WO2017198785A1 - Oxime derivatives useful as inhibitors of histone demethylase kdm4c - Google Patents

Oxime derivatives useful as inhibitors of histone demethylase kdm4c Download PDF

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WO2017198785A1
WO2017198785A1 PCT/EP2017/062026 EP2017062026W WO2017198785A1 WO 2017198785 A1 WO2017198785 A1 WO 2017198785A1 EP 2017062026 W EP2017062026 W EP 2017062026W WO 2017198785 A1 WO2017198785 A1 WO 2017198785A1
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pyridine
carbonimidoyl
methyl
hydroxy
carboxylic acid
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PCT/EP2017/062026
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French (fr)
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Mario Varasi
Manuela Villa
Paolo Trifiro'
Daniele Fancelli
Ciro Mercurio
Paola Vianello
Luca Sartori
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Ieo - Istituto Europeo Di Oncologia S.R.L.
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Publication of WO2017198785A1 publication Critical patent/WO2017198785A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/79—Acids; Esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/79—Acids; Esters
    • C07D213/803—Processes of preparation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to oxime derivatives, pharmaceutical compositions containing such compounds and to their use in therapy.
  • Histone lysine demethylases represent attractive targets for epigenetic drugs, since their expression and/or activities are often misregulated in cancer (Varier, R. A. et al. Biochim. Biophys. Acta. 201 1 , 1815, 75-89).
  • a lysine can be mono-, di-, and tri-methylated and each modification, even on the same amino acid, can exert different biological effects.
  • Histone lysine demethylases can be grouped into two major families with different enzymatic mechanisms (Anand, R. et al. J. Biol. Chem. 2007, 282, 35425-35429; Metzger, E. et al. Nat. Struct. Mol. Biol. 2007, 14, 252-254).
  • the first identified subfamily KDM1 (or lysine specific demethylases LSDs) includes KDM1A (also known as LSD1 ) and KDM1 B (also known as LSD2).
  • KDM1A and KDM1 B are both flavo-amino oxidases dependent proteins sharing a FAD coenzyme-binding motif, a SWIRM domain and an amine oxidase domain, all of which are integral to the enzymatic activity of KDM1 family members.
  • KDM1A and KDM1 B demethylation activity is limited to mono and dimethyl substrates, such as histone H3K4 me2/me and histone H3K9me2/me).
  • JmjC Jumonji C
  • This large class comprises 20 human enzymes grouped into five subfamilies: KDM2/7, KDM3, KDM4, KDM5, and KDM6 (Pedersen, M.T. et al. Trends Cell Biol. 2010, 20, 662- 671 ).
  • Jmj-type enzymes are involved in human pathological processes, including development, cancer, inflammation and metabolic diseases (Johansson, C. et a/ Epigenomics. 2014, 6, 89-120). Furthermore several KDMs are over expressed in multiple types of cancer cells (Hojfeldt, J. W. et al. Nat. Rev. Drug Discov. 2013, 12, 917-930).
  • KDM2A and KDM2B also known as FBXL1 1 and FBXL10, respectively
  • KDM2A and KDM2B also known as FBXL1 1 and FBXL10, respectively
  • FBXL1 1 and FBXL10 proteins
  • KDM2B has been found to be involved in hematological cancers, such as acute myeloid leukemia and myelodisplastic syndrome (He, J.
  • KDM7 subfamily includes three members, KDM7A (also known as KIAA1718), KDM7B (also PHF8) and KDM7C (also PHF2), all of them catalyze the demethylation of histone H3K9 and H3K27 mono- and di-methylated.
  • KDM7B has also been shown to demethylate the monomethyl of histone H4K20. It has been found that high expression of KDM7B is associated to an adverse prognosis in patients of laryngeal, hypopharyngeal squamous cell carcinoma (Zhu et al.Epigenomics. 2015;7(2):143-153) and non small cell lung cancer (Shen, Y. et al. Biochem. Biophys. Res.
  • KDM3A also JMJD1A
  • KDM3B also JMJD1 B
  • KDM3C also JMJD1 C
  • KDM3A has been published and specifically KDM3A has been reported to regulate tumor growth in hypoxia condition (Krieg, A.J. et al. Mol. Cell Biol. 2010, 30, 344-353). Moreover KDM3A appears to be highly expressed in renal cell carcinoma (Guo, X. et al. Neoplasma 201 1 , 58, 153-157), and to confers metastasis and chemo resistance in epithelial ovarian cancer (Pa, M. et ai. J Mol Hist (2015) 46:51 1-518). In addition KDM3A has been demonstrated to be a key estrogen regulator in breast cancer (Wade, M.A.
  • KDM3B has been found to have a role in leukemiogeneis (Kim, J.Y. et al. Mol. Cell. Biol. 2012, 32, 2917-2933).
  • KDM4 subfamily (also known as JMJD2A-D proteins) is composed by four proteins, KDM4A, KDM4B, KDM4C and KDM4Dall of them are capable of recognizing and demethylating histone H3K9me2/me3 and histone H3K36me2/me3 as well as histone H1 .4K26me3.
  • KDM4s proteins are also found to catalyze demethylation of non histone substrates and specifically trimethyl-lysine peptides of chromatin repressors such as WIZ, CDYL1 , CSB and G9a proteins (Ponnaluri, V.K. et al. Biochem. Biophys. Res. Commun. 2009, 390, 280-284).
  • KDM4E and KDM4F also known as JmjD2E and JmjD2F
  • JmjD2E and JmjD2F are known to exist but are currently considered being pseudogenes due to lack of intronic sequences in their genes.
  • KDM4A An involvement of KDM4A in inducing site- specific copy gain and re-replication of regions amplified in tumors highlights the important role of KDM4A in cancer development (Black, J.C. et al. Cell 2013, 154, 541 - 555).
  • KDM4A amplification and over expression has been reported in ovarian cancer, in squamous cell carcinoma (Ding, X. et al. Sci. Signal. 2013, 6(273): ra28.1 -13, SO-15; Black, J.C. et al. Cell 2013, 154, 541 -555) and in breast cancer (Berry, W.L. et al. Int.J. Oncol. 2012, 41 , 1701 -1706). Furthermore, KDM4A is associated to breast cancer progression (Li, L.L. et al. Breast Cancer Res. 2014, 16(3):R56) and to promotion of cellular transformation (Mallette, F.A. et al. Cell Rep. 2012, 2, 1233-1243).
  • KDM4B is over expressed in gastric cancer (Li, W. et al. Biochem. Biophys. Res. Commun. 201 1 , 416, 372-378), and it promotes hormonally responsive breast carcinogenesis (Shi, L. et al. Proc. Natl. Acad. Sci. U. S. A. 201 1 , 108, 7541 -7546).
  • KDM4B promotes epithelial-mesenchymal transition by cooperating with ⁇ -catenin and enhances gastric cancer metastasis (Zhao, L. et al. Clin. Cancer Res. 2013, 19, 6419-6429).
  • KDM4B a role of KDM4B in development of neuroblastoma has also been proposed (Yang, J. et al. J. Natl. Cancer Inst. 2015, 107(6), djv080).
  • KDM4C locus is amplified in esophageal squamous cell carcinoma, breast cancer, medulloblastoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL). Furthermore, high expression of the enzyme has been reported in prostate cancer.
  • KDM4C gene was found to be translocated in mucosa- associated lymphoid tissue lymphoma (Labbe, R.M et al. Am. J. Transl. Res. 2013 6, 1 - 15, Helin, K. et al. Nature 2013, 502, 480-488; Hojfeldt, J.W. et al. Nat. Rev. Drug Discov. 2013, 12, 917-930).
  • ectopic expression of KDM4C is able to transform immortalized mammary cells (MCF10A) conferring cancer stem cells characteristic, such as the capability to form mammospheres (Liu, G. et al. Oncogene. 2009, 28, 4491 -4500).
  • KDM4C down regulation in colon cancer cells strongly affects their sphere forming capacity and in vivo tumor formation ability, which are both characteristics of cancer stem cells suggesting that the inhibition of the enzyme could represent an effective therapeutic approach for colon cancer (Yamamoto, S. et al. Carcinogenesis 2013, 34, 2380-2388).
  • KDM4C has been found to have a critical role in acute myeloid leukemia. (Cheung, N. et al. Cancer Cell. 2016, 29, 32-48)
  • KDM4A, B and C proteins share more than 50% sequence identity, each contain JmjN, JmjC, two plant homeodomains (PHD) and two6.1 domains.
  • KDM4D is unique within the KDM4 family, since it lacks both the PHD and Vietnamese domains and thus is only half the size of KDM4A-C.
  • KDM4D has a different substrate specificity: it does not demethylate H3K36 due to several differences in its substrate binding cleft, yet has gained the ability to attack H1.4K26me2/me3. Also, KDM4D attacks H3K9me2 with similar efficiency as for H3K9me3 and may, albeit inefficiently, even demethylate H3K9me1 (Berry W.L.
  • KDM4D functions as a coactivator of the androgen receptor (Shin S et al. Biochem Biophys Res Commun. 2007, 359, 742-746) and stimulates p53-dependent gene expression (KiM TD et al. PLoS One. 2012;7:e34618.).
  • KDM4D in addition a role of KDM4D in DNA damage response has also recently been reported (Khoury-Haddad H. et al. Cell Cycle. 2015; 14, 950-958).
  • KDM5 subfamily (also known as Jarid) consists of four members: KDM5A (RBP2/JARID1A), KDM5B (also known as PLU1/JARID1 B), KDM5C (SMCX/JARID1 C), and KDM5D (SMCY/JARID1 D). All members catalyze the demethylation of histone H3K4me3/me2, which is a signature indicative of transcriptional activation, and hence KDM5 proteins are considered transcriptional co repressors. There are several evidences of an involvement of KDM5A in cancer. For instance, the loss of KDM5A has been reported to suppress the tumorigenesis in mice lacking Rb or Men 1 (Lin, W. et al. Proc. Natl.
  • KDM5A has been associated to promote lung tumorigenesis and cancer metastasis (Teng, Y.C. et al. Cancer Res. 2013, 73, 471 1 -4721 ) and the insurgency of a cell population not responsive to chemotherapeutic drugs (Sharma, S.V. et al. Cell 2010, 141 , 69-80).
  • KDM5A Over expression of KDM5A has also been reported in hepatocellular carcinoma and human gastric cancer (Liang, X. et al. PLoS One. 2013 Jul 29;8(7):e69784; Li, H. et al. Mol Cancer 2014, 13, 18). Moreover, an involvement of KDM5A in leukemiogenesis has been recently suggested by the identification of a translocation: NUP98/JARID1A(KDM5A) as recurrent abnormality in pediatric acute megakaryoblastic leukemia (de Rooij, J.D. et al. Leukemia. 2013, 27, 2280-2288.) High expression of KDM5B and its direct role in cancer progression has been reported for breast cancer (Yamane, K.
  • KDM5B expression has also been found in prostate, bladder, lung and gastric cancer (Xiang, Y. et al. Proc. Natl. Acad. Sci. U. S. A. 2007, 104, 19226-9231 ; Hayami, S. et al. Mol. Cancer 2010, 9, 59; Wang, Z. et al. Am. J. Cancer Res. 2015, 5, 87-100).
  • the overexpression of histone demethylase KDM5B has also been associated with progression of glioma cell growth (Dai, B. et al. Biochem.
  • KDM5B plays also an important role in chemioresistance and stem cell like phenotype of neuroblastoma cells (Kuo, Y.T. et al. PLoS One. 2015 May 7;10(5):e0125343).
  • KDM5B negatively regulates leukemogenesis in murine and human MLL-rearranged AML cells (Cancer Cell. 2015 Aug 10;28(2):198-209.).
  • KDM5C has been found to be over expressed in prostate cancer (Stein, J. et al. Am. J. Pathol. 2014, 184, 2430-2437) suggesting a possible role as oncogene.
  • the KDM6 family consists of KDM6A (also known as UTX), UTY and KDM6B (also known as JMJD3).
  • KDM6A and KDM6B are histone demethylases specific for H3K27me3/me2, whilst discordant data about the enzymatic activity has so far been reported for UTY (Walport, L.J. et al. J. Biol. Chem. 2014, 289, 18302-18313).
  • KDM6A ablation resulted in a significant decrease in the proliferation and invasiveness of breast cancer cells in vitro and in a mouse xenograft model (Kim, J.H. et al. Cancer Res. 2014, 74, 1705-1717).
  • a role in initiation and maintenance of T-ALL leukemia (Ntziachristos, P. et al. Nature 2014, 514, 513-517) as well a role in the development of Hodgkin's Lymphoma (Anderton, J.A. et al. Oncogene 201 1 , 30, 2037-2043) has also been demonstrated for KDM6B.
  • Inhibitors of histone demethylases and in particular of the potential oncogene KDM4C thus represent an innovative and novel approach for cancer therapy.
  • oxime derivatives of general formula (I) as described below are potent KDM4C inhibitors. As such they represent attractive therapeutic approach for cancer.
  • Ci-C 6 alkyl substituted by heterocyclyl, aryl or heteroaryl Ci-C 6 alkyl substituted by X-R 3 ; C 3 -C 7 -cycloalkyl; heterocyclyl, aryl or heteroaryl; is CH 2 OH, (CO)R 4 , CN, tetrazole, CH(OH)CF 3 or C(OH) 2 CF 3 is C1-C-6 alkyl; C 3 -C 7 cycloalkyl;
  • O or NH is hydrogen, OH, O-Ci-C 6 alkyl, NR 7 R 8 , or CF 3 ; p is 0 or 1 ;
  • R 5 , R 6 are, independently, hydrogen; Ci-C 6 alkyl; Ci-C 6 alkyl substituted by aryl, said aryl being optionally substituted by one ore more substituents independently selected from the group consisting of halogen, CN, or NH 2 ; Ci-C 6 alkyl substituted by C3-C7- cycloalkyl optionally substituted by halogen; or Ci-C 6 acyl;
  • R 7 , R 8 are, independently, hydrogen, OH, or Ci-C 6 alkyl; wherein the aryl or heteroaryl in R and R 3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxy; halogen; C-i- C 6 alkyl; Ci-C 6 alkyl substituted by NR 6 R 17 ; Ci-C 6 alkoxy; Ci-C 6 haloalkyl; Ci-C 6 haloalkoxy; NR 9 R 10 ; CONR 20 R 21 ; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C 6 alkyl; aryl; Ci-C 6 alkyl substituted by aryl or heteroaryl; Ci-C 6 alkoxy substituted by aryl or heteroaryl; phenyloxy, wherein the phenyl may be optionally substituted by halogen; C 2 -C 6 alkoxy substituted by NR 18 R 19 ; Ci-C
  • R 9 , R 0 are, independently, hydrogen; d-C 6 alkyl; or COR 15 ;
  • R , R 2 , R 3 are, independently, hydrogen or Ci-C 6 alkyl;
  • R 14 is C-i-C-6 alkyl or aryl;
  • R 5 is Ci-C 6 alkyl; or Ci-C 6 alkyl substituted by NH 2 ;
  • R 6 , R 7 , R 8 , R 9 , R 20 , R 2 are, independently, hydrogen, Ci-C 6 alkyl, Ci-C 6 alkyl substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH 2 ; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH 2 ; or stereoisomers or pharmaceutically acceptable salts thereof and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyridine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-is
  • the invention further provides compounds of formula (I)
  • R 2 is CH 2 OH, (CO)R 4 , CN, or tetrazole
  • R 3 is Ci-C 6 alkyl
  • R 4 is hydrogen, OH, 0-Ci-C 6 alkyl, or NR 7 R 8 ; p is O or l ;
  • R 5 , R 6 are, independently, hydrogen; Ci-C 6 alkyl; Ci-C 6 alkyl substituted by aryl; Ci- Ce alkyl substituted by C3-C7-cycloalkyl optionally substituted by halogen; or Ci-C 6 acyl;
  • R 7 , R 8 are, independently, hydrogen, OH, or Ci-C 6 alkyl; wherein the aryl or heteroaryl in R and R 3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, C1-C6 alkyl, Ci-C 6 alkoxy, Ci-C 6 haloalkyl, Ci-C 6 haloalkoxy, NR 9 R 10 , unsubstituted heterocyclyl, heterocyclyl substituted by C1-C6 alkyl, C1 -C6 alkyl substituted by aryl; or C1-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; and the heterocyclyl in R and R 3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of C1-C6 alkyl; C1 -C6 alkyl substituted by aryl; Ci
  • R , R 2 , R 3 are, independently, hydrogen or Ci-C 6 alkyl
  • R 4 is Ci-C 6 alkyl or aryl
  • R 5 is Ci-C 6 alkyl; or Ci-C 6 alkyl substituted by NH 2 ; and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyhdine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-isonicotinic acid.
  • R is Ci-C 6 alkyl; Ci-C 6 alkyl substituted by aryl;
  • aryl wherein the aryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, Ci- Ce alkyl, Ci-C 6 alkoxy; or Ci-C 6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen;
  • R 2 is CH 2 OH, COR 4 , CN, tetrazole, CH(OH)CF 3 or C(OH) 2 CF 3 ;
  • R 3 is Ci-C 6 alkyl;
  • R 4 is hydrogen, OH, 0-Ci-C 6 alkyl, NR 7 R 8 , or CF 3 ; p is O or l ; R 5 , R 6 are, independently, hydrogen; Ci-C 6 alkyl; Ci-C 6 alkyl substituted by aryl, said aryl being optionally substituted by halogen; or C-i-C-6 acyl;
  • R 7 , R 8 are, independently, hydrogen, OH, or Ci-C 6 alkyl; wherein the aryl in R or R 3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxyl; halogen; d-C-6 alkyl; C-i- C 6 alkyl substituted by NR 6 R 17 ; Ci-C 6 alkoxy; Ci-C 6 haloalkyl; Ci-C 6 haloalkoxy; NR 9 R 10 ; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C 6 alkyl; phenyl; Ci-C 6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; Ci- C-6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; phenyloxy, wherein the phenyl may be optionally substituted by halogen;
  • R 5 is Ci-C 6 alkyl substituted by NH 2 ;
  • R 6 , R 7 , R 8 , R 9 are, independently, hydrogen; Ci-C 6 alkyl; Ci-C 6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by phenyl, wherein the phenyl may be optionally substituted by halogen.
  • R is Ci-C 6 alkyl
  • C-i-C-6 alkyl substituted by aryl C-I -C6 alkyl substituted by heterocyclyl, wherein the heterocyclyl may be optionally substituted by C-i-C-6 alkyl;
  • R 2 is CH 2 OH, COR 4 , CN, or tetrazole
  • R 3 is Ci-C 6 alkyl
  • R 4 is hydrogen, OH, 0-Ci-C 6 alkyl, or NR 7 R 8 ; p is O or l ;
  • R 5 , R 6 are, independently, hydrogen; Ci-C 6 alkyl; Ci-C 6 alkyl substituted by aryl; or
  • R 7 , R 8 are, independently, hydrogen, OH, or Ci-C 6 alkyl; wherein the aryl in R 3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, Ci-C 6 alkyl, Ci-C 6 alkoxy, Ci-C 6 haloalkyl, Ci-C 6 haloalkoxy, NR 9 R 10 or heterocyclyl substituted by Ci-C 6 alkyl; and
  • R 9 , R 0 are, independently, hydrogen or COR 15 ;
  • R 5 is Ci-C 6 alkyl substituted by NH 2 .
  • aryl is phenyl or naphthyl.
  • Particularly preferred compounds of general formula (I) include:
  • the above mentioned compounds of the invention are for medical use or for use as medicament.
  • the above compounds are inhibitors of KDM4C.
  • the above compounds are inhibitors of KDM4C for medical use.
  • An inhibitor is a compound able to bind to KDM4C and blocks its activity or function.
  • the invention provides the compounds of general formula (I) for medical use, in particular for use in the treatment and/or prevention of cancer.
  • the compounds of general formula (I) are for the use in the treatment and/or prevention of leukemia, lymphoma, esophageal squamous cell carcinoma, breast cancer, medulloblastoma, prostate cancer, colon cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreas cancer or glioblastomas.
  • the glioblastomas are giant cell glioblastoma or gliosarcoma.
  • the leukemia or lymphoma are acute myeloid leukemia, mucosa-associated lymphoid tissue lymphoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL).
  • a further embodiment of the invention is a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of general formula (I) together with a pharmaceutically acceptable excipient and/or diluent.
  • the pharmaceutical composition may further comprise at least one further therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, anti-proliferative/antineoplastic agents, cytostatic agents, agents which inhibit cancer cell invasion, inhibitors of growth factor function, anti- angiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent.
  • the pharmaceutical composition may be for use in the treatment and/or prevention of cancer.
  • the invention provides a process for obtaining a compound of formula (I) as defined above, wherein R 2 is COOH, the process comprising the preparation of a compound of formula A3 by reaction of a compound of formula A1 with a compound of formula A2 in presence of an oxidant, a transition metal salt, and an acid, the preparation of a compound of formula A4 by reaction of a compound of formula A3 with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH2OH), and the final deprotection of a compound of formula A4 to obtain a compound of formula (I), as represented in Scheme A-IV:
  • aryl represents a mono or bicyclic aromatic ring system of, respectively, 6, 9 or 10 atoms.
  • Examples of such an aryl are phenyl, indenyl, indanyl and naphthyl and tetrahydronaphthalenyl.
  • Substituted aryl means that the hydrogen atoms on independently each carbon atom may be independently replaced by a substituent as defined herein above.
  • Heteroaryl represents a mono or bicyclic heteroaromatic ring system of, respectively, 5 to 10 members, which contains one, two, three or four heteroatoms selected from nitrogen, oxygen or sulphur and one to nine carbon atoms.
  • heteroaryls include, but are not limited to: pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, 1 ,2,3-oxadiazolyl, 1 ,3,4-thiadiazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, isoindolyl, benzo[b]furanyl, benzo[ib]thienyl, benzopyranyl, indazolyl, benzimidazolyl, purinyl, quinolyl, isoquinolyl, quinazolin
  • Heterocyclyl represents a mono, bicyclic or a spirocyclic saturated or partially saturated non-aromatic ring system of, respectively, 4 to 12 members, which contains one, two, or three heteroatoms selected from nitrogen, oxygen, and sulphur and three to eleven carbon atoms.
  • heterocycles include, but are not limited to: pyrrolidyl, pyrrolidinyl, piperidyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, benzodioxolyl, 2,3-dihydro-benzodioxinyl, benzoxazolyl, azetidyl, azepinyl, and diazapinyl.
  • bicyclic ring systems include, but are not limited to, 2-aza-bicyclo[2.2.1]heptanyl, 2,5-diaza-bicyclo[2.2.1]hept-2-yl or 8- azabicyclo[3.2.1]octanyl.
  • spirocyclic ring systems include, but are not limited to, 3,8-diazaspiro[4.5]decane.
  • C 1 -C6 alkyl refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, having from one to six carbon atoms.
  • the "C-i-C-6 alkyl” group is preferably a linear or branched Ci-C alkyl group, more preferably a Ci -C 2 alkyl group. Examples of Ci-C 6 alkyl include methyl, ethyl, n-propyl, isopropyl, butyl, te/t-butyl, pentyl, and hexyl.
  • C3-C7-cycloalkyl refers to a saturated monocyclic hydrocarbon ring system having three to seven carbon atoms.
  • Examples of C-3-C7-cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
  • d-C-6 alkoxy refers to a straight or branched O-C-i-C-6 alkyl, where alkyl is as defined herein.
  • the "C-i-C-6 alkoxy” group is preferably a linear or branched Ci-C 4 alkoxy group, more preferably a Ci-C 2 alkoxy group.
  • C-i-C-6 haloalkyi refers to a straight or branched hydrocarbon chain radical, which is substituted by one or more halogen atoms and having from one to six carbon atoms.
  • the "C-i-C-6 haloalkyi” group is preferably a linear or branched Ci-C 4 haloalkyi group, more preferably a Ci-C 2 haloalkyi group, being in particular CF 3 .
  • C1 -C6 haloalkoxy refers to a straight or branched O-C1-C6 haloalkyi, where haloalkyi is as defined herein.
  • the "C-i-C-6 haloalkoxy” group is preferably a linear or branched Ci-C haloalkoxy group, more preferably a C1 -C2 haloalkoxy group, being in particular OCF 3 , OCHF 2 or OCH 2 F.
  • Ci-C6 acyl refers to a straight or branched -(CO)-Ci -C6 alkyl, with C-i-C-6 alkyl as defined herein.
  • halogen refers to fluoro, chloro, bromo, or iodo.
  • Halogens are preferably fluorine, chlorine or bromine, being in particular fluorine or chlorine.
  • phenyloxy refers to a O-Phenyl.
  • aryloxy refers to a O-Aryl where aryl is as defined herein.
  • heteroaryloxy refers to a O-Heteroaryl where Heteroaryl is as defined herein.
  • Pharmaceutically acceptable salts comprise conventional non-toxic salts obtained by salification of a compound of formula (I) with inorganic acids (e.g. hydrochloric, hydrobromic, sulphuric, or phosphoric acids), or with organic acids (e.g. acetic, propionic, succinic, benzoic, sulfanilic, 2-acetoxy-benzoic, cinnamic, mandelic, salicylic, glycolic, lactic, oxalic, malic, maleic, malonic, fumaric, tartaric, citric, p-toluenesulfonic, methanesulfonic, ethanesulfonic, or naphthalensulfonic acids).
  • inorganic acids e.g. hydrochloric, hydrobromic, sulphuric, or phosphoric acids
  • organic acids e.g. acetic, propionic, succinic, benzoic, sulfanilic, 2-acetoxy-
  • solvated forms of the compounds of formula (I) with acceptable solvents such as water, EtOH and the like, including hydrates of the compounds, as well as mixtures of the hydrate- and keto- form of the compounds, are within the scope of the present invention.
