylthio)nicotinonitrile
0.81 mL (1.2 eq) of 4-difluoroaniline and 1.53 g (1.4 eq) of cesium(I) carbonate are added under nitrogen to a solution of 2 g (6.71 mmol) of 5-cyano-6-(methylthio)pyridin-2-yl trifluoromethanesulfonate in 30 mL of 1,4-dioxane. The medium is degassed for 5 minutes under argon before adding 0.25 g (0.06 eq) of de 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl and 0.08 g (0.04 eq) of (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one, palladium(II) complex. The reaction medium is stirred at 100° C. for 2 hours. After return to room temperature, ethyl acetate and brine are added. The organic phase is dried on anhydrous magnesium sulfate, filtered and evaporated. The residue obtained is purified on silica gel chromatography (eluent: cyclohexane/ethyl acetate 8:2 then 7:3) to yield 1.52 g (82%) of 6-(2,4-difluorophenylamino)-2-(methylthio)nicotinonitrile in the form of a white solid.
LCMS (1E, m/z): (M+1) 278.06.
1H NMR: δH ppm (400 MHz, DMSO): 9.57 (1H, s, NH), 7.73-7.86 (2H, m, CH), 7.28-7.44 (1H, m, CH), 7.02-7.18 (1H, m, CH), 6.60 (1H, d, CH), 2.41 (3H, s, CH3).
Example 12
N6-(2,4-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-3,6-diamine
769 mg (3.12 mmol) of m-chloroperbenzoic acid (mCPBA) is added under argon to a stiring solution of 786 mg (2.83 mmol) of 6-(2,4-difluorophenylamino)-2-(methylthio)nicotinonitrile in 25 mL of dichloromethane. The reaction medium is stirred 1 hour at room temperature before adding a fraction of ethyl acetate and washed this organic phase with a NaHCO3 saturated solution. The combined organic phases are dried on magnesium sulfate and dry evaporated. The crude reaction product is dissolved again in 10 mL of propanol and 2 equivalents of hydrazine hydrochloride in water are added. The mixture is heated at 90° C. for 6 hours before being diluted in water and extracted with ethyl acetate. The organic phase is dried on magnesium sulfate and dry evaporated before being purified by silica gel chromatography to yield 495 mg of N6-(2,4-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-3,6-diamine in the form of a yellow-orange solid (67%).
LCMS (1E, m/z): (M+1) 262.14.
1H NMR: δH ppm (400 MHz, DMSO): 11.40 (1H, s, NH), 8.76 (1H, s, NH), 8.15 (1H, m, CH), 7.81 (1H, d, CH), 7.28 (1H, m, CH), 7.06 (1H, m, CH), 6.55 (1H, d, CH), 5.24 (2H, s, NH2).
The following compound is obtained by a similar method:
















 















Compound

Masse


Ex.**
ArX
Y1
names
Yield
MH+





12-1




CH
N6-(3,5- difluoro- benzyl)-1H- pyrazolo[3,4- b]pyridine- 3,6-diamine
70%
276.15





**1H NMR, dmso-d6, Ex.: 12-1: 11.17 (1H, s, NH), 7.66 (1H, d, CH), 7.37 (1H, s, NH), 7.04 (3H, m, CH), 6.24 (1H, d, CH), 5.11 (2H, s, NH2), 4.52 (2H, s, CH2).






Example 12bis
N6-(2,4-difluorophenyl)-N6-methyl-1H-pyrazolo[3,4-b]pyridine-3,6-diamine




Example 12bis-a
6-((3,5-difluorophenyl)(methyl)amino)-2-(methylthio)nicotinonitrile
3.05 mL (5.04 mmol) of potassium 2-methylpropan-2-olate and then 286 μL (1.8 eq) of iodomethane are added dropwise under nitrogen to a solution of 700 mg (2.52 mmol) of 6-(2,4-difluorophenylamino)-2-(methylthio)nicotinonitrile in 20 mL of N,N-dimethyl formamide. The reaction medium is stirred at room temperature for 24 hours and then 126 μL (0.8 eq, 2.02 mmol) of iodomethane is added. The reaction medium is stirred at room temperature for 2 additional hours. Water is added and the product is extracted with ethyl acetate. The organic phase is dried on anhydrous magnesium sulfate, filtered, and evaporated to yield 660 mg (90%) of 6-((2,4-difluorophenyl)(methypamino)-2-(methylthio)nicotinonitrile in the form of a brown solid.
LCMS (IE, m/z): (M+1) 292.09.
1H NMR: δH ppm (400 MHz, DMSO): 7.74-7.80 (1H, m, CH), 7.55-7.63 (1H, m, CH), 7.43-7.52 (1H, m, CH), 7.18-7.27 (1H, m, CH), 6.16-6.30 (1H, m, CH), 3.43 (3H, s, CH3), 2.42 (3H, s, CH3).
Example 12bis
N6-(2,4-difluorophenyl)-N6-methyl-1H-pyrazolo[3,4-b]pyridine-3,6-diamine
452 mg (1.84 mmol) of mCPBA is added under argon to a stirring solution of 486 mg (1.67 mmol) of 6-((2,4-difluorophenyl)(methyl)amino)-2-(methylthio)nicotinonitrile in 15 mL of dichloromethane. The reaction medium is stirred 30 min at room temperature before adding an ethyl acetate fraction. The organic phase is washed with a NaHCO3 saturated solution, dried on magnesium sulfate and dry evaporated. The crude reaction product is dissolved again in 6 mL of propanol and 164 μL (3.38 mmol) of hydrazine hydrochloride in water is added. The mixture is heated at 90° C. for 6 hours before being diluted in water and extracted with ethyl acetate. The organic phase is dried on magnesium sulfate and dry evaporated before being purified by silica gel chromatography to yield 328 mg of N6-(2,4-difluorophenyl)-N6-methyl-1H-pyrazolo[3,4-b]pyridine-3,6-diamine in the form of a yellow-orange solid (70%).
LCMS (IE, m/z): (M+1) 276.15.
1H NMR: δH ppm (400 MHz, DMSO): 11.41 (1H, s, NH), 7.75 (1H, d, CH), 7.51-7.55 (1H, m, CH), 7.40-7.43 (1H, m, CH), 7.17-7.22 (1H, m, CH), 6.03 (1H, d, CH), 5.23 (2H, s, NH2), 3.28 (3H, s, CH3).
Example of Method C3
Example 12ter
6-(2,4-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine




Example 12ter-a
2-chloro-6-(2,4-difluorophenylthio)nicotinonitrile
A solution of 362 mg (1.05 eq) of potassium hydroxide in 10 mL of ethanol is added, under nitrogen, to a solution of 698 μL (6.16 mmol) of 2,4-difluorobenzenethiol in 30 mL of ethanol. The reaction medium is stirred at room temperature for 15 minutes and then cooled in ice before adding a solution of 1.015 g (0.95 eq) of 2,6-dichloronicotinonitrile in 30 mL of ethanol. The reaction medium is stirred for 2 hours at 0-5° C. 63 mL of a 0.1N HCl solution is added to stop the reaction. Water is added and the producted is extracted with ethyl acetate. The organic phase is dried on anhydrous magnesium sulfate, filtered and evaporated. The residue is purified by silica gel chromatography (eluent: cyclohexane/ethyl acetate 94:6) to yield 1.09 g (66%) of 2-chloro-6-(2,4-difluorophenylthio)-nicotinonitrile in the form of a white solid.
LCMS (IE, m/z): (M+1) 282.98.
1H NMR: δH ppm (400 MHz, DMSO): 8.24 (1H, d, CH), 7.77-7.85 (1H, m, CH), 7.52-7.63 (1H, m, CH), 7.25-7.35 (2H, m, CH), 2.41 (3H, s, CH3).
Example 12ter
6-(2,4-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine
0.561 mL (11.57 mmol) of hydrazine monohydrate is added under nitrogen to a stirring solution of 1.09 g (3.86 mmol) of 2-chloro-6-(2,4-difluorophenylthio)nicotinonitrile in 15 mL of propanol. The reaction medium is heated at 80° C. for 4 hours. A precipitate is obtained when the reaction medium is returned to room temperature. This precipitate is filtered and rinced with ethanol. The solid is dissolved in an ethyl acetate fraction and washed with a 1N HCl solution. The organic phase is dried on magnesium sulfate and dry evaporated to yield 420 mg (39%) of 6-(2,4-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine in the form of a yellow solid.
1H NMR: δH ppm (400 MHz, DMSO): 12.10 (1H, s, NH), 8.11 (1H, d, CH), 7.82-7.89 (1H, m, CH), 7.58-7.63 (1H, m, CH), 7.32-7.36 (1H, m, CH), 6.86 (1H, d, CH), 4.59 (2H, s, NH2).
The following compound is obtained by a similar method:
















 



















Mass


Ex.**
ArX
Y1
Rj
Compound names
Yield
MH+





12ter-1




CH
H
6-(2,4- difluorophenoxy)- 1H-pyrazolo[3,4- b]pyridin-3-amine
ND
263.06









Example 12quater
6-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-amine




17.35 mL of a 0.5M solution in THF of (3,5-difluorobenzyl)zinc chloride (8.58 mmol) is added under argon to a solution of 416 mg of palladium(II) chloride (510 mmol) and 883 mg of 2,6-dichloronicotinonitrile (5.1 mmol) in 2 mL of anhydrous THF. The reaction is refluxed for 7 hours, then cooled to room temperature. A 1N soda aqueous solution is added and the product is extracted with several successive ethyl acetate fractions. The organic phases are dried on magnesium sulfate and dry evaporated before being purified by silica gel chromatography to yield 680 mg of a mixture of 2-chloro-6-(3,5-difluorobenzyl)-nicotinonitrile and by-products wich is used without further purification in the following step.
The previous mixture is dissolved in 10 mL of isopropanol under stirring and 750 μL of 35% hydrazine hydrate is added. The solution is heated at 80° C. for 4 hours. The solvent is dry evaporated and the product is purified by silica gel chromatography (dichloromethane/methanol 9:1) to yield 290 mg of 6-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-amine (64%).
LCMS (IE, m/z): (M+1) 261.16.
1H NMR: δH ppm (400 MHz, DMSO): 11.82 (1H, s, NH), 8.01 (1H, d, CH), 6.99-7.04 (3H, m, CH), 6.91 (1H, d, CH), 5.49 (2H, s, NH2), 4.12 (2H, s, CH2).
Example of Method D1
Example 13
5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-amine




0.575 g (0.704 mmol) of (dppf)2PdCl2.CH2Cl2 and 28 ml (14.08 mmol) of 3,5-difluorobenzyl zinc (II) chloride are added to 1.5 g (7.04 mmol) of a solution of 5-bromo-1H-pyrazolo[3,4-b]pyridin-3-amine in 10 ml of tetrahydrofuran. The reaction medium is heated at 90° C. for 18 hours. After returning to room temperature, the reaction is hydrolyzed by slowly adding water at 0° C. After filtration of the precipitate formed, the solid is rinsed with tetrahydrofuran and the aqueous filtrate is extracted several times with ethyl acetate. The organic phases are combined, dried on magnesium sulfate and concentrated. The residue is purified by silica chromatography (95:4.5:0.5 and then 95:4:1 dichloromethane/methanol/ammonium as eluent) to yield 1.7 g (93%) of 5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-amine in the form of a beige solid.
LCMS (EI, m/z): (M+1) 261.41.
1H NMR: δH ppm (400 MHz, DMSO): 11.87 (1H, s, NH), 8.31 (1H, d, CHarom), 7.92 (1H, d, CHarom), 6.98-7.08 (3H, m, CHarom), 5.47 (2H, s, NH), 4.04 (2H, s, CH2).
The following compounds are obtained by a similar method:
















 














Ex.




Compound

Mass


**
ArX
Y4
W
Rj
names
Yield
MH+





13-1




CH
H
H
5-(3,5- difluoro- benzyl)-1H- pyrazolo[4,3- b]pyridin-3- amine
 8% 4 steps
261.1


 


13-2




N
H
H
5-(3,5- difluoro- benzyl)-1H- pyrazolo[3,4- b]pyrazin-3- amine
21% 3 steps
262.1





** 1H NMR: δH ppm (400 MHz, DMSO):


13-1: 11.61 (1H, sl, NH), 7.65 (1H, d, CHarom, J = 11.6 Hz), 7.20 (1H, d, CHarom, J = 11.2 Hz), 6.95-7.10 (3H, m, CHarom), 5.32 (2H, sl, NH2), 4.18 (2H, s, CH2).


13-2: 12.31 (1H, sl, NH), 8.44 (1H, s, CHarom), 7.03-7.08 (3H, m, CHarom), 5.61 (2H, sl, NH2), 4.25 (2H, s, CH2).






Examples of Method D2
Example 14
5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyrazin-3-amine




0.7 g (2.68 mmol) of 5-iodo-1H-pyrazolo[3,4-b]pyridine-3-amine, 0.74 g (5.36 mmol) of anhydrous potassium carbonate and 0.10 g of copper iodide (0.536 mmol) are mixed in a 50 ml round-bottom flask. 15 ml of propan-2-ol, 0.01 g (0.2 mmol) of polyethylene glycol and 0.43 g (2.95 mmol) of 3,5-difluorothiophenol are then added. The reaction mixture is heated at 80° C. for 2 hours. The solvent is evaporated and the solid formed is filtered, rinsed with water and then with pentane and dried in an oven at 50° C. to yield 0.75 g (100%) of 5-(3,5-diflurophenylthio)-1H-pyrazolo[3,4-b]pyrazin-3-amine in the form of a brown solid.
LCMS (EL m/z): (M+1) 280.03.
1H NMR: δH ppm (400 MHz, DMSO): 12.65 (1H, bs, NH), 8.52 (1H, s, CHarom), 7.18 (1H, t, CHarom), 7.05-7.18 (2H, m, CHarom), 5.89 (2H, s, NH).
The following derivatives were obtained according to the same method:
















 
















Ex.**
Ar
Rk
n
Y1, Y3, Y4
R3
Rj
Compound name
Yield
Mass MH+





14-1 




H
0
CH, CH, N
H
H
2-(3-amino-1H- pyrazolo[3,4- b]pyridin-5- ylthio)benzamide
ND
ND


 


14-2 








0
CH, CH, N
H
H
N-(5-(3,5- dimethylphenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
ND
ND


 


14-3 




H
0
CH, CH, N
H
H
5-(3,5- difluorophenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-amine
45%
(M + 1) 279.28


 


14-4 




H
0
CH, C—OMe, N
H
H
5-(2,5- dichlorophenylthio)- 6-methoxy-1H- pyrazolo[3,4- b]pyridin-3-amine
80%
ND


 


14-5 




H
0
CH, C—NH2, N
H
H
5-(2,5- dichlorophenylthio)- 1H-pyrazolo[3,4- b]pyridine-3,6- diamine
35%
ND


 


14-6 
 



H
0
CH, CH, N
H
tBu
1-tert-butyl-5-(3,5- difluorobenzylthio)- 1H-pyrazolo[3,4- b]pyridin-3-amine
ND
(M + 1) 293.08


 


14-7 
 



H
0
CH, CMe, N
H
H
5-(3,5- difluorophenylthio)- 6-methyl-1H- pyrazolo[3,4- b]pyridin-3-amine
ND
(M + 1) 293.06


 


14-8 
 



H
0
CH, C—OMe, N
H
H
5-(3,5- difluorophenylthio)- 6-methoxy-1H- pyrazolo[3,4- b]pyridin-3-amine
28%
(M + 1) 610.30


 


14-9 




H
0
CH, CH, N
H
tBu
1-tert-butyl-5-(3,5- difluorophenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-amine
79%
(M + 1) 335.26


 


14-10








0
CH, CH, N
H
H
N-(5-(2,5- dichlorophenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
31%
(M + 1) 612.37


 


14-11








0
CH, C—NH2, N
H
H
N-(6-amino-5-(3,5- difluorophenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
68%
ND





**1H NMR, DMSO-d6, Ex.:


14-3: 12.65 (1H, bs, NH), 8.52 (1H, s, CHarom), 7.18 (1H, t, CHarom), 7.05-7.18 (2H, m, CHarom), 5.89 (2H, s, NH).


14-6: 8.21 (2H, bs, CHarom), 7.07 (1H, m, CHarom), 6.90 (2H, m, CHarom), 6.27 (2H, bs, NH), 4.03 (2H, s, CH), 1.63 (9H, s, CH).


14-7: 12.16 (1H, bs, NH), 8.39 (1H, s, CHarom), 7.00-7.08 (1H, m, CHarom), 6.64-6.72 (2H, m, CHarom), 5.73 (2H, bs, NH2), 2.54 (3H, s, CH3).


14-9: 8.51 (1H, bs, CHarom), 8.35 (1H, bs, CHarom), 7.02 (1H, m, CHarom), 6.72 (2H, bs, CHarom), 6.52 (2H, bs, NH), 1.67 (9H, s, CH).


(ND: not determined).






Example 14bis
N-(5-(3,5-difluorophenylamino)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




A solution of 225 mg of N-(5-iodo-1-trityl-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide (0.25 mmol), 36 mg of difluoroaniline (0.275 mmol), 19 mg of R-(+)-2,2′-bis(diphenylphoshino)-1,1′-binaphtyle (0.030 mmol), 11 mg (0.013 mmol) of tris(dibenzylideneacetone)dipalladium(0) and 75 mg (0.75 mmol) of sodium tert-butoxide in 10 mL of THF is refluxed under argon overnight. The crude reaction medium is cooled, extracted with ethyl acetate and washed with water. The organic phase is dried on magnesium sulfate and purified by silica gel chromatography to yield N-(5-(3,5-difluorophenylamino)-1-trityl-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide which is used in the following step without further purification.
The product thus obtained is dissolved in 10 mL of dichloromethane at 0° C. and 56 mg (0.5 mmol) of TFA is added. The reaction medium is stirred for 4 hours. Water is added and the pH of the reaction medium is adjusted to 7 with a NaHCO3 solution. The aqueous phase is collected, basified (pH 9-10) with a concentrated K2CO3 solution and extracted with dichloromethane. The organic phase is collected, dried on magnesium sulfate and dry ccentrated to yield 40 mg of N-(5-(3,5-difluorophenylamino)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide.
LCMS (IE, m/z): (M+1) 562.12.
1H NMR: δH ppm (400 MHz, DMSO): 13.45 (1H, s1, NH), 10.47 (1H, s1, NH), 8.65 (1H, s, CHarom), 8.55 (1H, s, CHarom), 8.14 (1H, d, NH), 7.77 (1H, d, CHarom), 7.26 (2H, m, CHarom), 7.05 (1H, m, CHarom), 6.25 (1H, d, CHarom), 6.14 (1H, s, NH), 6.77 (1H, s, NH), 3.82-3.84 (2H, dt, CH), 3.72 (1H, m, CH), 3.47-3.52 (2H, m, CH), 3.28-3.34 (4H, m, CH), 2.43 (4H, m, CH), 2.23 (3H, s, CH3), 1.94-1.97 (2H, m, CH), 1.37-1.39 (2H, m, CH).
Examples of Method D3
Example 15
N-(5-((3,5-difluorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




0.94 mg (0.926 mmol) of triethylamine is added to 400 mg (0.712 mmol) of N-(5-iodo-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide, 67.8 mg (0.356 mmol) of CuI, and 50 mg (0.071 mmol) of Pd(PPh3)2Cl2 under argon in 12 ml of anhydrous dioxane under stirring. The reaction is heated for 3.5 hours at 100° C. The reaction mixture is diluted with 30 ml of water and extracted with ethyl acetate. The organic phase is dried on sodium sulfate, filtered and concentrated. The residue obtained is purified by silica gel chromatography (dichloromethane/methanol) to yield 152 mg of N-(5-((3,5-difluorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in the form of a yellow solid (yield=37%).
LCMS (EL in/z): (M+1) 572.17.
1H NMR: δH ppm (400 MHz, DMSO): 13.57 (1H, bs, NH), 10.56 (1H, bs, NH), 8.68 (1H, s, CHarom), 8.43 (1H, s, CHarom), 8.13 (1H, d, NH), 7.80 (1H, d, CHarom), 7.38 (2H, m, CHarom), 6.27 (1H, d, CHarom), 6.15 (1H, d, CHarom), 3.84-3.82 (2H, dt, CH), 3.70 (1H, m, CH), 3.45-3.50 (2H, m, CH), 3.21-3.33 (4H, m, CH), 2.42-2.46 (4H, m, CH), 2.28 (3H, s, CH3), 1.94-1.97 (2H, m, CH), 1.37-1.39 (2H, m, CH).
The following derivative was obtained according to the same method:
















 












Ex.**
ArX
Y1, Y3, Y4
Compound name
Yield
Mass MH+





15-1




N, CH, N
5-((3,5- difluorophenyl)ethynyl)- 1H-pyrazolo[3,4- b]pyrazin-3-amine
6% 6 steps
272.1





**1H NMR, dmso-d6, Ex.:


15-1: 12.71 (1H, sl, NH), 8.66 (1H, s, CHarom), 7.40-7.47 (3H, m, CHarom), 6.01 (2H, sl, NH2).