  • Certain compounds of formula (I) may exist in stereoisomeric forms (e.g. they may contain one or more asymmetric carbon atoms).
  • the individual stereoisomers (enantiomers and diastereomers) and mixtures of these are included within the scope of the present invention.
  • the present invention also covers the individual isomers of the compounds represented by formula (I) as mixtures with isomers thereof in which one or more chiral centres are inverted.
  • compounds of the invention may exist in tautomeric forms other than that shown in the formula and these are also included within the scope of the present invention.
  • the invention also includes all suitable isotopic variations of a compound of the invention.
  • isotopes that can be incorporated into compounds of the invention include isotopes such as 2 H, 3 H, 13 C, 14 C, 15 N, 17 O, 18 0, 31 P, 32 P, 35 S, 18 F and 36 CI, respectively.
  • Certain isotopic variations of the invention for example, those in which a radioactive isotope such as 3 H or 14 C is incorporated, are useful in drug and/or substrate tissue distribution studies. Further, substitution with isotopes such as deuterium 2 H, may afford certain therapeutic advantages resulting from greater metabolic stability.
  • Isotopic variations of the compounds of the invention can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the examples hereafter using appropriate isotopic variations of suitable reagents.
  • the invention also provides pharmaceutical compositions comprising one or more compounds of this invention and one or more pharmaceutically acceptable excipient and/or diluent.
  • the pharmaceutical compositions containing the active ingredient may be in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes, transdermal delivery devices.
  • R is as defined above for fomula (I);
  • PG is a protecting group chosen among those known in the art, for example methyl, ethyl etc..
  • Compounds of formula A1 and A2 are known compounds or can be prepared by known methods.
  • Compounds of formula A3 can be obtained by reacting a compound of formula A1 with a compound of formula A2 in presence of an oxidant, for instance 2-hydroperoxy- 2-methyl-propane, K 2 S 2 0 8 , or (NH 4 ) 2 S 2 08, a transition metal salt, for instance FeS0 or AgN0 3 , and an acid, for instance trifluoroacetic acid or H 2 S0 4 , in a suitable solvent, for instance acetonitrile, water or dichloroethane, at a temperature ranging from room temperature to the boiling point of the solvent.
  • an oxidant for instance 2-hydroperoxy- 2-methyl-propane
  • K 2 S 2 0 8 a compound of formula A2
  • a transition metal salt for instance FeS0 or AgN0 3
  • an acid for instance trifluoroacetic acid or H 2 S0 4
  • reaction can be carried out, in presence or absence of the transition metal salt, with tetrabutylammonium bromide (TBAB) instead of the acid.
  • TBAB tetrabutylammonium bromide
  • the reaction can be carried out under microwave irradiation.
  • a compound of formula A3, wherein R is methyl can be converted into a compound of A3, wherein R is benzyl, by reacting a compound of A3 with iodobenzene or bromobenzene in presence of a suitable catalyst, for instance palladium acetate, a suitable ligand, for instance 2-(2-dicyclohexylphosphanylphenyl)-A/,A/-dimethyl-aniline, and a suitable base, for instance potassium te/t-butoxide, in a suitable solvent, for instance THF, at a temperature ranging from room temperature to the boiling temperature of the solvent.
  • a suitable catalyst for instance palladium acetate
  • reaction is carried out under inert atmosphere.
  • a compound of formula A3, wherein R is methyl can be converted into a compound of A3, wherein R is 2-phenylethyl, by reacting a compound of A3 with benzaldehyde in presence of a base in a suitable solvent, for instance methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, providing a compound of A3, wherein R is phenylprop-2-enoyl.
  • a suitable solvent for instance methanol
  • the double bond can be reduced providing a compound of A3, wherein R is 2-phenylethyl, with 3,5-bis(ethoxycarbonyl)- 1 ,4-dihydro-2,6-dimethylpyridine in presence of silica gel in a suitable solvent, for instance toluene, at a temperature ranging from room temperature to the boiling temperature of the solvent.
  • a suitable solvent for instance toluene
  • compounds of formula A3, wherein R is methyl substituted by aryl or heteroaryl can be obtained by reacting a compound of formula A11 with a compound of formula A2, wherein R is aryl or heteroaryl, in presence of a suitable base, such as KOH or CS2CO3, in a suitable solvent, for instance methanol or isopropanol and THF, at a temperature ranging from -40°C to room temperature, preferably under inert atmosphere, providing a compound of formula A12, wherein R is aryl or heteroaryl.
  • a suitable base such as KOH or CS2CO3
  • a suitable solvent for instance methanol or isopropanol and THF
  • the compound of formula A12, wherein R is aryl or heteroaryl can be hydrolysed in presence of a suitable acid, such as HCI, providing a compound of formula A3 wherein R is methyl substituted by aryl or heteroaryl.
  • a suitable acid such as HCI
  • Compounds of formula A11 are known compounds or can be prepared by known methods (TeLe 39, 1998, 1717-1720, WO2005056550)
  • Compounds of formula A4 can be obtained by reacting a compound of formula A3 with hydroxylamine hydrochloride and a suitable base, for instance sodium acetate, or hydroxylamine (NH 2 OH) in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent.
  • a suitable base for instance sodium acetate, or hydroxylamine (NH 2 OH)
  • a suitable solvent for instance methanol, ethanol, a water/ethanol mixture
  • a compound of formula (I) can be obtained according to known methods, e.g. when the compound of formula A4 is a methyl, ethyl or te/t-butyl ester, by treatment of the ester with LiOH, NaOH or KOH in a suitable solvent, for example in ethanol/water, in THF, in THF/water, in ethanol/water, in methanol/water, or in a dioxane/ethanol/water mixture.
  • the hydrolysis may be carried out at a temperature ranging from 0°C to the boiling point of the solvent.
  • a compound of formula (I) can be obtained by treatment of a te/t-butyl ester derivative with TFA (trifluoroacetic acid) in a suitable solvent such as dichloromethane at a temperature ranging from 0°C to room temperature.
  • TFA trifluoroacetic acid
  • aryl is substituted by hydroxy
  • R , R 6 and R 7 are as defined above for fomula (I);
  • PG and PG are protecting groups chosen among those known in the art, for example methyl, ethyl, te/t-butyl, etc. for PG and 0-(tetrahydro-2H-pyran-2-yl), etc. for PG 1 .
  • a compound of formula A5, wherein R is methyl substituted by X-R 3 , with X and R 3 as defined above can be prepared first by treating a compound of formula A3, wherein R is methyl, with bromine or a bromine/HBr mixture in a suitable solvent, for instance acetic acid or methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, then with a protected hydroxylamine H2N-OPG 1 under the same conditions as with hydroxylamine as disclosed above in Scheme A-l and finally with HO-R 3 in presence of a base, for instance sodium hydride (NaH) or K 2 CO 3 , or KO- R 3 or H2NR 3 in a suitable solvent, for instance methanol, ethanol, a methanol/ethanol mixture, THF or DMF, at a temperature ranging from 0°C to room temperature.
  • a base for instance sodium hydride (NaH) or K 2 CO 3
  • a compound of formula A5, wherein R is methyl can be converted into a compound of A5, wherein R is benzyl, by reacting a compound of A5, wherein R is methyl, first with bromine or a bromine/HBr mixture in a suitable solvent, for instance acetic acid or methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, and then with phenylboronic acid in presence of a suitable catalyst, for instance palladium chloride, a suitable ligand, for instance cyclopentyl(diphenyl)phosphane, and a suitable Lewis acid, for instance cesium fluoride, in a suitable solvent, for instance THF, at a temperature ranging from room temperature to the boiling temperature of the solvent.
  • a suitable solvent for instance acetic acid or methanol
  • the reaction is carried out under microwave irradiation.
  • the reaction is carried out under inert atmosphere.
  • a compound of formula A5, wherein R is methyl can be converted at the same conditions into a compound of A5, wherein R is benzyl, wherein the benzyl is substituted by one or more substituents independently selected from the group consisting of halogen, C 1 -C6 alkyl, Ci-C 6 alkoxy, Ci-C 6 haloalkyl, and Ci-C 6 haloalkoxy.
  • a suitable base for instance K 2 CO 3 , Cs 2 CO 3 or NaH
  • a suitable solvent for instance DMF, aceton
  • a compound of formula A13 is a known compound or can be prepared by known methods.
  • a compound of formula A6 can be obtained according to known methods, e.g. when the PG group in A5 is methyl, ethyl or te/t-butyl, by treatment of the ester with LiOH, NaOH or KOH in a suitable solvent, for example in ethanol/water, in THF/water, in THF/ethanol/water, in methanol/water, or in a dioxane/ethanol/water mixture.
  • the hydrolysis may be carried out at a temperature ranging from 0°C to the boiling point of the solvent.
  • a compound of formula A6 can be obtained also by treatment of a te/t-butyl ester derivative with TFA (trifluoroacetic acid) in a suitable solvent such as dichloromethane at a temperature ranging from 0°C to room temperature.
  • TFA trifluoroacetic acid
  • a compound of formula A7 can be obtained according to known methods, e.g. in the case PG 1 is tetrahydropyranyl, the deprotection of the hydroxylamine, can be achieved by using HCI in aprotic solvents (such as THF, diethylether, dioxane or a methanol/dioxane mixture) at a temperature ranging from 0°C to room temperature.
  • aprotic solvents such as THF, diethylether, dioxane or a methanol/dioxane mixture
  • a compound of formula (I) can be obtained either by deprotection of the hydroxylamine of a compound of formula A6 under the same conditions as for obtaining a compound of formula A7, or by deprotection of the carboxylic acid of a compound of formula A7 under the same conditions as for obtaining a compound of formula A6.
  • R is as defined above for fomula (I); and R 2 is COR 4 , CN, CH(OH)CF 3 or C(OH) 2 CF 3 and wherein R 4 is O-Ci-C 6 alkyl, NR 7 R 8 , or CF 3 and R 7 and R 8 are as defined above.
  • Compounds of formula A8 are known compounds or can be prepared by known methods.
  • Compounds of formula A9 can be obtained by reacting a compound of formula A8 with a compound of formula A2 under the same conditions as for the reaction of a compound of formula A1 with a compound of formula A2 as outlined above in Scheme A-l.
  • Compounds of formula (I), wherein R 2 is COR 4 or CN and wherein R 4 is O-C 1 -C6 alkyl or NR 7 R 8 , can be obtained by reacting a compound of formula A9 with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH 2 OH), for instance sodium acetate, in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent.
  • a suitable solvent for instance methanol, ethanol, a water/ethanol mixture
  • compounds of formula (I), wherein R 2 is COR 4 or CN and wherein R 4 is O- C1-C6 alkyl or NR 7 R 8 can be obtained first by reacting a compound of formula A9 with a protected hydroxylamine H 2 N-OPG 1 under the same conditions as with hydroxylamine as disclosed above in Scheme A-l providing a compound of formula A10, and then by deprotection of the hydroxylamine according to known methods, e.g. in the case PG is tetrahydropyranyl, by using HCI in aprotic solvents (such as THF, diethylether or dioxane).
  • aprotic solvents such as THF, diethylether or dioxane
  • Compounds of formula (I), wherein R 2 is CH2OH can be obtained first by treating a compound of formula A10, wherein R 2 is -COOCH 3 , with a reducing agent, for instance LiAIH 4 or c// ' isobutylaluminum hydride, in an suitable solvent, for instance dichloromethane or THF, at a temperature ranging from -78°C to 0°C, and then by deprotection of the hydroxylamine according to known methods as described above.
  • a reducing agent for instance LiAIH 4 or c// ' isobutylaluminum hydride
  • an suitable solvent for instance dichloromethane or THF
  • the reduction step is carried out under inert atmosphere.
  • Compounds of formula (I), wherein R 2 is COR 4 and R 4 is hydrogen can be obtained first by reacting a compound of formula A10, wherein R 2 is -COOCH 3 , with a reducing agent, for instance LiAIH 4 or of/lsobutylaluminum hydride, in an suitable solvent, for instance dichloromethane, at -78°C, and then by deprotection of the hydroxylamine according to known methods.
  • a reducing agent for instance LiAIH 4 or of/lsobutylaluminum hydride
  • suitable solvent for instance dichloromethane
  • the obtained carboxylic acid can be treated with an amine HNR 7 R 8 with coupling agents, for instance benzotriazol-1 -yl- oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), benzotriazol-1 -yloxy- tris(dimethylamino)phosphonium hexafluorophosphate (BOP) or 1 -(3- dimethylaminopropyl)-3-ethylcarbodiimide (EDC), in the presence of a suitable base (e.g. triethylamine or di-isopropylethylamine) in a suitable solvent (e.g.
  • a suitable base e.g. triethylamine or di-isopropylethylamine
  • a suitable solvent e.g.
  • an activator of the condensation reaction such as HOBt (1 -hydroxybenzotriazole) or HOAt (1 -hydroxy-7-aza-benzotriazole)
  • HOBt 1-hydroxybenzotriazole
  • HOAt 1-hydroxy-7-aza-benzotriazole
  • the reaction can be carried out at room temperature for a period lasting between about 2 and 24 h.
  • the deprotection of the hydroxylamine can be carried out according to known methods as described above.
  • Compounds of formula (I), wherein R 2 is tetrazole can be obtained first by reacting a compound of formula A10, wherein R 2 is CN, with sodium azide (NaN 3 ) in presence of triethylamine hydrochloride in a suitable solvent, for instance dimethylacetamide, dimethylformamide or A/-methylpyrrolidone, at a temperature ranging from room temperature to the boiling temperature of the solvent, and then by deprotection of the hydroxylamine according to known methods.
  • a suitable solvent for instance dimethylacetamide, dimethylformamide or A/-methylpyrrolidone
  • the coupling with NaN 3 can be carried out under microwave irradiation.
  • a compound of general formula (I), wherein R 2 is CH(OH)CF 3 and R is methyl can be obtained by reacting a compound of formula A9, wherein R 2 is CH(OH)CF 3 and R is methyl, with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH 2 OH), for instance sodium acetate, in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent.
  • a suitable solvent for instance methanol, ethanol, a water/ethanol mixture
  • a compound of formula A14 is a known compound.
  • Compounds of general formula (I), wherein R 2 is (CO)R 4 and R 4 is CF 3 can be obtained first by reacting a compound of formula A10, wherein R 2 is CH(OH)CF 3 with an oxidizing agent, such as IBX in a suitable solvent, for instance EtOAc, at a temperature ranging from room temperature to the boiling temperature of the solvent and then by deprotection of the hydroxylamine according to known methods, e.g. in the case PG 1 is tetrahydropyranyl, by using HCI in aprotic solvents (such as THF, diethylether or dioxane).
  • aprotic solvents such as THF, diethylether or dioxane
  • the compounds of the present invention are useful in the prevention or treatment of tumor type diseases, including but not limited to: leukemia, lymphoma, esophageal squamous cell carcinoma, breast cancer, medulloblastoma, prostate cancer, colon cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreas cancer or glioblastomas.
  • the glioblastomas are giant cell glioblastoma or gliosarcoma.
  • the leukemia or lymphoma are acute myeloid leukemia, mucosa-associated lymphoid tissue lymphoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL).
  • the compounds of formula (I) can also be used in combination with additional agents, in particular anti-tumor and differentiating agents, either by separate administrations, or by including the two active principles in the same pharmaceutical formulation.
  • additional agents include:
  • histone deacetylase inhibitors for example, but not limited to SAHA, PXD101 , JNJ- 26481585, SB939, ITF-2357, LBH589, PCI-24781 , valproic acid, butyric acid, MS-275, MGCD0103 and FK-228);
  • retinoid receptor modulators such as 13-c/s-retinoic acid, 9-c/s-retinoic acid, bexarotene, alitretinoin, or tretinoin; vitamin D;
  • antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology such as alkylating agents (for example platin derivatives like c/s-platin, carboplatin, oxaliplatin, lobaplatin, satraplatin, nedaplatin, heptaplatin; nitrogen mustard such as chlorambucil, melphalan, chlormethine, cyclophosphamide, ifosfamide, trofosfamide, uramustine, bendamustine, estramustine; busulphan, temozolomide or nitrosoureas); antimetabolites (for example antifolates such as aminopterin, methotrexate, pemetrexed, raltitrexed); purines such as cladribine, clofarabine, fludarabine, mercaptopurine, pentostatin, thioguanine; pyrimidines like capecitabine, cylating agents
  • cytostatic agents such as antioestrogens (for example tamoxifen, toremifene, raloxifene, droloxifene and idoxifene), oestrogen receptor down regulators (for example fulvestrant), antiandrogens (for example bicalutamide, flutamide, nilutamide, liarozole or cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin or buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5-alpha-reductase such as finasteride;
  • antioestrogens for example tamoxifen, toremifene, raloxifene, droloxifene and idoxifen
  • agents which inhibit cancer cell invasion for example metalloproteinase inhibitors and inhibitors of urokinase plasminogen activator receptor function;
  • inhibitors of growth factor function for example growth factor antibodies, growth factor receptor antibodies (for example the anti-erbb2 antibody trastuzumab as well as ado-trastuzumab emtasine and pertuzumab, the anti-erbbl antibody cetuximab and panitumumab, the anti IGF1 R antibody figitumumab), farnesyl transferase inhibitors, MEK inhibitor(for example trametinib), tyrosine kinase inhibitors and serine/threonine kinase inhibitors, for example enzastaurin, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, nilotinib, sorafenib, sunitinib, afatinib, axinitinib, bosutinib, caboratinib, ceritinib, crizotin
  • antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, for example the anti-vascular endothelial cell growth factor antibody bevacizumab, ramucirumab, lenalidomide or thalidomide;
  • cell cycle inhibitors including for example CDK inhibitors (for example but not limited to flavopiridol, roscovitine, palbociclib and milciclib) and other inhibitors of cell cycle checkpoints; inhibitors of aurora kinase and other kinases involved in mitosis and cytokinesis regulation;
  • proteasome inhibitors for example lactacystin, bortezomib, epoxomicin and its analog carfilzomib
  • HSP90 inhibitors for example but not limited to AT-13387, KOS-953, KOS-1022, CNF-1010, CNF-2024, SNX 5422, STA-9090, NVP-HSP990, NVP-AUY922, PU-H17 and XL-888;
  • Selective COX-2 inhibitors for example celecoxib or parecoxib
  • non selective NSAIDs for example diclofenac, flurbiprofen, ibuprofen, ketoprofen, or naproxen
  • a compound of general formula (I) can be used in combination with radiation therapy.
  • a compound of general formula (I) may be administered in combination with standard chemotherapy combinations such as, but not restricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide, and paclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone);
  • CMF cyclophosphamide, methotrexate and 5-fluorouracil
  • CAF cyclophosphamide, doxorubicin and 5-fluorouracil
  • AC doxor
  • anti CD20 antibodies for example but not limited to ibritumomab tiuxetan,obinutuzumab,ofatumumab,rituximab and tositumomab
  • anti PD1 antibody for example nivolumab
  • anti CD30 antibodies such as brentuximab vedotin
  • anti CLTA-4 antibodies for example ipilimumab
  • antibodies targeting soluble and membrane- bound interleukin 6 as siltuximab for example but not limited to ibritumomab tiuxetan,obinutuzumab,ofatumumab,rituximab and tositumomab
  • anti CD30 antibodies such as brentuximab vedotin
  • anti CLTA-4 antibodies for example ipilimumab
  • m) epigenetic drugs such as other histone demethylase inhibitors (for example but not limited to KDM1A and EZH2 inhibitors), and bromodomain inhibitors (for example but not limited to GSK525762, OTX015, CPI-0610, TEN-010 and BAY1238097);
  • IDH-1/2 inhibitors such as AG-120, AG-221 and AG-881.
  • a compound of general formula (I) can be used in combination with radiation therapy.
  • a compound of general formula (I) may be administered in combination with standard chemotherapy combinations such as, but not restricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide, and paclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone).
  • CMF cyclophosphamide, methotrexate and 5-fluorouracil
  • CAF cyclophosphamide, doxorubicin and 5-fluorouracil
  • AC doxor
  • the invention also provides pharmaceutical compositions comprising one or more compounds of this invention and one or more pharmaceutically acceptable excipient and/or diluent.
  • the pharmaceutical compositions can be chosen on the basis of the treatment requirements.
  • Such compositions are prepared by blending and are suitably adapted to oral or parenteral administration, and as such can be administered in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable, or infusible liquid solutions, suspensions, or suppositories.
  • Tablets and capsules for oral administration are normally presented in unit dose form and contain conventional excipients such as binders, fillers (including cellulose, mannitol, lactose), diluents, tableting agents, lubricants (including magnesium stearate), detergents, disintegrants (e.g. polyvinylpyrrolidone and starch derivatives such as sodium glycolate starch), coloring agents, flavoring agents, and wetting agents (for example sodium lauryl sulfate).
  • excipients such as binders, fillers (including cellulose, mannitol, lactose), diluents, tableting agents, lubricants (including magnesium stearate), detergents, disintegrants (e.g. polyvinylpyrrolidone and starch derivatives such as sodium glycolate starch), coloring agents, flavoring agents, and wetting agents (for example sodium lauryl sulfate).
  • the oral solid compositions can be prepared by conventional methods of blending, filling or tableting.
  • the blending operation can be repeated to distribute the active principle throughout compositions containing large quantities of fillers. Such operations are conventional.
  • Oral liquid preparations can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or can be presented as a dry product for reconstitution with water or with a suitable vehicle before use.
  • Such liquid preparations can contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminium stearate gel, or hydrogenated edible fats; emulsifying agents, such as lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which can include edible oils), such as almond oil, fractionated coconut oil, oily esters such as esters of glycerine, propylene glycol, or ethyl alcohol; preservatives, such as methyl or propyl p- hydroxybenzoate or sorbic acid, and if desired, conventional flavoring or coloring agents.
  • Oral formulations also include conventional slow-release formulations such as enterically coated tablets or granules.
  • compositions for administration by inhalation can be delivered from an insufflator or a nebulizer pressurized pack.
  • parenteral administration fluid unit dosages can be prepared, containing the compound and a sterile vehicle.
  • the compound can be either suspended or dissolved, depending on the vehicle and concentration.
  • the parenteral solutions are normally prepared by dissolving the compound in a vehicle, sterilising by filtration, filling suitable vials and sealing.
  • adjuvants such as local anaesthetics, preservatives and buffering agents can also be dissolved in the vehicle.
  • the composition can be frozen after having filled the vials and removed the water under vacuum.
  • Parenteral suspensions are prepared in substantially the same manner, except that the compound can be suspended in the vehicle instead of being dissolved, and sterilized by exposure to ethylene oxide before suspending in the sterile vehicle.
  • a surfactant or wetting agent can be included in the composition to facilitate uniform distribution of the compound of the invention.
  • compositions may be tablets, lozenges, pastilles, or gel.
  • the compounds can be pharmaceutically formulated as suppositories or retention enemas, e.g. containing conventional suppositories bases such as cocoa butter, polyethylene glycol, or other glycerides, for a rectal administration.
  • suppositories or retention enemas e.g. containing conventional suppositories bases such as cocoa butter, polyethylene glycol, or other glycerides, for a rectal administration.
  • Topical formulations can contain for example ointments, creams, lotions, gels, solutions, pastes and/or can contain liposomes, micelles and/or microspheres.
  • ointments include oleaginous ointments such as vegetable oils, animal fats, semisolid hydrocarbons, emulsifiable ointments such as hydroxystearin sulfate, anhydrous lanolin, hydrophilic petrolatum, cetyl alcohol, glycerol monostearate, stearic acid, water soluble ointments containing polyethylene glycols of various molecular weights.
  • Creams are viscous liquids or semisolid emulsions, and contain an oil phase, an emulsifier and an aqueous phase.
  • the oil phase generally contains petrolatum and an alcohol such as cetyl or stearic alcohol.
  • Formulations suitable for topical administration to the eye also include eye drops, wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient.
  • transdermal delivery comprises conventional aqueous and nonaqueous vectors, such as creams, oils, lotions or pastes or can be in the form of membranes or medicated patches.
  • the compounds of the present invention may be employed alone as a sole therapy or in combination with other therapeutic agents (see the list of additional agents is as indicated previously and comprises also standard chemotherapeutic agents) for the treatment of the above-mentioned conditions.
  • the combination can be administered as separate compositions (simultaneous, sequential) of the individual components of the treatment or as a single dosage form containing both agents.
  • the active ingredients may be separately formulated into single-ingredient preparations of one of the above-described forms and then provided as combined preparations, which are given at the same time or different times, or may be formulated together into a two- or more- ingredient preparation.
  • Compounds of general formula (I) may be administered to a patient in a total daily dose of, for example, from 0.001 to 1000 mg/kg body weight daily. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose. The determination of optimum dosages for a particular patient is well known to one skilled in the art.
  • compositions are normally accompanied by written or printed instructions for use in the treatment in question.
  • ACN acetonitrile
  • ACN-d 3 deuterated acetonitrile
  • CDCI 3 deuterated chloroform
  • DCE dichloroethane
  • EtOK potassium ethylate
  • EtONa sodium ethylate
  • K 2 CO 3 potassium carbonate
  • KPS potassium persulfate
  • LiAIH 4 lithiumaluminum hydride
  • LiOH lithium hydroxide
  • NaHC0 3 sodium bicarbonate
  • NaOH sodium hydroxide
  • TBAB tetrabutylamonium bromide
  • TEA triethylamine
  • TFA trifluoroacetic acid
  • THF tetrahydrofurane
  • the 1 H-NMR spectra were acquired with a Varian 500 MHz instrument.
  • the chemical shifts are expressed in parts per million (ppm, ⁇ units).