Examples of Method E
The protocols comprising method E aim at functionalizing the exocyclic amine of the aminopyrazole rings by their reaction with an intermediate featuring an electrophilic function, optionally generated in situ, such as acid chloride, isocyanate, isothiocyanate or aldehyde.
PPreparation of the Reaction Intermediates
Example 16
2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methyl piperazin-1-yl)benzoic acid




Example 16a
Tert-butyl 4-(4-methylpiperazin-1-yl)-2-nitrobenzoate
This compound was previously described in WO 2008/74749.
5.28 ml (47.6 mmol) of 1-methylpiperazine is added to 4.1 g (17 mmol) of tert-butyl 4-fluoro-2-nitrobenzoate. The reaction mixture is stirred without solvent for 5 hours. 150 ml of water is added to the reaction mixture and it is stirred for 24 hours. The precipitate formed is filtered, rinsed with water and dried under vacuum to yield 4.9 g (90%) of tert-butyl 4-(4-methylpiperazin-1-yl)-2-nitrobenzoate in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 322.37.
1H NMR: δH ppm (400 MHz, DMSO): 7.69 (1H, d, CHarom), 7.30 (1H, d, CHarom), 7.20 (1H, dd, CHarom), 3.38 (4H, m, CH), 2.40 (4H, m, CH), 2.22 (3H, s, CH3), 1.45 (9H, s, CH3).
Example 16b
Tert-butyl 2-amino-4-(4-methylpiperazin-1-yl)benzoate
This compound was previously described in WO 2008/74749.
0.160 g (1.500 mmol) of palladium on carbon (10%) and 15.19 ml (150 mmol) of cyclohexene are added to a solution of 4.82 g (15 mmol) of tert-butyl 4-(4-methylpiperazin-1-yl)-2-nitrobenzoate in 100 ml of ethanol. The reaction mixture is heated at a temperature of 80° C. for 8 hours. The reaction mixture is filtered and then rinsed with ethanol to yield 4.2 g (yield=96%) of tert-butyl 2-amino-4-(4-methylpiperazin-1-yl)benzoate in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 292.39.
1H NMR: δH ppm (400 MHz, DMSO): 7.44 (1H, d, CHarom), 6.40 (2H, bs, NH2), 6.19 (1H, dd, CHarom), 6.12 (1H, d, CHarom), 3.17 (4H, m, CH), 2.40 (4H, m, CH), 2.22 (3H, s, CH3), 1.49 (9H, s, CH3).
Example 16c
Tert-butyl 2-(4,4-difluorocyclohexylamino)-4-(4-methylpiperazin-1-yl)benzoate
1.045 ml (13.57 mmol) of trifluoroacetic acid, 1 g (7.46 mmol) of 4,4-difluorocyclohexanone and 2.158 g (8.20 mmol) of tetramethylammonium triacetoxyborohydride are added to 1.521 g (5.22 mmol) of tert-butyl 2-amino-4-(4-methylpiperazin-1-yl)benzoate dissolved in 60 ml of dichloromethane. The reaction is left under stirring at room temperature for 24 hours. The solvent is evaporated and then the crude reaction product is redissolved in 30 ml of ethyl acetate. The solution is successively washed with 0.5 M HCl solution, 0.5 M soda solution and finally with saturated NaHCO3 solution. The organic phase is dried on sodium sulfate, filtered and concentrated to obtain 2.2 g of tert-butyl 2-(4,4-difluorocyclohexylamino)-4-(4-methylpiperazin-1-yl)benzoate in the form of a light brown gum (yield=72%).
LCMS (EI, m/z): (M+1) 410.3.
1H NMR: δH ppm (400 MHz, DMSO): 7.73 (1H, bs, NH), 7.58 (1H, m, CHarom), 7.77 (1H, m, CHarom), 6.09 (1H, bs, CHarom), 3.37 (4H, m, CH), 3.27 (4H, m, CH), 2.47 (4H, m, CH), 2.25 (3H, s, CH), 1.99 (4H, s, CH), 1.40 (9H, s, CH).
Example 16d
Tert-butyl 2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methylpiperazin-1-yl)benzoate
0.99 ml (6.98 mmol) of trifluoroacetic anhydride and 1.12 ml (8.06 mmol) of triethylamine are added to 2.2 g (5.3 mmol) of tert-butyl 2-(4,4-difluorocyclohexylamino)-4-(4-methylpiperazin-1-yl)benzoate dissolved in 40 ml of dichloromethane. The reaction is left under stirring at room temperature for 3 hours. The solvent is evaporated and then the crude reaction product is taken up in 30 ml of ethyl acetate. The solution is washed with saturated NaHCO3 solution. The organic phase is dried on sodium sulfate, filtered and concentrated to obtain 2.5 g of tert-butyl 2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methylpiperazin-1-yl)benzoate in the form of a light brown gum (yield=92%).
LCMS (EI, m/z): (M+1) 506.26.
1H NMR: δH ppm (400 MHz, DMSO): 7.84 (1H, m, CHarom), 7.09 (1H, m, CHarom), 6.89 (1H, bs, CHarom), 3.45-3.39 (8H, m, CH), 2.83 (4H, m, CH), 2.20 (4H, m, CH), 2.05 (3H, s, CH), 1.46 (9H, s, CH).
Example 16
2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methyl piperazin-1-yl)benzoic acid
7.62 ml (99 mmol) of trifluoroacetic acid is added to 2.5 g (4.95 mmol) of tert-butyl 2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methylpiperazin-1-yl)benzoate dissolved in 30 ml of dichloromethane. The reaction is left under stirring at room temperature overnight. The solvent is evaporated and then the crude reaction product is redissolved in 30 ml of ethyl acetate. The solvents are evaporated, the solid formed is redissolved in ethyl ether and the solvent is evaporated again. This operation is repeated three times until a light brown solid is obtained. 2.2 g of 2-(N-(4,4-difluorocyclohexyl)-2,2,2-trifluoroacetamido)-4-(4-methylpiperazin-1-yl)benzoic acid in the form of a trifluoroacetic salt is obtained (yield=79%).
LCMS (EI, m/z): (M+1) 450.1.
1H NMR: δH ppm (400 MHz, DMSO): 10.01 (1H, bs, OH), 7.92 (1H, m, CHarom), 7.13 (1H, m, CHarom), 7.01 (1H, bs, CHarom), 4.39 (1H, m, CH), 3.12-3.52 (8H, m, CH), 2.86 (3H, s, CH), 1.75-2.0 (8H, m, CH).
The following compounds are also obtained by this method:
4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid.
This compound was previously described in WO 2008/74749, WO 2009/13126 and WO 2010/69966.
LCMS (EI, m/z): (M+1) 416.40.
1H NMR: δH ppm (400 MHz, DMSO): 12.60 (1H, bs, OH), 10.08 (1H, bs, OH), 7.90 (1H, d, CHarom), 7.13 (1H, dd, CHarom), 6.90 (1H, d, CHarom), 4.40 (1H, m, CH), 4.10 (2H, m, CH), 3.70-3.90 (2H, m, CH), 3.59-3.62 (4H, m, CH), 3.30-3.32 (4H, m, CH), 2.87 (3H, s, CH3), 1.87-1.98 (1H, m, CH), 1.59-1.60 (1H, m, CH), 1.00-1.54 (2H, m, CH).
4-((3-(dimethylamino)propyl)(methyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid
This compound was previously described in WO 2009/13126 and WO 2008/74749.
Example 17
(S)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)-4-(3-(2,2,2-trifluoroacetamido)pyrrolidin-1-yl)benzoic acid




Example 17a
Tert-butyl(S)-4-(3-(tert-butoxycarbonylamino)pyrrolidin-1-yl)-2-(tetrahydro-2H-pyran-4ylamino)benzoate
This compound was obtained by reproducing example 16d using tert-butyl(S)-pyrrolidin-3-ylcarbamate.
Example 17b
(S)-4-(3-aminopyrrolidin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoic acid
19.7 ml (25 eq) of trifluoroacetic acid is added to a solution of 4.72 g (10.23 mmol) of tert-butyl(S)-4-(3-(tert-butoxycarbonylamino)pyrrolidin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoate in 100 ml of dichloromethane. The reaction medium is stirred at room temperature for 30 hours. The solvents are evaporated and the residue is redissolved in diethyl ether and triturated until a solid is obtained. The solid formed is filtered and dried under vacuum to yield 4.3 g (100%) of a yellow powder of (S)-4-(3-aminopyrrolidin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoic acid in the form of a trifluoroacetic acid salt.
LCMS (EI, m/z): (M+1) 306.22.
Example 17
(S)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)-4-(3-(2,2,2-trifluoroacetamido)pyrrolidin-1-yl)benzoic acid
1.74 ml (3.5 eq) of triethylamine and 1.6 ml (2.1 eq) of trifluoroacetic anhydride are added to a solution of 1.5 g (3.58 mmol) of (S)-4-(3-aminopyrrolidin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoic acid in the form of a trifluoroacetic acid salt in 40 ml of dichloromethane at 0° C. The reaction medium is stirred at room temperature for 24 hours. Water (10 ml) is added drop by drop and then the organic phase is washed with saturated sodium chloride solution, dried on magnesium sulfate, filtered and evaporated. The residue is purified by silica gel chromatography (96:4 dichloromethane/methanol as eluent) to yield 250 mg (14%) of (S)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)-4-(3-(2,2,2-trifluoroacetamido)pyrrolidin-1-yl)benzoic acid in the form of a yellow powder.
LCMS (EI, m/z): (M+1) 498.07.
Example 18
2-(2-fluoroethoxy)-4-(4-methylpiperazin-1-yl)benzoic acid
This compound can be prepared from the following intermediates.
Example 18a
Tert-butyl 4-fluoro-2-(2-fluoroethoxy)benzoate
Example 18b
Tert-butyl 2-(2-fluoroethoxy)-4-(4-methylpiperazin-1-yl)benzoate
The following compound was also obtained by this method:
2-(2-fluoroethoxy)-4-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)benzoic acid
Example 19
4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(2-fluoroethyl)-acetamido)-benzoic acid
This compound can be prepared from the following intermediates.
Example 19a
Tert-butyl 4-fluoro-2-(2-fluoroethylamino)benzoate
Example 19b
Tert-butyl 4-fluoro-2-(2,2,2-trifluoro-N-(2-fluoroethyl)acetamido)benzoate
Example 19c
Tert-butyl 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(2-fluoroethyl)-acetamido)-benzoate
The following compound was also obtained by this method:
4-((3-(dimethylamino)propyl)(methyl)amino)-2-(2,2,2-trifluoro-N-(2-fluoroethyl)acetamido)benzoic acid
Example 20
4-(1-methylpiperidin-4-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid hydrotrifloroacetate




This compound can be prepared from the following intermediates.
Example 20a
Tert-butyl 2-nitro-4-(pyridin-4-yl)benzoate
1.67 g of bis(triphenylphosphine)palladium(II)chloride (2.38 mmol) and 15.8 g of sodium carbonate (149 mmol) are added to a solution of 18 g of tert-butyl 4-bromo-2-nitrobenzoate (59.6 mmol) and 10.98 g of pyridine-4-ylboronic acid (89 mmol) in a mixture of 200 ml of dimethoxyethane and 100 mL of water. The reaction medium is heated at 100° C. for 24 hours and then concentrated under reduced pressure. The residue obtained is purified by flash chromatography (CH2Cl2/AcOEt: 100:0 to 70:30, 30 min) The product is isolated in the form of an oil which crystallizes to yield 14.64 g (82%) of crystals.
MS (m/z): (M+1) 301.0.
1H NMR: δH ppm (400 MHz, DMSO): 8.73 (2H, d, CHarom, J=6.0 Hz), 8.44 (1H, s, CHarom), 8.24 (1H, dd, CHarom, J=8.0 Hz), 7.97 (1H, d, CHarom, J=8.0 Hz), 7.85 (2H, dd, CHarom, J=4.4 Hz), 1.54 (9H, s).
Example 20b
4-(4-(tert-butoxycarbonyl)-3-nitrophenyl)-1-methylpyridinium iodide
7.55 mL od iodomethane (121 mmol) is added to a solution of 16.2 g of tert-butyl 2-nitro-4-(pyridin-4-yl)benzoate (60.6 mmol) in 20 mL of acetone. The reaction medium is heated at 60° C. for 4 hours and then at room temperature overnight. After dry concentration, 27 g of orange crystals are isolated (100%).
MS (m/z): (M+1) 315.0.
1H NMR: δH ppm (400 MHz, DMSO): 9.14 (2H, d, CHarom, J=6.4 Hz), 8.71 (1H, s, CHarom), 8.63 (2H, d, CHarom, J=6.4 Hz), 8.47 (1H, dd, CHarom, J=8.0 Hz), 8.08 (1H, d, CHarom, J=8.0 Hz), 4.37 (3H, s, CH), 1.54 (9H, s).
Example 20c
Tert-butyl 2-amino-4-(1-methylpiperidin-4-yl)benzoate
0.48 g of platine (IV) oxide (2.12 mmol) is added to a solution of 26.8 g of 4-(4-(tert-butoxycarbonyl)-3-nitrophenyl)-1-methylpyridinium iodide (60.6 mmol) in 200 mL of methanol placed in a reactor made in stainless steel. The reaction medium is brought under 5 bar of hydrogen for 24 h. The catalyst is filtered and the filtrate is concentrated under reduced pressure to yield 24.8 g (98%) of white crystals.
MS (m/z): (M+1) 291.1.
1H NMR: δH ppm (400 MHz, DMSO): 9.18 (1H, s, HI), 7.60 (1H, d, CHarom, J=8.4 Hz), 6.54-6.40 (3H, m, CHarom), 6.39 (1H, d, CHarom, J=8.0 Hz), 3.48-3.53 (2H, m, CH), 3.06 (2H, t, CH), 2.81 (3H, s, CH), 2.60-2.70 (1H, m, CH), 1.89-1.97 (2H, m, CH), 1.70-1.80 (2H, m, CH), 1.52 (9H, s).
Example 20d
Tert-butyl 4-(1-methylpiperidin-4-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoate
7.18 mL of 2,2,2-trifluoroacetic acid (93 mmol), 4.11 mg of dihydro-2H-pyran-4(3H)-one (44.5 mmol) and then 14.5 g of tetramethylammonium triacetoxyborohydride (53.8 mmol) are successively added to a solution of 15 g of tert-butyl 2-amino-4-(1-methylpiperidin-4-yl)benzoate in 200 mL of dichloromethane under stirring. The reaction medium is stirred at room temperature for 2 h and then taklen up with a 1N soda solution. The organic phase is isolated, dried on magnesium sulfate and then dried concentrated. The residue contained always HI. It is thus taken up in dichloromethane and washed with 100 mL of a 1H soda solution. The organic phase is decanted, dried on magnesium sulfate and dry concentrated to yield 14.6 g of a yellow solid (quantitative yield).
MS (m/z): (M+1) 375.2.
1H NMR: δH ppm (400 MHz, DMSO): 7.69 (1H, d, CHarom, J=8.4 Hz), 7.63 (1H, d, CHarom, J=7.6 Hz), 6.65 (1H, s, CHarom), 6.44 (1H, dd, CHarom, J=8.4 Hz), 3.74-3.86 (2H, m, CH), 3.66-3.71 (1H, m, CH), 3.51 (2H, t, CH), 3.05-3.12 (2H, m, CH), 2.6-2.5 (1H, m, CH), 2.42 (3H, s, CH), 2.30-2.40 (2H, m, CH), 1.89-1.97 (2H, m, CH), 1.64-1.77 (4H, m, CH), 1.52 (9H, s), 1.33-1.45 (2H, m, CH).
Example 20e
Tert-butyl 4-(1-methylpiperidin-4-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoate
6.35 mL of triethylamine and 5.50 mL of 2,2,2-trifluoroacetic anhydride (39.6 mmol) are added at 0° C. to a solution of 11.4 g of tert-butyl 4-(1-methylpiperidin-4-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzoate (30.4 mmol) in 240 mL of dichloromethane under stirring. The reaction medium is stirred at room temperature for 1 h and then 100 mL of water is added dropwise. The organic phase is decanted, dried on magnesium sulfate and dry concentrated. The residue is taken up in a mixture of ethanol/diethyl ether to yield a solid which is filtered on a fritted disc and 12.06 g of white crystals is isolated. The filtrate is concentrated (4.5 g) and then purified by flach chromatography on silica (CH2Cl2/meOH: 95:5 to 90:10, 20 min). The product obtained is recrysltallized in diethyl ether to yield 1.04 g of additional white crystals (global yield=74%).
MS (m/z): (M+1) 471.1.
1H NMR: δH ppm (400 MHz, DMSO): 9.45 (1H, s1, NH+), 7.96 (1H, d, CHarom, J=8 Hz), 7.51 (1H, d, CHarom, J=8 Hz), 7.31 (1H, s, CHarom), 4.6-4.5 (1H, m, CH), 3.90-3.75 (2H, m, CH), 3.5-3.35 (4H, m, CH), 3.1-2.85 (3H, m, CH), 2.79 (3H, s, CH3), 2.1-1.95 (3H, 3, CH), 1.9-1.75 (2H, m, CH), 1.55-1.40 (11H, m), 1.0-0.85 (1H, m, CH).
Example 20
4-(1-methylpiperidin-4-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid hydrotrifluoroacetate
6.33 mL of 2,2,2-trifluoroacetic acid (82 mmol) is added under stirring to a solution of 3.2 g of tert-butyl 4-(1-methylpiperidin-4-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoate (5.47 mmol) (in the form of a salt of trifluoroacetic acid) in 30 mL of dichloromethane. The reaction medium is stirred at room temperature for 16 h, and then evaporated under reduced pressure. The residue is taken up in ethanol, and the white solid formed is filtered on a fritted disc to yield 1.61 g (53%) of white crystals.
MS (m/z): (M+1) 415.1.
1H NMR: δH ppm (400 MHz, DMSO): 13.39 (1H, s1, COOH), 9.46 (1H, s1, COOH du TFA), 7.99 (1H, d, CHarom, J=8.4 Hz), 7.49 (1H, d, CHarom, J=8.4 Hz), 7.30 (1H, s, CHarom), 4.53 (1H, m, CH), 3.74-3.86 (2H, m, CH), 3.35-3.45 (5H, m, CH), 2.90-3.01 (3H, m, CH), 2.76 (3H, s, CH), 1.65-2.04 (5H, m, CH), 1.44-1.54 (2H, m, CH).
Example 21
1-(4-isothiocyanatophenyl)-4-methylpiperazine
This compound was prepared by adapting the method described in EP1215208.
The following compound was also obtained by this method:
tert-butyl 2-isothiocyanato-5-(4-methylpiperazin-1-yl)phenylcarbamate.
Example 22
Tert-butyl 2-isocyanato-5-(4-methylpiperazin-1-yl)phenylcarbamate
This compound can be prepared from the following intermediates.
Example 22a
Tert-butyl 5-(4-methylpiperazin-1-yl)-2-nitrophenylcarbamate
Example 22b
Tert-butyl 2-amino-5-(4-methylpiperazin-1-yl)phenylcarbamate
Example 22
Tert-butyl 2-isocyanato-5-(4-methylpiperazin-1-yl)phenylcarbamate
Example 23
4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzaldehyde




Example 23a
(4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)phenyl)methanol
500 mg (1.060 mmol) of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid dissolved in 5 ml of tetrahydrofuran is added at 0° C. to a suspension of 201 mg (5.30 mmol) of LiAlH4 in 9 ml of tetrahydrofuran. The reaction mixture is stirred at 0° C. for 1 hour and then at room temperature for 3 hours. The reaction mixture is cooled at 0° C. and then, drop by drop, 200 μl water, then 200 μl of soda solution (15% by weight) and finally 1 ml of water are added. The reaction mixture is stirred at room temperature for 2 hours and then filtered and rinsed with tetrahydrofuran. The filtrate is concentrated to yield 250 mg (yield=77%) of (4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)phenyl)methanol in the form of a white solid.
LCMS (EI, m/z): (M+1) 306.14.
1H NMR: δH ppm (400 MHz, DMSO): 6.85 (1H, d, CHarom), 6.20 (1H, d, CHarom), 6.10 (1H, d, CHarom), 4.95 (1H, bs, OH), 4.87 (1H, d, NH), 4.37 (2H, d, CH2), 3.83-3.86 (2H, m, CH), 3.56 (1H, m, CH), 3.46-3.56 (3H, m, CH), 3.45 (1H, m, CH), 3.05-3.07 (4H, m, CH), 2.41-2.44 (4H, m, CH), 2.21 (3H, s, CH3), 1.89-1.92 (2H, m, CH).
Example 23
4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzaldehyde
85 mg (0.982 mmol) of manganese dioxide is added at room temperature to a solution of (4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)phenyl)methanol (100 mg, 0.327 mmol) in a mixture of ethyl acetate (10 ml) and dichloromethane (9 ml). The reaction mixture is placed in an ultrasonic bath for 5 hours. The reaction mixture is filtered, the solvents are evaporated and the crude product is purified by chromatography to yield 50.0 mg (yield-50.3%) of (4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzaldehyde in the form of a white solid.
LCMS (EI, m/z): (M+1) 304.19.
1H NMR: δH ppm (400 MHz, DMSO): 9.43 (1H, d, CH), 7.32 (1H, d, CHarom), 6.36 (1H, d, CHarom), 6.08 (1H, d, CHarom), 3.94-3.99 (2H, m, CH), 3.77 (1H, m, CH), 3.61-3.63 (2H, m, CH), 3.42-3.45 (4H, m, CH), 2.57-2.60 (4H, m, CH), 2.36 (3H, s, CH3), 2.04-2.08 (2H, m, CH), 1.51-1.60 (2H, m, CH).
Example 24
2-(4-(4-methylpiperazin-1-yl)phenyl)acetic acid




Example 24a
2,2,2-trichloro-1-(4-(4-methylpiperazin-1-yl)phenyl)ethanol
1.0 ml (10.00 mmol) of trichloroacetic acid and, in small portions, 1.854 g. (10 mmol) of sodium 2,2,2-trichloroacetate are added at room temperature to a solution of 1.362 g (6.67 mmol) of 4-(4-methylpiperazine-1-yl)benzaldehyde in 13.5 ml of dimethylformamide. The reaction mixture is stirred for 3 hours at room temperature. The solvent is concentrated and the crude reaction product extracted with ethyl acetate. The organic phase is washed using saturated sodium bicarbonate solution. The organic phases are combined, dried on magnesium sulfate and then concentrated to yield 1.760 g (yield=82%) of 2,2,2-trichloro-1-(4-(4-methylpiperazine-1-yl)phenyl)ethanol in the form of a white solid.
LCMS (EI, m/z): (M+1) 324.04.
1H NMR: δH ppm (400 MHz, DMSO): 7.41 (2H, d, CHarom), 7.02 (1H, bs, OH), 6.90 (2H, d, CHarom), 5.08 (1H, bs, CH), 3.14-3.16 (4H, m, CH), 2.42-2.47 (4H, m, CH), 2.21 (3H, s, CH3).
Example 24
2-(4-(4-methylpiperazin-1-yl)phenyl)acetic acid
0.559 g (14.77 mmol) of sodium borohydride is added quickly to 2.294 g (7.35 mmol) of dibenzyl diselenide in 28 ml of ethanol. The reaction mixture is stirred at room temperature for 1 hour. 2.266 g (7 mmol) of 2,2,2-trichloro-1-(4-(4-methylpiperazine-1-yl)phenyl)ethanol and 1.680 g (42.0 mmol) of sodium hydroxide are then added. The reaction mixture is stirred at 35° C. for 24 hours. The solvent is concentrated and the crude product extracted with ethyl acetate after adding a pH 5 aqueous phase. The organic phases are combined, dried on magnesium sulfate and then concentrated to yield 2-(4-(4-methylpiperazine-1-yl)phenyl)acetic acid which is used without additional purification.
LCMS (EI, m/z): (M+1) 235.294.
Example 25
2-(4-(4-methylpiperazin-1-yl)-2-nitrophenyl)acetic acid
This compound can be prepared from the following intermediates.
Example 25a
Diethyl 2-(4-fluoro-2-nitrophenyl)malonate
Example 25b
Diethyl 2-(4-(4-methylpiperazin-1-yl)-2-nitrophenyl)malonate
Example of Method E1
Example 26
N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide




0.95 ml (11.21 mmol) of oxalyl chloride and 2 drops of anhydrous dimethylformamide are added to 2.97 g (5.61 mmol) of a solution of 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid in 95 ml of dichloromethane. The reaction mixture is stirred for 2 hours at room temperature. The solvents are evaporated, the solid formed is taken up in toluene and the solvent evaporated. This operation is repeated three times until a white solid is obtained. The acid chloride is dissolved in 35 ml of anhydrous tetrahydrofuran at −20° C. and then the solution formed is added to a solution containing 1.56 g (5.61 mmol) of 5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine and 3.71 ml (21.30 mmol) of N-ethyl-N-isopropylpropan-2-amine in 30 ml of anhydrous tetrahydrofuran. The reaction mixture is stirred for 3 hours at −20° C. and then overnight at room temperature. The precipitate obtained is filtered and rinsed with tetrahydrofuran and water and then dried to yield 2 g (53%) of N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide.
LCMS (EI, m/z): (M+1) 676.20.
1H NMR: δH ppm (400 MHz, DMSO): 13.66 (1H, bs, NH), 11.08 (1H, bs, NH), 8.61 (1H, s, CHarom), 8.46 (1H, s, CHarom), 7.83 (1H, d, CHarom), 7.05-7.10 (2H, m, CHarom), 6.83-6.89 (3H, m, CHarom), 4.39-4.44 (1H, m, CH), 3.83-3.85 (1H, m, CH), 3.69-3.72 (1H, m, CH), 3.59-3.62 (1H, m, CH), 3.30-3.32 (4H, m, CH2), 2.30-2.44 (4H, m, CH2), 2.27 (3H, s, CH3), 1.87-1.90 (1H, m, CH), 1.59-1.60 (1H, m, CH), 1.49-1.50 (1H, m, CH), 1.20-1.40 (1H, m, CH).
The following derivatives were obtained according to the same method:
















 
























Y1,


Mass


Ex.**
Y
R1
R2
n
W
Rj
Y4
Compound names
Yield
MH+





26-1 












0
H
H
CH, N
N-(5-(2- carbamoylphenylthio)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-2 












0
H
H
CH, N
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
ND


 


26-3 
I








0
H
H
CH, N
N-(5-iodo-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-4 




H
NO2
1
H
H
CH, N
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-2-(4- nitrophenyl)acetamide
ND
442.21


 


26-5 












0
H
H
CH, N
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-6 












0
H
H
CH, N
N-(5-(4,4- difluorocyclohexyl)- 2,2,2- trifluoroacetamido)-N-(5- (3,5- difluorophenylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1- yl)benzamide
ND
ND


 


26-7 












0
H
H
CH, N
N-(5-(3,5- difluorobenzyl)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
22%
676.2 


 


26-8 




H




1
H
H
CH, N
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-2-(4-(4- methylpiperazin-1- yl)phenyl)acetamide
ND
495.17


 


26-9 
I








0
OMe
H
CH, N
N-(5-iodo-6-methoxy- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
688.18


 


26-10
I








0
NH2
H
CH, N
N-(6-amino-5-iodo-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
673.06


 


26-11












1
H
H
CH, N
(S)-N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)-4-(3- (2,2,2- trifluoroacetamido) pyrrolidin-1- yl)benzamide
ND
ND


 


26-12




H




0
H
H
CH, N
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)benzamide
46%
481.38


 


26-13












0
H
H
N, CH
N-(5-(3,5- difluorobenzyl)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
35%
658.1 


 


26-14












0
H
H
N, CH
N-(5-(3,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
63%
671.1 


 


26-15












0
H
H
N, CH
N-(5-(3,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
673.1 


 


26-16












0
H
H
N, CH
N-(5-(2,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
62%
674.2 


 


26-17












0
H
H
N, CH
N-(5-(2,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
673.3 


 


26-18












0
H
H
N, CH
N-(5-(2,5- dichlorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-19












0
H
H
N, CH
N-(5-(2,5- dichlorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-20












0
H
H
N, CH
N-(5-(5-chloro-2- (trifluoromethyl) benzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
55%
740.2 


 


26-21












0
H
H
N, CH
N-(5-(5-chloro-2- (trifluoromethyl) benzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
739.3 


 


26-22












0
H
H
N, CH
4-(4-methylpiperazin-1- yl)-N-(5-(pyridin-3- ylmethoxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
90%
639.2 


 


26-23












0
H
H
N, CH
4-(1-methylpiperazin-4- yl)-N-(5-(pyridin-3- ylmethoxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
638.2 


 


26-24












0
H
H
N, CH
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-25












0
H
H
N, CH
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
50%
ND


 


26-26












0
H
H
N, CH
N-(5-(2,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-27












0
H
H
N, CH
N-(5-(2,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-28












0
H
H
N, CH
N-(5-(3,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-29












0
H
H
N, CH
N-(5-(3,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-30












0
H
H
N, CH
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-31












0
H
H
N, CH
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-32












0
H
H
N, CH
4-(4-methylpiperazin-1- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)-N-(5-(2- (trifluoromethyl) phenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)benzamide
ND
ND


 


26-33












0
H
H
N, CH
4-(1-methylpiperidin-4- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)-N-(5-(2- (trifluoromethyl) phenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)benzamide
ND
ND


 


26-34












0
H
H
N, CH
N-(5-(3,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-35












0
H
H
N, CH
N-(5-(3,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
73%
ND


 


26-36












0
H
H
N, CH
N-(5-(2,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-37












0
H
H
N, CH
N-(5-(2,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-38












0
H
H
N, CH
N-(5-(2,5- dichlorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-39












0
H
H
N, CH
N-(5-(2,5- dichlorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin- 4-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
ND
ND


 


26-40












0
H
H
N, CH
N-(5-(3,5- difluorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
79%
659.2 


 


26-41












0
H
H
N, CH
N-(5-(3,5- difluorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
658.2 


 


26-42












0
H
H
N, CH
N-(5-(2,5- difluorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
659.2 


 


26-43












0
H
H
N, CH
N-(5-(3,5- difluorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
658.2 


 


26-44












0
H
H
N, CH
N-(5-(2,5- dichlorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
26%
691.2 


 


26-45












0
H
H
N, CH
N-(5-(2,5- dichlorophenylamino)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
98%
692.2 


 


26-46












0
H
H
N, N
N-(5-(3,5- difluorobenzyl)-1H- pyrazolo[3,4-b]pyrazin- 3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-47












0
H
H
N, N
N-(5-(3,5- difluorobenzyl)-1H- pyrazolo[3,4-b]pyrazin- 3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-48












0
H
H
N, N
N-(5-((3,5- difluorophenyl)ethynyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-((3- (dimethylamino)propyl) (methyl)amino)-2-(2,2,2- trifluoro-N-(tetrahydro- 2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-49












0
H
H
N, N
N-(5-((3,5- difluorophenyl)ethynyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-50












0
H
H
N, N
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyrazin- 3-yl)-4-((3- (dimethylamino)propyl) (methyl)amino)-2-(2,2,2- trifluoro-N-(tetrahydro- 2H-pyran-4- yl)acetamido)benzamide
43%
693.2 


 


26-51












0
H
H
N, N
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[3,4-b]pyrazin- 3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-52












0
H
H
N, CH
4-(4-methylpiperazin-1- yl)-2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)-N-(5-(2- (trifluoromethyl) phenylthio)-1H- pyrazolo[3,4-b]pyrazin- 3-yl)benzamide
66%
709.1 


 


26-53












0
H
H
N, CH
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
28%
708.2 


 


26-54












0
H
H
N, CH
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
74%
707.2 


 


26-55












0
H
H
N, CH
N-(5-(2,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-56












0
H
H
N, CH
N-(5-(2,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-57












0
H
H
N, CH
N-(5-(3,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-58












0
H
H
N, CH
N-(5-(3,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-59












0
H
H
N, CH
N-(5-(2,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-60












0
H
H
N, CH
N-(5-(2,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-61












0
H
H
N, CH
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
94%
ND


 


26-62












0
H
H
N, CH
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
99%
ND


 


26-63












0
H
H
N, CH
N-(5-(2,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
60%
ND


 


26-64












0
H
H
N, CH
N-(5-(2,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
36%
ND


 


26-65












0
H
H
N, CH
N-(5-(2,5- difluorobenzylsulfinyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
ND


 


26-66












0
H
H
N, CH
N-(5-(2,5- dichlorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
52%
ND


 


26-67












0
H
H
N, CH
N-(5-(2,5- dichlorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
90%
ND


 


26-68












0
H
H
N, CH
N-(5-(2,5- dichlorobenzylsulfinyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
50%
ND


 


26-69
I








0
H
C(Ph)3
CH, N
N-(5-iodo-1-trityl-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
67%
900.23


 


26-70












0
H
H
CH, N
N-(5-(3,5- difluorophenyl- sulfonamido)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(2,2,2-trifluoro- N-(tetrahydro-2H-pyran- 4- yl)acetamido)benzamide
11%
ND





**1H NMR, dmso-d6, Ex.:


26-4: 13.64 (1H, sl, NH), 11.26 (1H, sl, NH), 8.68 (1H, d, CHarom), 8.58 (1H, d, CHarom), 8.20 (2H, d, CHarom), 7.64 (2H, d, CHarom), 7.03 (1H, m, CHarom), 6.78 (2H, m, CHarom), 3.95 (2H, m, CH2).


26-8: 13.59 (1H, sl, NH), 11.05 (1H, sl, NH), 8.68 (1H, d, CHarom), 8.57 (1H, d, CHarom), 7.19 (2H, d, CHarom), 6.99-7.08 (1H, m, CHarom), 6.88 (2H, d, CHarom), 6.75-6.79 (2H, m, CHarom), 3.61 (2H, m, CH2), 3.07-3.09 (4H, m, CH), 2.41-2.44 (4H, m, CH), 2.20 (3H, s, CH3).


26-9: 13.17 (1H, sl, NH), 10.90 (1H, sl, NH), 8.55 (1H, s, CHarom), 7.79 (1H, d, CHarom), 7.07 (1H, dd, CHarom), 6.90 (1H, d, CHarom), 4.40-4.50 (1H, m, CH), 3.96 (3H, s, CH3), 3.82-3.89 (1H, m, CH), 3.74-3.80 (1H, m, CH), 3.34-3.41 (2H, m, CH), 3.28-3.33 (4H, m, 2 * CH2), 2.43-2.47 (4H, m, 2 * CH2), 2.23 (3H, s, CH3), 1.85-1.92 (1H, m, CH), 1.58-1.63 (1H, m, CH), 1.45-1.53 (1H, m, CH), 1.22-1.33 (1H, m, CH).


26-10: 12.48 (1H, sl, NH), 10.72 (1H, sl, NH), 8.30 (1H, s, CHarom), 7.77 (1H, d, CHarom), 7.06 (1H, dd, CHarom), 6.88 (1H, d, CHarom), 6.40 (2H, sl, NH2), 4.40-4.50 (1H, m, CH), 3.82-3.89 (1H, m, CH), 3.74-3.80 (1H, m, CH), 3.34-3.41 (2H, m, CH), 3.28-3.33 (4H, m, 2 * CH2), 2.43-2.47 (4H, m, 2 * CH2), 2.23 (3H, s, CH3), 1.85-1.92 (1H, m, CH), 1.58-1.65 (1H, m, CH), 1.45-1.55 (1H, m, CH), 1.22-1.34 (1H, m, CH).


(ND: not determined).


26-14: 12.99 (1H, sl, NH), 10.25 (1H, s, NH), 7.96 (1H, d, CHarom, J = 9.2 Hz), 7.90-7.80 (1H, m, CHarom), 7.23-7.16 (3H, m, CHarom), 7.12-7.08 (1H, m, CHarom), 6.96 (1H, d, CHarom, J = 8.8 Hz), 6.87 (1H, s, CHarom), 5.31 (2H, s), 4.49-4.42 (1H, m), 3.86-3.75 (2H, m), 3.45 (1H, m), 3.37 (1H, m), 3.35 (4H, s), 2.42 (4H, s), 2.22 (3H, s), 1.90-1.75 (2H, m), 1.53-1.49 (1H, m), 1.31-1.25 (1H, m).


26-16: 13.00 (1H, s, NH), 10.27 (1H, s, NH), 7.95 (1H, d, CHarom, J = 8.8 Hz), 7.89-7.84 (1H, m, CHarom), 7.50-7.40 (1H, m, CHarom), 7.35-7.20 (2H, m, CHarom), 7.12-7.09 (1H, m, CHarom), 6.94 (1H, d, CHarom, J = 8.8 Hz), 6.87 (1H, s, CHarom), 5.30 (2H, s), 4.52-4.43 (1H, m), 3.85-3.75 (2H, m), 3.46-3.43 (1H, m), 3.36 (5H, s), 2.45 (4H, s), 2.22 (3H, s), 1.92-1.82 (2H, m), 1.60-1.52 (1H, m), 1.33-1.26 (1H, m).


26-20: 13.01 (1H, s, NH), 10.22 (1H, s, NH), 7.97 (1H, d, CHarom, J = 8.8 Hz), 7.90-7.78 (3H, m, CHarom), 7.68-7.64 (1H, m, CHarom), 7.12-7.08 (1H, m, CHarom), 6.97 (1H, d, CHarom, J = 8.8 Hz), 6.85 (1H, s, CHarom), 5.43 (2H, s), 4.45-4.40 (1H, m), 3.86-3.70 (2H, m), 3.46-3.42 (1H, m), 3.30-3.28 (5H, m), 2.46 (4H, s), 2.23 (3H, s), 1.90 (1H, d, J = 11.2 Hz), 1.77 (1H, d, J = 11.2 Hz), 1.58-1.50 (1H, m), 1.30-1.20 (1H, m).