  • the coupling constants are expressed in Hertz (Hz) and the splitting patterns are described as s (singlet), bs (broad signal), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet).
  • the LC-MS analyses were carried out on a Waters Acquity UPLC or Waters Acquity UPLC H-Class linked to with a SQD Single quadrupole (Waters) using an Acquity UPLC BEH C18 (50 x 2.1 mm, 1.7 ⁇ ) or Acquity UPLC HSS T3 (50 x 2.1 mm, 1.8 ⁇ ) column.
  • Phase A was composed by either Milli-Q water/ACN 95/5 + 0.07% formic acid or Milli-Q water + 0.07% formic acid; Phase B by ACN + 0.05% formic acid; flow rate: 0.6 mL/min; UV detection (DIODE array) from 210 to 400 nm; ESI+ detection in the 100-2000 m/z range. The yields were calculated assuming that products were 100% pure if not stated otherwise.
  • ntermediate 42 2-[(Z)-C-(3-methoxy-phenyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
  • the reaction mixture was heated at reflux for about 20 h and then under microwave irradiation at 135°C for about 6 h.
  • the two obtained products, the E-isomer and the Z-isomer of the oxime, were separated by flash chromatography (eluent hexane:EtOAc from 92:8 to 70:30).
  • Example 1 methyl 2-[(E)-A -hydroxy-C-methyl-carbonimidoyl]pyridine-4- carboxylate
  • Example 2 methyl 2-[(E)-/V-hydroxy-C-phenyl-carbonimidoyl]pyridine-4- carboxylate and Example 3: methyl 2-[(Z)-/V-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate
  • Example 3 0.018 g (28%) of methyl 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylate (Example 2) and 0.010 g (16%) of methyl 2-[(Z)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 3) were prepared according to the procedure described for Example 1 , starting from 0.060 g (0.25 mmol) of methyl 2- benzoylpyridine-4-carboxylate (Intermediate 4). The reaction gave two products, the £- isomer and the Z-isomer of the oxime.
  • Example 4 methyl 2-[(E)-/V-hydroxy-C-cyclopropyl-carbonimidoyl]pyridine-4- carboxylate and Example 5: methyl 2-[(Z)-A -hydroxy-C-cyclopropyl- carbonimidoyl]pyridine-4-carboxylate
  • Example 5 0.035 g (54%) of methyl 2-[(E)-A/-hydroxy-C-cyclopropyl-carbonimidoyl]pyridine-4- carboxylate (Example 4) and 0.012 g (19%) of methyl 2-[(Z)-N-hydroxy-C-cyclopropyl- carbonimidoyl]pyridine-4-carboxylate (Example 5) were prepared according to the procedure described for Example 1 , starting from 0.060 g (0.29 mmol) of methyl 2- (cyclopropanecarbonyl)pyridine-4-carboxylate (Intermediate 5). The reaction gave two products, the E-isomer and the Z-isomer of the oxime.
  • Example 4 (E- isomer): 1 H NMR (ACN-d 3 ) ⁇ (ppm): 9.12 (bs, 1 H), 8.79-8.61 (m, 1 H), 8.12-7.98 (m, 1 H), 7.86-7.69 (m, 1 H), 3.95 (s, 3 H), 2.59-2.41 (m, 1 H), 1 .31-0.80 (m, 4 H); MS (ESI): m/z: 221 [M+H] + ;
  • Example 6 0.046 g (70.5%) of methyl 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 6) was prepared according to the procedure described for Example 1 , starting from 0.060 g (0.10 mmol) of methyl 2-propanoylpyridine-4-carboxylate (Intermediate 6).
  • Example 7 methyl 2-[(E)-/V-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine- 4-carboxylate and Example 8: methyl 2-[(Z)-/V-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate
  • Example 8 0.041 g (43%) of methyl 2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate (Example 7) and 0.035 g (37%) of methyl 2-[(Z)-N-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyhdine-4-carboxylate (Example 8) were prepared according to the procedure described for Example 1 , starting from 0.090 g (0.33 mmol) of methyl 2-(2-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 7) and using MeOH as solvent.
  • Example 8 (Z-isomer): 1 H NMR (DMSO-d 6 ) ⁇ (ppm): 1 1 .83 (s, 1 H), 8.76-8.61 (m, 1 H), 8.34-8.21 (m, 1 H), 7.83-7.
  • Example 9 methyl 2-[(E)-/V-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine- 4-carboxylate and Example 10: methyl 2-[(Z)-/V-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate
  • Example 10 0.022 g (42%) of methyl 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate (Example 9) and 0.027 g (51 %) of methyl 2-[(Z)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 10) were prepared according to the procedure described for Example 1 , starting from 0.050 g (0.18 mmol) of methyl 2-(3-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 9) and using 2 ml_ of MeOH as solvent.
  • Example 11 0.055 g (70%) of te/t-butyl 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylate (Example 11 ) was prepared according to the procedure described for Example 1 , starting from 0.075 g (0.25 mmol) of te/t-butyl 2-(2-phenylacetyl)pyridine-4- carboxylate (Intermediate 10).
  • Example 13 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid (Example 13) was prepared according to the procedure described for Example 12, starting from 0.018 g (0.070 mmol) of methyl 2-[(E)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 2).
  • Example 14 2-[(Z)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid
  • Example 14 was prepared according to the procedure described for Example 12, starting from 0.010 g (0.040 mmol) of methyl 2-[(Z)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 3).
  • Example 15 2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 15) was prepared according to the procedure described for Example 12, starting from 0.035 g (0.59 mmol) of methyl 2-[(E)-C-cyclopropyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 4).
  • Example 16 2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
  • Example 5 2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
  • Example 17 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 17) was prepared according to the procedure described for Example 12, starting from 0.039 g (0.19 mmol) of methyl 2-[(E)-C-ethyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 6).
  • Example 18 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
  • Example 18 was prepared according to the procedure described for Example 12, starting from 0.020 g (0.070 mmol) of methyl 2-[(E)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 9).
  • Example 19 2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 19) was prepared according to the procedure described for Example 12, starting from 0.025 g (0.090 mmol) of methyl 2-[(Z)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 10).
  • Example 20 2-[(E)-N-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
  • Example 20 was prepared according to the procedure described for Example 12, starting from 0.040 g (0.14 mmol) of methyl 2-[(E)-N-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 7).
  • Example 21 2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
  • Example 21 was prepared according to the procedure described for Example 12, starting from 0.034 g (0.12 mmol) of methyl 2-[(Z)-A/-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 8).
  • Example 22 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 22) was prepared according to the procedure described for Example 12, starting from 0.045 g (0.14 mmol) of te/t-butyl 2-[(E)-C-benzyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 11 ).
  • Example 25 2-[(Z)-A -hydroxy-C-(2- methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid trifluoroacetate
  • Example 25 2-[(Z)-A/-hydroxy-C-(2- methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid trifluoroacetate (Example 25) was prepared according to the procedure described for Example 24, starting from 0.020 9 (0.046 mmol) 2-[(Z)-C-[2-(fert- butoxycarbonyl(methyl)amino)ethoxymethyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 19).
  • Example 26 2-[(Z)-C-[2-(benzylamino)ethoxymethyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 26) was prepared according to the procedure described for Example 24, starting from 0.013 g (0.025 mmol) of 2-[(Z)-C-[2-(benzyl(te/t-butoxycarbonyl)amino)ethoxymethyl]-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 21 ) and using 2 M HCI in Et 2 0 and DCM as solvent.
  • Example 27 2-[(Z)-A/-hydroxy-C-(phenoxymethyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 27) was prepared according to the procedure described for Example 24, starting from 0.017 g (0.048 mmol) of 2-[(Z)-C-(phenoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 24) and using DCM as solvent.
  • Example 28 2-[(Z)-C-(ethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4- carboxylic acid
  • Example 28 was prepared according to the procedure described for Example 24, starting from 0.015 g (0.049 mmol) of 2-[(Z)-C-(ethoxymethyl)-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 18) and using 2 M HCI in Et 2 0 and DCM as solvent.
  • Example 29 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide hydrochloride (Example 29) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.095 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxamide (Intermediate 25) and 25 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 6 days.
  • Example 30 A -ethyl-2-[(E)-A -hydroxy-C-methyl-carbonimidoyl]pyridine-4- carboxamide hydrochloride
  • Example 30 0.018 g (85%) of A/-ethyl-2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyndine-4- carboxamide hydrochloride (Example 30) was prepared according to the procedure described for Example 24, starting from 0.026 g (0.089 mmol) of N-ethyl-2-[(E)-C-methyl- N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxamide (Intermediate 26) and 20 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 24 h.
  • Example 31 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbohydroxamic acid (Example 31 ) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.1 1 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]-A/-tetrahydropyran-2-yloxy-pyridine-4-carboxamide (Intermediate 27) and 20 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 18 h.

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Abstract

The present invention relates to oxime derivatives (Formula I), wherein R1 and R2 are as defined in the specification, pharmaceutical compositions containing such compounds and to their use in therapy.

Description

Oxime Derivatives Useful as Inhibitors of Histone Demethylase KDM4C
Field of the Invention
The present invention relates to oxime derivatives, pharmaceutical compositions containing such compounds and to their use in therapy.
Background of the invention
Alterations in the structural and functional states of chromatin, mainly determined by post- translational modification of histone components, are involved in the pathogenesis of a variety of diseases. These reversible modifications contribute to the dynamicity of chromatin remodeling and are tightly controlled by opposing activities of enzyme families. The enzymatic processes governing these post-translational modifications on the nucleosomes have become potential targets for the so-called epigenetic therapies (Portela, A. et ai. Nat. Biotechnol. 2010, 28, 1057-1068). The discovery of an increasing number of histone lysine demethylases has highlighted the dynamic nature of the regulation of histone methylation, a key chromatin modification that is involved in eukaryotic genome and gene regulation. Histone lysine demethylases represent attractive targets for epigenetic drugs, since their expression and/or activities are often misregulated in cancer (Varier, R. A. et al. Biochim. Biophys. Acta. 201 1 , 1815, 75-89). A lysine can be mono-, di-, and tri-methylated and each modification, even on the same amino acid, can exert different biological effects.
Histone lysine demethylases can be grouped into two major families with different enzymatic mechanisms (Anand, R. et al. J. Biol. Chem. 2007, 282, 35425-35429; Metzger, E. et al. Nat. Struct. Mol. Biol. 2007, 14, 252-254). The first identified subfamily KDM1 (or lysine specific demethylases LSDs) includes KDM1A (also known as LSD1 ) and KDM1 B (also known as LSD2). KDM1A and KDM1 B are both flavo-amino oxidases dependent proteins sharing a FAD coenzyme-binding motif, a SWIRM domain and an amine oxidase domain, all of which are integral to the enzymatic activity of KDM1 family members. KDM1A and KDM1 B demethylation activity is limited to mono and dimethyl substrates, such as histone H3K4 me2/me and histone H3K9me2/me). On the other hand, we find the large protein family of Jumonji C (JmjC) domain-containing proteins, wherein the demethylation reaction is carried out by JmjC domain proteins and wherein a conserved JmjC domain, the presence of Fe (II) and a-ketoglutarate is required to allow the removal of mono-, di-, and trimethylated lysines. This large class comprises 20 human enzymes grouped into five subfamilies: KDM2/7, KDM3, KDM4, KDM5, and KDM6 (Pedersen, M.T. et al. Trends Cell Biol. 2010, 20, 662- 671 ). Many of the human Jmj-type enzymes are involved in human pathological processes, including development, cancer, inflammation and metabolic diseases (Johansson, C. et a/ Epigenomics. 2014, 6, 89-120). Furthermore several KDMs are over expressed in multiple types of cancer cells (Hojfeldt, J. W. et al. Nat. Rev. Drug Discov. 2013, 12, 917-930).
The KDM2 subfamily is constituted by two proteins, KDM2A and KDM2B (also known as FBXL1 1 and FBXL10, respectively), which catalyze the demethylation of H3K36me2/me1 histone chromatin marks. Evidences of an involvement of KDM2A in development and progression in breast and lung cancer have been published (Rizwani, W. etal. PLoS One. 2014 Jul 16;9(7):e100888.; Wagner, K.W. et al. J. Clin. Invest. 2013, 123, 5231 -5246). KDM2B has been found to be involved in hematological cancers, such as acute myeloid leukemia and myelodisplastic syndrome (He, J. et al. Blood 201 1 , 1 17, 3869-3880, Ueda, T. et al. Blood 2015, 125, 3437-3446; Karoopongse, E. et al. PLoS One. 2014 Sep 16;9(9):e107817), and in solid tumors, such as pancreatic adenocarcinoma cancer and breast cancer (Tzatsos, A. et al. J. Clin. Invest. 2013, 123, 727-739, Kottakis, F. et al. Cancer Res. 2014, 74, 3935-3946).
KDM7 subfamily includes three members, KDM7A (also known as KIAA1718), KDM7B (also PHF8) and KDM7C (also PHF2), all of them catalyze the demethylation of histone H3K9 and H3K27 mono- and di-methylated. In addition, KDM7B has also been shown to demethylate the monomethyl of histone H4K20. It has been found that high expression of KDM7B is associated to an adverse prognosis in patients of laryngeal, hypopharyngeal squamous cell carcinoma (Zhu et al.Epigenomics. 2015;7(2):143-153) and non small cell lung cancer (Shen, Y. et al. Biochem. Biophys. Res. Commun. 2014, 451 , 1 19-125 as well as in prostate (Ma,Q. ei a/ Onco Targets Ther. 2015 Aug 10;8:1979-1988) The KDM3 subfamily includes three proteins, KDM3A (also JMJD1A), KDM3B (also JMJD1 B) and KDM3C (also JMJD1 C), of which evidences of histone lysine demethylation activities versus histone H3K9me2/me have been reported for KDM3A and KDM3B (Brauchle, M. et al. PLoS One. 2013 Apr 1 1 ;8(4):e60549.) The involvement of KDM3A in cancer has been published and specifically KDM3A has been reported to regulate tumor growth in hypoxia condition (Krieg, A.J. et al. Mol. Cell Biol. 2010, 30, 344-353). Moreover KDM3A appears to be highly expressed in renal cell carcinoma (Guo, X. et al. Neoplasma 201 1 , 58, 153-157), and to confers metastasis and chemo resistance in epithelial ovarian cancer (Pa, M. et ai. J Mol Hist (2015) 46:51 1-518). In addition KDM3A has been demonstrated to be a key estrogen regulator in breast cancer (Wade, M.A. et ai. Nucleic Acids Res. 2015, 43, 196-207) as well as to be a tumor promoter in Ewing Sarcoma (Parrish, J.K. et al. Oncogene 2015, 34, 257-262). KDM3B has been found to have a role in leukemiogeneis (Kim, J.Y. et al. Mol. Cell. Biol. 2012, 32, 2917-2933).
The KDM4 subfamily (also known as JMJD2A-D proteins) is composed by four proteins, KDM4A, KDM4B, KDM4C and KDM4Dall of them are capable of recognizing and demethylating histone H3K9me2/me3 and histone H3K36me2/me3 as well as histone H1 .4K26me3. KDM4s proteins are also found to catalyze demethylation of non histone substrates and specifically trimethyl-lysine peptides of chromatin repressors such as WIZ, CDYL1 , CSB and G9a proteins (Ponnaluri, V.K. et al. Biochem. Biophys. Res. Commun. 2009, 390, 280-284). Further two members of the human KDM4 subfamily (KDM4E and KDM4F, also known as JmjD2E and JmjD2F) are known to exist but are currently considered being pseudogenes due to lack of intronic sequences in their genes (Katoh, Y. et al. Int. J. Mol. Med. 2007, 20, 269-273). An involvement of KDM4A in inducing site- specific copy gain and re-replication of regions amplified in tumors highlights the important role of KDM4A in cancer development (Black, J.C. et al. Cell 2013, 154, 541 - 555). Moreover, KDM4A amplification and over expression has been reported in ovarian cancer, in squamous cell carcinoma (Ding, X. et al. Sci. Signal. 2013, 6(273): ra28.1 -13, SO-15; Black, J.C. et al. Cell 2013, 154, 541 -555) and in breast cancer (Berry, W.L. et al. Int.J. Oncol. 2012, 41 , 1701 -1706). Furthermore, KDM4A is associated to breast cancer progression (Li, L.L. et al. Breast Cancer Res. 2014, 16(3):R56) and to promotion of cellular transformation (Mallette, F.A. et al. Cell Rep. 2012, 2, 1233-1243). KDM4B is over expressed in gastric cancer (Li, W. et al. Biochem. Biophys. Res. Commun. 201 1 , 416, 372-378), and it promotes hormonally responsive breast carcinogenesis (Shi, L. et al. Proc. Natl. Acad. Sci. U. S. A. 201 1 , 108, 7541 -7546). In addition, KDM4B promotes epithelial-mesenchymal transition by cooperating with β-catenin and enhances gastric cancer metastasis (Zhao, L. et al. Clin. Cancer Res. 2013, 19, 6419-6429). Recently, a role of KDM4B in development of neuroblastoma has also been proposed (Yang, J. et al. J. Natl. Cancer Inst. 2015, 107(6), djv080). Several data suggest that KDM4C could play an important role in cancer (Berry, W.L. et al. Cancer Res. 2013, 73, 2936-2942). KDM4C locus is amplified in esophageal squamous cell carcinoma, breast cancer, medulloblastoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL). Furthermore, high expression of the enzyme has been reported in prostate cancer. In addition, the KDM4C gene was found to be translocated in mucosa- associated lymphoid tissue lymphoma (Labbe, R.M et al. Am. J. Transl. Res. 2013 6, 1 - 15, Helin, K. et al. Nature 2013, 502, 480-488; Hojfeldt, J.W. et al. Nat. Rev. Drug Discov. 2013, 12, 917-930). Importantly, ectopic expression of KDM4C is able to transform immortalized mammary cells (MCF10A) conferring cancer stem cells characteristic, such as the capability to form mammospheres (Liu, G. et al. Oncogene. 2009, 28, 4491 -4500). Furthermore, KDM4C down regulation in colon cancer cells strongly affects their sphere forming capacity and in vivo tumor formation ability, which are both characteristics of cancer stem cells suggesting that the inhibition of the enzyme could represent an effective therapeutic approach for colon cancer (Yamamoto, S. et al. Carcinogenesis 2013, 34, 2380-2388). In addition, KDM4C has been found to have a critical role in acute myeloid leukemia. (Cheung, N. et al. Cancer Cell. 2016, 29, 32-48)
The KDM4A, B and C proteins, share more than 50% sequence identity, each contain JmjN, JmjC, two plant homeodomains (PHD) and two Tudor domains. KDM4D is unique within the KDM4 family, since it lacks both the PHD and Tudor domains and thus is only half the size of KDM4A-C. KDM4D has a different substrate specificity: it does not demethylate H3K36 due to several differences in its substrate binding cleft, yet has gained the ability to attack H1.4K26me2/me3. Also, KDM4D attacks H3K9me2 with similar efficiency as for H3K9me3 and may, albeit inefficiently, even demethylate H3K9me1 (Berry W.L. et ai. Cancer Res. 2013, 73, 2936-2942). KDM4D functions as a coactivator of the androgen receptor (Shin S et al. Biochem Biophys Res Commun. 2007, 359, 742-746) and stimulates p53-dependent gene expression (KiM TD et al. PLoS One. 2012;7:e34618.). In addition a role of KDM4D in DNA damage response has also recently been reported (Khoury-Haddad H. et al. Cell Cycle. 2015; 14, 950-958).
KDM5 subfamily (also known as Jarid) consists of four members: KDM5A (RBP2/JARID1A), KDM5B (also known as PLU1/JARID1 B), KDM5C (SMCX/JARID1 C), and KDM5D (SMCY/JARID1 D). All members catalyze the demethylation of histone H3K4me3/me2, which is a signature indicative of transcriptional activation, and hence KDM5 proteins are considered transcriptional co repressors. There are several evidences of an involvement of KDM5A in cancer. For instance, the loss of KDM5A has been reported to suppress the tumorigenesis in mice lacking Rb or Men 1 (Lin, W. et al. Proc. Natl. Acad. Sci. U. S. A. 201 1 , 108, 13379-13386), and genomic amplification of KDM5A have been reported in breast cancer (Hou, J. et al. Am. J. Transl. Res. 2012, 4, 247-256). In addition, KDM5A has been associated to promote lung tumorigenesis and cancer metastasis (Teng, Y.C. et al. Cancer Res. 2013, 73, 471 1 -4721 ) and the insurgency of a cell population not responsive to chemotherapeutic drugs (Sharma, S.V. et al. Cell 2010, 141 , 69-80). Over expression of KDM5A has also been reported in hepatocellular carcinoma and human gastric cancer (Liang, X. et al. PLoS One. 2013 Jul 29;8(7):e69784; Li, H. et al. Mol Cancer 2014, 13, 18). Moreover, an involvement of KDM5A in leukemiogenesis has been recently suggested by the identification of a translocation: NUP98/JARID1A(KDM5A) as recurrent abnormality in pediatric acute megakaryoblastic leukemia (de Rooij, J.D. et al. Leukemia. 2013, 27, 2280-2288.) High expression of KDM5B and its direct role in cancer progression has been reported for breast cancer (Yamane, K. et al. Mol. Cell 2007, 25, 801 -812; Yamamoto, S. et al. Cancer Cell 2014, 25, 762-777). Similarly, the up-regulation of KDM5B expression has also been found in prostate, bladder, lung and gastric cancer (Xiang, Y. et al. Proc. Natl. Acad. Sci. U. S. A. 2007, 104, 19226-9231 ; Hayami, S. et al. Mol. Cancer 2010, 9, 59; Wang, Z. et al. Am. J. Cancer Res. 2015, 5, 87-100). The overexpression of histone demethylase KDM5B has also been associated with progression of glioma cell growth (Dai, B. et al. Biochem. Biophys. Res. Commun. 2014, 454, 221 -227) and with poor prognosis and chemotherapy resistance in epithelial ovarian cancer (Wang, L. et al. Tumour Biol. 2015, 36, 2465-2472). Moreover, data indicate that KDM5B plays also an important role in chemioresistance and stem cell like phenotype of neuroblastoma cells (Kuo, Y.T. et al. PLoS One. 2015 May 7;10(5):e0125343). In addition, there are indications that KDM5B negatively regulates leukemogenesis in murine and human MLL-rearranged AML cells (Cancer Cell. 2015 Aug 10;28(2):198-209.). KDM5C has been found to be over expressed in prostate cancer (Stein, J. et al. Am. J. Pathol. 2014, 184, 2430-2437) suggesting a possible role as oncogene. The KDM6 family consists of KDM6A (also known as UTX), UTY and KDM6B (also known as JMJD3). KDM6A and KDM6B are histone demethylases specific for H3K27me3/me2, whilst discordant data about the enzymatic activity has so far been reported for UTY (Walport, L.J. et al. J. Biol. Chem. 2014, 289, 18302-18313). Recently it has been reported that KDM6A ablation resulted in a significant decrease in the proliferation and invasiveness of breast cancer cells in vitro and in a mouse xenograft model (Kim, J.H. et al. Cancer Res. 2014, 74, 1705-1717). Concomitantly, a role in initiation and maintenance of T-ALL leukemia (Ntziachristos, P. et al. Nature 2014, 514, 513-517) as well a role in the development of Hodgkin's Lymphoma (Anderton, J.A. et al. Oncogene 201 1 , 30, 2037-2043) has also been demonstrated for KDM6B. Inhibitors of histone demethylases and in particular of the potential oncogene KDM4C thus represent an innovative and novel approach for cancer therapy.
US patent application US201 10003827 discloses HIV protease inhibitors of general formula
Figure imgf000007_0001
and wherein (A/-hydroxy-C-methyl-carbonimidoyl)pyridine-4-carbaldehyde was used as intermediate in the synthesis. Description of the Invention
In the present invention it was surprisingly found that oxime derivatives of general formula (I) as described below are potent KDM4C inhibitors. As such they represent attractive therapeutic approach for cancer.
According to the present invention there are provided compounds of formula (I)
Figure imgf000008_0001
(I)
Ci-C6 alkyl;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; Ci-C6 alkyl substituted by X-R3 ; C3-C7-cycloalkyl; heterocyclyl, aryl or heteroaryl; is CH2OH, (CO)R4, CN, tetrazole, CH(OH)CF3 or C(OH)2CF3 is C1-C-6 alkyl; C3-C7 cycloalkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl;
O or NH; is hydrogen, OH, O-Ci-C6 alkyl, NR7R8, or CF3; p is 0 or 1 ;
R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl, said aryl being optionally substituted by one ore more substituents independently selected from the group consisting of halogen, CN, or NH2; Ci-C6 alkyl substituted by C3-C7- cycloalkyl optionally substituted by halogen; or Ci-C6 acyl;
R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl or heteroaryl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxy; halogen; C-i- C6 alkyl; Ci-C6 alkyl substituted by NR 6R17; Ci-C6 alkoxy; Ci-C6 haloalkyl; Ci-C6 haloalkoxy; NR9R10; CONR20R21; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C6 alkyl; aryl; Ci-C6 alkyl substituted by aryl or heteroaryl; Ci-C6 alkoxy substituted by aryl or heteroaryl; phenyloxy, wherein the phenyl may be optionally substituted by halogen; C2-C6 alkoxy substituted by NR18R19; Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; or C1-C6 alkyl substituted by aryloxy or heteroaryloxy, wherein the aryl or heteroaryl on the C1-C6 alkyl substituted by aryl or heteroaryl, or on the C1-C6 alkoxy substituted by aryl or heteroaryl, or on the C1-C6 alkyl substituted by aryloxy or heteroaryloxy may indipendently and optionally be substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of aryl or heteroaryl, wherein the aryl or the heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl or heteroaryl, wherein the aryl or the heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; or Ci-C6 alkyl substituted by OR11 or NR 2R13; or (CO)R 4 ; and
R9, R 0 are, independently, hydrogen; d-C6 alkyl; or COR15;
R , R 2, R 3 are, independently, hydrogen or Ci-C6 alkyl; R14 is C-i-C-6 alkyl or aryl;
R 5 is Ci-C6 alkyl; or Ci-C6 alkyl substituted by NH2;
R 6, R 7, R 8, R 9, R20, R2 are, independently, hydrogen, Ci-C6 alkyl, Ci-C6 alkyl substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; or stereoisomers or pharmaceutically acceptable salts thereof and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyridine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-isonicotinic acid.
The invention further provides compounds of formula (I)
Figure imgf000010_0001
wherein:
Ci-C6 alkyl;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl;
Ci-C6 alkyl substituted by X-R3 ;
C3-C7-cycloalkyl; heterocyclyl, aryl or heteroaryl; R2 is CH2OH, (CO)R4, CN, or tetrazole; R3 is Ci-C6 alkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, or NR7R8; p is O or l ;
R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl; Ci- Ce alkyl substituted by C3-C7-cycloalkyl optionally substituted by halogen; or Ci-C6 acyl;
R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl or heteroaryl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, C1-C6 alkyl, Ci-C6 alkoxy, Ci-C6 haloalkyl, Ci-C6 haloalkoxy, NR9R10, unsubstituted heterocyclyl, heterocyclyl substituted by C1-C6 alkyl, C1 -C6 alkyl substituted by aryl; or C1-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of C1-C6 alkyl; C1 -C6 alkyl substituted by aryl; Ci-Ce alkyl substituted by OR11 or NR 2R13; or (CO)R 4 ; and R9, R 0 are, independently, hydrogen; Ci-C6 alkyl; or COR15;
R , R 2, R 3 are, independently, hydrogen or Ci-C6 alkyl
R 4 is Ci-C6 alkyl or aryl;
R 5 is Ci-C6 alkyl; or Ci-C6 alkyl substituted by NH2; and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyhdine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-isonicotinic acid.
Preferably,
R is Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl;
C-I -C6 alkyl substituted by heterocyclyl, wherein the heterocyclyl may be optionally substituted by Ci-C6 alkyl;
Ci-C6 alkyl substituted by X-R3;
C3-C7-cycloalkyl; aryl, wherein the aryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, Ci- Ce alkyl, Ci-C6 alkoxy; or Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen;
R2 is CH2OH, COR4, CN, tetrazole, CH(OH)CF3 or C(OH)2CF3; R3 is Ci-C6 alkyl;
C3-C7 cycloalkyl
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, NR7R8, or CF3; p is O or l ; R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl, said aryl being optionally substituted by halogen; or C-i-C-6 acyl;
R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl in R or R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxyl; halogen; d-C-6 alkyl; C-i- C6 alkyl substituted by NR 6R17; Ci-C6 alkoxy; Ci-C6 haloalkyl; Ci-C6 haloalkoxy; NR9R10; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C6 alkyl; phenyl; Ci-C6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; Ci- C-6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; phenyloxy, wherein the phenyl may be optionally substituted by halogen; or C-2- C6 alkoxy substituted by NR18R19 and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of C-i-C-6 alkyl or aryl, wherein the aryl is optionally substituted by halogen; R9, R 0 are, independently, hydrogen or COR15;
R 5 is Ci-C6 alkyl substituted by NH2;
R 6, R 7, R 8, R 9 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by phenyl, wherein the phenyl may be optionally substituted by halogen.
Still preferably,
R is Ci-C6 alkyl;
C-i-C-6 alkyl substituted by aryl; C-I -C6 alkyl substituted by heterocyclyl, wherein the heterocyclyl may be optionally substituted by C-i-C-6 alkyl;
C1-C-6 alkyl substituted by X-R3; C3-C7-cycloalkyl; aryl, wherein the aryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, Ci- Ce alkyl, Ci-C6 alkoxy; or Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen;
R2 is CH2OH, COR4, CN, or tetrazole;
R3 is Ci-C6 alkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, or NR7R8; p is O or l ;
R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl; or
Ci-C6 acyl;
R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl in R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, Ci-C6 alkyl, Ci-C6 alkoxy, Ci-C6 haloalkyl, Ci-C6 haloalkoxy, NR9R10 or heterocyclyl substituted by Ci-C6 alkyl; and
R9, R 0 are, independently, hydrogen or COR15;
R 5 is Ci-C6 alkyl substituted by NH2.
In a further preferred embodiment, aryl is phenyl or naphthyl.
Particularly preferred compounds of general formula (I) include:
methyl 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyndine-4-carboxylate;
methyl 2-[(Z)-N-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; te/t-butyl 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[A/-hydroxy-C-[(4-methylpiperazin-1 -yl)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-(A/-hydroxy-C-(methoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-(A/-hydroxy-C-(2-methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-[C-[2-(benzylamino)ethoxymethyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-(A/-hydroxy-C-(phenoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-(C-(ethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide;
A/-ethyl-2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbohydroxamic acid; 1 - [4-(1 H-tetrazol-5-yl)-2-pyridyl]ethanone oxime;
1 -[4-(hydroxymethyl)-2-pyridyl]ethanone oxime;
2- [(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyndine-4-carbonitrile;
methyl 2-(A/-hydroxy-C-(phenoxymethyl)carbonimidoyl)pyridine-4-carboxylate;
methyl 2-[(E)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(Z)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-(C-(benzyloxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid;
2-[C-[[3-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-(C-(2-acetamidoethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-[(2-methoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-[(3-methoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-(anilinomethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-N-hydroxy-C-(methylaminomethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(2-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-[[4-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3-aminophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-[(4-aminophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(4-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-pyridyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(pyrimidin-2-yloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-A/-hydroxy-C-(1 -naphthyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-naphthyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid; 2-[(Z)-N-hydroxy-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]carbonimidoyl]pyridine^ 4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]carbonimidoyl]pyridine- 4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[3-(trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyndine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[(4-methoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
methyl 2-[(E)-A/-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-A/-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[3-(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyndine-4- carboxylic acid;
2-[(Z)-C-[2-[(4-fluorophenyl)methylamino]ethoxymethyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid hydrochloride;
2-[(E)-C-[(4-fluoroanilino)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
2-[(Z)-N-hydroxy-C-[(4-morpholinophenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]carbonimidoyl]pyridine- 4-carboxylic acid 2,2,2-trifluoroacetic acid; 2-[(Z)-N-hydroxy-C-[[4-(1 -piperidyl)phenoxy]methyl]carbonimidoyl]pyndine-4-carboxylic acid hydrochloride;
2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-[(4-phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[(3-phenylphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[(3-phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(4-bromophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-[(4-phenylphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-(cyclopentoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-(cyclobutoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-(propoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-N-hydroxy-C-[(4-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-N-hydroxy-C-[(4-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid;
ethyl 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid;
ethyl 2-[(E)-N-hydroxy-C-[(4-phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-N-hydroxy-C-[(4-phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-N-hydroxy-C-[(3-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylate;
ethyl 2-[(E)-N-hydroxy-C-[(3-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3-phenylphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-N-hydroxy-C-[(3-phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-N-hydroxy-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]carbonimidoyl]pyridine-4- carboxylic acid hydrochloride;
ethyl 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride;
1 -[4-(2,2,2-trifluoro-1 ,1 -dihydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride;
2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl-amino]ethoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
methyl 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; Ethyl 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; Ethyl 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-[2-(4-phenyl-1 -piperidyl)ethoxymethyl]carbonimidoyl]pyridine-4- carboxylic acid hydrochloride
or stereoisomers or pharmaceutically acceptable salts thereof.
Preferably, the above mentioned compounds of the invention are for medical use or for use as medicament. Preferably, the above compounds are inhibitors of KDM4C. Still preferably, the above compounds are inhibitors of KDM4C for medical use. An inhibitor is a compound able to bind to KDM4C and blocks its activity or function.
In another embodiment, the invention provides the compounds of general formula (I) for medical use, in particular for use in the treatment and/or prevention of cancer. Preferably, the compounds of general formula (I) are for the use in the treatment and/or prevention of leukemia, lymphoma, esophageal squamous cell carcinoma, breast cancer, medulloblastoma, prostate cancer, colon cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreas cancer or glioblastomas. Still preferably the glioblastomas are giant cell glioblastoma or gliosarcoma. Still preferably the leukemia or lymphoma are acute myeloid leukemia, mucosa-associated lymphoid tissue lymphoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL).
A further embodiment of the invention is a pharmaceutical composition comprising a compound of general formula (I) together with a pharmaceutically acceptable excipient and/or diluent. The pharmaceutical composition may further comprise at least one further therapeutic agent, preferably selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, anti-proliferative/antineoplastic agents, cytostatic agents, agents which inhibit cancer cell invasion, inhibitors of growth factor function, anti- angiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent. The pharmaceutical composition may be for use in the treatment and/or prevention of cancer.
In a further embodiment, the invention provides a process for obtaining a compound of formula (I) as defined above, wherein R2 is COOH, the process comprising the preparation of a compound of formula A3 by reaction of a compound of formula A1 with a compound of formula A2 in presence of an oxidant, a transition metal salt, and an acid, the preparation of a compound of formula A4 by reaction of a compound of formula A3 with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH2OH), and the final deprotection of a compound of formula A4 to obtain a compound of formula (I), as represented in Scheme A-IV:
Figure imgf000021_0001
A1 A2 A3 A4 I
Scheme A-IV wherein R is as defined above and PG is a protecting group, preferably PG is methyl or ethyl.
In the present invention, "aryl" represents a mono or bicyclic aromatic ring system of, respectively, 6, 9 or 10 atoms. Examples of such an aryl are phenyl, indenyl, indanyl and naphthyl and tetrahydronaphthalenyl. Substituted aryl means that the hydrogen atoms on independently each carbon atom may be independently replaced by a substituent as defined herein above.
"Heteroaryl" represents a mono or bicyclic heteroaromatic ring system of, respectively, 5 to 10 members, which contains one, two, three or four heteroatoms selected from nitrogen, oxygen or sulphur and one to nine carbon atoms. Examples of said heteroaryls include, but are not limited to: pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, 1 ,2,3-oxadiazolyl, 1 ,3,4-thiadiazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, isoindolyl, benzo[b]furanyl, benzo[ib]thienyl, benzopyranyl, indazolyl, benzimidazolyl, purinyl, quinolyl, isoquinolyl, quinazolinyl, and quinoxalinyl. Substituted heteroaryl, means that the hydrogen atoms on independently each carbon atom or heteroatom may be independently replaced by a substituent as defined herein above. "Heterocyclyl" represents a mono, bicyclic or a spirocyclic saturated or partially saturated non-aromatic ring system of, respectively, 4 to 12 members, which contains one, two, or three heteroatoms selected from nitrogen, oxygen, and sulphur and three to eleven carbon atoms. Examples of such heterocycles include, but are not limited to: pyrrolidyl, pyrrolidinyl, piperidyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, benzodioxolyl, 2,3-dihydro-benzodioxinyl, benzoxazolyl, azetidyl, azepinyl, and diazapinyl. Examples of bicyclic ring systems include, but are not limited to, 2-aza-bicyclo[2.2.1]heptanyl, 2,5-diaza-bicyclo[2.2.1]hept-2-yl or 8- azabicyclo[3.2.1]octanyl. Examples of spirocyclic ring systems include, but are not limited to, 3,8-diazaspiro[4.5]decane.
The term "C1 -C6 alkyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, having from one to six carbon atoms. The "C-i-C-6 alkyl" group is preferably a linear or branched Ci-C alkyl group, more preferably a Ci -C2 alkyl group. Examples of Ci-C6 alkyl include methyl, ethyl, n-propyl, isopropyl, butyl, te/t-butyl, pentyl, and hexyl.
The term "C3-C7-cycloalkyl" refers to a saturated monocyclic hydrocarbon ring system having three to seven carbon atoms. Examples of C-3-C7-cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
The term "d-C-6 alkoxy" refers to a straight or branched O-C-i-C-6 alkyl, where alkyl is as defined herein. The "C-i-C-6 alkoxy" group is preferably a linear or branched Ci-C4 alkoxy group, more preferably a Ci-C2 alkoxy group.
The term "C-i-C-6 haloalkyi" refers to a straight or branched hydrocarbon chain radical, which is substituted by one or more halogen atoms and having from one to six carbon atoms. The "C-i-C-6 haloalkyi" group is preferably a linear or branched Ci-C4 haloalkyi group, more preferably a Ci-C2 haloalkyi group, being in particular CF3.
The term "C1 -C6 haloalkoxy" refers to a straight or branched O-C1-C6 haloalkyi, where haloalkyi is as defined herein. The "C-i-C-6 haloalkoxy" group is preferably a linear or branched Ci-C haloalkoxy group, more preferably a C1 -C2 haloalkoxy group, being in particular OCF3, OCHF2 or OCH2F.
The term "Ci-C6 acyl" refers to a straight or branched -(CO)-Ci -C6 alkyl, with C-i-C-6 alkyl as defined herein.
The term "halogen" refers to fluoro, chloro, bromo, or iodo. "Halogens" are preferably fluorine, chlorine or bromine, being in particular fluorine or chlorine.
The term "phenyloxy" refers to a O-Phenyl. The term "aryloxy" refers to a O-Aryl where aryl is as defined herein. The term "hetereoaryloxy" refers to a O-Heteroaryl where Heteroaryl is as defined herein.
Pharmaceutically acceptable salts comprise conventional non-toxic salts obtained by salification of a compound of formula (I) with inorganic acids (e.g. hydrochloric, hydrobromic, sulphuric, or phosphoric acids), or with organic acids (e.g. acetic, propionic, succinic, benzoic, sulfanilic, 2-acetoxy-benzoic, cinnamic, mandelic, salicylic, glycolic, lactic, oxalic, malic, maleic, malonic, fumaric, tartaric, citric, p-toluenesulfonic, methanesulfonic, ethanesulfonic, or naphthalensulfonic acids). For reviews on suitable pharmaceutical salts see Berge S. M. et al., J. Pharm. Sci. 1977, 66, 1 -19; Gould P. L. Int. J. Pharm 1986, 33, 201 -217; and Bighley et al. Encyclopedia of Pharmaceutical Technology, Marcel Dekker Inc, New York 1996, Volume 13, page 453-497. Other salts, which are not pharmaceutically acceptable, for example the trifluoroacetate salt, may be useful in the preparation of compounds of this invention and these form a further aspect of the invention. The invention includes within its scope all possible stoichiometric and non-stoichiometric forms of the salts of the compounds of formula (I).
In addition, solvated forms of the compounds of formula (I) with acceptable solvents such as water, EtOH and the like, including hydrates of the compounds, as well as mixtures of the hydrate- and keto- form of the compounds, are within the scope of the present invention.
Certain compounds of formula (I) may exist in stereoisomeric forms (e.g. they may contain one or more asymmetric carbon atoms). The individual stereoisomers (enantiomers and diastereomers) and mixtures of these are included within the scope of the present invention. The present invention also covers the individual isomers of the compounds represented by formula (I) as mixtures with isomers thereof in which one or more chiral centres are inverted. Likewise, it is understood that compounds of the invention may exist in tautomeric forms other than that shown in the formula and these are also included within the scope of the present invention.
The invention also includes all suitable isotopic variations of a compound of the invention. Examples of isotopes that can be incorporated into compounds of the invention include isotopes such as 2H, 3H, 13C, 14C, 15N, 17O, 180, 31P, 32P, 35S, 18F and 36CI, respectively. Certain isotopic variations of the invention, for example, those in which a radioactive isotope such as 3H or 14C is incorporated, are useful in drug and/or substrate tissue distribution studies. Further, substitution with isotopes such as deuterium 2H, may afford certain therapeutic advantages resulting from greater metabolic stability. Isotopic variations of the compounds of the invention can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the examples hereafter using appropriate isotopic variations of suitable reagents.
The invention also provides pharmaceutical compositions comprising one or more compounds of this invention and one or more pharmaceutically acceptable excipient and/or diluent. The pharmaceutical compositions containing the active ingredient may be in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable or infusible liquid, solutions, suspensions, emulsions, suppositories, ointments, creams, lotions, gels, pastes, transdermal delivery devices.
Compounds of general formula (I), wherein R2 is COOH may be prepared according to Scheme A-l:
Figure imgf000025_0001
A11 A2 A12
Scheme A-l
wherein R is as defined above for fomula (I); PG is a protecting group chosen among those known in the art, for example methyl, ethyl etc..
Compounds of formula A1 and A2 are known compounds or can be prepared by known methods. Compounds of formula A3 can be obtained by reacting a compound of formula A1 with a compound of formula A2 in presence of an oxidant, for instance 2-hydroperoxy- 2-methyl-propane, K2S208, or (NH4)2S208, a transition metal salt, for instance FeS0 or AgN03, and an acid, for instance trifluoroacetic acid or H2S04, in a suitable solvent, for instance acetonitrile, water or dichloroethane, at a temperature ranging from room temperature to the boiling point of the solvent. Alternatively, the reaction can be carried out, in presence or absence of the transition metal salt, with tetrabutylammonium bromide (TBAB) instead of the acid. Optionally, the reaction can be carried out under microwave irradiation. A compound of formula A3, wherein R is methyl, can be converted into a compound of A3, wherein R is benzyl, by reacting a compound of A3 with iodobenzene or bromobenzene in presence of a suitable catalyst, for instance palladium acetate, a suitable ligand, for instance 2-(2-dicyclohexylphosphanylphenyl)-A/,A/-dimethyl-aniline, and a suitable base, for instance potassium te/t-butoxide, in a suitable solvent, for instance THF, at a temperature ranging from room temperature to the boiling temperature of the solvent. Preferably, the reaction is carried out under inert atmosphere. A compound of formula A3, wherein R is methyl, can be converted into a compound of A3, wherein R is 2-phenylethyl, by reacting a compound of A3 with benzaldehyde in presence of a base in a suitable solvent, for instance methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, providing a compound of A3, wherein R is phenylprop-2-enoyl. Subsequently, the double bond can be reduced providing a compound of A3, wherein R is 2-phenylethyl, with 3,5-bis(ethoxycarbonyl)- 1 ,4-dihydro-2,6-dimethylpyridine in presence of silica gel in a suitable solvent, for instance toluene, at a temperature ranging from room temperature to the boiling temperature of the solvent.
Alternatively compounds of formula A3, wherein R is methyl substituted by aryl or heteroaryl, can be obtained by reacting a compound of formula A11 with a compound of formula A2, wherein R is aryl or heteroaryl, in presence of a suitable base, such as KOH or CS2CO3, in a suitable solvent, for instance methanol or isopropanol and THF, at a temperature ranging from -40°C to room temperature, preferably under inert atmosphere, providing a compound of formula A12, wherein R is aryl or heteroaryl. Subsequently, the compound of formula A12, wherein R is aryl or heteroaryl can be hydrolysed in presence of a suitable acid, such as HCI, providing a compound of formula A3 wherein R is methyl substituted by aryl or heteroaryl. Compounds of formula A11 are known compounds or can be prepared by known methods (TeLe 39, 1998, 1717-1720, WO2005056550) Compounds of formula A4 can be obtained by reacting a compound of formula A3 with hydroxylamine hydrochloride and a suitable base, for instance sodium acetate, or hydroxylamine (NH2OH) in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent.
A compound of formula (I) can be obtained according to known methods, e.g. when the compound of formula A4 is a methyl, ethyl or te/t-butyl ester, by treatment of the ester with LiOH, NaOH or KOH in a suitable solvent, for example in ethanol/water, in THF, in THF/water, in ethanol/water, in methanol/water, or in a dioxane/ethanol/water mixture. The hydrolysis may be carried out at a temperature ranging from 0°C to the boiling point of the solvent. Alternatively, when the compound of formula A4 is a te/t-butyl ester, a compound of formula (I) can be obtained by treatment of a te/t-butyl ester derivative with TFA (trifluoroacetic acid) in a suitable solvent such as dichloromethane at a temperature ranging from 0°C to room temperature.
Alternatively, compounds of general formula (I), wherein R2 is COOH may be prepared according to Scheme A-l I:
Figure imgf000027_0001
A5 wherein R1 is methyl
substituted by aryl, wherein
aryl is substituted by hydroxy
Scheme A-ll
wherein R , R 6 and R 7 are as defined above for fomula (I); PG and PG are protecting groups chosen among those known in the art, for example methyl, ethyl, te/t-butyl, etc. for PG and 0-(tetrahydro-2H-pyran-2-yl), etc. for PG1.
Compounds of formula A3 can be prepared as described in Scheme A-l above. A compound of A3 can be treated with a protected hydroxylamine H2N-OPG1 under the same conditions as with hydroxylamine as disclosed above in Scheme A-l providing a compound of formula A5.
Alternatively, a compound of formula A5, wherein R is methyl substituted by X-R3, with X and R3 as defined above, can be prepared first by treating a compound of formula A3, wherein R is methyl, with bromine or a bromine/HBr mixture in a suitable solvent, for instance acetic acid or methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, then with a protected hydroxylamine H2N-OPG1 under the same conditions as with hydroxylamine as disclosed above in Scheme A-l and finally with HO-R3 in presence of a base, for instance sodium hydride (NaH) or K2CO3, or KO- R3 or H2NR3 in a suitable solvent, for instance methanol, ethanol, a methanol/ethanol mixture, THF or DMF, at a temperature ranging from 0°C to room temperature. Preferably, the reaction is carried out under inert atmosphere.
Alternatively, a compound of formula A5, wherein R is methyl, can be converted into a compound of A5, wherein R is benzyl, by reacting a compound of A5, wherein R is methyl, first with bromine or a bromine/HBr mixture in a suitable solvent, for instance acetic acid or methanol, at a temperature ranging from room temperature to the boiling temperature of the solvent, and then with phenylboronic acid in presence of a suitable catalyst, for instance palladium chloride, a suitable ligand, for instance cyclopentyl(diphenyl)phosphane, and a suitable Lewis acid, for instance cesium fluoride, in a suitable solvent, for instance THF, at a temperature ranging from room temperature to the boiling temperature of the solvent. Optionally, the reaction is carried out under microwave irradiation. Preferably, the reaction is carried out under inert atmosphere. A compound of formula A5, wherein R is methyl, can be converted at the same conditions into a compound of A5, wherein R is benzyl, wherein the benzyl is substituted by one or more substituents independently selected from the group consisting of halogen, C1 -C6 alkyl, Ci-C6 alkoxy, Ci-C6 haloalkyl, and Ci-C6 haloalkoxy.
Alternatively a compound of formula A5, wherein R is methyl substituted by aryl, wherein the aryl is substituted by hydroxy, can be converted into a compound of formula A5, wherein R is methyl substituted by aryl, wherein the aryl is substituted by C-2-C-6 alkoxy substituted by NR16R17, wherein R 6 and R 7 are as defined above for formula (I), by reaction with a compound of formula A13, wherein R 6 and R 7 are as defined above for formula (I), X is a suitable leaving group such as chlorine or bromine, n=1 -5, in presence of a suitable base, for instance K2CO3, Cs2CO3 or NaH, in a suitable solvent, for instance DMF, acetonitrile, acetone, at a temperature ranging from 0°C to the boiling point of the solvent. A compound of formula A13 is a known compound or can be prepared by known methods. A compound of formula A6 can be obtained according to known methods, e.g. when the PG group in A5 is methyl, ethyl or te/t-butyl, by treatment of the ester with LiOH, NaOH or KOH in a suitable solvent, for example in ethanol/water, in THF/water, in THF/ethanol/water, in methanol/water, or in a dioxane/ethanol/water mixture. The hydrolysis may be carried out at a temperature ranging from 0°C to the boiling point of the solvent. Alternatively, when the compound of formula A5 is a te/t-butyl ester, a compound of formula A6 can be obtained also by treatment of a te/t-butyl ester derivative with TFA (trifluoroacetic acid) in a suitable solvent such as dichloromethane at a temperature ranging from 0°C to room temperature.
A compound of formula A7 can be obtained according to known methods, e.g. in the case PG1 is tetrahydropyranyl, the deprotection of the hydroxylamine, can be achieved by using HCI in aprotic solvents (such as THF, diethylether, dioxane or a methanol/dioxane mixture) at a temperature ranging from 0°C to room temperature.
Finally, a compound of formula (I) can be obtained either by deprotection of the hydroxylamine of a compound of formula A6 under the same conditions as for obtaining a compound of formula A7, or by deprotection of the carboxylic acid of a compound of formula A7 under the same conditions as for obtaining a compound of formula A6.
Compounds of general formula (I), wherein R2 is COR4, CN, CH(OH)CF3 or C(OH)2CF3 and wherein R4 is O-C1-C6 alkyl, NR7R8, or CF3, may be prepared according to Scheme A-lll:
Figure imgf000030_0001
Figure imgf000030_0002
A10
Scheme A-lll
wherein R is as defined above for fomula (I); and R2 is COR4, CN, CH(OH)CF3 or C(OH)2CF3 and wherein R4 is O-Ci-C6 alkyl, NR7R8, or CF3 and R7 and R8 are as defined above.
Compounds of formula A8 are known compounds or can be prepared by known methods. Compounds of formula A9 can be obtained by reacting a compound of formula A8 with a compound of formula A2 under the same conditions as for the reaction of a compound of formula A1 with a compound of formula A2 as outlined above in Scheme A-l.
Compounds of formula (I), wherein R2 is COR4 or CN and wherein R4 is O-C1 -C6 alkyl or NR7R8, can be obtained by reacting a compound of formula A9 with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH2OH), for instance sodium acetate, in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent.