In certain cases, the major product of these reactions corresponds to the disubstituted product characterized by the additional functionalization of the pyrazole ring. In these cases, this product is isolated and transformed into a monosubstituted product by treatment with a base as described below.
Example 27
N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




Example 27a
N-(5-(3,5-difluorophenylthio)-1-(4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl)-1H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide
1.51 ml (17.90 mmol) of oxalyl chloride and 2 drops of anhydrous dimethylformamide are added to 4.74 g (8.95 mmol) of a solution of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid in 60 ml of dichloromethane. The reaction mixture is stirred for 2 hours at room temperature. The solvents are evaporated, the solid formed is taken up in toluene and the solvent is evaporated; this operation is repeated three times until a white solid is obtained.
The acid chloride is added at 0° C. in small fractions to 1 g (3.58 mmol) of 5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyrazine-3-amine dissolved in 15 ml of pyridine. The reaction mixture is stirred at 25° C. overnight at room temperature. After evaporation of the solvent, the residue is purified by silica gel chromatography (90:10 dichloromethane/methanol and then 90:9:1 and then 90:5:5 dichloromethane/methanol/ammonium as eluent) to yield N-(5-(3,5-difluorophenylthio)-1-(4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl)-1H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide.
LCMS (EI, m/z): (M+1) 1074.64.
Example 27
N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide
0.27 ml (1.95 mmol) of triethylamine is added to 0.21 g (0.19 mmol) of a solution of N-(5-(3,5-difluorophenylthio)-1-(4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl)-1H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide in 5 ml of methanol. The reaction medium is heated at 65° C. for 4 hours, and then overnight at room temperature. After evaporation of the solvent, the product is extracted several times with ethyl acetate. The organic phases are combined, washed with saturated sodium bicarbonate solution, dried on magnesium sulfate and concentrated. The residue is purified by silica gel chromatography (95:4:1 dichloromethane/methanol/ammonium as eluent) to yield 0.065 g (57%) of N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyrazine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in the form of a yellow solid.
LCMS (EI, m/z): (M−1) 579.21.
1H NMR: δH ppm (400 MHz, DMSO): 13.95 (1H, bs, NH), 10.25 (1H, bs, NH), 8.62 (1H, s, CHarom), 8.27 (1H, d, NH), 7.80 (1H, d, CHarom), 7.17-7.27 (3H, m, CHarom), 6.27 (1H, d, CHarom), 6.12 (1H, d, CHarom), 3.79-3.82 (2H, m, CH), 3.67 (1H, m, CH), 3.45-3.50 (2H, m, CH), 3.26-3.29 (4H, m, CH), 2.42-2.44 (4H, m, CH), 2.22 (3H, s, CH3), 1.90-1.93 (2H, m, CH), 1.31-1.36 (2H, m, CH).
The following compounds were obtained by the same method:
















 

















Ex.**
Y
R1
R2
Y1
n
W
Rj
Compound names
Yield
Mass





27-1












CH
0
H
H
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
18.6%
(M + H) 612.13


 


27-2












N
0
H
H
N-(5-(3,5- difluorophenylsulfinyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
(M + Na) 619.6 


 


27-3












N
0
H
H
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
(M + H) 613.5 


 


27-4
H








CH
0




H
N-(6-(3,5- difluorobenzyl)-1H- pyrazolo[3,4-b]pyridin- 3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  24%
(M + H) 562.00


 


27-5
H








CH
0




H
N-(6-(3,5- difluorobenzylamino)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
(M − H) 275.1 









Reactions carried out in pyridine often make it possible to modify the regioisomer distribution of the products. The following example is characteristic of a reaction of this type.
Example 27-bis
N-(5-(N-(3,5-difluorophenyl)sulfamoyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide




0.224 ml (2.63 mmol) of oxalyl chloride and 2 drops of anhydrous dimethylformamide are added to 0.697 g (1.316 mmol) of a solution of 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid in 20 ml of dichloromethane. The reaction mixture is stirred for 2 hours at room temperature. The solvents are evaporated, the solid formed is redissolved in toluene and the solvent is evaporated. This operation is repeated three times until a white solid is obtained.
The acid chloride is dissolved in 5 ml of anhydrous pyridine and then the solution formed is added to a solution of 0.214 g (0.658 mmol) of 3-amino-N-(3,5-difluorophenyl)-1H-pyrazolo[3,4-b]pyridine-5-sulfonamide in 5 ml of pyridine at 0° C. The reaction mixture is stirred for 3 hours at 0° C., and then overnight at room temperature. The pyridine is evaporated and the crude reaction product is redissolved in toluene and then dry concentrated. The reaction mixture is diluted with saturated NaHCO3 solution and extracted with ethyl acetate. The organic phase is dried on MgSO4, filtered and concentrated and the crude product is used directly in the deprotection reaction with no purification or characterization.
The following compounds were obtained by the same method:
















 

















Ex.**
Y
R1
R2
Y1
n
W
Rj
Compound names
Yield
Mass





27bis-1
H








CH
0




H
N-(6-(2,4-difluorophenylthio)- 1H-pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin-1-yl)- 2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
676.18 (M + H)


 


27bis-2
H








CH
0




H
N-(6-(2,4- difluorophenylamino)-1H- pyrazolo[3,4-b]pyridin-3-yl)-4- (4-methylpiperazin-1-yl)-2- (2,2,2-trifluoro-N-(tetrahydro- 2H-pyran-4- yl)acetamido)benzamide
28%
657.13 (M − H)


 


27bis-3
H








CH
0




H
N-(6-((2,4- difluorophenyl)(methyl)amino)- 1H-pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin-1-yl)- 2-(2,2,2-trifluoro-N- (tetrahydro-2H-pyran-4- yl)acetamido)benzamide
ND
671.05 (M − H)









Example of Method E2
Example 28
5-(3,5-difluorophenylthio)-N-(4-(4-methylpiperazin-1-yl)benzyl)-1H-pyrazolo[3,4-b]pyridin-3-amine




41.5 μl of trifluoroacetic acid (0.539 mmol) and, in small fractions, 129 mg (0.611 mmol) of sodium triacetoxyborohydride are added to a solution of 100 mg (0.35 mmol) of 5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine and 81 mg (0.395 mmol) of 4-(4-methylpiperazin-1-yl)benzaldehyde in 20 ml of a 1:1 mixture of dichloromethane and tetrahydrofuran. The reaction medium is stirred for 16 hours at room temperature. An additional fraction of 125 μl of trifluoroacetic acid and 388 mg of sodium triacetoxyborohydride are added and the reaction medium is stirred for an additional 24 hours. The solvent is then concentrated and the reaction medium extracted with ethyl acetate and washed using saturated sodium bicarbonate solution. The organic phases are combined, dried on magnesium sulfate and then concentrated to yield a yellow oil. A trituration of this oil in methanol leads to the isolation of 135 mg of a yellow solid.
LCMS (EI, m/z): (M+1) 467.57.
1H NMR: δH ppm (400 MHz, DMSO): 12.43 (1H, bs, NH), 8.49 (1H, d, CHarom), 8.47 (1H, d, CHarom), 7.25 (2H, d, CHarom), 7.03-7.08 (1H, m, CHarom), 6.89 (2H, d, CHarom), 6.76-6.77 (3H, m, NH and CHarom), 4.34 (2H, d, CH), 3.08 (4H, m, CH), 2.44 (4H, m, CH), 2.21 (3H, s, CH3).
The following derivative was obtained according to the same method:


















 
















Ex.**
ArX
R1
R2
n
W
Rj
Compound name
Yield
Mass MH+





28-1




NO2




0
H
H
5-(3,5- difluorophenylthio)- N-(4-(4- methylpiperazin-1- yl)-2-nitrobenzyl)- 1H-pyrazolo[3,4- b]pyridin-3-amine
91%
512.16





**1H NMR, DMSO-d6, Ex.:


28-1: 12.43 (1H, bs, NH), 8.49 (1H, d, CHarom), 8.47 (1H, d, CHarom), 7.51 (1H, d, CHarom), 7.45 (1H, m, CHarom), 7.27 (1H, m, CHarom), 7.03-7.08 (1H, m, CHarom), 7.00 (1H, t, NH), 6.77-6.80 (2H, m, CHarom), 4.63 (2H, d, CH), 3.19-3.21 (4H, m, CH), 2.42-2.45 (4H, m, CH), 2.21 (3H, s, CH3).






Example of Method E3
Example 29
1-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)thiourea




0.507 g (2.17 mmol) of 1-(4-isothiocyanatophenyl)-4-methylpiperazine is added at 25° C. to 0.540 g (2.17 mmol) of 3,5-difluorophenylthio-1H-pyrazolo[3,4-b]pyridin-3-amine dissolved in 12 ml of anhydrous dimethylacetamide. The mixture is left under stirring for 15 hours at 85° C. The reaction is treated by adding 20 ml of water and then is extracted with ethyl acetate. The organic phase is dried on sodium sulfate, filtered and concentrated. The product is purified by silica chromatography (15:1 dichloromethane/methanol as eluent) to yield 0.156 g (yield-15%) of 1-(1-tert-butyl-5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)thiourea in the form of a light brown solid.
LCMS (EI, m/z): (M+1) 512.08.
1H NMR: δH ppm (400 MHz, DMSO): 13.69 (1H, bs, NH), 11.50 (1H, bs, NH), 11.19 (1H, bs, NH), 8.96 (1H, d, CHarom), 8.66 (1H, d, CHarom), 7.41 (2H, d, CHarom), 7.10 (1H, ddd, CHarom), 6.95 (2H, d, CHarom), 6.89 (2H, bd, CHarom), 3.13-3.16 (4H, m, CH), 2.45-2.47 (4H, m, CH), 2.23 (3H, s, CH).
Example 29-bis
1-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)urea




0.048 g (1.19 mmol) of sodium hydride is added at 0° C. to 0.200 g (0.598 mmol) of 1-tert-butyl-5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine dissolved in 10 ml of anhydrous dimethylacetamide. The reaction is left under stirring for 10 minutes. 0.130 g (0.598 mmol) of 1-(4-isocyanatophenyl)-4-methylpiperazine is then added at 0° C. The mixture is left under stirring for 3 hours at room temperature. The reaction is treated by adding 20 ml of water drop by drop at 0° C. and then is extracted with ethyl acetate. The organic phase is dried on sodium sulfate, filtered and concentrated. The product is purified by silica chromatography to yield 0.150 g (yield=45%) of 1-(1-tert-butyl-5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)urea in the form of a light brown solid.
LCMS (EL m/z): (M+1) 552.21.
1H NMR: δH ppm (400 MHz, DMSO): 8.92 (1H, bs, NH), 8.58 (1H, bs, NH), 8.51 (1H, bs, CHarom), 8.30 (1H, bs, CHarom), 7.31 (2H, d, CHarom), 7.05 (1H, m, CHarom), 6.83-6.85 (2H, m, CHarom), 3.03-3.08 (4H, m, CH), 2.45-2.48 (4H, m, CH), 2.21 (3H, s, CH), 1.76 (9H, s, CH).
A solution of 0.150 g (0.272 mmol) of 1-(1-tert-butyl-5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)urea dissolved in 20 ml of TFA (trifluoroacetic acid) is refluxed for 3 hours. The solvent is evaporated and the crude reaction product is diluted with saturated NaHCO3 solution and extracted with ethyl acetate. The organic phase is dried on MgSO4, filtered and concentrated. The solid obtained is triturated in methanol, filtered and dried. 110 mg (82%) of 1-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-3-(4-(4-methylpiperazin-1-yl)phenyl)urea in the form of a beige solid is obtained.
LCMS (EI, m/z): (M+1): 496.06.
1H NMR: δH ppm (400 MHz, DMSO): 10.85 (1H, bs, NH), 9.57 (1H, bs, NH), 8.57 (1H, bs, CHarom), 8.30 (1H, bs, CHarom), 7.39 (2H, d, CHarom), 6.99 (1H, m, CHarom), 6.89 (2H, d, CHarom), 6.70 (2H, bd, CHarom), 3.03-3.08 (4H, m, CH), 2.45-2.48 (4H, m, CH), 2.21 (3H, s, CH).
Examples of Method F
Examples of Method F1: Deprotection
Example 30
N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyridine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




9.08 ml (65.1 mmol) of triethylamine is added to 2 g (2.96 mmol) of a solution of N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide in 65 ml of methanol. The reaction medium is heated at 65° C. for 2 hours, and then overnight at room temperature. The precipitate formed is filtered, rinsed with pentane, with water and then with diethyl ether, and then is dried under vacuum to yield 0.73 g (43%) of (N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyridine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in the form of a white solid.
LCMS (EI, m/z): (M+1) 580.23.
1H NMR: δH ppm (400 MHz, DMSO): 13.59 (1H, bs, NH), 10.56 (1H, bs, NH), 8.61 (1H, S, CHarom), 8.50 (1H, s, CHarom), 8.17 (1H, d, NH), 7.80 (1H, d, CHarom), 7.07 (1H, m, CHarom), 6.86 (2H, m, CHarom), 6.23 (1H, d, CHarom), 6.13 (1H, d, CHarom), 3.79-3.82 (2H, dt, CH), 3.60 (1H, m, CH), 3.45-3.50 (2H, m, CH), 3.21-3.33 (4H, m, CH), 2.42-2.46 (4H, m, CH), 2.22 (3H, s, CH3), 1.91-1.94 (2H, m, CH), 1.35-1.38 (2H, m, CH).
The following derivatives were obtained according to the same method:




















Ex.





Y1, Y2, 






**
ArX
R1
R2
(U)n
V
Y3, Y4
R3
Compound name
Yield
Mass







30-1 












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(2- carbamoylphenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
ND
ND


 


30-2 
I








n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-iodo-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
ND
ND


 


30-3 












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
(M + 1) 626.14


 


30-4 












n = 0
CO
CH,  CXAr,  CH, N
H
2-(4,4- difluorocyclohexylamino)- N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)benzamide
ND
ND


 


30-5 












n = 0
CO
CH,  CXAr,  COMe,  N
H
N-(5-(3,5- difluorophenylthio)-6- methoxy-1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
610.20


 


30-6 
I








n = 0
CO
CH,  CXAr,  COMe,  N
H
N-(5-iodo-6-methoxy-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
ND
592.12


 


30-7 
I








n = 0
CO
CH,  CXAr,  CNH2,  N
H
N-(6-amino-5-iodo-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
ND
(M − 1) 574.87


 


30-8 












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(N-(3,5- difluorophenyl)sulfamoyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  30% (2 steps)
(M + 1) 627.20


 


30-9 












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(3,5-difluorobenzyl)- 1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  20%
(M + 1) 562.42


 


30-10












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(3,5- difluorobenzylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
20.6%
(M + 1) 594.11


 


30-11












n = 0
CO
CH,  CXAr,  CH, N
H
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
59.4% (2 steps
(M + 1) 579.11


 


30-12












n = 0
CO
CH,  CXAr,  CCH3,  N
H
N-(5-(3,5- difluorophenylthio)-6- methyl-1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  45%
(M + 1) 579.11


 


30-13












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5-difluorobenzyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  70%
562.27


 


30-14












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  55%
578.27


 


30-15












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  49% 2 steps
577.27


 


30-16












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  61%
578.27


 


30-17












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  54% 2 steps
577.27


 


30-18












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  10% 2 steps
610.01


 


30-19












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  40% 2 steps
609.03


 


30-20












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(5-chloro-2- (trifluoromethyl) benzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  64%
644.24


 


30-21












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(5-chloro-2- (trifluoromethyl) benzyloxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  39% 2 steps
643.24


 


30-22












n = 0
CO
N,  ArXC,  CH,  CH
H
4-(4-methylpiperazin-1- yl)-N-(5-(pyridin-3- ylmethoxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  30%
543.28


 


30-23












n = 0
CO
N,  ArXC,  CH,  CH
H
4-(1-methylpiperidin-4-yl)- N-(5-(pyridin-3- ylmethoxy)-1H- pyrazolo[4,3-b]pyridin-3- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  67% 2 steps
542.29


 


30-24












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  65% 2 steps
580.23


 


30-25












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  56% 2 steps
579.23


 


30-26












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  27% 2 steps
580.23


 


30-27












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  54% 2 steps
579.23


 


30-28












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  31% 2 steps
612.17


 


30-29












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  43% 2 steps
611.17


 


30-30












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  31% 2 steps
612.20


 


30-31












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  57% 2 steps
611.18


 


30-32












n = 0
CO
N,  ArXC,  CH,  CH
H
4-(4-methylpiperazin-1- yl)-2-(tetrahydro-2H- pyran-4-ylamino)-N-(5-(2- (trifluoromethyl) phenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)benzamide
  38% 2 steps
612.24


 


30-33












n = 0
CO
N,  ArXC,  CH,  CH
H
4-(1-methylpiperidin-4-yl)- 2-(tetrahydro-2H-pyran-4- ylamino)-N-(5-(2- (trifluoromethyl) phenylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)benzamide
  57% 2 steps
611.24


 


30-34












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  21% 2 steps
594.25


 


30-35












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  91%
593.25


 


30-36












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide 2,2,2- trifluoroacetate
  86% 2 steps
594.25


 


30-37












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  55% 2 steps
593.25


 


30-38












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  24% 2 steps
626.19


 


30-39












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzylthio)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  50% 2 steps
625.19


 


30-40












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  70%
563.27


 


30-41












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  27% 2 steps
562.27


 


30-42












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  37% 2 steps
563.27


 


30-43












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  41% 2 steps
562.27


 


30-44












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  74%
595.21


 


30-45












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylamino)-1H- pyrazolo[4,3-b]pyridin-3- yl)-4-(1-methylpiperidin-4- yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  33%
594.21


 


30-46












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-(3,5-difluorobenzyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  12% 2 steps
563.07


 


30-47












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-(3,5-difluorobenzyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  15% 2 steps
562.04


 


30-48












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-((3,5- difluorophenyl)ethynyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-((3- (dimethylamino)propyl) (methyl)amino)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  12% 2 steps
589.24


 


30-49












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-((3,5- difluorophenyl)ethynyl)- 1H-pyrazolo[3,4- b]pyrazin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  16% 2 steps
572.07


 


30-50












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyrazin-3- yl)-4-((3- (dimethylamino)propyl) (methyl)amino)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  27%
597.26


 


30-51












n = 0
CO
N,  ArXC,  CH, N
H
N-(5-(2,5- dichlorophenylthio)-1H- pyrazolo[3,4-b]pyrazin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  23% 2 steps
613.16


 


30-52












n = 0
CO
N,  ArXC,  CH, N
H
4-(4-methylpiperazin-1- yl)-2-(tetrahydro-2H- pyran-4-ylamino)-N-(5-(2- trifluoromethyl) phenylthio)-1H- pyrazolo[3,4-b]pyrazin-3- yl)benzamide
  64%
613.23


 


30-53












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  64%
612.22


 


30-54












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  48%
611.22


 


30-55












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  50% 2 steps
612.22


 


30-56












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  28% 2 steps
611.22


 


30-57












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  29% 2 steps
644.16


 


30-58












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  28% 2 steps
643.17


 


30-59












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  21% 2 steps
644.16


 


30-60












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  36% 2 steps
643.17


 


30-61












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  66%
626.24


 


30-62












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  67%
625.24


 


30-63












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  74%
626.24


 


30-64












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  58%
625.24


 


30-65












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- difluorobenzylsulfinyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide 2,2,2- trifluoroacetate
  31% 2 steps
629.24


 


30-66












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  55%
658.18


 


30-67












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  57%
657.18


 


30-68












n = 0
CO
N,  ArXC,  CH,  CH
H
N-(5-(2,5- dichlorobenzylsulfinyl)- 1H-pyrazolo[4,3- b]pyridin-3-yl)-4-(1- methylpiperidin-4-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  20%
641.19


 


30-69












n = 0
CO
CH,  ArXC,  CH, N
H
N-(5-(3,5- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-((3- (dimethylamino)propyl) (methyl)amino)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
  57%
596.26


 


30-70












n = 0
CO
CH,  CH,  ArXC,  N
H
N-(6-(2,4- difluorophenylamino)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  33%
561.19 (M − 1)


 


30-71












n = 0
CO
CH,  CH,  ArXC,  N
H
N-(6-((2,4- difluorophenyl)(methyl) amino)-1H-pyrazolo[3,4- b]pyridin-3-yl)-4-(4- methylpiperazin-1-yl)-2- (tetrahydro-2H-pyran-4- ylamino)benzamide
ND
577.26 (M + 1)


 


30-72












n = 0
CO
CH,  ArXC,  CH, N
H
N-(5-(3,5- difluorophenyl- sulfonamido)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  74%
627.19 (M + 1)


 


30-73












n = 0
CO
CH,  CH,  ArXC,  N
H
N-(6-(2,4- difluorophenylthio)-1H- pyrazolo[3,4-b]pyridin-3- yl)-4-(4-methylpiperazin- 1-yl)-2-(tetrahydro-2H- pyran-4- ylamino)benzamide
  42% (2 steps
(M + 1) 580.23





** 1H NMR, DMSO-d6, Ex.:


30-3: 13.86 (1H, bs, NH), 10.70 (1H, bs, NH), 8.67 (2H, bs, CHarom), 8.10 (1H, d, NH), 7.77 (1H, d, CHarom), 7.22 (1H, m, CHarom), 6.95 (2H, d, CHarom), 6.26 (1H, d, CHarom), 6.16 (1H, bs, CHarom), 4.85 (2H, bs, CH), 3.82-3.86 (2H, dt, CH), 3.70 (1H, m, CH), 3.47-3.53 (2H, m, CH), 3.28-3.32 (4H, m, CH), 2.42-2.46 (2H, m, CH), 2.20 (3H, s, CH3), 1.94-1.98 (2H, m, CH), 1.34-1.41 (2H, m, CH).;


30-5: 13.25 (1H, bs, NH), 10.48 (1H, bs, NH), 8.42 (1H, s, CHarom), 8.11 (1H, d, NH), 7.76 (1H, d, CHarom), 7.00-7.10 (1H, m, CHarom), 6.79-6.87 (2H, m, CHarom), 6.23 (1H, dd, CHarom), 6.12 (1H, d, CHarom), 3.94 (3H, s, CH3), 3.75-3.83 (2H, m, CH), 3.63-3.71 (1H, m, CH), 3.43-3.52 (2H, m, CH), 3.22-3.32 (4H, m, 2 * CH2), 2.36-2.48 (4H, m, 2 * CH2), 2.22 (3H, s, CH3), 1.88-1.97 (2H, m, CH), 1.32-1.42 (2H, m, CH).