Alternatively, compounds of formula (I), wherein R2 is COR4 or CN and wherein R4 is O- C1-C6 alkyl or NR7R8, can be obtained first by reacting a compound of formula A9 with a protected hydroxylamine H2N-OPG1 under the same conditions as with hydroxylamine as disclosed above in Scheme A-l providing a compound of formula A10, and then by deprotection of the hydroxylamine according to known methods, e.g. in the case PG is tetrahydropyranyl, by using HCI in aprotic solvents (such as THF, diethylether or dioxane). Compounds of formula (I), wherein R2 is CH2OH, can be obtained first by treating a compound of formula A10, wherein R2 is -COOCH3, with a reducing agent, for instance LiAIH4 or c//'isobutylaluminum hydride, in an suitable solvent, for instance dichloromethane or THF, at a temperature ranging from -78°C to 0°C, and then by deprotection of the hydroxylamine according to known methods as described above. Preferably, the reduction step is carried out under inert atmosphere.
Compounds of formula (I), wherein R2 is COR4 and R4 is hydrogen can be obtained first by reacting a compound of formula A10, wherein R2 is -COOCH3, with a reducing agent, for instance LiAIH4 or of/lsobutylaluminum hydride, in an suitable solvent, for instance dichloromethane, at -78°C, and then by deprotection of the hydroxylamine according to known methods. Preferably, the reduction step is carried out under inert atmosphere. Compounds of formula (I), wherein R2 is COR4 and R4 is NR7R8 and R7 and R8 are as defined above, can be obtained first by hydrolyzing a compound of formula A10, wherein R2 is -COOCH3, with LiOH, NaOH or KOH in a suitable solvent, for example in ethanol/water, in THF/water, in THF/ethanol/water, in methanol/water, or in a dioxane/ethanol/water mixture. Then, the obtained carboxylic acid can be treated with an amine HNR7R8 with coupling agents, for instance benzotriazol-1 -yl- oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), benzotriazol-1 -yloxy- tris(dimethylamino)phosphonium hexafluorophosphate (BOP) or 1 -(3- dimethylaminopropyl)-3-ethylcarbodiimide (EDC), in the presence of a suitable base (e.g. triethylamine or di-isopropylethylamine) in a suitable solvent (e.g. tetrahydrofuran, dichloromethane or DMF). Generally, an activator of the condensation reaction, such as HOBt (1 -hydroxybenzotriazole) or HOAt (1 -hydroxy-7-aza-benzotriazole), can be added to the reaction mixture. The reaction can be carried out at room temperature for a period lasting between about 2 and 24 h. Finally, the deprotection of the hydroxylamine can be carried out according to known methods as described above.
Compounds of formula (I), wherein R2 is tetrazole, can be obtained first by reacting a compound of formula A10, wherein R2 is CN, with sodium azide (NaN3) in presence of triethylamine hydrochloride in a suitable solvent, for instance dimethylacetamide, dimethylformamide or A/-methylpyrrolidone, at a temperature ranging from room temperature to the boiling temperature of the solvent, and then by deprotection of the hydroxylamine according to known methods. Optionally, the coupling with NaN3 can be carried out under microwave irradiation.
Compounds of general formula (I), wherein R2 is CH(OH)CF3, can be obtained first by reacting a compound of formula A10, wherein R2 is COR4 and R4 is hydrogen, with trimethyl(trifluoromethyl)silane (CF3SiMe3) in presence of a base such as K2CO3 in a suitable solvent, for instance DMF, at a temperature ranging from room temperature to the boiling temperature of the solvent and then deprotecting the hydroxylamine and the alcohol, according to known methods, e.g. in the case PG1 is tetrahydropyranyl, by using HCI in aprotic solvents (such as THF, diethylether or dioxane).
Alternatively, a compound of general formula (I), wherein R2 is CH(OH)CF3 and R is methyl, can be obtained by reacting a compound of formula A9, wherein R2 is CH(OH)CF3 and R is methyl, with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH2OH), for instance sodium acetate, in a suitable solvent, for instance methanol, ethanol, a water/ethanol mixture, at a temperature ranging from room temperature to the boiling point of the solvent. A compound of formula A9, wherein R2 is CH(OH)CF3 and R is methyl, can be obtained from a compound of formula A14, wherein R2 is COR4 and R4 is hydrogen, by known methods. A compound of formula A14 is a known compound. Compounds of general formula (I), wherein R2 is (CO)R4 and R4 is CF3, can be obtained first by reacting a compound of formula A10, wherein R2 is CH(OH)CF3 with an oxidizing agent, such as IBX in a suitable solvent, for instance EtOAc, at a temperature ranging from room temperature to the boiling temperature of the solvent and then by deprotection of the hydroxylamine according to known methods, e.g. in the case PG1 is tetrahydropyranyl, by using HCI in aprotic solvents (such as THF, diethylether or dioxane). In the case it is necessary to protect a chemical group of a compound of the present invention and/or an intermediate thereof, before carrying out one of the aforedescribed reactions, said chemical group may be protected and deprotected according to known methods. A thorough discussion for suitable protecting groups and the means for protection/deprotection steps can be found for example in Greene and Wuts (Greene, T.W.; Wuts, P.G.M. "Protective Groups in Organic Synthesis", John Wiley & Sons Inc., 1991 ) or in Kocienski (Kocienski, P.J. "Protecting Groups", George Thieme Verlag, 1994). Salification of the compounds of formula (I), and preparation of compounds of formula (I), free of their salts, may be carried out by known conventional methods.
In view of the above described mechanism of action, the compounds of the present invention are useful in the prevention or treatment of tumor type diseases, including but not limited to: leukemia, lymphoma, esophageal squamous cell carcinoma, breast cancer, medulloblastoma, prostate cancer, colon cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreas cancer or glioblastomas. Preferably, the glioblastomas are giant cell glioblastoma or gliosarcoma. Still preferably, the leukemia or lymphoma are acute myeloid leukemia, mucosa-associated lymphoid tissue lymphoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL).
The compounds of formula (I) can also be used in combination with additional agents, in particular anti-tumor and differentiating agents, either by separate administrations, or by including the two active principles in the same pharmaceutical formulation. Non- exhaustive examples of suitable additional agents include:
a) histone deacetylase inhibitors (for example, but not limited to SAHA, PXD101 , JNJ- 26481585, SB939, ITF-2357, LBH589, PCI-24781 , valproic acid, butyric acid, MS-275, MGCD0103 and FK-228);
b) retinoid receptor modulators such as 13-c/s-retinoic acid, 9-c/s-retinoic acid, bexarotene, alitretinoin, or tretinoin; vitamin D;
c) antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (for example platin derivatives like c/s-platin, carboplatin, oxaliplatin, lobaplatin, satraplatin, nedaplatin, heptaplatin; nitrogen mustard such as chlorambucil, melphalan, chlormethine, cyclophosphamide, ifosfamide, trofosfamide, uramustine, bendamustine, estramustine; busulphan, temozolomide or nitrosoureas); antimetabolites (for example antifolates such as aminopterin, methotrexate, pemetrexed, raltitrexed); purines such as cladribine, clofarabine, fludarabine, mercaptopurine, pentostatin, thioguanine; pyrimidines like capecitabine, cytarabine, fluorouracil, floxuridine, gemcitabine; azacitidine, decitabine; cytosine arabinoside or hydroxyurea; antitumour antibiotics (for example anthracyclines like aclarubicin, amrubicin, daunomycin, doxorubicin, epirubicin, idarabicin, valrubicin, zorubicine; mitoxantrone; or antibiotics from streptomyces like actinomycin, bleomycin, mitomycin, or plicamycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine or vinorelbine; taxoids like docetaxel, paclitaxel or tesetaxel; epothilones like ixabepilone) and topoisomerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide; amsacrine, camptothecin, irinotecan, rubitecan, and topotecan);
d) cytostatic agents such as antioestrogens (for example tamoxifen, toremifene, raloxifene, droloxifene and idoxifene), oestrogen receptor down regulators (for example fulvestrant), antiandrogens (for example bicalutamide, flutamide, nilutamide, liarozole or cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin or buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5-alpha-reductase such as finasteride;
e) agents which inhibit cancer cell invasion (for example metalloproteinase inhibitors and inhibitors of urokinase plasminogen activator receptor function);
f) inhibitors of growth factor function, for example growth factor antibodies, growth factor receptor antibodies (for example the anti-erbb2 antibody trastuzumab as well as ado-trastuzumab emtasine and pertuzumab, the anti-erbbl antibody cetuximab and panitumumab, the anti IGF1 R antibody figitumumab), farnesyl transferase inhibitors, MEK inhibitor(for example trametinib), tyrosine kinase inhibitors and serine/threonine kinase inhibitors, for example enzastaurin, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, nilotinib, sorafenib, sunitinib, afatinib, axinitinib, bosutinib, caboratinib, ceritinib, crizotinib, dafrafenib, ibrutinib, idelalisib, lenvatinib, pazopanib, regorafenib, ruxolinitinib, vandetanib, vefuramenib, midostaurin, everolimus, sirolimus or temsirolimus;
g) antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, for example the anti-vascular endothelial cell growth factor antibody bevacizumab, ramucirumab, lenalidomide or thalidomide;
h) cell cycle inhibitors including for example CDK inhibitors (for example but not limited to flavopiridol, roscovitine, palbociclib and milciclib) and other inhibitors of cell cycle checkpoints; inhibitors of aurora kinase and other kinases involved in mitosis and cytokinesis regulation;
i) proteasome inhibitors (for example lactacystin, bortezomib, epoxomicin and its analog carfilzomib);
j) HSP90 inhibitors (for example but not limited to AT-13387, KOS-953, KOS-1022, CNF-1010, CNF-2024, SNX 5422, STA-9090, NVP-HSP990, NVP-AUY922, PU-H17 and XL-888);
k) Selective COX-2 inhibitors (for example celecoxib or parecoxib), or non selective NSAIDs (for example diclofenac, flurbiprofen, ibuprofen, ketoprofen, or naproxen).
In another aspect, a compound of general formula (I) can be used in combination with radiation therapy. In yet another aspect, a compound of general formula (I) may be administered in combination with standard chemotherapy combinations such as, but not restricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide, and paclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone);
I) anti CD20 antibodies (for example but not limited to ibritumomab tiuxetan,obinutuzumab,ofatumumab,rituximab and tositumomab), anti PD1 antibody (for example nivolumab), anti CD30 antibodies such as brentuximab vedotin, anti CLTA-4 antibodies (for example ipilimumab) and antibodies targeting soluble and membrane- bound interleukin 6 as siltuximab;
m) epigenetic drugs such as other histone demethylase inhibitors (for example but not limited to KDM1A and EZH2 inhibitors), and bromodomain inhibitors (for example but not limited to GSK525762, OTX015, CPI-0610, TEN-010 and BAY1238097);
n) IDH-1/2 inhibitors such as AG-120, AG-221 and AG-881.
In another aspect, a compound of general formula (I) can be used in combination with radiation therapy. In yet another aspect, a compound of general formula (I) may be administered in combination with standard chemotherapy combinations such as, but not restricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide, doxorubicin and 5-fluorouracil), AC (doxorubicin and cyclophosphamide), FEC (5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC (doxorubicin, cyclophosphamide, and paclitaxel), or CMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone). The invention also provides pharmaceutical compositions comprising one or more compounds of this invention and one or more pharmaceutically acceptable excipient and/or diluent. The pharmaceutical compositions can be chosen on the basis of the treatment requirements. Such compositions are prepared by blending and are suitably adapted to oral or parenteral administration, and as such can be administered in the form of tablets, capsules, oral preparations, powders, granules, pills, injectable, or infusible liquid solutions, suspensions, or suppositories.
Tablets and capsules for oral administration are normally presented in unit dose form and contain conventional excipients such as binders, fillers (including cellulose, mannitol, lactose), diluents, tableting agents, lubricants (including magnesium stearate), detergents, disintegrants (e.g. polyvinylpyrrolidone and starch derivatives such as sodium glycolate starch), coloring agents, flavoring agents, and wetting agents (for example sodium lauryl sulfate).
The oral solid compositions can be prepared by conventional methods of blending, filling or tableting. The blending operation can be repeated to distribute the active principle throughout compositions containing large quantities of fillers. Such operations are conventional.
Oral liquid preparations can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or can be presented as a dry product for reconstitution with water or with a suitable vehicle before use. Such liquid preparations can contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminium stearate gel, or hydrogenated edible fats; emulsifying agents, such as lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which can include edible oils), such as almond oil, fractionated coconut oil, oily esters such as esters of glycerine, propylene glycol, or ethyl alcohol; preservatives, such as methyl or propyl p- hydroxybenzoate or sorbic acid, and if desired, conventional flavoring or coloring agents. Oral formulations also include conventional slow-release formulations such as enterically coated tablets or granules.
Pharmaceutical preparation for administration by inhalation can be delivered from an insufflator or a nebulizer pressurized pack. For parenteral administration fluid unit dosages can be prepared, containing the compound and a sterile vehicle. The compound can be either suspended or dissolved, depending on the vehicle and concentration. The parenteral solutions are normally prepared by dissolving the compound in a vehicle, sterilising by filtration, filling suitable vials and sealing. Advantageously, adjuvants such as local anaesthetics, preservatives and buffering agents can also be dissolved in the vehicle. To increase the stability, the composition can be frozen after having filled the vials and removed the water under vacuum. Parenteral suspensions are prepared in substantially the same manner, except that the compound can be suspended in the vehicle instead of being dissolved, and sterilized by exposure to ethylene oxide before suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent can be included in the composition to facilitate uniform distribution of the compound of the invention.
For buccal or sublingual administration the compositions may be tablets, lozenges, pastilles, or gel.
The compounds can be pharmaceutically formulated as suppositories or retention enemas, e.g. containing conventional suppositories bases such as cocoa butter, polyethylene glycol, or other glycerides, for a rectal administration.
Another means of administering the compounds of the invention regards topical treatment. Topical formulations can contain for example ointments, creams, lotions, gels, solutions, pastes and/or can contain liposomes, micelles and/or microspheres. Examples of ointments include oleaginous ointments such as vegetable oils, animal fats, semisolid hydrocarbons, emulsifiable ointments such as hydroxystearin sulfate, anhydrous lanolin, hydrophilic petrolatum, cetyl alcohol, glycerol monostearate, stearic acid, water soluble ointments containing polyethylene glycols of various molecular weights. Creams, as known to formulation experts, are viscous liquids or semisolid emulsions, and contain an oil phase, an emulsifier and an aqueous phase. The oil phase generally contains petrolatum and an alcohol such as cetyl or stearic alcohol. Formulations suitable for topical administration to the eye also include eye drops, wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient.
A further method of administering the compounds of the invention regards transdermal delivery. Typical transdermal formulations comprise conventional aqueous and nonaqueous vectors, such as creams, oils, lotions or pastes or can be in the form of membranes or medicated patches.
A reference for the formulations is the book by Remington ("Remington: The Science and Practice of Pharmacy", Lippincott Williams & Wilkins, 2000).
The compounds of the present invention may be employed alone as a sole therapy or in combination with other therapeutic agents (see the list of additional agents is as indicated previously and comprises also standard chemotherapeutic agents) for the treatment of the above-mentioned conditions. The combination can be administered as separate compositions (simultaneous, sequential) of the individual components of the treatment or as a single dosage form containing both agents. When the compounds of this invention are in combination with others active ingredients, the active ingredients may be separately formulated into single-ingredient preparations of one of the above-described forms and then provided as combined preparations, which are given at the same time or different times, or may be formulated together into a two- or more- ingredient preparation.
Compounds of general formula (I) may be administered to a patient in a total daily dose of, for example, from 0.001 to 1000 mg/kg body weight daily. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose. The determination of optimum dosages for a particular patient is well known to one skilled in the art.
As is common practice, the compositions are normally accompanied by written or printed instructions for use in the treatment in question.
The following non limiting examples and biological data are presented and reference to the following figures is made in order to further illustrate the invention.
1. CHEMICAL SYNTHESIS
Unless otherwise indicated, commercially available reagents and solvents (HPLC grade) were used without further purification.
Specifically, the following abbreviations may have been used in the descriptions of the experimental methods:
NMR (Nuclear Magnetic Resonance) 1H (proton)
MHz (Megahertz) Hz (Hertz) HPLC (High Performance Liquid LC-MS (Liquid Chromatography Mass
Chromatography) Spectrum)
s (seconds) min (minutes) h (hours) mg (milligrams) g (grams) it (microlitres) mL (millilitres) mmol (millimoles) nm (nanometers) μΜ (micromolar)
M (molarity) NOE (Nuclear Overhauser Effect)
RP-SPE (reverse phase-solid phase
r.t. (room temperature) extraction)
ACN (acetonitrile) ACN-d3 (deuterated acetonitrile)
AcOH (acetic acid) AcONa (sodium acetate)
BOP ((Benzotriazol-1 - yloxy)tris(dimethylamino)phosphonium n-BuOH (n-butanol)
hexafluorophosphate
CDCI3 (deuterated chloroform) DCE (dichloroethane)
DCM (dichloromethane) DMA (dimethylacetamide)
DMSO-de (deuterated dimethyl
DMF (dimethylformamide)
sulfoxide)
Et20 (diethyl ether) EtOAc (ethyl acetate)
EtOK (potassium ethylate) EtONa (sodium ethylate)
HBr (hydrobromic acid) HCI (hydrochloric acid)
K2CO3 (potassium carbonate) KPS (potassium persulfate)
LiAIH4 (lithiumaluminum hydride) LiOH (lithium hydroxide)
MeOH (methanol) MeONa (sodium methylate)
NaH (sodium hydride) NH4CI (ammonium chloride)
NaHC03 (sodium bicarbonate) NaOH (sodium hydroxide)
PyBOP ((Benzotriazol-1 -
Na2S0 (sodium sulphate) yloxy)tripyrrolidinophosphonium hexafluorophosphate
TBAB (tetrabutylamonium bromide) TEA (triethylamine)
TFA (trifluoroacetic acid) THF (tetrahydrofurane) Except where indicated otherwise, all temperatures are expressed in °C (degrees centigrade) or K (Kelvin).
The 1H-NMR spectra were acquired with a Varian 500 MHz instrument. The chemical shifts are expressed in parts per million (ppm, δ units). The coupling constants are expressed in Hertz (Hz) and the splitting patterns are described as s (singlet), bs (broad signal), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet).
The LC-MS analyses were carried out on a Waters Acquity UPLC or Waters Acquity UPLC H-Class linked to with a SQD Single quadrupole (Waters) using an Acquity UPLC BEH C18 (50 x 2.1 mm, 1.7 μηι) or Acquity UPLC HSS T3 (50 x 2.1 mm, 1.8 μηι) column. Phase A was composed by either Milli-Q water/ACN 95/5 + 0.07% formic acid or Milli-Q water + 0.07% formic acid; Phase B by ACN + 0.05% formic acid; flow rate: 0.6 mL/min; UV detection (DIODE array) from 210 to 400 nm; ESI+ detection in the 100-2000 m/z range. The yields were calculated assuming that products were 100% pure if not stated otherwise.
Intermediate 1 : methyl 2-acetylpyridine-4-carboxylate
Figure imgf000040_0001
4.5 g (33 mmol) of methyl isonicotinate (Aldrich, Cat No. M52950-100G) and 18 g (66 mmol) of ferrous sulfate heptahydrate were dissolved in 100 mL of ACN and 2.8 mL (36 mmol) of TFA. 9.1 mL (66 mmol) of 70% te/t-butyl hydroperoxyde and and 3.7 mL (66 mmol) of acetaldehyde were added. The mixture was heated at reflux with stirring for about 3 h. The mixture was allowed to cool down to r.t. and the volatiles were removed under vacuum. The residue was taken up in 1 M aqueous NaOH and extracted with EtOAc. The organic phase was dried over Na2S04, concentrated and the residue was purified by column chromatography (eluent n-hexane:EtOAc 9:1 ) providing methyl 2- acetylpyridine-4-carboxylate (Intermediate 1 ) as a beige solid (3.61 g, 61 %). 1H NMR (CDCIs) δ (ppm): 8.85 (m, 1 H), 8.57 (m, 1 H), 8.04 (m, 1 H), 4.00 (s, 3 H), 2.77 (s, 3 H); MS (ESI): m/z: 180 [M+H]+ Intermediate 2: ferf-butyl 2-acetylpyridine-4-carboxylate
Figure imgf000041_0001
3.7 g (60%) of te/t-butyl 2-acetylpyridine-4-carboxylate (Intermediate 2) was prepared according to the procedure described for Intermediate 1 , starting from 5.0 g (28 mmol) of te/t-butyl isonicotinate and 9.39 ml_ (167 mmol) of acetaldehyde. 1H NMR (CDCI3) δ (ppm): 8.82 (m, 1 H), 8.49 (m, 1 H), 8.00 (m, 1 H), 2.76 (s, 3 H), 1.63 (s, 9 H); MS (ESI): m/z: 222 [M+H]+.
Intermediate 3: 2-acetylpyridine-4-carbonitrile
Figure imgf000041_0002
1.6 g (46%) of 2-acetylpyridine-4-carbonitrile (Intermediate 3) was prepared according to the procedure described for Intermediate 1 , starting from 2.5 g (24 mmol) of 4- cyanopyridine (Aldrich, Cat No C95005-100G) and 8.1 ml_ (164 mmol) of acetaldehyde. 1H NMR (CDCI3) δ (ppm): 8.98-8.78 (m, 1 H), 8.35-8.20 (m, 1 H), 7.82-7.60 (m, 1 H), 2.74 (s, 3 H); MS (ESI): m/z: 147 [M+H]+.
Intermediate 4: methyl 2-benzoylpyridine-4-carboxylate
Figure imgf000041_0003
0.075 g (21 %) of methyl 2-benzoylpyridine-4-carboxylate (Intermediate 4) was prepared according to the procedure described for Intermediate 1 , starting from 0.20 g (1.5 mmol) of methyl isonicotinate and 0.88 ml_ (8.8 mmol) of benzaldehyde. 1 H NMR (CDCI3) δ (ppm): 8.95-8.82 (m, 1 H), 8.63-8.53 (m, 1 H), 8.13-7.99 (m, 3 H), 7.70-7.45 (m, 3 H), 4.02 (s, 3 H); MS (ESI): m/z: 242 [M+H]+.
Intermediate 5: methyl 2-(cyclopropanecarbonyl)pyridine-4-carboxylate
Figure imgf000042_0001
0.075 g (25%) of methyl 2-(cyclopropanecarbonyl)pyridine-4-carboxylate (Intermediate 5) was prepared according to the procedure described for Intermediate 1 , starting from 0.20 g (1 .5 mmol) of methyl isonicotinate and 0.41 g (5.8 mmol) of cyclopropanaldehyde. 1H NMR (CDCIs) δ (ppm): 8.98-8.80 (m, 1 H), 8.65-8.44 (m, 1 H), 8.05 (dd, J=1.7, 5.1 Hz, 1 H), 4.00 (s, 3 H), 3.59-3.32 (m, 1 H), 1.41-1.03 (m, 4 H); MS (ESI): m/z: 206 [M+H]+. Intermediate 6: methyl 2-propanoylpyridine-4-carboxylate
Figure imgf000042_0002
0.13 g (31 %) of methyl 2-propanoylpyridine-4-carboxylate (Intermediate 6) was prepared according to the procedure described for Intermediate 1 , starting from 0.30 g (2.2 mmol) of methyl isonicotinate and 0.76 g (13 mmol) of propanaldehyde. 1H NMR (CDCI3) δ (ppm): 8.84 (dd, J=1.0, 4.9 Hz, 1 H), 8.61-8.51 (m, 1 H), 8.04 (dd, J=1.5, 4.9 Hz, 1 H), 4.00 (s, 3 H), 3.27 (q, J=7.3 Hz, 2 H), 1.25 (t, J=7.3 Hz, 3 H); MS (ESI): m/z: 194 [M+H]+. Intermediate 7: methyl 2-(2-methoxybenzoyl)pyridine-4-carboxylate
Figure imgf000042_0003
A mixture of 0.200 g (1.46 mmol) of methyl isonicotinate, 0.79 g (5.8 mmol) of 2- methoxybenzaldehyde, 0.10 g (0.40 mmol) of TBAB, and 0.79 g (2.9 mmol) of KPS in 15 ml_ of DCE was stirred at 1 10°C for about 20 h. After cooling down to r.t, saturated NaHC03 was added, and the mixture was extracted with EtOAc. The combined organic layers were dried over Na2S04 and concentrated under reduced pressure. The crude product was purified by flash chromatography (eluent n-hexane:EtOAc from 95:5 to 80:20), followed by trituration in Et.20 to afford methyl 2-(2-methoxybenzoyl)pyridine-4- carboxylate (Intermediate 7, 0.09 g, 23%). 1H NMR (CDCI3) δ (ppm): 8.84-8.76 (m, 1 H), 8.53-8.46 (m, 1 H), 8.03-7.97 (m, 1 H), 7.62-7.57 (m, 1 H), 7.56-7.49 (m, 1 H), 7.14- 7.06 (m, 1 H), 7.03-6.95 (m, 1 H), 4.01 (s, 3 H), 3.64 (s, 3 H); MS (ESI): m/z: 272 [M+H]+. Intermediate 8: methyl 2-(4-acetoxybenzoyl)pyridine-4-carboxylate
Figure imgf000043_0001
A mixture of 0.515 g (3.76 mmol) of methyl isonicotinate, 2.41 g (17.7 mmol) of 4- acetoxybenzaldehyde, 0.355 g (1.10 mmol) of TBAB, and 2.00 g (7.40 mmol) of KPS in 20 ml_ of DCE was stirred at 1 10°C for about 20 h. After cooling to r.t, saturated NaHC03 was added, and the mixture was extracted with EtOAc. The combined organic layers were dried over Na2SO and concentrated under reduced pressure. The crude product was purified by flash chromatography (eluent n-hexane: EtOAc from 92:8 to 82:18), followed by trituration in Et2O to afford methyl 2-(4-acetoxybenzoyl)pyridine-4-carboxylate (Intermediate 8) as a colourless solid (0.1 1 g, 10%). 1H NMR (CDCI3) δ (ppm): 8.92-8.84 (m, 1 H), 8.63-8.52 (m, 1 H), 8.22-8.12 (m, 2 H), 8.07 (dd, J=1.5, 4.9 Hz, 1 H), 7.32-7.19 (m, 2 H), 4.02 (s, 3 H), 2.35 (s, 3 H); MS (ESI): m/z: 300 [M+H]+.
Intermediate 9: methyl 2-(3-methoxybenzoyl)pyridine-4-carboxylate
Figure imgf000043_0002
0.057 g (14%) of methyl 2-(3-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 9) was prepared according to the procedure described for Intermediate 8, starting from 0.200 g (1 .46 mmol) of methyl isonicotinate and 0.794 g (5.83 mmol) of 3- methoxybenzaldehyde. 1H NMR (DMSO-d6) δ (ppm): 8.98-8.89 (m, 1 H), 8.38-8.29 (m, 1 H), 8.10 (dd, J=1.7, 5.1 Hz, 1 H), 7.57-7.43 (m, 3 H), 7.31-7.23 (m, 1 H), 3.95 (s, 3 H), 3.80 (s, 3 H); MS (ESI): m/z: 272 [M+H]+.
Intermediate 10: ferf-butyl 2-(2-phenylacetyl)pyridine-4-carboxylate
Figure imgf000044_0001
0.24 g (1 .1 mmol) of te/t-butyl 2-acetylpyridine-4-carboxylate (Intermediate 2) and 0.1 1 ml_ (0.98 mmol) of iodobenzene were dissolved in 4 ml_ of dry THF under argon atmosphere. 0.004 g (0.02 mmol) of palladium acetate, 0.02 g (0.04 mmol) of 2-(2- dicyclohexylphosphanylphenyl)-A/,A/-dimethyl-aniline and 0.24 g (2.2 mmol) of potassium te/t-butylate were subsequently added. Argon was bubbled into the mixture for about 10 min and the mixture was heated at 80°C for about 2 h. After cooling down to r.t, the reaction mixture was slowly quenched with AcOH, filtered through a thin pad of Celite®, and washed thoroughly with EtOAc. The combined organic layers were washed with saturated NaHC03, brine, dried and concentrated. The residue was purified by flash chromatography (eluent: n-hexane: EtOAc from 98:2 to 80:20) to afford te/t-butyl 2-(2- phenylacetyl)pyridine-4-carboxylate (Intermediate 10) as an orange oil (0.08 g, 24%). 1H NMR (CDCIs) δ (ppm): 8.93-8.78 (m, 1 H), 8.56-8.37 (m, 1 H), 8.00 (dd, J=1.5, 4.9 Hz, 1 H), 7.38-7.18 (m, 5 H), 4.57 (s, 2 H), 1.68-1.53 (m, 9 H); MS (ESI): m/z: 298 [M+H]+. Intermediate 11 : methyl 2-(2-bromoacetyl)pyridine-4-carboxylate
Figure imgf000044_0002
1.00 g (5.58 mmol) of methyl 2-acetylpyridine-4-carboxylate (Intermediate 1 ) was suspended in 0.450 mL (28 mmol; 33% in AcOH) of HBr. The mixture was stirred at r.t. for about 5 min, then 0.302 mL (5.86 mmol) of bromine were added quickly; stirring was continued for about 2 min, the mixture was cooled to 0°C, water was added and the aqueous phase was extracted with EtOAc. The combined organic layers were dried and concentrated. The resulting crude methyl 2-(2-bromoacetyl)pyridine-4-carboxylate (Intermediate 11 , 1 .34 g, 65%) was used in the next steps without any further purification. MS (ESI): m/z: 258 [M+H]+.
Intermediate 12: methyl 2-Z-(C-(bromomethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate
Figure imgf000045_0001
1 .32 g (3.58 mmol) of methyl 2-(2-bromoacetyl)pyridine-4-carboxylate (Intermediate 11 ) was dissolved in 15 mL of dry MeOH under nitrogen in a sealed cap vial. The resulting solution was added drop wise to a solution of 0.41 g (3.58 mmol) of O-tetrahydropyran- 2-ylhydroxylamine in 15 mL of dry MeOH at 0°C. The mixture was stirred 5 min at 0°C, then 6 h at r.t.. Water was added and the mixture was extracted with DCM. The combined organic layers were dried, concentrated and the crude product was purified by flash chromatography (eluent n-hexane:EtOAc from 98:2 to 80:20) to afford 0.68 g (53%) of methyl 2-Z-(C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4- carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81-8.77 (m, 1 H), 8.57-8.55 (m, 1 H), 7.89-7.85 (m, 1 H), 5.62-5.58 (m, 1 H), 4.73 (s, 2 H), 4.06-3.95 (m, 4 H), 3.74- 3.68 (m, 1 H), 2.02-1 .90 (m, 3 H), 1 .79-1 .60 (m, 3 H); MS (ESI): m/z: 379 [M+Na]+. Intermediate 13: methyl 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000046_0001
0.075 g (96%) of methyl 2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 13) was prepared according to the procedure described for Intermediate 12, starting from 0.050 g (0.28 mmol) of methyl 2- acetylpyridine-4-carboxylate (Intermediate 1 ) and 0.045 g (0.39 mmol) of O- tetrahydropyran-2-ylhydroxylamine and heating the reaction mixture at reflux for about 20 h. 1H NMR (CDCIs) δ (ppm): 8.86-8.62 (m, 1 H), 8.57-8.39 (m, 1 H), 7.88-7.64 (m, 1 H), 5.56-5.40 (m, 1 H), 4.05-3.57 (m, 5 H), 2.45 (s, 3 H), 1.97-1.62 (m, 6 H); MS (ESI): m/z: 301 [M+Na]+.
Intermediate 14: 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000046_0002
A solution of 0.075 g (0.27 mmol) of methyl 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 13) and 0.016 g (0.67 mmol) of LiOH in 7 ml_ of THF and 1 .4 ml_ of water was stirred at r.t. for about 48 h. THF was removed under vacuum, the residue diluted with water and washed with DCM. The aqueous phase was brought to pH 3 with AcOH, while cooling in an ice/water bath, then it was extracted with DCM:n-BuOH 8:2. The combined organic layers were evaporated, the residue was treated with Et2O at r.t., filtered and dried under vacuum providing 0.055 g (77%) of 2- [(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 14) as a colourless solid. 1H NMR (CDCI3) δ (ppm): 8.82-8.67 (m, 1 H), 8.63-8.47 (m, 1 H), 7.82 (dd, J=1 .2, 5.1 Hz, 1 H), 5.69-5.54 (m, 1 H), 4.07-3.71 (m, 2 H), 2.47 (s, 3 H), 2.04-1.55 (m, 6 H); MS (ESI): m/z: 263 [M-H]".
Intermediate 15: 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbonitrile
Figure imgf000047_0001
0.283 g (84%) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carbonitrile (Intermediate 15) was prepared according to the procedure described for Intermediate 12, starting from 0.200 g (1.37 mmol) of 2-acetylpyridine-4-carbonitrile (Intermediate 3) and 0.232 g (1 .98 mmol) of O-tetrahydropyran-2-ylhydroxylamine. 1H NMR (CDCI3) δ (ppm): 8.84-8.64 (m, 1 H), 8.35-8.19 (m, 1 H), 7.47 (dd, J=1.5, 5.4 Hz, 1 H), 5.54-5.41 (m, 1 H), 4.02-3.61 (m, 2 H), 2.41 (s, 3 H), 1.97-1.58 (m, 6 H); MS (ESI): m/z: 162 [M-THP]+.
Intermediate 16: methyl 2-[(E)-C-[(4-methylpiperazin-1 -yl)methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000047_0002
0.04 g (0.1 1 mmol) of methyl 2-Z-(C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12) were added at 0°C to a suspension of 0.01 1 g (0.1 1 mmol) of 4-methyl piperazine in 1.2 mL of dry MeOH. After stirring for about 30 min, a further portion of 0.01 1 g (0.1 1 mmol) of 4-methyl piperazine was added and the mixture stirred for additional 1 h. The mixture was diluted with DCM and water and the aqueous phase was extracted with DCM/MeOH 90:10. The combined organic layers were dried, concentrated and the crude product was purified by flash chromatography (eluent DCM:MeOH 97:3) to afford 0.085 g (83%) of methyl 2-[(E)-C-[(4- methylpiperazin-1 -yl)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 16). 1H NMR (CDCI3) δ (ppm): 8.79-8.68 (m, 1 H), 8.46-8.36 (m, 1 H), 7.86-7.75 (m, 1 H), 5.57-5.46 (m, 1 H), 4.09-3.88 (m, 4 H), 3.74-3.61 (m, 1 H), 2.98-1.42 (m, 19 H); MS (ESI): m/z: 377 [M+H]+.
Intermediate 17: methyl 2-[(Z)-C-(methoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000048_0001
A solution of 0.04 g (0.1 1 mmol) of methyl 2-Z-(C-(bromomethyl)-A/-tetrahydropyran-2- yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12) in 1 ml_ of dry MeOH was added at 0°C and under nitrogen atmosphere to 0.026 ml_ of MeONa 25% wt. in MeOH. The reaction mixture was allowed to warm to r.t, stirred at r.t. for about 3 h and then quenched with 5% citric acid. The mixture was extracted with DCM, the combined organic layers were dried, concentrated and the crude mixture was purified by flash chromatography (eluent n-hexane:EtOAc from 95:5 to 60:40) to afford 0.019 g (55%) of methyl 2-[(Z)-C-(methoxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 17). 1H NMR (CDCI3) δ (ppm): 8.81-8.77 (m, 1 H), 8.47-8.44 (m, 1 H), 7.85-7.80 (m, 1 H), 5.55-5.51 (m, 1 H), 4.83 (s, 2 H), 3.99-3.90 (m, 4 H), 3.73- 3.65 (m, 1 H), 3.41 (s, 3 H), 1.96-1.84 (m, 3 H), 1.79-1.54 (m, 3 H); MS (ESI): m/z: 309 [M+H]+.
Intermediate 18: 2-[(Z)-C-(ethoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000049_0001
0.018 g (42%) of 2-[(Z)-C-(ethoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 18) was prepared according to the procedure described for Intermediate 17, starting from 0.050 g (0.14 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12) and 1 .5 g (4.3 mmol) of EtOK. The reaction was quenched after the addition of EtOK at 0°C. 1H NMR (CDCI3) δ (ppm): 8.79 (m, 1 H), 8.50-8.44 (m, 1 H), 7.82 (m, 1 H), 5.59 (bs, 1 H), 4.86 (s, 2 H), 4.01-3.91 (m, 1 H), 3.80-3.70 (m, 1 H), 3.62 (q, J=1 .0 Hz, 2 H), 1 .98-1 .82 (m, 3 H), 1 .79-1 .57 (m, 3 H), 1 .17 (t, J=1 .0 Hz, 3 H); MS (ESI): m/z: 307[M-H]-.
Intermediate 19: 2-[(Z)-C-[2-(ferf-butoxycarbonyl(methyl)amino)ethoxymethyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000049_0002
A solution of 0.04 g (0.1 1 mmol) of methyl 2-Z-(C-(bromomethyl)-A/-tetrahydropyran-2- yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12) in 0.5 mL of dry THF was added drop wise at 0°C and under nitrogen atmosphere to a suspension of 0.024 g (0.13 mmol) of te/t-butyl A/-(2-hydroxyethyl)-A/-methyl-carbamate and 0.0054 g (0.13 mmol) of NaH (60% mineral oil) in 0.5 mL of dry THF. Once the addition was complete, the reaction mixture was quenched with 5% citric acid and extracted with DCM and DCM/MeOH 90: 10. The combined organic layers were dried, concentrated and the crude product was purified by flash chromatography (eluent DCM:MeOH 97:3) to afford 0.022 g (27%) of 2- [(Z)-C-[2-(te/t-butoxycarbonyl(methyl)amino)ethoxymethyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 19). MS (ESI): m/z: 436 [M-H]". Intermediate 20: 2-[(Z)-C-[2-(ferf-butoxycarbonylamino)ethoxymethyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000050_0001
0.042 g (58%) of 2-[(Z)-C-[2-(te/t-butoxycarbonylamino)ethoxymethyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 20) was prepared according to the procedure described for Intermediate 19, starting from 0.040 g (0.1 1 mmol) of 2-Z-(C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine- 4-carboxylate (Intermediate 12). The reaction was carried out at r.t. for about 20 h. 1H NMR (DMSO-de) δ (ppm): 14.12-13.56 (m, 1 H), 8.81 (m, 1 H), 8.64-8.63 (m, 1 H), 8.27- 8.13 (m, 1 H), 7.92-7.79 (m, 1 H), 6.75-6.60 (m, 1 H), 5.46-5.42 (m, 1 H), 4.76-4.68 (m, 1 H), 3.82-3.71 (m, 1 H), 3.61-3.52 (m, 1 H), 3.44 (t, J=6.0 Hz, 2 H), 3.37-3.32 (m, 2 H), 1.88-1.70 (m, 3 H), 1.68-1.47 (m, 3 H), 1.41-1.30 (m, 9 H); MS (ESI): m/z: 424 [M+H]+. Intermediate 21 : 2-[(Z)-C-[2-(benzyl(ferf-butoxycarbonyl)amino)ethoxymethyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000050_0002
0.03 g (35%) of 2-[(Z)-C-[2-(benzyl(te/t-butoxycarbonyl)amino)ethoxymethyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 21 ) was prepared according to the procedure described for Intermediate 19, starting from 0.060 g (0.168 mmol) of 2-Z-(C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). The reaction was carried out at r.t. for about 48. 1 H NMR (CDCI3) δ (ppm): 8.82 (m, 1 H), 8.52-8.44 (m, 1 H), 7.94-7.86 (m, 1 H), 7.31-7.08 (m, 5 H), 5.59-5.55 (m, 1 H), 4.95-4.85 (m, 2 H), 4.36 (s, 2 H), 3.99- 3.89 (m, 1 H), 3.79-3.59 (m, 3 H), 3.47-3.19 (m, 2 H), 1 .97-1 .55 (m, 6 H), 1 .51 -1 .34 (m, 9 H); MS (ESI): m/z: 514 [M+H]+.
Intermediate 22: methyl 2-[(Z)-C-(phenoxymethyl)-/V-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000051_0001
A mixture of 0.06 g (0.17 mmol) of methyl 2-Z-(C-(bromomethyl)-A/-tetrahydropyran-2- yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12), 0.017 g (0.18 mmol) of phenol and 0.046 g (0.34 mmol) of K2C03 in 1 mL of dry DMF was stirred at r.t. for about 3 h. Saturated aqueous NH4CI was added and the mixture was concentrated to dryness. The residue was taken up in DCM and water. The aqueous phase was extracted with DCM and the combined organic layers were dried, concentrated and the crude product was purified by flash chromatography (eluent DCM:EtOAc 99: 1 ) to afford 0.033 g (53%) of methyl 2-[(Z)-C-(phenoxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 22). 1 H NMR (CDCI3) δ (ppm): 8.80 (m, 1 H), 8.49-8.47 (m, 1 H), 7.89 (m, 1 H), 7.30-7.24 (m, 2 H), 6.99-6.91 (m, 3 H), 5.59-5.55 (m, 1 H), AB System: VA=5.47, VB=5.44, JAB= 1 1 .2 Hz, 3.98 (s, 3 H), 3.97-3.90 (m, 1 H), 3.73-3.65 (m, 1 H), 1 .95-1 .52 (m, 6 H); MS (ESI): m/z: 371 [M+H]+.
Intermediate 23: 2-[(Z)-C-(methoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000052_0001
A solution of 0.018 g (0.0060 mmol) of methyl 2-[(Z)-C-(methoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 17) and 0.003 g (0.1 mmol) of LiOH in 1 mL of THF and 0.3 mL of water was stirred at r.t. for about 1 h. THF was removed under vacuum, the residue was treated with NH4CI and DCM. The aqueous phase was acidified with 5% citric acid and the product was extracted with DCM. The combined organic layers were dried and concentrated to afford about 0.017 g (98%) of 2-[(Z)-C-(methoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 23). MS (ESI): m/z: 293 [M-H]. Intermediate 24: 2-[(Z)-C-(phenoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000052_0002
0.020 g (69%) of 2-[(Z)-C-(phenoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 24) was prepared according to the procedure described for Intermediate 23, starting from 0.030 g (0.0080 mmol) of methyl 2-[(Z)-C-(phenoxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 22). 1H NMR (CDCI3) δ (ppm): 8.81 (m, 1 H), 8.52 (bs, 1 H), 7.88 (dd, J=1 .2, 4.9 Hz, 1 H), 7.34-7.18 (m, 2 H), 7.03-6.87 (m, 3 H), 5.66-5.61 (m, 1 H), 4.01-3.92 (m, 1 H), 3.80-3.71 (m, 1 H), 1 .97-1 .50 (m, 6 H); MS (ESI): m/z: 357 [M+H]+. Intermediate 25: 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxamide
Figure imgf000053_0001
0.035 mL (0.25 mmol) of TEA, 0.065 g (0.13 mmol) of PyBOP and 0.010 g (0.13 mmol) of ammonium carbonate were subsequently added under an argon atmosphere to a solution of 0.033 g (0.13 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 14) in 0.7 mL of dry THF. The mixture was stirred at r.t. for about 24 h, then 0.013 g (0.025 mmol) of PyBOP and 0.002 g (0.025 mmol) of ammonium carbonate were added and stirring was continued for further 20 h. The reaction mixture was poured into a saturated aqueous NaHCO3 solution and extracted with DCM. The combined organic layers were dried, concentrated and the crude mixture was purified by chromatography (eluent: DCM: MeOH from 99:1 to 90:10) to afford 0.0257 g (78%) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxamide (Intermediate 25) as a colourless solid. 1H NMR (CDCIs) δ (ppm): 8.84-8.64 (m, 1 H), 8.31-8.14 (m, 1 H), 7.79-7.60 (m, 1 H), 6.30 (bs, 1 H), 5.77 (bs, 1 H), 5.54-5.36 (m, 1 H), 4.05-3.10 (m, 3 H), 2.45 (s, 3 H), 2.01-1.54 (m, 5 H); MS (ESI): m/z: 286 [M+Na]+.
Intermediate 26: A -ethyl-2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxamide
Figure imgf000053_0002
0.027 g (73%) of N-ethyl-2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxamide (Intermediate 26) was prepared according to the procedure described for Intermediate 25, starting from 0.033 g (0.125 mmol) of 2-[(E)-C- methyl-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 14) and 0.075 ml_ (0.15 mmol) of ethyl amine. 1H NMR (CDCI3) δ (ppm): 8.84-8.59 (m, 1 H), 8.28-8.16 (m, 1 H), 7.82-7.64 (m, 1 H), 6.37 (bs, 1 H), 5.60-5.40 (m, 1 H), 4.04-3.83 (m, 1 H), 3.77-3.41 (m, 3 H), 2.46 (s, 3 H), 2.05-1.53 (m, 6 H), 1.38-1.15 (m, 3 H); MS (ESI): m/z: 292 [M+H]+.
Intermediate 27: 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy-carbonimidoyl]-A - tetrahydropyran-2-yloxy-pyridine-4-carboxamide
Figure imgf000054_0001
0.043 g (57%) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]-/V- tetrahydropyran-2-yloxy-pyridine-4-carboxamide (Intermediate 27) was prepared according to the procedure described for Intermediate 25, starting from 0.055 g (0.21 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 14), 0.037 g (0.31 mmol) of O-tetrahydropyran-2-ylhydroxylamine and using 0.13 g (0.29 mmol) of BOP as the condensating agent. 1H NMR (CDCI3) δ (ppm): 9.16 (bs, 1 H), 8.82-8.61 (m, 1 H), 8.26-8.06 (m, 1 H), 7.72-7.59 (m, 1 H), 5.48 (bs, 1 H), 5.1 1 (bs, 1 H), 4.12-3.56 (m, 4 H), 2.44 (s, 3 H), 2.05-1.51 (m, 12 H); MS (ESI): m/z: 364 [M+H]+.
Intermediate 28: A -tetrahydropyran-2-yloxy-1 -[4-(1 H-tetrazol-5-yl)-2- pyridyl]ethanimine
Figure imgf000055_0001
A solution of 0.16 g (0.65 mmol) of 2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbonitrile (Intermediate 15), 0.064 g (0.98 mmol) of sodium azide and 0.13 g (0.98 mmol) of triethylamine hydrochloride in 20 ml_ of DMA was heated under microwave irradiation at 160°C for about 4 h. Twice further 0.064 g (0.98 mmol) of sodium azide and 0.13 g (0.98 mmol) of triethylamine hydrochloride were added and the mixture was heated under microwave irradiation at 160°C for about 2 h. While cooling in an ice/water bath, the reaction was quenched by carefully adding saturated aqueous NaHC03. The mixture was extracted with DCM; the aqueous phase was brought to pH 3 with AcOH, and extracted with DCM/n-BuOH 8:2. The combined organic layers were concentrated and the crude was purified by chromatography (eluent DCM :MeOH: AcOH from 100:0:0 to 95:5:0.5), followed by treatment with EtOAc (decanted) and Et2O/MeOH, to afford 0.025 g (13%) of A/-tetrahydropyran-2-yloxy-1 -[4-(1 H-tetrazol-5-yl)-2- pyridyl]ethanimine (Intermediate 28) as a beige solid, upon filtration of the resulting suspension. 1H NMR (DMSO-d6) δ (ppm): 8.91-8.77 (m, 1 H), 8.53-8.43 (m, 1 H), 8.02 (dd, J=1.7, 5.1 Hz, 1 H), 5.57-5.33 (m, 1 H), 3.88-3.52 (m, 2 H), 2.36 (s, 3 H), 1.87-1.45 (m, 6 H); MS (ESI): m/z: 287 [M-H].
Intermediate 29: 2-[(E)-C-methyl-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbaldehyde and Intermediate 30: [2-[(E)-C-methyl-/V- tetrahydropyran-2-yloxy-carbonimidoyl]-4-pyridyl]methanol
Figure imgf000056_0001
0.44 ml_ (0.44 mmol) of 1 M LiAIH4 in THF was added drop wise to a solution of 0.1 g (0.36 mmol) of methyl 2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 13) in 2.8 ml_ of dry DCM at -78°C. After 20 min of stirring at -78°C the reaction mixture was quenched with NH4CI, water was added and the two phases separated. The aqueous phase was extracted with DCM and the combined organic layers were dried, concentrated and the two products were separated by chromatography (eluent n-hexane:EtOAc from 95:5 to 35:65) to afford 0.035 g (39%) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carbaldehyde (Intermediate 29); 1H NMR (CDCI3) δ (ppm): 10.1 1 (s, 1 H), 8.91-8.75 (m,
1 H), 8.47-8.36 (m, 1 H), 7.70 (dd, J=1.5, 5.4 Hz, 1 H), 5.56-5.46 (m, 1 H), 4.02-3.62 (m,
2 H), 2.47 (s, 3 H), 2.01-1.57 (m, 6 H); MS (ESI): m/z:165 [M-THP]+; and 0.018 g (20%) of [2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy-carbonimidoyl]-4-pyridyl]methanol (Intermediate 30): 1H NMR (CDCI3) δ (ppm): 8.67-8.52 (m, 1 H), 8.04-7.90 (m, 1 H), 7.44-7.33 (m, 1 H), 5.56-5.42 (m, 1 H), 4.79 (s, 2 H), 4.01-3.57 (m, 2 H), 2.46 (s, 3 H), 1.99-1.58 (m, 6 H); MS (ESI): m/z: 251 [M+H]+.
Intermediate 31 : methyl 2-(4-methoxybenzoyl)pyridine-4-carboxylate
Figure imgf000056_0002
0.156 g (39%) of methyl 2-(4-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 31 ) was prepared according to the procedure described for Intermediate 8, starting from 0.200 g (1 .46 mmol) of methyl isonicotinate and 0.794 g (5.83 mmol) of 4- methoxybenzaldehyde. 1H NMR (CDCI3) δ (ppm): 8.93-8.83 (m, 1 H), 8.57-8.50 (m, 1 H), 8.17-8.10 (m, 2 H), 8.09-7.99 (m, 1 H), 7.04-6.94 (m, 2 H), 4.02 (s, 3 H), 3.91 (s, 3 H); MS (ESI): m/z: 272 [M+H]+.
Intermediate 32: methyl 2-[(E)-C-(p-tolylmethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000057_0001
0.038 g (0.28 mmol) of p-tolylboronic acid, 0.085 g (0.56 mmol) of cesium fluoride, 0.01 g (0.01 mmol) of (bisdiphenylphosphino)ferrocene dichloro palladium(ll) and 0.05 g (0.14 mmol) of methyl 2-[(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 12) were charged in a dried flask, evacuated and back filled with nitrogen. The mixture was stirred under microwave irradiation at 1 10°C for about 1 h, then cooled to r.t, filtered through a Celite pad, and washed with DCM. The solvents were removed under reduced pressure and the residue was purified by flash chromatography (eluent cyclohexane:EtOAc 9:1 ) to yield 0.025 g (49%) of methyl 2-[(E)-C-(p-tolylmethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 32) as a yellow oil. 1H NMR (CDCI3) δ (ppm): 8.79-8.75 (m, 1 H), 8.53-8.48 (m, 1 H), 7.82-7.77 (m, 1 H), 7.22 (m, 2 H), 7.03 (m, 2 H), 5.57-5.52 (m, 1 H), 4.48-4.33 (m, 2 H), 3.95 (s, 3 H), 3.81-3.72 (m, 1 H), 3.67- 3.59 (m, 1 H), 2.28 (s, 3 H), 1.93-1.82 (m, 3 H), 1.71-1.56 (m, 3 H); MS (ESI): m/z: 369 [M+H]+.