30-6: 13.10 (1H, bs, NH), 10.38 (1H, bs, NH), 8.56 (1H, s, CHarom), 8.12 (1H, d, NH), 7.75 (1H, d, CHarom), 6.23 (1H, dd, CHarom), 6.14 (1H, d, CHarom), 3.97 (3H, s, CH3), 3.80-3.86 (2H, m, CH), 3.62-3.74 (1H, m, CH), 3.40-3.55 (2H, m, CH), 3.22-3.32 (4H, m, 2 * CH2), 2.36-2.48 (4H, m, 2 * CH2), 2.23 (3H, s, CH3), 1.90-1.99 (2H, m, CH), 1.32-1.45 (2H, m, CH).


30-7: 12.43 (1H, bs, NH), 10.22 (1H, bs, NH), 8.32 (1H, s, CHarom), 8.13 (1H, d, NH), 7.73 (1H, d, CHarom), 6.37 (2H, bs, NH2), 6.22 (1H, dd, CHarom), 6.13 (1H, d, CHarom), 3.78-3.86 (2H, m, CH), 3.65-3.74 (1H, m, CH), 3.44-3.54 (2H, m, CH), 3.22-3.32 (4H, m, 2 * CH2), 2.36-2.48 (4H, m, 2 * CH2), 2.23 (3H, s, CH3), 1.90-1.99 (2H, m, CH), 1.32-1.45 (2H, m, CH).


30-8: 13.79 (1H, bs, NH), 10.91 (1H, bs, NH), 10.69 (1H, bs, NH), 8.83 (1H, s, CHarom), 8.76 (1H, s, CHarom), 8.18 (1H, d, NH), 7.80 (1H, d, CHarom), 6.82-6.75 (3H, m, CHarom), 6.26 (1H, d, CHarom), 6.15 (1H, d, CHarom), 3.87-3.82 (2H, dt, CH), 3.72 (1H, m, CH), 3.54-3.47 (2H, m, CH), 3.32-3.29 (4H, m, CH), 2.42-2.46 (4H, m, CH), 2.28 (3H, s, CH3), 1.97-1.95 (2H, m, CH), 1.43-1.36 (2H, m, CH).


30-13: 12.99 (1H, s, NH), 9.92 (1H, s, NH), 8.38 (1H, d, NH, J = 7.6 Hz), 7.92 (1H, d, CHarom, J = 8.4 Hz), 7.84 (1H, CHarom, J = 9.2 Hz), 7.32 (1H, d, CHarom, J = 8.4 Hz), 7.07-7.00 (3H, m, CHarom), 6.26 (1H, d, CHarom, J = 8.8 Hz), 6.14 (1H, s, CHarom), 4.21 (2H, s), 3.82-3.72 (2H, m), 3.69-3.63 (1H, m), 3.48 (2H, t), 3.28 (4H, s), 2.46 (4H, s), 2.25 (3H, s), 2.00-1.90 (2H, m), 1.37-1.26 (2H, m).


30-14: 12.96 (1H, sl, NH), 9.84 (1H, s, NH), 8.34 (1H, d, NH, J = 7.6 Hz), 7.96 (1H, d, CHarom, J = 9.2 Hz), 7.81 (1H, d, CHarom, J = 8.8 Hz), 7.25 (1H, s, CHarom), 7.23 (1H, s, CHarom), 7.17 (1H, t, CHarom), 6.96 (1H, d, CHarom, J = 9.2 Hz), 6.25 (1H, d, CHarom, J = 7.6 Hz), 6.14 (1H, s, CHarom), 5.35 (2H, s), 3.82-3.77 (2H, m), 3.67 (1H, sl), 3.46 (2H, t), 3.29 (4H, s), 2.50 (4H, s), 2.29 (3H, s), 1.93-1.88 (2H, m), 1.35-1.25 (2H, m).


30-15: 13.01 (1H, sl, NH), 10.11 (1H, sl, NH), 7.99 (1H, sl, NH), 7.97 (1H, d, CHarom, J = 9.2 Hz), 7.84 (1H, d, CHarom, J = 8.4 Hz), 7.25-7.14 (3H, m, CHarom), 6.97 (1H, d, CHarom, J = 8.8 Hz), 6.67 (1H, sl, CHarom), 6.51 (1H, d, CHarom, J = 8.0 Hz), 5.35 (2H, s, CHarom), 3.83-3.78 (2H, m), 3.68-3.63 (1H, m), 3.47 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.19 (3H, s), 2.00-1.87 (4H, m), 1.75-1.65 (4H, m), 1.34-1.28 (2H, m).


30-16: 12.95 (1H, sl, NH), 9.85 (1H, s, NH), 8.33 (1H, d, NH, J = 7.6 Hz), 7.95 (1H, d, CHarom, J = 8.8 Hz), 7.81 (1H, d, CHarom, J = 8.8 Hz), 7.48 (1H, q, CHarom), 7.31-7.20 (2H, m, CHarom), 6.93 (1H, d, CHarom, J = 9.2 Hz), 6.25 (1H, d, CHarom, J = 9.2 Hz), 6.14 (1H, s, CHarom), 5.35 (2H, s), 3.81-3.76 (2H, m), 3.68 (1H, sl), 3.47 (2H, t), 3.26 (4H, s), 2.44 (4H, s), 2.29 (3H, s), 1.94-1.88 (2H, m), 1.36-1.27 (2H, m).


30-17: 13.06 (1H, sl, NH), 10.12 (1H, sl, NH), 7.93 (1H, sl, NH), 7.86 (2H, d, CHarom, J = 8.4 Hz), 7.51-7.44 (1H, m, CHarom), 7.30-7.20 (2H, m, CHarom), 6.90 (1H, sl, CHarom), 6.64 (1H, sl, CHarom), 6.49 (1H, sl, CHarom), 5.37 (2H, s, CHarom), 3.83-3.76 (2H, m), 3.68-3.63 (1H, m), 3.46 (2H, t), 2.86 (2H, d, J = 10.4 Hz), 2.44-2.38 (1H, m), 2.19 (3H, s), 1.99-1.90 (4H, m), 1.75-1.65 (4H, m), 1.40-1.30 (2H, m).


30-18: 12.94 (1H, sl, NH), 9.81 (1H, s, NH), 8.32 (1H, d, CHarom, J = 7.7 Hz), 7.96 (1H, d, CHarom, J = 9 Hz), 7.81 (1H, d, CHarom, J = 9 Hz), 7.71 (1H, d, NH), 7.51 (1H, d, CHarom, J = 8.6 Hz), 7.43 (1H, dd, CHarom, J = 8.6 Hz), 6.97 (1H, d, CHarom, J = 8.6 Hz), 6.24 (1H, d, CHarom, J = 8.9 Hz), 6.13 (1H, s, CHarom), 5.39 (2H, s), 3.82-3.74 (2H, m), 3.72-3.62 (1H, m), 3.46 (2H, t), 3.28-3.22 (4H, m), 2.46-2.40 (4H, m), 2.22 (3H, s), 1.95-1.87 (2H, m), 1.37-1.26 (2H, m).


30-19: 13.01 (1H, sl, NH), 10.09 (1H, s, NH), 7.97 (2H, d, CHarom, J = 9 Hz), 7.83 (1H, d, CHarom, J = 8.2 Hz), 7.71 (1H, dd, NH), 7.50 (1H, d, CHarom, J = 7.4 Hz), 7.43 (1H, dd, CHarom, J = 8.6 Hz), 6.98 (1H, d, CHarom, J = 9 Hz), 6.67 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 8.2 Hz), 5.38 (2H, s), 3.84-3.75 (2H, m), 3.72-3.62 (1H, m), 3.46 (2H, t), 2.86 (2H, d), 2.43 (1H, m), 2.19 (3H, s), 1.99-1.88 (4H, m), 1.74-1.64 (4H, m), 1.38-1.26 (2H, m).


30-20: 12.97 (1H, sl, NH), 9.82 (1H, s, NH), 8.32 (1H, d, NH, J = 8.0 Hz), 7.97 (1H, d, CHarom, J = 8.8 Hz), 7.87 (1H, s, CHarom), 7.80-7.76 (2H, m, CHarom), 7.64 (1H, d, CHarom, J = 8.4 Hz), 6.96 (1H, d, CHarom, J = 8.8 Hz), 6.24 (1H, d, CHarom, J = 8.8 Hz), 6.13 (1H, s, CHarom), 5.47 (2H, s), 3.81-3.76 (2H, m), 3.66 (1H, sl), 3.46 (2H, t), 3.26 (4H, s), 2.43 (4H, s), 2.29 (3H, s), 1.93-1.88 (2H, m), 1.35-1.25 (2H, m).


30-21: 13.03 (1H, s, NH), 10.08 (1H, s, NH), 8.00-7.95 (2H, m, CHarom), 7.87-7.75 (3H, m, CHarom), 7.63 (1H, d, CHarom, J = 8.4 Hz), 6.97 (1H, d, CHarom, J = 8.8 Hz), 6.67 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 8.0 Hz), 5.47 (2H, s, CHarom), 3.83-3.76 (2H, m), 3.68-3.64 (1H, m), 3.47 (2H, t), 2.87 (2H, d, J = 10.4 Hz), 2.45-2.40 (1H, m), 2.20 (3H, s), 2.00-1.87 (4H, m), 1.74-1.65 (4H, m), 1.36-1.25 (2H, m).


30-22: 12.93 (1H, s, NH), 9.86 (1H, s, NH), 8.70 (1H, s, CHarom), 8.51 (1H, dd, CHarom, J = 5.2 Hz), 8.38 (1H, d, NH, J = 8.0 Hz), 7.96-7.90 (2H, m, CHarom), 7.84 (1H, d, CHarom, J = 8.8 Hz), 7.73-7.33 (1H, m, CHarom), 6.91 (1H, d, CHarom, J = 8.8 Hz), 6.27 (1H, d, CHarom, J = 8.8 Hz), 6.15 (1H, s, CHarom), 5.35 (2H, s), 3.83-3.77 (2H, m), 3.70-3.64 (1H, m), 3.47 (2H, t), 3.59 (4H, s), 2.59 (4H, s), 2.34 (3H, s), 1.95-1.88 (2H, m), 1.40-1.28 (2H, m).


30-23: 13.03 (1H, s, NH), 10.17 (1H, s, NH), 8.70 (1H, s, CHarom), 8.52 (1H, dd, CHarom, J = 4.8 Hz), 8.06 (1H, d, NH, J = 7.6 Hz), 7.96 (1H, d, CHarom, J = 8.8 Hz), 7.94-7.88 (2H, m, CHarom), 7.37-7.34 (1H, m, CHarom), 6.93 (1H, d, CHarom, J = 9.2 Hz), 6.69 (1H, s, CHarom), 6.52 (1H, d, CHarom, J = 8.0 Hz), 5.36 (2H, s, CHarom), 3.83-3.79 (2H, m), 3.68-3.64 (1H, m), 3.46 (2H, t), 3.25-3.15 (2H, m), 2.65-2.55 (3H, m), 2.54 (3H, s), 2.00-1.85 (6H, m), 1.41-1.28 (2H, m).


30-24: 13.21 (1H, s, NH), 10.00 (1H, s, NH), 8.30 (1H, d, NH, J = 7.6 Hz), 8.00 (1H, d, CHarom, J = 8.8 Hz), 7.79 (1H, d, CHarom, J = 9.2 Hz), 7.33 (1H, d, CHarom, J = 8.8 Hz), 7.26-7.16 (3H, m, CHarom), 6.24 (1H, d, CHarom, J = 8.8 Hz), 6.13 (1H, s, CHarom), 4.06-3.99 (2H, m), 3.67 (1H, sl), 3.47 (2H, t), 3.28 (4H, s), 2.47 (4H, s), 2.25 (3H, s), 1.94-1.88 (2H, m), 1.37-1.26 (2H, m).


30-25: 13.26 (1H, s, NH), 10.28 (1H, s, NH), 8.02 (1H, d, CHarom, J = 8.8 Hz), 7.97 (1H, d, NH, J = 7.6 Hz), 7.83 (1H, d, CHarom, J = 8.0 Hz), 7.34 (1H, d, CHarom, J = 8.8 Hz), 7.27-7.17 (3H, m, CHarom), 6.68 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 8.0 Hz), 3.85-3.78 (2H, m), 3.71-3.65 (1H, m), 3.47 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.48-2.40 (1H, m), 2.19 (3H, s), 1.98-1.88 (4H, m), 1.74-1.66 (4H, m), 1.36-1.27 (2H, m).


30-26: 13.12 (1H, s, NH), 9.95 (1H, s, NH), 8.32 (1H, d, NH, J = 7.6 Hz), 7.93 (1H, d, CHarom, J = 8.8 Hz), 7.79 (1H, d, CHarom, J = 8.8 Hz), 7.73 (1H, t, CHarom), 7.52-7.40 (2H, m, CHarom), 7.12 (1H, d, CHarom, J = 8.8 Hz), 6.25 (1H, d, CHarom, J = 8 .8 Hz), 6.13 (1H, s, CHarom), 3.83-3.77 (2H, m), 3.69 (1H, sl), 3.48 (2H, t), 3.28 (4H, s), 2.44 (4H, s), 2.27 (3H, s), 1.96-1.89 (2H, m), 1.37-1.27 (2H, m).


30-27: 13.17 (1H, s, NH), 10.21 (1H, s, NH), 7.99-7.92 (2H, m, CHarom et NH), 7.81 (1H, d, CHarom, J = 8.4 Hz), 7.77-7.70 (1H, m, CHarom), 7.51-7.40 (2H, m, CHarom), 7.13 (1H, dd, CHarom, J = 8.8 Hz), 6.69 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 8.4 Hz), 3.85-3.78 (2H, m), 3.72-3.67 (1H, m), 3.48 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.47-2.40 (1H, m), 2.20 (3H, s), 1.96-1.87 (4H, m), 1.75-1.65 (4H, m), 1.38-1.28 (2H, m).


30-28: 13.31 (1H, sl, NH), 9.95 (1H, sl, NH), 8.31 (1H, d, NH, J = 7.6 Hz), 7.99 (1H, d, CHarom, J = 7.6 Hz), 7.78 (1H, d, CHarom, J = 9.2 Hz), 7.58-7.49 (3H, m, CHarom), 7.31 (1H, d, CHarom, J = 8.8 Hz), 6.24 (1H, d, CHarom, J = 8.8 Hz), 6.10 (1H, s, CHarom), 3.83-3.76 (2H, m), 3.70-3.60 (1H, m), 3.45 (2H, t), 3.21 (4H, s), 2.43 (4H, s), 2.22 (3H, s), 1.94-1.86 (2H, m), 1.38-1.28 (2H, m).


30-29: 13.26 (1H, s, NH), 10.25 (1H, s, NH), 8.01 (1H, d, CHarom, J = 8.8 Hz), 7.94 (1H, d, NH, J = 7.6 Hz), 7.82 (1H, d, CHarom, J = 8.4 Hz), 7.59-7.54 (3H, m, CHarom), 7.32 (1H, d, CHarom, J = 8.8 Hz), 6.67 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 7.6 Hz), 3.84-3.78 (2H, m), 3.71-3.62 (1H, m), 3.47 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.45-2.41 (1H, m), 2.19 (3H, s), 1.96-1.90 (4H, m), 1.74-1.68 (4H, m), 1.34-1.27 (2H, m).


30-30: 13.23 (1H, s, NH), 9.98 (1H, s, NH), 8.29 (1H, d, NH, J = 7.6 Hz), 8.01 (1H, d, CHarom, J = 8.8 Hz), 7.79 (1H, d, CHarom, J = 8.8 Hz), 7.62 (1H, d, CHarom, J = 8.4 Hz), 7.52 (1H, s, CHarom), 7.44 (1H, d, CHarom, J = 7.6 Hz), 7.24 (1H, d, CHarom, J = 8.4 Hz), 6.25 (1H, d, CHarom, J = 8.0 Hz), 6.12 (1H, s, CHarom), 3.82-3.75 (2H, m), 3.73-3.67 (1H, m), 3.47 (2H, t), 3.27 (4H, s), 2.43 (4H, s), 2.22 (3H, s), 1.95-1.87 (2H, m), 1.35-1.28 (2H, m).


30-31: 13.28 (1H, s, NH), 10.25 (1H, s, NH), 8.02 (1H, d, CHarom, J = 8.8 Hz), 7.95 (1H, d, NH, J = 7.6 Hz), 7.81 (1H, d, CHarom, J = 8.0 Hz), 7.61 (1H, d, CHarom, J = 8.4 Hz), 7.56 (1H, s, CHarom), 7.43 (1H, dd, CHarom, J = 8.4 Hz), 7.25 (1H, d, CHarom, J = 8.8 Hz), 6.68 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 7.2 Hz), 3.84-3.78 (2H, m), 3.69-3.61 (1H, m), 3.47 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.47-2.41 (1H, m), 2.20 (3H, s), 2.00-1.90 (4H, m), 1.76-1.69 (4H, m), 1.40-1.30 (2H, m).