Intermediate 33: 2-[(E)-C-(p-tolylmethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000058_0001
0.014 g (63%) of 2-[(E)-C-(p-tolylmethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 33) was prepared according to the procedure described for Intermediate 14, starting from 0.023 g (0.062 mmol) of methyl 2- [(E)-C-(p-tolylmethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 32) with the only difference that a mixture of MeOH (1 mL) and water (0.5 mL) was used as reaction solvent. 1 H NMR (CDCI3) δ (ppm): 8.81-8.75 (m, 1 H), 8.57- 8.52 (m, 1 H), 7.85-7.79 (m, 1 H), 7.22 (m, 2 H), 7.04 (m, 2 H), 5.67-5.59 (m, 1 H), 4.49- 4.33 (m, 2 H), 3.83-3.66 (m, 2 H), 2.28 (s, 3 H), 1 .96-1 .83 (m, 3 H), 1 .76-1 .55 (m, 3 H); MS (ESI): m/z: 355 [M+H]+.
Intermediate 34: methyl 2 [(Z)-C-[[3-[3-(ferf- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000058_0002
0.074 g (68%) of methyl 2 [(Z)-C-[[3-[3-(fert- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 34) was prepared according to the procedure described for Intermediate 22, starting from 0.070 g (0.196 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12) and te/t-butyl N-[3-[(3-hydroxyphenyl)amino]-3-oxo-propyl]carbamate. 1H NMR (CDCIs) δ (ppm): 8.77 (m, 1 H), 8.47-8.43 (m, 1 H), 7.88-7.83 (m, 1 H), 7.65 (bs,
1 H), 7.26-7.1 1 (m, 3 H), 6.73-6.68 (m, 1 H), 5.58-5.53 (m, J=3.6, 3.6 Hz, 1 H), 5.42 (s,
2 H), 5.17 (bs, 1 H), 4.01-3.93 (m, 4 H), 3.73-3.66 (m, J=1 1.5 Hz, 1 H), 3.49 (s, 2 H), 2.59 (s, 2 H), 1.97-1.54 (m, 6 H), 1.44 (s, 9 H); MS (ESI): m/z: 557 [M+H]+.
Intermediate 35: 2-[(Z)-C-(2-acetamidoethoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000059_0001
0.026 g (36%) of 2-[(Z)-C-(2-acetamidoethoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 35) was prepared according to the procedure described for Intermediate 19, starting from 0.070 g (0.196 mmol) of 2- [(Z)-C- (bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
(Intermediate 12) and A/-acetylethanolamine (Fluorochem, cat. Nbr. 094790), with the only difference that the reaction was carried out at r.t. for about 3 h. 1H NMR (CDCI3) δ (ppm): 8.68 (bs, 2 H), 8.40 (bs, 1 H), 7.80 (bs, 1 H), 6.51 (bs, 1 H), 5.48 (bs, 1 H), 4.85 (s, 2 H), 3.95-3.86 (m, 1 H), 3.71-3.64 (m, 1 H), 3.63-3.57 (m, 2 H), 3.43-3.33 (m, 2 H), 1.96-1.75 (m, 6 H), 1.73-1.54 (m, 3 H); MS (ESI): m/z: 366 [M+H]+.
Intermediate 36: 2-[(Z)-C-[[3-[3-(ferf- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid o O H
Figure imgf000060_0001
O
0.045 g (71 %) of 2-[(Z)-C-(2-acetamidoethoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 36) was prepared according to the procedure described for Intermediate 23, starting from 0.065 g (0.1 17 mmol) of methyl 2 [(Z)-C-[[3-[3-(te/ -butoxycarbonylamino)propanoylamino]phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 34). 1H NMR (CDCIs) δ (ppm): 8.76 (m, 1 H), 8.49-8.40 (m, 1 H), 7.89-7.81 (m, 2 H), 7.25-7.08 (m, 3 H), 6.72 (s, 1 H), 5.62-5.53 (m, 1 H), 5.40 (bs, 2 H), 5.25 (bs, 1 H), 3.97 (s, 1 H), 3.76- 3.67 (m, 1 H), 3.50 (q, J=6.0 Hz, 2 H), 2.60 (bs, 2 H), 1.96-1.54 (m, 6 H), 1.44 (bs, 9 H); MS (ESI): m/z: 543 [M+H]+.
Intermediate 37: 2-[(Z)-C-(benzyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000060_0002
0.018 g (25%) of 2-[(Z)-C-(benzyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 37) was prepared according to the procedure described for Intermediate 19, starting from 0.070 g (0.20 mmol) of 2- [(Z)-C- (bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
(Intermediate 12) with the only difference that the reaction was carried out at r.t. for about 48 h. 1H NMR (CDCI3) δ (ppm): 8.79-8.73 (m, 1 H), 8.48-8.42 (m, 1 H), 7.89-7.79 (m, 1 H), 7.34-7.20 (m, 5 H), 5.56-5.50 (m, 1 H), 4.92 (s, 2 H), 4.61 (s, 2 H), 3.94-3.83 (m, 1 H), 3.71-3.60 (m, 1 H), 1 .92-1 .53 (m, 6 H); MS (ESI): m/z: 371 [M+H]+.
Intermediate 38: 2-[(Z)-C-(phenethyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000061_0001
0.020 g (27%) of 2-[(Z)-C-(phenethyloxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 38) was prepared according to the procedure described for Intermediate 19, starting from 0.070 g (0.20 mmol) of 2- [(Z)-C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
(Intermediate 12) with the only difference that the reaction was carried out at r.t. for about 24 h. 1H NMR (CDCI3) δ (ppm): 8.80-8.76 (m, 1 H), 8.45-8.41 (m, 1 H), 7.85-7.81 (m, 1 H), 7.24-7.1 1 (m, 5 H), 5.61-5.57 (m, 1 H), 4.92 (s, 2 H), 3.98-3.87 (m, 1 H), 3.81-3.70 (m, 3 H), 2.89-2.83 (m, 2 H), 1 .96-1 .52 (m, 6 H); MS (ESI): m/z: 383 [M-H].
Intermediate 39: methyl 2-[(Z)-C-(2-methoxy-phenyloxymethyl)-/V-tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000061_0002
0.10 g (92%) of methyl 2-[(Z)-C-(2-methoxy-phenyloxymethyl)-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 39) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4- carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80-8.74 (m, 1 H), 8.48-8.42 (m, 1 H), 7.86-7.78 (m, 1 H), 7.09-7.02 (m, 1 H), 6.96-6.81 (m, 3 H), 5.55-5.42 (m, 3 H), 3.97 (s, 3 H), 3.92-3.83 (m, 1 H), 3.75 (s, 3 H), 3.68-3.61 (m, 1 H), 1.87-1.78 (m, 3 H), 1.70-1.59 (m, 3 H); MS (ESI): m/z: 401 [M+H]+.
Intermediate 40: 2-[(Z)-C-(2-methoxy-phenyloxymethyl)-A -tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000062_0001
0.086 g (99%) of 2-[(Z)-C-(2-methoxy-phenyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 40) was prepared according to the procedure described for Intermediate 23, starting from 0.090 g (0.22 mmol) of methyl 2- [(Z)-C-(2-methoxy-phenyloxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylate (Intermediate 39). MS (ESI): m/z: 385[M-H]\
Intermediate 41 : methyl 2-[(Z)-C-(3-methoxy-phenyloxymethyl)-A -tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000062_0002
0.10 g (91 %) of methyl 2-[(Z)-C-(3-methoxy-phenyloxymethyl)-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 41 ) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4- carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.79-8.73 (m, 1 H), 8.49-8.43 (m, 1 H), 7.86-7.78 (m, 1 H), 7.21-7.10 (m, 1 H), 6.62-6.39 (m, 3 H), 5.59-5.52 (m, 1 H), 5.47-5.34 (m, 2 H), 3.99-3.89 (m, 4 H), 3.76 (s, 3 H), 3.72-3.65 (m, 1 H), 1 .90-1 .79 (m, 3 H), 1 .73-1 .54 (m, 3 H); MS (ESI): m/z: 401 [M+H]+.
ntermediate 42: 2-[(Z)-C-(3-methoxy-phenyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000063_0001
2-[(Z)-C-(3-methoxy-phenyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 42) was prepared according to the procedure described for Intermediate 23, starting from 0.090 g (0.22 mmol) of methyl 2- [(Z)-C-(3-methoxy-phenyloxymethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylate (Intermediate 41 ). MS (ESI): m/z: 385[M-H]".
Intermediate 43: 2-[(E)-C-(anilinomethyl)-/V-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000063_0002
methyl 2 (E)-C-(anilinomethyl)^-tetrahvdropyran-2-yloxy-carbonimidoyllpyricline-4- carboxylate
0.070 g (0.20 mmol) of methyl 2-Z-(C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12) was added to a suspension of 1 .03 mL (1 1 .3 mmol) of aniline in 1 .2 mL of dry MeOH at 0°C. The mixture was stirred at 0°C for about 1 .5 h, then at r.t. for about 18 h, diluted with water, extracted with 9: 1 DCM:MeOH. The combined organic layers were dried over Na2S04, the solvent was evaporated and the residue purified by flash chromatography (eluent DCM:EtOAc 9:1 ) to yield 0.035 g (48%) of methyl 2-[(E)-C-(anilinomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate. 1H NMR (CDCI3) δ (ppm): 8.78-8.72 (m, 1 H), 8.45-8.41 (m, 1 H), 7.83-7.80 (m, 1 H), 7.14-7.09 (m, 2 H), 6.76-6.64 (m, 3 H), 5.55- 5.50 (m, 1 H), 4.80-4.62 (AB System: VA=1 1942.08, VB=1 1891.45, JAB= 1 1912073.5 Hz), 3.99-3.91 (m, 4 H), 3.74-3.66 (m, 1 H), 1.81-1.58 (m, 6 H); MS (ESI): m/z: 370 [M+H]+.
2-i(E)-C-(anilinomethyl)-N-tetrahvdropyran-2-yloxy-carbonimidoyllpyridi
acid
0.022 g (76%) of 2-[(E)-C-(anilinomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 43) was prepared according to the procedure described for Intermediate 23, starting from 0.030 g (0.081 mmol) of methyl 2- [(E)-C-(anilinomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate. 1H NMR (CDCI3) δ (ppm): 8.78-8.70 (m, 1 H), 8.48-8.42 (m, 1 H), 7.83-7.76 (m, 1 H), 7.19-7.12 (m, 2 H), 6.87-6.71 (m, 3 H), 5.62-5.54 (m, 1 H), 4.79-4.67 (AB System: VA=4.75, VB=4.71 , JAB= 14.8 Hz), 3.96-3.87 (m, 1 H), 3.78-3.71 (m, 2 H), 1.81-1.59 (m, 6 H); MS (ESI): m/z: 356 [M+H]+.
Intermediate 44: 2-[(E)-C-(methylaminomethyl)-/V-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000064_0001
0.030 g (37%) of 2-[(E)-C-(methylaminomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 44) was prepared according to the procedure described for Intermediate 19, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12) and 0.073 g (0.56 mmol) of fert-butyl N-methylcarbamate. 1H NMR (CDCI3) δ (ppm): 8.84-8.79 (m, 1 H), 8.51-8.47 (m, 1 H), 7.88-7.82 (m, 1 H), 5.63-5.57 (m, 1 H), 4.86 (s, 2 H), 4.02-3.92 (m, 1 H), 3.82-3.72 (m, 1 H), 3.42 (s, 3 H), 2.01-1 .58 (m, 6 H); MS (ESI): m/z: 293[M+H]+.
Intermediate 45: methyl 2-[(Z)-C-[(2-fluorophenoxy)methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000065_0001
0.057 g (87%) of methyl 2-[(Z)-C-[(2-fluorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 45) was prepared according to the procedure described for Intermediate 22, starting from 0.06 g (0.17 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1 H NMR (CDCI3) δ (ppm): 8.79-8.74 (m, 1 H), 8.51-8.47 (m, 1 H), 7.86- 7.83 (m, 1 H), 7.17-7.09 (m, 1 H), 7.07-7.00 (m, 2 H), 6.95-6.87 (m, 1 H), 5.56-5.53 AB System: VA=5.5, VB=5.49, JAB= 6 Hz, 5.52-5.45 (m, 2 H), 3.97 (s, 3 H), 3.93-3.86 (m, 1 H), 3.70-3.63 (m, 1 H), 1 .93-1 .50 (m, 6 H); MS (ESI): m/z: 389[M+H]+.
Intermediate 46: 2-[(Z)-C-[(2-fluorophenoxy)methyl]-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000065_0002
0.045 g (87%) of 2-[(Z)-C-[(2-fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 46) was prepared according to the procedure described for Intermediate 23, starting from 0.056 g (0.14 mmol) of methyl 2- [(Z)-C-[(2-fluorophenyloxy)methyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridi carboxylate (Intermediate 45). 1H NMR (DMSO-d6) δ (ppm): 13.87 (bs, 1 H), 8.83-8.80 (m, 1 H), 8.27 (s, 1 H), 7.88-7.85 (m, 1 H), 7.35-7.30 (m, 1 H), 7.21 -7.1 1 (m, 2 H), 6.99- 6.93 (m, 1 H), 5.50-5.47 (m, 1 H), 5.39 (s, 2 H), 3.76-3.69 (m, 1 H), 3.57-3.50 (m, 1 H), 1 .81-1 .39 (m, 6 H); MS (ESI): m/z: 375[M+H]+.
Intermediate 47: methyl 2-[(Z)-C-[[4-[3-(ferf- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000066_0001
0.050 g (62%) of methyl 2-[(Z)-C-[[4-[3-(fert- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 47) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4 -carboxylate (Intermediate 12). 1 H NMR (CDCI3) δ (ppm): 8.79-8.74 (m, 1 H), 8.46-8.43 (m, 1 H), 7.86- 7.83 (m, 1 H), 7.40 (s, 2 H), 6.93 (s, 2 H), 5.58-5.53 (m, 1 H), 5.45-5.35 (AB System: VA=5.42, VB=5.4, JAB= 5.5 Hz), 4.00-3.89 (m, 4 H), 3.73-3.64 (m, 1 H), 3.54-3.42 (m, 2 H), 2.62-2.54 (m, 2 H), 1 .95-1 .53 (m, 6 H), 1 .48-1 .39 (m, 9 H); MS (ESI): m/z: 557[M+H]+.
Intermediate 48: 2-[(Z)-C-[[4-[3-(ferf- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000067_0001
0.035 g (65%) of 2-[(Z)-C-[[4-[3-(tert- butoxycarbonylamino)propanoylamino]phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 48) was prepared according to the procedure described for Intermediate 23, starting from 0.055 g (0.10 mmol) of methyl 2- [(Z)-C-[[4-[3-(te/ -butoxycarbonylamino)propanoylamino]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 47). 1H NMR (DMSO-de) δ (ppm): 14.09-13.56 (m, 1 H), 9.86-9.68 (m, 1 H), 8.83-8.79 (m, 1 H), 8.26- 8.23 (m, 1 H), 7.88-7.83 (m, 1 H), 7.50-7.45 (m, 2 H), 6.94-6.88 (m, 2 H), 6.85 (bs, 1 H), 5.50-5.45 (m, 1 H), 5.27 (s, 2 H), 3.81-3.72 (m, 1 H), 3.56 (s, 1 H), 3.22-3.15 (m, 2 H), 2.45-2.38 (m, 2 H), 1.85-1.41 (m, 6 H), 1.37 (s, 9 H); MS (ESI): m/z: 543[M+H]+.
Intermediate 49: methyl 2-[(Z)-C-[[3-(ferf-butoxycarbonylamino)phenoxy]methyl]- /V-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000067_0002
0.080 g (74%) of methyl 2-[(Z)-C-[[3-(te/t-butoxycarbonylamino)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 49) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2-[(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80- 8.75 (m, 1 H), 8.43-8.40 (m, 1 H), 7.85-7.81 (m, 1 H), 7.17-6.99 (m, 2 H), 6.73-6.55 (m, 2 H), 5.56-5.50 (m, 1 H), 5.46-5.35 (m, 2 H), 4.09-4.02 (m, 1 H), 3.96 (s, 3 H), 3.74-3.65 (m, 1 H), 2.00-1.57 (m, 6 H), 1.54-1 .49 (m, 9 H); MS (ESI): m/z: 486[M+H]+.
Intermediate 50: 2-[(Z)-C-[[3-(ferf-butoxycarbonylamino)phenoxy]methyl]-A - tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000068_0001
0.052 g (76%) of 2-[(Z)-C-[[3-(te/t-butoxycarbonylamino)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 50) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.14 mmol) of methyl 2-[(Z)-C-[[3-(te/t-butoxycarbonylamino)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 49). 1H NMR (DMSO-de) δ (ppm): 13.86 (bs, 1 H), 9.40-9.23 (m, 1 H), 8.86-8.74 (m, 1 H), 8.32-8.19 (m, 1 H), 7.92-7.75 (m, 1 H), 7.17-7.10 (m, 2 H), 7.03-6.97 (m, 1 H), 6.63-6.58 (m, 1 H), 5.50-5.46 (m, 1 H), 5.30-5.21 (AB System: VA=5.27, VB=5.25, JAB= 10.5 Hz), 3.81- 3.72 (m, 1 H), 3.62-3.49 (m, 1 H), 1.82-1.42 (m, 15 H); MS (ESI): m/z: 472[M+H]+.
Intermediate 51 : methyl 2-[(Z)-C-[[4-(ferf-butoxycarbonylamino)phenoxy]methyl]- /V-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000068_0002
0.074 g (68%) of methyl 2-[(Z)-C-[[4-(te/t-butoxycarbonylamino)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 51 ) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1 H NMR (CDCI3) δ (ppm): 8.79- 8.75 (m, 1 H), 8.46-8.42 (m, 1 H), 7.86-7.80 (m, 1 H), 7.26-7.21 (m, 2 H), 6.93-6.87 (m, 2 H), 6.32 (bs, 1 H), 5.57-5.52 (m, 1 H), 5.44-5.34 (AB System: VA=5.41 , VB=5.37, JAB= 1 1 Hz), 4.00-3.87 (m, 4 H), 3.73-3.64 (m, 1 H), 1 .93-1 .55 (m, 6 H), 1 .51 (s, 9 H); MS (ESI): m/z: 486[M+H]+.
Intermediate 52: 2-[(Z)-C-[[4-(ferf-butoxycarbonylamino)phenoxy]methyl]-A - tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000069_0001
0.054 g (93%) of 2-[(Z)-C-[[4-(te/t-butoxycarbonylamino)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 52) was prepared according to the procedure described for Intermediate 23, starting from 0.060 g (0.12 mmol) of methyl 2-[(Z)-C-[[4-(te/t-butoxycarbonylamino)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 51 ). 1H NMR (DMSO-de) δ (ppm): 13.85 (bs, 1 H), 9.14 (bs, 1 H), 8.88-8.73 (m, 1 H), 8.28-8.18 (m, 1 H), 7.92-7.80 (m, 1 H), 7.37-7.26 (m, 2 H), 6.92-6.82 (m, 2 H), 5.52-5.42 (m, 1 H), 5.25 (s, 2 H), 3.82-3.69 (m, 1 H), 3.61 -3.48 (m, 1 H), 1 .84-1 .37 (m, 15 H); MS (ESI): m/z: 472[M+H]+.
Intermediate 53: methyl 2-[(Z)-C-[(4fluorophenoxy)methyl]-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000070_0001
0.050 g (57%) of methyl 2-[(Z)-C-[(4-fluorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 53) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.79-8.75 (m, 1 H), 8.47-8.45 (m, 1 H), 7.85-7.81 (m, 1 H), 6.98-6.88 (m, 4 H), 5.56-5.52 (m, 1 H), 5.42-5.34 (AB System: VA=5.39, VB=5.37, JAB= 10.7 Hz), 3.99-3.88 (m, 4 H), 3.73-3.64 (m, 1 H), 1 .95-1 .48 (m, 6 H); MS (ESI): m/z: 389[M+H]+.
Intermediate 54: 2-[(Z)-C-[(4fluorophenoxy)methyl]-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000070_0002
0.038 g (88%) of 2-[(Z)-C-[(4fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 54) was prepared according to the procedure described for Intermediate 23, starting from 0.045 g (0.12 mmol) of methyl 2- [(Z)-C-[(4-fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 53). 1H NMR (DMSO-d6) δ (ppm): 13.87 (bs, 1 H), 8.83-8.78 (m, 1 H), 8.27-8.24 (m, 1 H), 7.88-7.83 (m, 1 H), 7.14-7.08 (m, 2 H), 7.03-6.97 (m, 2 H), 5.50-5.46 (m, 1 H), 5.29 (s, 2 H), 3.80-3.70 (m, 1 H), 3.60-3.52 (m, 1 H), 1 .83-1 .39 (m, 6 H); MS (ESI): m/z: 375[M+H]+. Intermediate 55: methyl 2-[(Z)-C-[(3-fluorophenoxy)methyl]-A -tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000071_0001
0.052 g (60%) of methyl 2-[(Z)-C-[(3-fluorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 55) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80-8.74 (m, 1 H), 8.49-8.44 (m, 1 H), 7.86-7.81 (m, 1 H), 7.24-7.16 (m, 1 H), 6.78-6.62 (m, 3 H), 5.59-5.54 (m, 1 H), 5.46- 5.36 (AB System: VA=5.42, VB=5.4, JAB= 1 1 Hz), 4.00-3.88 (m, 4 H), 3.73-3.64 (m, 1 H), 1 .95-1 .51 (m, 6 H); MS (ESI): m/z: 389[M+H]+.
Intermediate 56: 2-[(Z)-C-[(3-fluorophenoxy)methyl]-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000071_0002
0.025 g (86%) of 2-[(Z)-C-[(3-fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 56) was prepared according to the procedure described for Intermediate 23, starting from 0.030 g (0.077 mmol) of methyl 2- [(Z)-C-[(3-fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 55). 1H NMR (DMSO-d6) δ (ppm): 13.87 (bs, 1 H), 8.83-8.79 (m, 1 H), 8.28-8.24 (m, 1 H), 7.88-7.84 (m, 1 H), 7.35-7.26 (m, 1 H), 6.95-6.89 (m, 1 H), 6.86-6.74 (m, 2 H), 5.52-5.47 (m, 1 H), 5.33 (s, 2 H), 3.81 -3.72 (m, 1 H), 3.60-3.52 (m, 1 H), 1 .84-1 .41 (m, 6 H); MS (ESI): m/z: 375[M+H]+.
Intermediate 57: methyl 2-[(Z)-C-(2-pyridyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000072_0001
0.10 g (97%) of methyl 2-[(Z)-C-(2-pyndyloxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 57) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.74-8.70 (m, 1 H), 8.51-8.47 (m, 1 H), 7.86-7.80 (m, 1 H), 7.49-7.44 (m, 1 H), 7.29-7.23 (m, 1 H), 6.54-6.47 (m, 1 H), 6.13- 6.07 (m, 1 H), 5.57-5.26 (m, 3 H), 3.96 (s, 3 H), 3.71-3.57 (m, 2 H), 1 .83-1 .53 (m, 6 H); MS (ESI): m/z: 394[M+Na]+.
Intermediate 58: 2-[(Z)-C-(pyrimidin-2-yloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000072_0002
0.10 g (96%) of methyl 2-[(Z)-C-(pyrimidin-2-yloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 58) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxyl^ (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.74-8.70 (m, 1 H), 8.51-8.47 (m, 1 H), 7.86-7.80 (m, 1 H), 7.49-7.44 (m, 1 H), 7.29-7.23 (m, 1 H), 6.54-6.47 (m, 1 H), 6.13- 6.07 (m, 1 H), 5.57-5.26 (m, 3 H), 3.96 (s, 3 H), 3.71-3.57 (m, 2 H), 1 .83-1 .53 (m, 6 H); MS (ESI): m/z: 373[M+H]+.
Intermediate 59: methyl 2-[(Z)-C-(1 -naphthyloxymethyl)-/V-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000073_0001
0.068 g (58%) of methyl 2-[(Z)-C-(1 -naphthyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 59) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81-8.71 (m, 1 H), 8.53-8.39 (m, 1 H), 8.10-8.00 (m, 1 H), 7.87-7.79 (m, 1 H), 7.79-7.73 (m, 1 H), 7.49-7.31 (m, 4 H), 7.09- 6.97 (m, 1 H), 5.66-5.53 (m, 3 H), 4.01-3.61 (m, 5 H), 1 .91 -1 .45 (m, 6 H); MS (ESI): m/z: 421 [M+H]+.
Intermediate 60: 2-[(Z)-C-(1 -naphthyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000073_0002
0.020 g (41 %) of 2-[(Z)-C-(1 -naphthyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 60) was prepared according to the procedure described for Intermediate 23, starting from 0.050 g (0.12 mmol) of methyl 2- [(Z)-C-(1 -naphthyloxymethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 59). 1H NMR (DMSO-d6) δ (ppm): 13.87 (bs, 1 H), 8.86-8.73 (m, 1 H), 8.36-8.24 (m, 1 H), 7.92-7.77 (m, 3 H), 7.54-7.32 (m, 4 H), 7.23-7.12 (m, 1 H), 5.77-5.45 (m, 3 H), 3.80-3.48 (m, 2 H), 1 .52 (bs, 6 H); MS (ESI): m/z: 407[M+H]+.