30-32: 13.16 (1H, s, NH), 9.95 (1H, s, NH), 8.33 (1H, d, NH, J = 8.0 Hz), 7.93 (1H, d, CHarom, J = 8.8 Hz), 7.89 (1H, d, CHarom, J = 9.2 Hz), 7.79 (1H, d, CHarom, J = 9.2 Hz), 7.70-7.63 (2H, m, CHarom), 7.60 (1H, t, CHarom), 6.97 (1H, d, CHarom, J = 8.8 Hz), 6.25 (1H, d, CHarom, J = 9.2 Hz), 6.14 (1H, s, CHarom), 3.83-3.78 (2H, m), 3.68 (1H, sl), 3.48 (2H, t), 3.28 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.95-1.90 (2H, m), 1.38-1.28 (2H, m).


30-33: 13.21 (1H, s, NH), 10.22 (1H, s, NH), 7.99 (1H, d, NH, J = 7.6 Hz), 7.94 (1H, d, CHarom, J = 9.2 Hz), 7.89 (1H, d, CHarom, J = 7.2 Hz), 7.82 (1H, d, CHarom, J = 8.4 Hz), 7.71-7.57 (3H, m, CHarom), 6.98 (1H, d, CHarom, J = 8.8 Hz), 6.69 (1H, s, CHarom), 6.52 (1H, d, CHarom, J = 8.0 Hz), 3.85-3.79 (2H, m), 3.72-3.62 (1H, m), 3.48 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.47-2.41 (1H, m), 2.19 (3H, s), 2.00-1.90 (4H, m), 1.76-1.69 (4H, m), 1.40-1.30 (2H, m).


30-34: 13.07 (1H, s, NH), 10.11 (1H, s, NH), 8.32 (1H, d, NH, J = 7.6 Hz), 7.90-7.85 (2H, m, CHarom), 7.22 (1H, d, CHarom, J = 8.8 Hz), 7.19 (1H, s, CHarom), 7.17 (1H, s, CHarom), 7.03 (1H, t, CHarom), 6.30 (1H, d, CHarom, J = 8.4 Hz), 6.19 (1H, s, CHarom), 4.43 (2H, s), 4.02 (2H, sl), 3.80-3.74 (2H, m), 3.67 (1H, sl), 3.44 (2H, t), 3.10 (4H, s), 2.84 (3H, s), 1.89-1.84 (2H, m), 1.30-1.14 (4H, m).


30-35: 13.08 (1H, s, NH), 10.28 (1H, s, NH), 7.96 (1H, d, NH, J = 7.6 Hz), 7.88 (1H, d, CHarom, J = 8.8 Hz), 7.86 (1H, d, CHarom, J = 6.8 Hz), 7.22 (1H, d, CHarom, J = 8.8 Hz), 7.18 (1H, s, CHarom), 7.17 (1H, s, CHarom), 7.02 (1H, t, CHarom), 6.66 (1H, s, CHarom), 6.51 (1H, d, CHarom, J = 8.4 Hz), 4.43 (2H, s), 3.80-3.74 (2H, m), 3.64 (1H, sl), 3.44 (2H, t), 2.89-2.84 (2H, m), 2.43 (1H, sl), 2.20 (3H, s), 1.98-1.95 (2H, m), 1.89-1.84 (2H, m), 1.72-1.69 (4H, m), 1.29-1.20 (2H, m).


30-36: 13.10 (1H, sl, NH), 10.11 (1H, s, NH), 9.73 (1H, sl, COOH), 8.34 (1H, sl, NH), 7.92-7.86 (2H, m, CHarom), 7.47-7.40 (1H, m, CHarom), 7.23 (1H, d, CHarom, J = 8.8 Hz), 7.20-7.13 (1H, m, CHarom), 7.11-7.05 (1H, m, CHarom), 6.31 (1H, dd, CHarom, J = 9.2 Hz), 6.20 (1H, s, CHarom), 4.41 (2H, s), 4.04 (2H, d, J = 8.8 Hz), 3.81-3.75 (2H, m), 3.70-3.66 (1H, m), 3.51 (2H, d, J = 11.2 Hz), 3.44 (2H, t), 3.16-2.97 (4H, m), 2.87 (3H, s), 1.91-1.84 (2H, m), 1.34-1.22 (2H, m).


30-37: 13.09 (1H, s, NH), 10.29 (1H, s, NH), 7.97 (1H, d, NH, J = 7.6 Hz), 7.90-7.86 (2H, m, CHarom), 7.47-7.41 (1H, m, CHarom), 7.23 (1H, d, CHarom, J = 8.8 Hz), 7.19-7.13 (1H, m, CHarom), 7.11-7.05 (1H, m, CHarom), 6.67 (1H, s, CHarom), 6.52 (1H, d, CHarom, J = 8.0 Hz), 4.41 (2H, s), 3.79-3.74 (2H, m), 3.66-3.62 (1H, m), 3.44 (2H, t), 2.86 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.19 (3H, s), 2.00-1.85 (4H, m), 1.74-1.65 (4H, m), 1.33-1.23 (2H, m).


30-38: 13.02 (1H, s, NH), 10.04 (1H, s, NH), 8.28 (1H, d, NH, J = 8.0 Hz), 7.88-7.84 (2H, m, CHarom), 7.74 (1H, s, CHarom), 7.43 (1H, d, CHarom, J = 8.8 Hz), 7.29 (1H, dd, CHarom, J = 8.4 Hz), 7.22 (1H, d, CHarom, J = 8.8 Hz), 6.25 (1H, dd, CHarom, J = 9.2 Hz), 6.12 (1H, s, CHarom), 4.50 (2H, s), 3.78-3.74 (2H, m), 3.66-3.62 (1H, m), 3.44 (2H, t), 3.26 (4H, s), 2.43 (4H, s), 2.22 (3H, s), 1.91-1.84 (2H, m), 1.35-1.23 (2H, m).


30-39: 13.09 (1H, s, NH), 10.32 (1H, s, NH), 8.28 (1H, d, NH, J = 8.0 Hz), 7.90 (2H, D, CHarom), 7.74 (1H, s, CHarom), 7.43 (1H, d, CHarom, J = 8.4 Hz), 7.29 (1H, dd, CHarom, J = 8.4 Hz), 7.25 (1H, d, CHarom, J = 8.8 Hz), 6.67 (1H, s, CHarom), 6.54 (1H, dd, CHarom, J = 8.4 Hz), 4.51 (2H, s), 3.79-3.76 (2H, m), 3.70-3.64 (1H, m), 3.44 (2H, t), 2.95-2.92 (2H, m), 2.52-2.51 (1H, m), 2.27 (3H, s), 2.13-2.01 (2H, m), 1.90-1.87 (2H, m) 1.77-1.69 (4H, m), 1.32-1.24 (2H, m).


30-40: 12.77 (1H, s, NH), 9.86 (1H, s, NH), 9.60 (1H, s, NH), 8.40 (1H, d, NH, J = 7.6 Hz), 7.86 (1H, d, CHarom, J = 8.8 Hz), 7.83 (1H, d, CHarom, J = 9.2 Hz), 7.56 (2H, d, CHarom, J = 8.8 Hz), 6.93 (1H, d, CHarom, J = 9.2 Hz), 6.55 (1H, t, CHarom), 6.23 (1H, dd, CHarom, J = 9.2 Hz), 6.13 (1H, s, CHarom), 3.82-3.75 (2H, m), 3.69-3.61 (1H, m), 3.46 (2H, t), 3.27 (4H, s), 2.44 (4H, s), 2.29 (3H, s), 1.96-1.88 (2H, m), 1.38-1.26 (2H, m).


30-41: 12.85 (1H, sl, NH), 10.13 (1H, s, NH), 9.62 (1H, s, NH), 8.03 (1H, d, NH, J = 7.2 Hz), 7.90-7.84 (2H, m, CHarom), 7.57 (2H, dd, CHarom, J = 10.4 Hz), 6.95 (1H, d, CHarom, J = 8.8 Hz), 6.68 (1H, s, CHarom), 6.60-6.50 (2H, m, CHarom), 3.83-3.78 (2H, m), 3.68-3.63 (1H, m), 3.46 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.20 (3H, s), 2.00-1.92 (4H, m), 1.75-1.65 (4H, m), 1.37-1.27 (2H, m).


30-42: 12.77 (1H, s, NH), 9.87 (1H, s, NH), 9.02 (1H, s, NH), 8.80-8.72 (1H, m, CHarom), 8.41 (1H, d, NH, J = 7.6 Hz), 7.86 (1H, d, CHarom, J = 8.8 Hz), 7.83 (1H, d, CHarom, J = 9.2 Hz), 7.28 (1H, d, CHarom, J = 9.2 Hz), 7.22-7.15 (2H, m, CHarom), 6.63-6.57 (1H, m, CHarom), 6.23 (1H, d, CHarom, J = 8.8 Hz), 6.13 (1H, s, CHarom), 3.83-3.75 (2H, m), 3.70-3.64 (1H, m), 3.46 (2H, t), 3.27 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.95-1.88 (2H, m), 1.39-1.26 (2H, m).


30-43: 12.84 (1H, s, NH), 10.13 (1H, s, NH), 9.05 (1H, sl, NH), 8.81-8.74 (1H, m, CHarom), 8.05 (1H, d, J = 7.2 Hz), 7.89-7.84 (2H, m, CHarom), 7.30 (1H, d, CHarom, J = 8.8 Hz), 7.23-7.15 (1H, m, CHarom), 6.67 (1H, s, CHarom), 6.64-6.58 (1H, m, CHarom), 6.51 (1H, d, CHarom, J = 8.4 Hz), 3.83-3.76 (2H, m), 3.68-3.64 (1H, m), 3.47 (2H, t), 2.89 (2H, d, J = 10.8 Hz), 2.45-2.40 (1H, m), 2.21 (3H, s), 2.01-1.91 (4H, m), 1.74-1.66 (4H, m), 1.38-1.27 (2H, m).


30-44: 12.80 (1H, s, NH), 10.16 (1H, s, NH), 8.89 (1H, s, CHarom), 8.52 (1H, s, NH), 8.34 (1H, d, NH, J = 7.6 Hz), 7.89 (1H, d, CHarom, J = 9.2 Hz), 7.81 (1H, d, CHarom, J = 9.2 Hz), 7.41 (1H, d, CHarom, J = 8.8 Hz), 7.35 (1H, d, CHarom, J = 9.2 Hz), 6.89 (1H, dd, CHarom, J = 8.4 Hz), 6.21 (1H, d, CHarom, J = 9.2 Hz), 6.11 (1H, s, CHarom), 3.83-3.75 (2H, m), 3.66-3.60 (1H, m), 3.46 (2H, t), 3.25 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.95-1.87 (2H, m), 1.37-1.26 (2H, m).


30-45: 12.86 (1H, s, NH), 10.10 (1H, s, NH), 8.91 (1H, s, CHarom), 8.54 (1H, s, NH), 8.00 (1H, d, NH, J = 7.6 Hz), 7.90 (1H, d, CHarom, J = 9.2 Hz), 7.85 (1H, d, CHarom, J = 8.0 Hz), 7.41 (1H, d, CHarom, J = 8.4 Hz), 7.37 (1H, d, CHarom, J = 9.2 Hz), 6.88 (1H, dd, CHarom, J = 8.4 Hz), 6.64 (1H, s, CHarom), 6.48 (1H, d, CHarom, J = 8.4 Hz), 3.83-3.77 (2H, m), 3.67-3.60 (1H, m), 3.47 (2H, t), 2.88 (2H, d, J = 11.2 Hz), 2.45-2.38 (1H, m), 2.21 (3H, s), 2.00-1.87 (4H, m), 1.75-1.65 (4H, m), 1.37-1.26 (2H, m).


30-46: 13.74 (1H, sl, NH), 10.14 (1H, s, NH), 8.62 (1H, s, CHarom), 8.33 (1H, d, NH), 7.81 (1H, d, CHarom, J = 8.7 Hz), 7.12-7.03 (3H, m, CHarom), 6.26 (1H, d, CHarom, J = 8.8 Hz), 6.13 (1H, s, CHarom), 4.31 (2H, s), 4.14-4.07 (4H, m), 3.68 (1H, sl), 3.28 (4H, s), 2.43 (4H, s), 2.23 (3H, s), 1.92 (2H, d, J = 12.4 Hz), 1.38-1.26 (2H, m).


30-47: 13.80 (1H, sl, NH), 10.41 (1H, s, NH), 8.64 (1H, s, CHarom), 8.02 (1H, d, NH), 7.85 (1H, d, CHarom, J = 8.1 Hz), 7.12-7.03 (3H, m, CHarom), 6.69 (1H, s, CHarom), 6.52 (1H, d, CHarom, J = 8.1 Hz), 4.30 (2H, s), 3.81 (2H, d, J = 11.1 Hz), 3.68 (1H, sl), 3.48 (2H, t), 2.87 (2H, d, J = 10.5 Hz), 2.47-2.39 (1H, sl), 2.19 (3H, s), 2-1.88 (4H, m), 1.76-1.66 (4H, m), 1.39-1.27 (2H, m).


30-48: 13.99 (1H, sl, NH), 10.17 (1H, s, NH), 8.34 (1H, s, CHarom), 8.29 (1H, dl, NH), 7.78 (1H, d, CHarom, J = 8.9 Hz), 7.54-7.41 (3H, m, CHarom), 6.07 (1H, d, CHarom, J = 8.9 Hz), 5.87 (1H, s, CHarom), 3.82 (2H, dl), 3.62 (1H, sl), 3.51-3.37 (4H, m), 2.97 (3H, s), 2.28-2.19 (2H, m), 2.15 (6H, s), 2-1.90 (2H, m), 1.71-1.61 (2H, m), 1.42-1.28 (2H, m).


30-49: 14.06 (1H, sl, NH), 10.56 (1H, s, NH), 8.85 (1H, s, CHarom), 7.97 (1H, sl, NH), 7.85 (1H, d, CHarom, J = 8.1 Hz), 7.50-7.40 (3H, m, CHarom), 6.71 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 8.1 Hz), 3.83-3.76 (2H, m), 3.70 (1H, sl), 3.48 (2H, t), 2.88 (2H, d, J = 10.6 Hz), 2.48-2.40 (1H, m), 2.20 (3H, s), 2.01-1.89 (4H, m), 1.76-1.66 (4H, m), 1.40-1.28 (2H, m).


30-50: 13.94 (1H, sl, NH), 10.11 (1H, sl, NH), 8.59 (1H, s, CHarom), 8.30 (1H, sl, NH), 7.76 (1H, d, CHarom, J = 9.2 Hz), 7.27-7.13 (3H, m, CHarom), 6.04 (1H, dd, CHarom, J = 9.2 Hz), 5.85 (1H, s, CHarom), 3.87-3.76 (2H, m), 3.66-3.55 (1H, m), 3.49-3.26 (4H, m), 2.96 (3H, s), 2.22 (2H, t), 2.14 (6H, s), 1.97-1.89 (2H, m), 1.69-1.60 (2H, q), 1.40-1.28 (2H, m).


30-51: 13.95 (1H, sl, NH), 10.17 (1H, sl, NH), 8.54 (1H, s, CHarom), 8.28 (1H, sl, NH), 7.78 (1H, d, CHarom, J = 8.8 Hz), 7.59 (1H, d, CHarom, J = 9.2 Hz), 7.42-7.38 (2H, m, CHarom), 6.23 (1H, d, CHarom, J = 8.0 Hz), 6.11 (1H, s, CHarom), 3.82-3.77 (2H, m), 3.66 (1H, sl), 3.46 (2H, t), 3.26 (4H, s), 2.43 (4H, s), 2.22 (3H, s), 1.92-1.88 (2H, m), 1.34-1.24 (2H, m).


30-52: 13.97 (1H, sl, NH), 10.20 (1H, s, NH), 8.38 (1H, s, CHarom), 8.27 (1H, d, NH), 7.88 (1H, d, CHarom, J = 7.2 Hz), 7.78 (1H, d, CHarom, J = 9.2 Hz), 7.66-7.55 (3H, m, CHarom), 6.26 (1H, dd, CHarom, J = 9.2 Hz), 6.13 (1H, s, CHarom), 3.85-3.76 (2H, m), 3.75-3.63 (1H, m), 3.48 (2H, t), 3.37-3.26 (4H, m), 2.61-2.52 (4H, m), 2.32 (3H, sl), 1.96-1.88 (2H, m), 1.39-1.26 (2H, m).


30-53: 13.64 (1H, s, NH), 10.20 (1H, s, NH), 8.30 (1H, d, CHarom, J = 8.8 Hz), 8.23 (1H, d, CHarom, J = 8.0 Hz), 8.19 (1H, d, CHarom, J = 8.8 Hz), 7.81 (1H, d, CHarom, J = 9.2 Hz), 7.75-7.65 (3H, m, CHarom), 6.28 (1H, dd, CHarom, J = 8.8 Hz), 6.14 (1H, s, CHarom), 3.83-3.77 (2H, m), 3.70-3.64 (1H, m), 3.48 (2H, t), 3.29 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.95-1.89 (2H, m), 1.38-1.26 (2H, m).


30-54: 13.64 (1H, sl, NH), 10.48 (1H, sl, NH), 8.32 (1H, d, CHarom, J = 8.8 Hz), 8.19 (1H, d, CHarom, J = 8.8 Hz), 7.91 (1H, sl, NH), 7.85 (1H, d, CHarom, J = 8.4 Hz), 7.77-7.65 (3H, m, CHarom), 6.71 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 8.4 Hz), 3.86-3.80 (2H, m), 3.71-3.64 (1H, m), 3.48 (2H, t), 2.89 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.21 (3H, s), 2.00-1.90 (4H, m), 1.75-1.65 (4H, m), 1.38-1.27 (2H, m).


30-55: 13.64 (1H, s, NH), 10.16 (1H, s, NH), 8.29 (1H, d, CHarom, J = 8.8 Hz), 8.24 (1H, d, NH, J = 7.6 Hz), 8.17 (1H, d, CHarom, J = 8.8 Hz), 8.09 (1H, t, CHarom), 7.88-7.85 (1H, m, CHarom), 7.81 (1H, d, CHarom, J = 9.2 Hz), 7.67 (1H, q, CHarom), 6.28 (1H, d, CHarom, J = 8.8 Hz), 6.14 (1H, s, CHarom), 3.83-3.75 (2H, m), 3.72-3.67 (1H, m), 3.48 (2H, t), 3.29 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.96-1.89 (2H, m), 1.35-1.28 (2H, m).


30-56: 13.67 (1H, s, NH), 10.43 (1H, s, NH), 8.31 (1H, d, CHarom, J = 8.8 Hz), 8.18 (1H, d, CHarom, J = 8.8 Hz), 8.13-8.05 (1H, m, CHarom), 7.92 (1H, d, NH, J = 7.6 Hz), 7.90-7.82 (2H, m, CHarom), 7.66 (1H, q, CHarom), 6.71 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 8.4 Hz), 3.85-3.80 (2H, m), 3.73-3.65 (1H, m), 3.49 (2H, t), 2.89 (2H, d, J = 11.2 Hz), 2.48-2.42 (1H, m), 2.21 (3H, s), 1.99-1.90 (4H, m), 1.76-1.68 (4H, m), 1.37-1.27 (2H, m).


30-57: 13.66 (1H, s, NH), 10.17 (1H, s, NH), 8.30 (1H, d, CHarom, J = 8.8 Hz), 8.24-8.16 (2H, m, CHarom et NH), 8.03-7.97 (3H, m, CHarom), 7.81 (1H, d, CHarom, J = 9.2 Hz), 6.28 (1H, d, CHarom, J = 7.2 Hz), 6.14 (1H, s, CHarom), 3.83-3.77 (2H, m), 3.71-3.67 (1H, m), 3.48 (2H, t), 3.29 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.96-1.89 (2H, m), 1.34-1.28 (2H, m).


30-58: 13.71 (1H, s, NH), 10.45 (1H, s, NH), 8.32 (1H, d, CHarom, J = 9.2 Hz), 8.22 (1H, d, CHarom, J = 8.8 Hz), 8.02-7.96 (3H, m, CHarom), 7.86-7.81 (1H, m, NH), 7.83 (1H, d, CHarom), 6.71 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 7.6 Hz), 3.85-3.78 (2H, m), 3.72-3.65 (1H, m), 3.48 (2H, t), 2.88 (2H, d, J = 11.2 Hz), 2.48-2.44 (1H, m), 2.21 (3H, s), 1.97-1.87 (4H, m), 1.76-1.70 (4H, m), 1.36-1.28 (2H, m).


30-59: 13.69 (1H, s, NH), 10.04 (1H, s, NH), 8.34 (1H, d, NH, J = 8.8 Hz), 8.26-8.16 (3H, m, CHarom), 7.81 (1H, dd, CHarom, J = 8.4 Hz), 7.74 (1H, d, CHarom, J = 9.2 Hz), 7.66 (1H, d, CHarom, J = 8.4 Hz), 6.24 (1H, dd, CHarom, J = 9.2 Hz), 6.10 (1H, s, CHarom), 3.82-3.76 (2H, m), 3.68-3.62 (1H, m), 3.48 (2H, t), 3.27 (4H, s), 2.43 (4H, s), 2.22 (3H, s), 1.93-1.86 (2H, m), 1.31-1.21 (2H, m).


30-60: 13.74 (1H, s, NH), 10.31 (1H, s, NH), 8.35 (1H, d, CHarom, J = 8.8 Hz), 8.25 (1H, d, CHarom, J = 8.8 Hz), 8.21 (1H, s, CHarom), 7.85 (1H, d, NH, J = 7.2 Hz), 7.81 (1H, dd, CHarom, J = 8.8 Hz), 7.76 (1H, d, CHarom, J = 8.0 Hz), 7.66 (1H, d, CHarom, J = 8.8 Hz), 6.67 (1H, s, CHarom), 6.50 (1H, d, CHarom, J = 8.0 Hz), 3.85-3.78 (2H, m), 3.68-3.62 (1H, m), 3.48 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.46-2.40 (1H, m), 2.20 (3H, s), 1.97-1.87 (4H, m), 1.75-1.67 (4H, m), 1.32-1.24 (2H, m).


30-61: 13.61 (1H, s, NH), 10.32 (1H, s, NH), 8.71 (1H, d, NH, J = 8.0 Hz), 8.21 (1H, d, CHarom, J = 8.8 Hz), 7.87 (1H, d, CHarom, J = 9.2 Hz), 7.80 (1H, d, CHarom, J = 8.8 Hz), 7.17 (1H, t, CHarom), 7.05-7.02 (2H, m, CHarom), 6.29 (1H, d, CHarom, J = 9.2 Hz), 6.14 (1H, s, CHarom), 4.93 (2H, s), 3.74-3.68 (3H, m), 3.43 (2H, t), 3.29 (4H, s), 2.44 (4H, s), 2.88 (3H, s), 1.90-1.84 (2H, m), 1.28-1.20 (2H, m).


30-62: 13.67 (1H, sl, NH), 10.59 (1H, s, NH), 8.23 (1H, d, CHarom, J = 8.8 Hz), 8.10 (1H, d, NH, J = 7.6 Hz), 7.92 (1H, d, CHarom, J = 8.0 Hz), 7.82 (1H, d, CHarom, J = 8.8 Hz), 7.17 (1H, t, CHarom), 7.05-7.02 (2H, m, CHarom), 6.71 (1H, s, CHarom), 6.56 (1H, d, CHarom, J = 8.0 Hz), 4.94 (2H, s), 3.77-3.70 (3H, m), 3.43 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.20 (3H, s), 1.98-1.91 (2H, m), 1.89-1.95 (2H, m), 1.75-1.67 (4H, m), 1.30-1.20 (2H, m).


30-63: 13.63 (1H, sl, NH), 10.28 (1H, s, NH), 8.37 (1H, d, NH, J = 8.0 Hz), 8.24 (1H, d, CHarom, J = 8.8 Hz), 7.88-7.82 (2H, m, CHarom), 7.24-7.17 (3H, m, CHarom), 6.29 (1H, d, CHarom, J = 9.2 Hz), 6.14 (1H, s, CHarom), 4.87 (2H, s), 3.75-3.70 (3H, m), 3.43 (2H, t), 3.28 (4H, s), 2.45 (4H, s), 2.23 (3H, s), 1.90-1.85 (2H, m), 1.32-1.20 (2H, m).


30-64: 13.69 (1H, sl, NH), 10.55 (1H, s, NH), 8.26 (1H, d, CHarom, J = 8.8 Hz), 8.05 (1H, d, NH, J = 7.6 Hz), 7.90 (1H, d, CHarom, J = 8.4 Hz), 7.86 (1H, d, CHarom, J = 8.8 Hz), 7.24-7.15 (3H, m, CHarom), 6.70 (1H, s, CHarom), 6.56 (1H, d, CHarom, J = 8.0 Hz), 4.88 (2H, s), 3.80-3.65 (3H, m), 3.43 (2H, t), 2.87 (2H, d, J = 11.2 Hz), 2.46-2.40 (1H, m), 2.20 (3H, s), 2.00-1.86 (4H, m), 1.75-1.67 (4H, m), 1.29-1.23 (2H, m).


30-65: 13.49 (1H, sl, NH), 10.45 (1H, s, NH), 9.31 (1H, sl, COOH), 8.21 (1H, d, CHarom, J = 8.8 Hz), 8.06 (1H, sl, NH), 7.92 (1H, d, CHarom, J = 8.4 Hz), 7.57 (1H, d, CHarom, J = 8.8 Hz), 7.17-7.11 (2H, m, CHarom), 6.96-6.91 (1H, m, CHarom), 6.67 (1H, s, CHarom), 6.53 (1H, d, CHarom, J = 8.0 Hz), 4.51 (1H, d, J = 13.2 Hz), 4.20 (1H, d, J = 13.2 Hz), 3.81-3.76 (2H, m), 3.71-3.62 (1H, m), 3.56-3.41 (4H, m), 3.08 (2H, t), 2.83 (3H, s), 2.45-2.40 (1H, m), 2.07-2.00 (2H, m), 1.95-1.86 (4H, m), 1.41-1.29 (2H, m).


30-66: 13.62 (1H, sl, NH), 10.22 (1H, sl, NH), 8.36 (1H, d, NH, J = 7.6 Hz), 8.23 (1H, d, CHarom, J = 8.8 Hz), 7.85 (1H, d, CHarom, J = 9.2 Hz), 7.80 (1H, d, CHarom, J = 8.8 Hz), 7.48 (1H, s, CHarom), 7.45-7.37 (2H, m, CHarom), 6.29 (1H, d, CHarom, J = 7.2 Hz), 6.14 (1H, s, CHarom), 4.97 (2H, s), 3.76-3.70 (3H, m), 3.44 (2H, t), 3.28 (4H, s), 2.44 (4H, s), 2.23 (3H, s), 1.91-1.86 (2H, m), 1.30-1.24 (2H, m).


30-67: 13.67 (1H, sl, NH), 10.49 (1H, s, NH), 8.25 (1H, d, CHarom, J = 8.8 Hz), 8.02 (1H, d, NH, J = 7.2 Hz), 7.89 (1H, d, CHarom, J = 8.0 Hz), 7.82 (1H, d, CHarom, J = 8.8 Hz), 7.49 (1H, t, CHarom), 7.45-7.35 (2H, m, CHarom), 6.70 (1H, s, CHarom), 6.56 (1H, d, CHarom, J = 8.0 Hz), 4.97 (2H, s), 3.78-3.64 (3H, m), 3.44 (2H, t), 2.88 (2H, d, J = 11.2 Hz), 2.45-2.40 (1H, m), 2.20 (3H, s), 1.98-1.86 (4H, m), 1.76-1.66 (4H, m), 1.32-1.22 (2H, m).


30-68: 13.46 (1H, s, NH), 10.36 (1H, s, NH), 8.21 (1H, d, CHarom, J = 8.8 Hz), 8.00 (1H, d, NH, J = 7.6 Hz), 7.86 (1H, d, CHarom, J = 8.4 Hz), 7.59 (1H, d, CHarom, J = 8.8 Hz), 7.43-7.33 (2H, m, CHarom), 7.28 (1H, s, CHarom), 6.69 (1H, s, CHarom), 6.54 (1H, d, CHarom, J = 7.6 Hz), 4.58 (1H, d, J = 12.8 Hz), 4.30 (1H, d, J = 12.8 Hz), 3.78-3.75 (2H, m), 3.70-3.65 (1H, m), 3.46 (2H, t), 2.92-2.88 (2H, m), 2.45-2.40 (1H, m), 2.24 (3H, s), 2.05-1.95 (2H, m), 1.93-1.89 (2H, m), 1.77-1.70 (4H, m), 1.34-1.24 (2H, m).


(ND; not determined).






Example 30-bis
(S)-4-(3-aminopyrrolidin-1-yl)-N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




876 μl (20 eq) of triethylamine is added to a solution of 238 mg (0.314 mmol) of (S)—N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl) acetamido)-4-(3-(2,2,2-trifluoroacetamido)pyrrolidin-1-yl)benzamide in 6 ml of methanol. The reaction medium is stirred at 65° C. for 4 hours. After returning to room temperature, 8 ml of n-butanol and 260 mg (6 eq) of potassium carbonate are added. The reaction medium is stirred at 80° C. for 24 hours. After returning to room temperature, the solvents are evaporated, water is added and the product is extracted with dichloromethane. The organic phase is washed with saturated sodium chloride solution, dried on magnesium sulfate, filtered and evaporated. The residue is purified by silica gel chromatography (8:2 dichloromethane/methanol as eluent) to yield 87 mg (yield=49%) of (S)-4-(3-aminopyrrolidin-1-yl)-N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyrazine-3-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in the form of a brown powder.
LCMS (EI, m/z): (M+1) 566.24.
1H NMR: δH ppm (400 MHz, DMSO): 10.46 (1H, bs, NH), 8.60 (1H, s, CHarom), 8.50 (1H, s, CHarom), 8.26 (1H, d, NH), 7.78 (1H, d, CHarom), 7.08 (1H, t, CHarom), 6.86 (2H, d, CHarom), 5.86 (1H, dd, CHarom), 5.71 (1H, d, CHarom), 3.80-3.88 (2H, m, CH), 3.63-3.70 (2H, m, CH), 3.40-3.55 (5H, m, CH), 3.01-3.08 (1H, m, CH), 2.08-2.13 (1H, m, CH), 1.92-1.99 (2H, m, CH3), 1.76-1.82 (1H, m, CH), 1.30-1.41 (2H, m, CHpyranone).
Examples of Method F2
Reduction
Example 31
N-(5-(3,5-difluorophenethyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




10 mg of 10% Pd/C is added to 100 mg (0.175 mmol) of N-(5-(3,5-difluorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in solution in a mixture of 10 ml of tetrahydrofuran and 5 ml of methanol before placing the reaction medium under an atmosphere of hydrogen. The reaction mixture is stirred for 12 hours at room temperature and then filtered on Celite and concentrated. 62 mg (yield=60%) of N-(5-(3,5-difluorophenethyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide are isolated in the form of a white solid.
LCMS (EI, m/z): (M+1) 576.23.
1H NMR: δH ppm (400 MHz, DMSO): 13.14 (1H, bs, NH), 10.32 (1H, bs, NH), 8.40 (1H, d, CHarom), 8.22 (1H, d, NH), 7.96 (1H, d, CHarom), 7.80 (1H, d, CHarom), 7.03-6.98 (3H, m, CHarom), 6.23 (1H, d, CHarom), 6.16 (1H, bs, CHarom), 3.84-3.81 (2H, dt, CH), 3.70 (1H, m, CH), 3.52-3.46 (2H, m, CH), 3.04-2.93 (4H, m, CH), 2.59-2.69 (4H, m, CH), 2.42-2.46 (4H, m, CH), 2.38 (3H, s, CH3), 1.96-1.93 (2H, m, CH), 1.40-1.33 (2H, m, CH).
The following derivative was obtained according to the same method:
















 

















Ex.**
ArX
R1
R2
Y1
n
W
Rj
Compound names
Yield
Mass





31-1












N
0
H
H
N-(5-(3,5-difluorophenethyl)-1H- pyrazolo[3,4-b]pyrazin-3-yl)-4-(4- methylpiperazin-1-yl)-2-(tetrahydro- 2H-pyran-4-ylamino)benzamide
47%
577.07 (M + H)





**1H NMR, dmso-d6, Ex.: 31-1: 13.68 (1H, sl, NH), 10.11 (1H, s, NH), 8.52 (1H, s, CHarom), 8.35 (1H, dl, NH), 7.82 (1H, d, CHarom, J = 9 Hz), 7.05-6.97 (3H, m, CHarom), 6.27 (1H, dd, CHarom), 6.14 (1H, s, CHarom), 3.83-3.76 (2H, m), 3.74-3.64 (1H, m), 3.47 (2H, t), 3.32-3.20 (6H, m), 3.07 (2H, dd), 2.44 (4H, dd), 2.23 (3H, s), 1.91 (2H, d), 1.38-1.27 (2H, m).






Example 32
5-(3,5-difluorophenylthio)-N-(4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzyl)-1H-pyrazolo[3,4-b]pyridin-3-amine




100 mg (0.173 mmol) of N-(5-(3,5-difluorophenylthio)-1-H-pyrazolo[3,4-b]pyridine-3-yl)-4-(4-methylpiperazine-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide is added, in small fractions, to a solution of 19.64 mg (0.518 mmol) of LiAlH4 in 3 ml of anhydrous tetrahydrofuran under argon at 0° C. The reaction mixture is heated at 90° C. for 15 hours. An additional portion of 20 mg of LiAlH4 is then added and the reaction medium stirred at 90° C. for 5 hours. 45 μl of water at 0° C. is then added to the reaction mixture, followed by 45 μl of sodium hydroxide (15% wt) and finally 120 μl of water. The reaction mixture is stirred at 25° C. for 1 hour and then filtered on Dicalite. After evaporation of the solvents, the crude product is purified by chromatography. 16.80 mg (17%) of 5-(3,5-difluorophenylthio)-N-(4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzyl)-1H-pyrazolo[3,4-b]pyridin-3-amine in the form of a yellow solid is obtained.
LCMS (EI, m/z): (M+1) 566.68.
1H NMR: δH ppm (400 MHz, DMSO): 12.57 (1H, bs, NH), 8.45 (2H, d, CHarom), 6.97-7.06 (2H, m CHarom), 6.73-6.75 (2H, m, CHarom), 6.65 (1H, t, NH), 6.13-6.19 (2H, m, CHarom), 4.98 (1H, d, NH), 4.30 (2H, m, CH2), 3.73-3.77 (2H, m, CH), 3.60 (1H, m, CH), 3.45-3.50 (2H, m, CH), 3.04 (4H, m, CH), 2.42 (4H, m, CH), 2.18 (3H, s, CH3), 1.80-1.83 (2H, m, CH), 1.27-1.32 (2H, m, CH).
The following derivatives were obtained according to the same method:




















Ex.**
ArX
R1
R2
(U)n
V
Y1, Y2, Y3, Y4
R3
Compound name
Yield
Mass







32-1












n = 0
CH2
N, ArXC, CH, N
H
5-(3,5-difluorophenylthio)-N- (4-(4-methylpiperazin-1-yl)- 2-(tetrahydro-2H-pyran-4- ylamino)benzyl)-1H- pyrazolo[4,3-b]pyrazin-3- amine
1%
567.3









Example 33
2-(4-aminophenyl)-N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-]pyridin-3-yl)acetamide




A solution of 152 mg (2.72 mmol) of iron and 70 mg (1.3 mmol) of ammonium chloride in 100 μl of water is added to a solution of 0.24 g (0.544 mmol) of N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)-2-(4-nitrophenyl)acetamide in 10 ml of a 2:1 ethanol/water mixture. Several drops of acetic acid are added to this mixture and it is heated at 60° C. for 4 hours. After cooling and concentration of the solvents, the crude reaction product is extracted with ethyl acetate and is washed with saturated sodium bicarbonate solution. The organic phases are combined, dried on magnesium sulfate and then concentrated. The crude product is purified by silica gel chromatography (DCM/MeOH) to yield 11 mg (4%) of 2-(4-aminophenyl)-N-(5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide in the form of a brown solid.
LCMS (EI, m/z): (M+1) 412.09.
1H NMR: δH ppm (400 MHz, DMSO): 13.60 (1H, bs, NH), 10.96 (1H, bs, NH), 8.68 (1H, d, CHarom), 8.55 (1H, d, CHarom), 7.06 (1H, m, CHarom), 6.98 (2H, CHarom), 6.79 (2H, m, CHarom), 6.50 (2H, m, CHarom), 4.92 (2H, s, NH), 3.51 (2H, m, CH2).
Examples of Method F3
Sulfide Oxidation
Example 34
5-(3,5-difluorophenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-3-amine




A solution of 663 mg (1.078 mmol) of oxone in 1.1 ml of water is added to a solution of 300 mg (1.078 mmol) of 5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyridin-3-amine in 10 ml of a 1:1 mixture of tetrahydrofuran and methanol at 0° C. The reaction mixture is stirred at room temperature for 16 hours. An additional portion of 663 mg of oxone at 0° C. is then added and the reaction medium stirred at room temperature for 24 hours. The solvents are evaporated and the reaction medium is diluted with sodium bicarbonate solution, extracted with ethyl acetate, dried on MgSO4 and then concentrated to yield 340 mg (81%) of 5-(3,5-difluorophenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-3-amine in the form of a yellow solid.
LCMS (EI, m/z): (M+1) 311.03.
1H NMR: δH ppm (400 MHz, DMSO): 12.72 (1H, bs, NH), 8.92 (1H, d, CHarom), 8.84 (1H, d, CHarom), 7.89-8.01 (1H, d, CHarom), 7.62-7.80 (2H, m, CHarom), 6.06 (2H, bs, NH).
The following compounds were also obtained by this method:
















 














Ex.**
ArX
Q
Y1, Y4
W
Compound name
Yield
Mass MH+





34-2




H
CH, N
H
5-(3,5- difluorobenzylsulfonyl)- 1H-pyrazolo[3,4- b]pyridin-3-amine
ND
(M + 1) 325.07


 


34-3








N, CH
H
tert-butyl 5-(3,5- dichlorophenylsulfonyl)- 1H-pyrazolo[4,3- b]pyridin-3-ylcarbamate
ND
ND





**1H NMR, DMSO-d6, Ex.: 33-2: 12.64 (1H, bs, NH), 8.56 (1H, d, CHarom), 8.49 (1H, d, CHarom), 7.24 (1H, ddd, CHarom), 6.94 (2H, bd, CHarom), 6.03 (2H, bs, NH), 4.80 (2H, s, CH).


(ND: not determined).






Alternatively, a protection step can be carried out before the oxidation reaction, followed by a deprotection step which can lead to the preparation of the corresponding sulfones or sulfoxides.
Example 34-bis
5-(3,5-difluorophenylsulfinyl)-1H-pyrazolo[4,3-b]pyrazin-3-amine




0.55 mL of triethylamine and 22 mg of 4-dimethylaminopyridine are added under argon to a solution of 500 mg (1.790 mmol) of 5-(3,5-difluorophenylthio)-1H-pyrazolo[3,4-b]pyrazin-3-amine in 10 mL of tetrahydrofurane. The solution is stirred at 0° C. and 0.915 mL of di-tert-butyl dicarbonate is added and the reaction medium is stirred overnight. An aqueous fraction is added to the reaction medium which is then extracted with ethyl acetate. The organic phases are dried on MgSO4 and concentrated in vacuum to give a crude product which is used in the oxidation step without further purification. The crude product obtained is dissolved in 10 mL of a 1:1 mixture of tetrahydrofurane and methanol at 0° C. and then a solution of 1.103 g (1.794 mmol) of oxone in 2 mL of water is added. The reaction medium is stirred at room temperature for 16 hours. An additional portion of 550 mg of oxone is then added and the reaction medium is stirred at room temperature for 5 hours. The solvents are evaporated and the reaction medium is diluted with a sodium bicarbonate solution, extracted with ethyl acetate, dried on magnesium sulfate and concentrated to lead to a mixture of the corresponding sulfone and sulfoxide which are used without further purification in the deprotection step.
0.373 mL of TFA in 4 mL of anhydrous THF is added at 0° C. to a solution of 600 mg of the previously obtained mixture in 6 mL of dichloromethane. The mixture is stirred 1 hour at room temperature and an additional portion of 4 equivalents of TFA in 4 mL of THF is added. After 1 hour of stirring, this operation is repeated and the reaction medium is stirred for a total time of 3 h 45. The solvents are evaporated and the reaction medium is diluted with a potassium carbonate solution, extracted with ethyl acetate, dried on magnesium sulfate and concentrated to yield a 1:1 mixture of 5-(3,5-difluorophenylsulfonyl)-1H-pyrazolo[3,4-b]pyrazin-3-amine and 5-(3,5-difluorophenyl sulfinyl)-1H-pyrazolo[3,4-b]pyrazin-3-amine. This mixture is used in the following steps without further purification.
The following compounds were also obtained by this method:
















 












Ex.**
ArX
Y1, Y4
Compound names
Yield
Mass MH+





34bis-1




CH, N
5-(2,5-difluorophenylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
58% 3 steps
(M + 1) 310.9


 


34bis-2




CH, N
5-(3,5-dichlorophenylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
38% 3 steps
(M + 1) 342.8


 


34bis-3




CH, N
5-(2,5-dichlorophenylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
41% 3 steps
(M + 1) 342.9


 


34bis-4




CH, N
5-(3,5-difluorobenzylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
58% 3 steps
(M + 1) 325.0


 


34bis-5




CH, N
5-(2,5-difluorobenzylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
45% 3 steps
(M + 1) 325.0


 


34bis-6




CH, N
5-(2,5-difluorobenzylsulfinyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
 5% 3 steps
(M + 1) 308.9


 


34bis-7




CH, N
5-(2,5-dichlorobenzylsulfonyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
 3% 3 steps
ND


 


34bis-8




CH, N
5-(2,5-dichlorobenzylsulfinyl)-1H- pyrazolo[4,3-b]pyridin-3-amine
18% 3 steps
ND





**1H NMR, DMSO-d6, Ex.: 34bis-1: 12.31 (1H, sl, NH), 8.08-8.18 (1H, m, CHarom), 8.05 (1H, d, CHarom, J = 11.6 Hz), 7.97 (1H, d, CHarom, J = 11.6 Hz), 7.87-7.93 (1H, m, CHarom), 7.64-7.76 (1H, m, CHarom), 5.81 (2H, sl, NH2). 34bis-2: 12.32 (1H, sl, NH), 7.94-8.11 (5H, m, CHarom), 5.85 (2H, sl, NH2). 34bis-3: 12.34 (1H, sl, NH), 8.27 (1H, s, CHarom), 8.12 (1H, d, CHarom, J = 11.6 Hz), 8.01 (1H, d, CHarom, J = 11.6 Hz), 7.82-7.89 (1H, m, CHarom), 7.67 (1H, d, CHarom, J = 11.2 Hz), 5.70 (2H, sl, NH2). 34bis-4: 12.28 (1H, sl, NH), 7.89 (1H, d, CHarom, J = 8.8 Hz), 7.68 (1H, d, CHarom, J = 8.8 Hz), 7.21 (1H, m, CHarom), 6.91-6.97 (2H, m, CHarom), 5.87 (2H, s, NH2), 4.94 (2H, s, CH). 34bis-5: 12.28 (1H, sl, NH), 7.89 (1H, d, CHarom, J = 8.8 Hz), 7.68 (1H, d, CHarom, J = 8.8 Hz), 7.20-7.25 (2H, m, CHarom), 7.10-7.15 (1H, m, CHarom), 5.84 (2H, s, NH2), 4.87 (2H, s, CH). 34bis-6: 12.04 (1H, s, NH), 7.87 (1H, d, CHarom, J = 8.8 Hz), 7.40 (1H, d, CHarom, = 8.8 Hz), 7.10-7.25 (2H, m, CHarom), 6.90-6.97 (1H, m, CHarom), 5.61 (2H, s, NH2), 4.47 (1H, d, CH, J = 13.2 Hz), 4.18 (1H, d, CH, J = 13.2 Hz). 34bis-7: 12.28 (1H, s, NH), 7.89 (1H, d, CHarom, J = 8.8 Hz), 7.64 (1H, d, CHarom, J = 8.8 Hz), 7.40-7.50 (3H, m, CHarom), 5.81 (2H, s, NH2), 4.96 (2H, s, CH).






Example of Method F4
Demethylation
Example 35
N-(5-(3,5-difluorophenylthio)-6-hydroxy-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide




443 μl (3 eq) of a solution of 1 M boron tribromide in dichloromethane is added to a solution of 90 mg (0.148 mmol) of N-(5-(3,5-difluorophenylthio)-6-methoxy-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide (example 18) in 4 ml of 1,2-dichloroethane at 0° C. The reaction medium is stirred at 60° C. for 3 hours and then cooled in an ice bath before adding methanol. The solvents are evaporated and the residue is redissolved in a mixture of methanol and ethyl acetate. The solid formed is filtered, redissolved in 3 ml of tetrahydrofuran and is added to 1 N soda solution. The reaction medium is stirred for 18 hours at room temperature. The pH of the solution is adjusted to 8-9 and the aqueous phase is extracted with ethyl acetate. The organic phase is dried on magnesium sulfate and the crude product is purified on a silica gel column (dichloromethane/methanol as eluent) to yield 21 mg (24%) of N-(5-(3,5-difluorophenylthio)-6-hydroxy-1H-pyrazolo[3,4-b]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(tetrahydro-2H-pyran-4-ylamino)benzamide in the form of a yellow powder.
LCMS (EI, m/z): (M+1) 596.13.
1H NMR: δH ppm (400 MHz, DMSO): 12.96 (1H, broad flat singlet), 12.02 (1H, broad flat singlet), 10.64 (1H, bs, NH), 8.46 (1H, bs), 8.09 (1H, bs), 7.72 (1H, d, CHarom), 6.97-7.10 (1H, m, CHarom), 6.60-6.74 (2H, m, CHarom), 6.28 (1H, dd, CHarom), 6.13 (1H, d, CHarom), 3.80-3.90 (2H, m, CHpyranone), 3.65-3.77 (1H, m, CHpyranone), 3.50 (2H, t, CHpyranone), 3.25-3.32 (4H, m, 2*CH2), 2.37-2.45 (4H, m, 2*CH2), 2.22 (3H, s, CH3), 1.91-2.00 (2H, m, CHpyranone), 1.28-1.43 (2H, m, CHpyranone).
II. Biological Tests of the Compounds According to the Invention
Test for Measuring Inhibition of ALK Kinase:
A ViewPlate microplate (Packard) is incubated with 0.1 mg/ml GST-PLCγ1 substrate (purified recombinant form) in phosphate buffer (PBS, pH 7.4) (100 μl/well) for one hour under stirring. The plate is then saturated with blocking solution comprising 5% bovine serum albumin (BSA) (Sigma) in PBS buffer, pH 7.4.
After having added a compound according to the invention to the desired final concentration (typical range between 30 μM and 10 nM), the reaction is carried out by adding 180 ng/ml ALK to a reaction buffer comprised of 13 mM Tris, pH 7.5 (Sigma); 6.5 mM MgCl2 (Merck); 0.65 mM dithiothreitol (DTT) (Acros); 39 mM sodium β-glycerophosphate (TCI); 0.65 mM sodium orthovanadate (Sigma); and 250 μM ATP (Sigma). Incubation is carried out for 30 minutes at 30° C. under stirring.
After three washings under stirring in 0.1% PBS/Tween-20 buffer (Sigma), an anti-phosphotyrosine antibody, coupled with HRP (UBI) diluted to 1/1000 in 5 mg/ml PBS/BSA buffer, is incubated for one hour with stirring. After three new washings in 0.1% PBS/Tween-20, the wells are incubated for two minutes with 100 μl of SuperSignal ELISA mixture (Pierce).
The signal is read in luminescence mode using a luminometer (SpectraMax M5e, Molecular Devices).
IC50s are determined by nonlinear regression on the basis of a sigmoidal dose/response relationship model, wherein the Hill coefficient is left variable, carried out on the GraphPad software package according to the algorithm provided.
Test for Measuring Inhibition of Cell (Karpas 299) Proliferation:
The antiproliferative activities of the compounds according to the invention were measured by the ATPlite technique (Perkin Elmer).
Nonadherent human anaplastic large-cell lymphoma cells (Karpas 299) are inoculated in 96-well plates (300,000 cells/ml) at day 1, at a concentration compatible with logarithmic growth for the 72 hours required for the evaluation of the compounds. All of the cells are treated at day 1 and then placed in an incubator at 37° C. under an atmosphere of 5% CO2. Cell viability is evaluated at day 4 by assaying released ATP, which is characteristic of viable cells. IC50s are determined by nonlinear regression on the basis of a sigmoidal dose/response relationship model, wherein the Hill coefficient is left variable, carried out on the GraphPad software package according to the algorithm provided.
The results of these two tests obtained with the compounds of the invention are indicated below:













Enzymatic inhibition of
Karpas 299 cell proliferation


Molecule
ALK (IC50, μM)
inhibition (IC50, μM)

















30-1
0.020
2.2


30
0.001
0.16


30-9
0.001
ND


28
0.036
ND


31
0.017
0.83


30-5
0.066
2.6





(ND: not determined)






Pharmacological Activity In Vivo
The molecules described and tested exhibit marked antitumor activity in vivo which is expressed, in an unexpected manner, by a particularly wide therapeutic index, thus suggesting that these compounds are particularly well tolerated. This was demonstrated by evaluating the effects in vivo of the compounds on a human anaplastic large-cell lymphoma (ALCL) tumor model. The compounds were administered orally at various doses on a daily schedule to mice with ALCL tumors grafted subcutaneously. Tumor size was measured regularly during the study and the animals were weighed several times per week in order to identify any adverse effects. A compound is declared active if it induces inhibition of ALCL tumor growth by at least 58%. Several compounds of the present invention, in particular molecules 30 and 30-9, induced 100% inhibition of tumor growth, with no adverse effects, which corresponds to complete regression of the tumors.
Furthermore, the molecules described and tested have general pharmacological properties which seem quite favorable. Notably, they accumulate within the experimental tumors in a long-lasting manner, after their administration in vivo. To that end, ALCL tumors were grafted subcutaneously in the mice and then when the tumors reached a size of approximately 70-130 mm3 the compounds were administered orally in a therapeutically-active dose. The tumors were removed at various times after the administration of the compounds and then ground. The presence of the compounds within the ALCL tumors sampled was then investigated by assays using chromatography with UV and mass.
Test for Measuring Inhibition of a Panel of Kinases:
These kinases are produced by Millipore and are screened according to the manufacturer's protocols.
The results are presented in the table below:















Enzyme inhibition (IC50, nM)











Kinases
30-9
30















ALK
15
7



Abl
38
18



c-Src
9
4

## Claims

1. A compound of formula (I):



or a pharmaceutically acceptable salt of same, a tautomer of same, or a stereoisomer or mixture of stereoisomers of same in any proportions, wherein:
Y1 represents a CH group
Y4 represents a nitrogen atom,
Y2 represents a C—X—Ar group and Y3 represents a C—W group,
Ar represents an aryl or heteroaryl group optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, NR24SO2R25, and R26NR27R28 and/or optionally fused to a heterocycle,
X represents a divalent group selected from O, S, S(O), S(O)2, NR4, S(NR4), S(O)(NR4), S(O)2(NR4), NR4S, NR4S(O), NR4S(O)2, CH2, CH2S, CH2S(O), CH2S(O)2, SCH2, S(O)CH2, S(O)2CH2, CH2CH2, CH═CH, C≡C, CH2O, OCH2, NR4CH2, and CH2NR4,
W represents an R5, SR5, OR5 or NR5R6 group,
U represents a CH2 or NH group, where one or more hydrogen atoms which may be replaced by a (C1-C6)alkyl group,
V represents C(O), C(S) or CH2,
n represents 0 or 1,
R1 represents a hydrogen atom, or an OR7 or NR7R8 group,
R2 represents a hydrogen atom, an optionally substituted heterocycle, NO2, OR9 or NR9R10,
R3, R4, R11, R12, R13, R14, R15, R16, R17, R18, R19, R20, R21, R22, R23, R24, R25, R27 and R28 each represent, independently of each other, a hydrogen atom or a (C1-C6)alkyl group,
R5 and R6 each represent, independently of each other, a hydrogen atom or a (C1-C6)alkyl, optionally substituted aryl or optionally substituted benzyl group,
R7, R8, R9 and R10 each represent, independently of each other, a hydrogen atom or an optionally substituted (C1-C6)alkyl or (C3-C12)cycloalkyl group or an optionally substituted heterocycle, and
R26 represents a (C1-C6)alkyl group,
wherein the heterocycle of R2, R7, R8, R9 and R10, the benzyl and aryl groups of R5 and R6, and the (C1-C6)alkyl and (C3-C12)cycloalkyl groups of R7, R8, R9 and R10 are independently optionally substituted with one or more substituents selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23, NR24SO2R25, and R26NR27R28.

2. The compound according to claim 1, wherein Ar represents an aryl group optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R18, SO2NR19R20, COR21, NR22COR23 and NR24SO2R25; or a pyridinyl group.

3. The compound according to claim 2, wherein Ar represents a phenyl group optionally substituted by one or more groups selected from a halogen atom, (C1-C6)alkyl, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)halothioalkoxy, CN, NO2, OR11, SR12, NR13R14, CO2R15, CONR16R17, SO2R15, SO2NR19R20, COR21, NR22COR23 and NR24SO2R25; or a pyridinyl group.

4. The compound according to claim 2, wherein Ar represents a group selected from the following groups:

5. The compound according to claim 1, wherein:
R3 represents H,
U represents CH2 or NH,
V represents C(O) or C(S), and
n is 0 or 1.

6. The compound according to claim 5, wherein V represents C(O).

7. The compound according to claim 5, wherein n is 0.

8. The compound according to claim 1, wherein X represents a divalent group selected from S, S(O), S(O)2, NR4, CH2, CH2S, CH2S(O), CH2S(O)2, CH2O, CH2NR4, NHS(O)2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, OCH2, NR4CH2, CH2CH2, CH═CH, and C≡C, wherein the first atom of these groups is bound to atom C of chain C—X—Ar.

9. The compound according to claim 8, wherein X represents a divalent group selected from S, S(O), S(O)2, NR4, CH2, SCH2, S(O)CH2, S(O)2CH2, S(O)2NH, CH2CH2, C≡C, OCH2, and NR4CH2, wherein the first atom of these groups is bound to atom C of chain C—X—Ar.

10. The compound according to claim 9, wherein X represents a divalent group selected from S, S(O)2, CH2, SCH2, S(O)2CH2, S(O)2NH, CH2CH2, and C≡C, wherein the first atom of these groups is bound to atom C of chain C—X—Ar.

11. The compound according to claim 1, wherein R1 represents a hydrogen atom or an NR7R8 group, with R7 representing a hydrogen atom and R8 representing an optionally substituted (C3-C12)cycloalkyl group or an optionally substituted heterocycle.

12. The compound according to claim 11, wherein R1 represents one of the following groups:

13. The compound according to claim 1, wherein R2 represents NO2, NR9R10 or a heterocycle optionally substituted by (C1-C6)alkyl or NH2.

14. The compound according to claim 13, wherein R2 represents one of the following groups:

15. The compound according to claim 1, wherein W represents an R5, SR5, OR5 or NR5R6 group, with R5 and R6 representing, independently of each other, a hydrogen atom or a (C1-C6)alkyl group.

16. The compound according to claim 1, selected from the following compounds:

17. A pharmaceutical composition comprising at least one compound of formula (I) according to claim 1 and at least one pharmaceutically acceptable excipient.

18. A method for treating an anaplastic large-cell lymphoma comprising the administration to a person in need thereof of an effective amount of a compound according to claim 1.

19. A method for inhibiting anaplastic lymphoma kinase, Abelson kinase and/or c-Src kinase activity comprising the administration to a person in need thereof of an effective amount of a compound according to claim 1.

20. A method for the preparation of a compound of formula (I) according to claim 1, wherein V represents C(O) or C(S), comprising the successive steps:
(a1) coupling between a compound of formula (A):




wherein Y1, Y2, Y3 and Y4 are as defined in claim 1, and R29 represents a hydrogen atom or an N-protecting group,
with a compound of formula (B):




wherein R1, R2, U and n are as defined in claim 1, V represents C(O) or C(S), R30 represents OH or a leaving group selected from the group consisting of Cl, Br, I, OSO2CH3, OSO2CF3 and OSO2C6H4-p-CH3,
to yield a compound of formula (C):




wherein Y1, Y2, Y3, Y4, R1, R2, U and n are as defined in claim 1, R29 is as defined above and V represents C(O) or C(S),
(b1) when R29 group represents an N-protecting group then deprotection of the nitrogen atom carrying the R29 group to yield a compound of formula (I) with V represents C(O) or C(S), and optionally substitution of the nitrogen atom bound to V of the compound of formula (C) obtained in the preceding step with an R3 group other than H,
(c1) optionally reacting the compound of formula (I) with a pharmaceutically acceptable organic or inorganic acid or a pharmaceutically acceptable organic or inorganic base to form a pharmaceutically acceptable salt of same.

21. A method for the preparation of a compound of formula (I) according to claim 1, wherein V represents CH2, comprising the successive steps:
(a2) reductive amination of a compound of formula (A):




wherein Y1, Y2, Y3 and Y4 are as defined in claim 1, and R29 represents a hydrogen atom or an N-protecting group,
and an aldehyde of formula (D):




wherein R1, R2, U and n are as defined in claim 1,
to yield a compound of formula (E):




wherein Y1, Y2, Y3, Y4, R1, R2, U and n are as defined in claim 1 and R29 is as defined above,
(b2) when R29 group represents an N-protecting group then deprotection of the nitrogen atom carrying the R29 group and optionally substitution of the nitrogen atom bound to V with an R3 group other than H of the compound of formula (E) obtained in the preceding step to yield a compound of formula (I) with V represents CH2, and
(c2) optionally reacting the compound of formula (I) with a pharmaceutically acceptable organic or inorganic acid or a pharmaceutically acceptable organic or inorganic base to form a pharmaceutically acceptable salt of same.

22. A method for the preparation of a compound of formula (I) according to claim 1 wherein V represents C(O) or C(S), n represents 1 and U represents NH, comprising the successive steps:
(a3) coupling between a compound of formula (A):




wherein Y1, Y2, Y3 and Y4 are as defined in claim 1, and R29 represents a hydrogen atom or an N-protecting group,
and a compound of formula (F):




wherein R1 and R2 are as defined in claim 1 and Z represents O or S,
to yield a compound of formula (G):




wherein Y1, Y2, Y3, Y4, R1 and R2 are as defined in claim 1, R29 is as defined above and Z is as defined above,
(b3) when R29 group represents an N-protecting group then deprotection of the nitrogen atom carrying the R29 group and optionally substitution of the nitrogen atom bound to V with an R3 group other than H of the compound of formula (G) obtained in the preceding step to yield a compound of formula (I) with V represents C(O) or C(S), n represents 1 and U represents NH, and
(c3) optionally reacting the compound of formula (I) with a pharmaceutically acceptable organic or inorganic acid or a pharmaceutically acceptable organic or inorganic base to form a pharmaceutically acceptable salt of same.