Intermediate 61 : methyl 2-[(Z)-C-(2-naphthyloxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000074_0001
0.088 g (75%) of methyl 2-[(Z)-C-(2-naphthyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 61 ) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.83-8.74 (m, 1 H), 8.52-8.41 (m, 1 H), 7.90-7.63 (m, 4 H), 7.51-7.31 (m, 3 H), 7.16-7.04 (m, 1 H), 5.63-5.46 (m, 3 H), 4.04- 3.64 (m, 5 H), 1 .96-1 .51 (m, 6 H); MS (ESI): m/z: 421 [M+H]+.
Intermediate 62: 2-[(Z)-C-(2-naphthyloxymethyl)-/V-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000074_0002
0.067 g (99%) of 2-[(Z)-C-(2-naphthyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 62) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.17 mmol) of methyl 2- [(Z)-C-(2-naphthyloxymethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 61 ). 1H NMR (DMSO-d6) δ (ppm): 13.87 (bs, 1 H), 8.91-8.68 (m, 1 H), 8.35-8.18 (m, 1 H), 7.94-7.71 (m, 4 H), 7.55-7.29 (m, 3 H), 7.1 1 (dd, J=2A, 8.8 Hz, 1 H), 5.58-5.34 (m, 3 H), 3.90-3.48 (m, 2 H), 1 .84-1 .38 (m, 6 H); MS (ESI): m/z: 407[M+H]+.
Intermediate 63: methyl 2-(4-hydroxybenzoyl)pyridine-4-carboxylate
Figure imgf000075_0001
0.85 g (1 1 mmol) of ammonium acetate was added to a suspension of 0.33 g (1 .1 mmol) of methyl 2-(4-acetoxybenzoyl)pyridine-4-carboxylate (Intermediate 8) in 35 ml_ of a mixture of MeOH:water (4: 1 , v:v) and the resulting mixture was stirred at r.t. for about 16 h. 0.30 g (3.9 mmol) of ammonium acetate was added and stirring was continued for additional 4 h. Then, the solvents were removed under vacuum, the residue taken up with water and extracted with EtOAc and the combined organic layers were dried over Na2S04. EtOAc was removed and the raw material was purified by flash chromatography (eluent hexane: EtOAc from 90:10 to 70:30) to give 0.15 g (66%) of methyl 2-(4- hydroxybenzoyl)pyridine-4-carboxylate (Intermediate 63) as a colourless solid. 1 H NMR (CDCIs) δ (ppm): 8.97-8.84 (m, 1 H), 8.57-8.52 (m, 1 H), 8.14-8.07 (m, 1 H), 8.07-8.01 (m, 2 H), 6.95-6.87 (m, 2 H), 4.03 (s, 3 H); MS (ESI): m/z: 258[M+H]+.
Intermediate 64: methyl 2-[(E)-C-(4-hydroxyphenyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate and Intermediate 65 : methyl 2-[(Z)-C-(4- hydroxyphenyl)-/V-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000076_0001
0.062 g (33%) of methyl 2-[(E)-C-(4-hydroxyphenyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 64) and 0.082 (43%) of methyl 2-[(Z)- C-(4-hydroxyphenyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 65) was prepared according to the procedure described for Intermediate 12, starting from 0.15 g (0.58 mmol) of methyl 2-(4-hydroxybenzoyl)pyridine-4- carboxylate (Intermediate 63) and 1 .1 g (9.2 mmol) of O-tetrahydropyran-2- ylhydroxylamine. The reaction mixture was heated at reflux for about 20 h and then under microwave irradiation at 135°C for about 6 h. The two obtained products, the E-isomer and the Z-isomer of the oxime, were separated by flash chromatography (eluent hexane:EtOAc from 92:8 to 70:30). Intermediate 64 (E-isomer): 1H NMR (CDCI3) δ (ppm): 8.71-8.82 (m, 1 H) 8.41 (s, 1 H) 7.81-7.89 (m, 1 H) 7.33-7.41 (m, 2 H) 6.78-6.86 (m, 2 H) 5.49-5.56 (m, 1 H) 3.99 (s, 3 H) 3.86-3.95 (m, 1 H) 3.65-3.74 (m, 1 H) 1 .47-1 .89 (m, 6 H); MS (ESI): m/z: 357 [M+H]+. Intermediate 65 (Z-isomer): 1 H NMR (CDCI3) δ (ppm): 8.86-8.94 (m, 1 H), 8.15 (s, 1 H), 7.88-7.93 (m, J=1 .70, 5.10 Hz, 1 H), 7.28-7.37 (m, 2 H), 6.69-6.78 (m, 2 H), 5.44-5.52 (m, 1 H), 4.01 (s, 3 H), 3.84-3.93 (m, 1 H), 3.63-3.73 (m, 1 H), 1 .43-1 .80 (m, 6 H); MS (ESI): m/z: 357 [M+H]+.
Intermediate 66: methyl 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000077_0001
0.032 g (0.23 mmol) of K2C03 was added to a solution of 0.050 g (0.14 mmol) of methyl 2-[(E)-C-(4-hydroxyphenyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 64) and 0.021 ml_ (0.17 mmol) of 1 -(bromomethyl)-4-fluoro- benzene in 2.5 ml_ of a mixture of ACN:acetone (1 .5:1 , v;v). The mixture was stirred at r.t. for about 20 h and at 40°C for about 2 h. Then the solution was filtered and the filtrate was evaporated providing a yellow residue, which was treated with Et20 and filtered to give 0.044g (68%) of methyl 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 66). 1H NMR (CDCI3) δ (ppm): 8.78-8.72 (m, 1 H), 8.42-8.36 (m, 1 H), 7.86-7.81 (m, 1 H), 7.51-7.46 (m, 2 H), 7.46-7.41 (m, 2 H), 7.14-7.07 (m, 2 H), 7.06-7.00 (m, 2 H), 5.56-5.50 (m, 1 H), 5.08 (s, 2 H), 3.98 (s, 3 H), 3.95-3.87 (m, 1 H), 3.74-3.65 (m, 1 H), 1.89-1.49 (m, 6 H); MS (ESI): m/z: 465[M+H]+.
Intermediate 67: 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A -tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000077_0002
0.028 g (74%) of 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 67) was prepared according to the procedure described for Intermediate 23, starting from 0.039 g (0.084 mmol) of methyl 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 66). 1H NMR (DMSO-de) δ (ppm): 8.65-8.77 (m, 1 H) 8.22 (s, 1 H) 7.77-7.89 (m, 1 H) 7.48-7.58 (m, 2 H) 7.33-7.45 (m, 2 H) 7.17-7.30 (m, 2 H) 7.01-7.14 (m, 2 H) 5.36-5.45 (m, 1 H), 5.14 (s, 2 H) 3.65-3.79 (m, 1 H) 3.50-3.64 (m, 1 H) 1.32-1.85 (m, 6 H); MS (ESI): m/z: 449[M+H]+.
Intermediate 68: methyl 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000078_0001
0.047 g (36%) of methyl 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 68) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (DMSO-de) δ (ppm): 8.96-8.74 (m, 1 H), 8.27-8.19 (m, 1 H), 7.89 (dd, J=1.5, 5.4 Hz, 1 H), 7.13-6.95 (m, 1 H), 6.56-6.33 (m, 3 H), 5.53-5.44 (m, 1 H), 5.36-5.21 (m, 2 H), 3.92 (s, 3 H), 3.82-3.50 (m, 2 H), 3.1 1-2.99 (m, 4 H), 2.45-2.30 (m, 4 H), 2.19 (s, 3 H), 1.85-1.40 (m, 6 H); MS (ESI): m/z: 469[M+H]+.
Intermediate 69: 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000078_0002
0.044 g (65%) of 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 69) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.15 mmol) of methyl 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 68). 1H NMR (DMSO-de) δ (ppm): 8.84-8.61 (m, 1 H), 8.29-8.10 (m, 1 H), 7.83 (dd, J=1.5, 5.4 Hz, 1 H), 7.14-6.97 (m, J=8.1 , 8.1 Hz, 1 H), 6.58-6.34 (m, 3 H), 5.54-5.39 (m, 1 H), 5.33-5.18 (m, 2 H), 3.86-3.49 (m, 2 H), 3.22-2.50 (m, 8 H), 2.30 (s, 3 H), 1 .81-1.39 (m, 6 H); MS (ESI): m/z: 455[M+H]+.
Intermediate 70: methyl 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000079_0001
0.053 g (54%) of methyl 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 70) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (DMSO-de) δ (ppm): 8.87-8.75 (m, 1 H), 8.25-8.12 (m, 1 H), 7.94-7.82 (m, 1 H), 7.09-6.75 (m, 4 H), 5.49- 5.42 (m, 1 H), 5.39-5.26 (m, 2 H), 3.93 (s, 3 H), 3.79-3.49 (m, 2 H), 2.81-2.66 (m, 4 H), 2.21-2.00 (m, 7 H), 1.85-1.42 (m, 6 H); MS (ESI): m/z: 469[M+H]+.
Intermediate 71 : 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000080_0001
0.042 g (96%) of 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 71 ) was prepared according to the procedure described for Intermediate 23, starting from 0.045 g (0.096 mmol) of methyl 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 70). 1H NMR (DMSO-de) δ (ppm): 8.82-8.63 (m, 1 H), 8.26-8.13 (m, 1 H), 7.83 (dd, J=1.5, 4.9 Hz, 1 H), 7.1 1-6.70 (m, 4 H), 5.51-5.24 (m, 3 H), 3.81-3.45 (m, 2 H), 2.91-2.08 (m, 1 1 H), 1.87-1.38 (m, 6 H); MS (ESI): m/z: 455[M+H]+.
Intermediate 72: methyl 2-[(Z)-A -tetrahydropyran-2-yloxy-C-[[3-
(trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate
Figure imgf000080_0002
0.055 g (45%) of methyl 2-[(Z)-A/-tetrahydropyran-2-yloxy-C-[[3- (trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate (Intermediate 72) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.79- 8.75 (m, 1 H), 8.50-8.45 (m, 1 H), 7.88-7.82 (m, 1 H), 7.41-7.34 (m, 1 H), 7.30-7.10 (m, 3 H), 5.59-5.53 (m, 1 H), 5.53-5.40 (m, 2 H), 3.99-3.87 (m, 4 H), 3.73-3.65 (m, 1 H), 1.89-1.59 (m, 6 H); MS (ESI): m/z: 439[M+H]+. Intermediate 73: methyl 2-[(Z)-C-[(4-methoxyphenoxy)methyl]-A -tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000081_0001
0.056 g (50%) of methyl 2-[(Z)-C-[(4-methoxyphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 73) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4- carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.83-8.72 (m, 1 H), 8.49-8.38 (m, 1 H), 7.88-7.77 (m, 1 H), 6.98-6.69 (m, 4 H), 5.60-5.49 (m, 1 H), 5.44-5.28 (m, 2 H), 4.03-3.86 (m, 4 H), 3.81 -3.59 (m, 4 H), 1 .94-1 .53 (m, 6 H); MS (ESI): m/z: 401 [M+H]+. Intermediate 74: methyl 2-[(E)-3-phenylprop-2-enoyl]pyridine-4-carboxylate
Figure imgf000081_0002
0.97 mL (9.83 mmol) of piperidine and 0.56 mL (9.88 mmol) of AcOH was added to a stirred solution of 0.91 mL (7.89 mmol) of benzaldehyde and 1 .6 g (8.9 mmol) of Intermediate 1 in 16 mL of MeOH; the resulting mixture was stirred at reflux for about 5 h, cooled to r.t, then with an ice bath. The resulting precipitated was filtered, washed with little cold MeOH, followed by Et2O and dried to give 0.57 g (35%) of the title compound as a light brown solid. 1H NMR (CDCI3) δ (ppm): 8.95-8.88 (m, 1 H), 8.77-8.67 (m, 1 H), 8.32 (d, J=15.7 Hz, 1 H), 8.07 (dd, J=1 .5, 4.9 Hz, 1 H), 8.00 (d, J=16.1 Hz, 1 H), 7.80- 7.69 (m, 2 H), 7.51-7.40 (m, 3 H), 4.01 (s, 3 H); MS (ESI): m/z: 268[M+H]+.
Intermediate 75: methyl 2-(3-phenylpropanoyl)pyridine-4-carboxylate
Figure imgf000082_0001
0.66 g of silica gel was added to a stirred solution of 0.075 g (0.28 mmol) of methyl 2-[(E)- 3-phenylprop-2-enoyl]pyridine-4-carboxylate (Intermediate 74) and 0.13 g (0.50 mmol) of 3,5-bis(ethoxycarbonyl)-1 ,4-dihydro-2,6-dimethylpyridine in 2.5 ml_ of toluene. After stirring at 70°C in the dark for about 17 h, the silica gel was removed by filtration and the solution was concentrated providing about 0.70 g of raw material, which was purified by flash chromatography (eluent: n-hexane:DCM:EtOAc 20:5:1 ) to yield 0.42 g (56%) of methyl 2-(3-phenylpropanoyl)pyridine-4-carboxylate. 1H NMR (CDCI3) δ (ppm): 8.77-8.87 (m, 1 H), 8.57 (s, 1 H), 7.99-8.07 (m, 1 H), 7.25-7.35 (m, 4 H), 7.10-7.25 (m, 1 H), 4.00 (s, 3 H), 3.55-3.66 (m, 2 H), 3.05-3.15 (m, 2 H); MS (ESI): m/z: 270[M+H]+.
Intermediate 76: methyl 2-[(Z)-A -tetrahydropyran-2-yloxy-C-[[3-
(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate
Figure imgf000082_0002
0.078 g (77%) of methyl 2-[(Z)-A/-tetrahydropyran-2-yloxy-C-[[3- (trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate (Intermediate 76) was prepared according to the procedure described for Intermediate 22, starting from 0.80 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.83- 8.70 (m, 1 H), 8.54-8.36 (m, 1 H), 7.85 (dd, J=1.2, 5.1 Hz, 1 H), 7.36-7.14 (m, 1 H), 6.98- 6.72 (m, 3 H), 5.64-5.48 (m, 1 H), 5.56-5.34 (m, 2 H), 4.05-3.60 (m, 5 H), 1.95-1.48 (m, 6 H); MS (ESI): m/z: 455[M+H]+. Intermediate 77: 2-[(Z)-A -tetrahydropyran-2-yloxy-C-[[3-
(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000083_0001
0.045 g (66%) of 2-[(Z)-N-tetrahydropyran-2-yloxy-C-[[3-
(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 77) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.15 mmol) of methyl 2-[(Z)-N-tetrahydropyran-2-yloxy-C-[[3- (trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate (Intermediate 76). 1 H NMR (DMSO-d6) δ (ppm): 13.90 (bs, 1 H), 8.86-8.72 (m, 1 H), 8.30-8.20 (m, 1 H), 7.86 (dd, J=1 .5, 4.9 Hz, 1 H), 7.47-7.36 (m, 1 H), 7.12-6.87 (m, 3 H), 5.54-5.44 (m, 1 H), 5.37 (s, 2 H), 3.82-3.49 (m, 2 H), 1 .83-1 .34 (m, 6 H); MS (ESI): m/z: 439[M-H]\ Intermediate 78: 2-[(Z)-C-[2-[tert-butoxycarbonyl-[(4- fluorophenyl)methyl]amino]ethoxymethyl]-/V-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000083_0002
0.026 g (44%) of 2-[(Z)-C-[2-[tert-butoxycarbonyl-[(4- fluorophenyl)methyl]amino]ethoxymethyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 78) was prepared according to the procedure described for Intermediate 19, starting from 0.040 g (0.1 1 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 20 h. 1H NMR (DMSO-de) δ (ppm): 8.80-8.74 (m, 1 H), 8.49-8.42 (m, 1 H), 7.85-7.79 (m, 1 H), 7.16-7.09 (m, 2 H), 6.99-6.91 (m, 2 H), 5.56 (bs, 1 H), 4.87 (bs, 2 H), 4.42 (s, 2 H), 3.98-3.88 (m, 1 H), 3.77- 3.57 (m, 3 H), 3.39-3.20 (m, 2 H), 2.00-1 .55 (m, 9 H), 1 .26 (s, 9 H); MS (ESI): m/z: 530 [M-H]-.
Intermediate 79: 2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-A - tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000084_0001
0.023 g (28%) of 2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 79) was prepared according to the procedure described for Intermediate 19, starting from 0.080 g (0.22 mmol) of 2-Z- (C-(bromomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). The reaction was carried out at r.t. for about 20 h. 1 H NMR (CDCI3) δ (ppm): 8.76-8.71 (m, 1 H), 8.43-8.38 (m, 1 H), 7.81-7.75 (m, 1 H), 5.56-5.49 (m, 1 H), 5.01 (s, 2 H), 4.33 (s, 2 H), 3.99-3.89 (m, 1 H), 3.76-3.67 (m, 1 H), 2.97 (s, 6 H), 1 .97- 1 .55 (m, 6 H); MS (ESI): m/z: 366 [M+H]+.
Intermediate 80: 2-[(Z)-C-(3-pyridylmethoxymethyl)-A -tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000084_0002
2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 80) was prepared according to the procedure described for Intermediate 19, starting from of 2-Z-(C-(bromomethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate (Intermediate 12). The reaction was carried out at r.t. for about 3 h. MS (ESI): m/z: 372 [M+H]+.
Intermediate 81 : 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000085_0001
0.040 g (51 %) of 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 81 ) was prepared according to the procedure described for Intermediate 19, starting from 0.080 g (0.22 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 3 h. 1H NMR (DMSO-d6) δ (ppm): 1 1 .99 (bs, 1 H), 8.73 - 8.60 (m, 1 H), 8.23 - 8.08 (m, 1 H), 7.82 - 7.65 (m, 1 H), 5.47 - 5.34 (m, 1 H), 4.85 - 4.66 (m, 2 H), 3.81 - 3.74 (m, 1 H), 3.69 - 3.53 (m, 3 H), 2.58 - 2.53 (m, 2 H), 2.40 - 2.34 (m, 6 H), 1 .95 - 1 .31 (m, 6 H); MS (ESI): m/z: 352 [M+H]+. Intermediate 82: methyl 2-[(E)-C-[(4-fluoroanilino)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000085_0002
0.040 g (46%) of methyl 2-[(E)-C-[(4-fluoroanilino)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 82) was prepared according to the procedure described for Intermediate 16, starting from 0.080 g (0.22 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 24 h. 1H NMR (CDCI3) δ (ppm): 8.77 - 8.73 (m, 1 H), 8.44 - 8.41 (m, 1 H), 7.84 - 7.80 (m, 1 H), 6.90 - 6.77 (m, 2 H), 6.67 - 6.59 (m, 2 H), 5.54 - 5.48 (m, 1 H), 4.73 - 4.58 (m, 2 H), 4.00 - 3.90 (m, 4 H), 3.74 - 3.66 (m, 1 H), 2.01 - 1.56 (m, 6 H); MS (ESI): m/z: 388 [M+H]+.
Intermediate 83: 2-[(E)-C-[(4-fluoroanilino)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000086_0001
0.020 g (56%) of 2-[(E)-C-[(4-fluoroanilino)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 83) was prepared according to the procedure described for Intermediate 23, starting from 0.037 g (0.096 mmol) of methyl 2- [(E)-C-[(4-fluoroanilino)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 82). 1H NMR (DMSO-d6) δ (ppm): 13.79 (bs, 1 H), 8.85 - 8.81 (m, 1 H), 8.14 - 8.10 (m, 1 H), 7.85 - 7.81 (m, 1 H), 6.89 - 6.82 (m, 2 H), 6.66 - 6.59 (m, 2 H), 5.69 - 5.56 (m, 1 H), 5.46 - 5.41 (m, 1 H), 4.57 - 4.44 (m, 2 H), 3.78 (s, 1 H), 3.61 - 3.53 (m, 1 H), 1.89 - 1.42 (m, 6 H); MS (ESI): m/z: 373 [M-H]".
Intermediate 84: methyl 2-[(Z)-C-[(4-morpholinophenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000087_0001
0.12 g (93%) of methyl 2-[(Z)-C-[(4-morpholinophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 84) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.72 (m, 1 H), 8.48 - 8.39 (m, 1 H), 7.82 (dd, J = 1.5, 4.9 Hz, 1 H), 6.97 - 6.78 (m, 4 H), 5.58 - 5.49 (m, 1 H), 5.44 - 5.32 (m, 2 H), 4.02 - 3.62 (m, 9 H), 3.06 (bs, 4 H), 1.93 - 1.51 (m, 6 H); MS (ESI): m/z: 456 [M+H]+. Intermediate 85: 2-[(Z)-C-[(4-morpholinophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000087_0002
0.090 g (85%) of 2-[(Z)-C-[(4-morpholinophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 85) was prepared according to the procedure described for Intermediate 23, starting from 0.1 1 g (0.24 mmol) of methyl 2- [(Z)-C-[(4-morpholinophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 84). 1H NMR (DMSO-d6) δ (ppm): 13.83 (bs, 1 H), 8.85 - 8.76 (m, 1 H), 8.26 - 8.17 (m, 1 H), 7.85 (dd, J = 1.7, 5.1 Hz, 1 H), 6.92 - 6.77 (m, 4 H), 5.50 - 5.42 (m, 1 H), 5.28 - 5.16 (m, 2 H), 3.80 - 3.47 (m, 6 H), 3.01 - 2.92 (m, 4 H), 1.84 - 1.41 (m, 6 H); MS (ESI): m/z: 442 [M+H]+. Intermediate 86: methyl 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000088_0001
0.098 g (99%) of methyl 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 86) was prepared according to the procedure described for Intermediate 22, starting from 0.070 g (0.20 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.84 - 8.69 (m, 1 H), 8.51 - 8.36 (m, 1 H), 7.83 (dd, J = 1.2, 5.1 Hz, 1 H), 7.45 - 6.99 (m, 5 H), 6.66 - 6.50 (m, 3 H), 5.60 - 5.50 (m, 1 H), 5.47 - 5.29 (m, 2 H), 4.97 (s, 3 H), 3.97 (s, 4 H), 1.95 - 1.45 (m, 6 H); MS (ESI): m/z: 495 [M+H]+.
Intermediate 87: 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylic acid
Figure imgf000088_0002
0.070 g (86%) of 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 87) was prepared according to the procedure described for Intermediate 23, starting from 0.085 g (0.17 mmol) of methyl 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 86). 1H NMR (DMSO-de) δ (ppm): 13.87 (bs, 1 H), 8.86 - 8.76 (m, 1 H), 8.28 - 8.14 (m, 1 H), 7.90 - 7.80 (m, 1 H), 7.52 - 7.43 (m, 2 H), 7.26 - 7.1 1 (m, 3 H), 6.69 - 6.51 (m, 3 H), 5.52 - 5.42 (m, 1 H), 5.29 (s, 2 H), 5.03 (s, 2 H), 3.82 - 3.47 (m, 2 H), 1.82 - 1.37 (m, 6 H); MS (ESI): m/z: 481 [M+H]+.
Intermediate 88: methyl 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000089_0001
0.082 g (86%) of methyl 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 88) was prepared according to the procedure described for Intermediate 22, starting from 0.070 g (0.20 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.69 (m, 1 H), 8.49 - 8.40 (m, 1 H), 7.83 (dd, J = 1.5, 4.9 Hz, 1 H), 7.46 - 7.08 (m, 6 H), 6.68 - 6.56 (m, 3 H), 5.61 - 5.48 (m, 1 H), 5.46 - 5.31 (m, 2 H), 5.01 (s, 3 H), 4.02 - 3.56 (m, 4 H), 1.92 - 1.46 (m, 6 H); MS (ESI): m/z: 477 [M+H]+.
Intermediate 89: 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylic acid
Figure imgf000089_0002
0.048 g (69%) of 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 89) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.15 mmol) of methyl 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 88). 1H NMR (DMSO-de) δ (ppm): 13.88 (bs, 1 H), 8.86 - 8.71 (m, 1 H), 8.28 - 8.19 (m, 1 H), 7.86 (dd, J = 1.5, 4.9 Hz, 1 H), 7.45 - 7.09 (m, 6 H), 6.69 - 6.52 (m, 3 H), 5.51 - 5.42 (m, 1 H), 5.29 (s, 2 H), 5.05 (s, 2 H),
3.81 - 3.46 (m, 2 H), 1.81 - 1.36 (m, 6 H); MS (ESI): m/z: 463 [M+H]+.
Intermediate 90: methyl 2-[(Z)-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000090_0001
0.065 g (50%) of methyl 2-[(Z)-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 90) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.74 (m, 1 H), 8.48 - 8.39 (m, 1 H), 7.84 - 7.78 (m, 1 H), 6.95 - 6.80 (m, 4 H), 5.57 - 5.51 (m, 1 H), 5.43 - 5.31 (m, 2 H), 4.00 - 3.86 (m, 4 H), 3.72 - 3.61 (m, 1 H), 3.13 (bs, 3 H), 2.74 - 2.52 (m, 4 H), 2.48 - 2.27 (m, 4 H), 1.86 - 1.57 (m, 6 H); MS (ESI): m/z: 469 [M+H]+. Intermediate 91 : 2-[(Z)-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylic acid
Figure imgf000090_0002
0.040 g (68%) of 2-[(Z)-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 91 ) was prepared according to the procedure described for Intermediate 23, starting from 0.060 g (0.13 mmol) of methyl 2-[(Z)-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 90). MS (ESI): m/z: 455 [M+H]+. Intermediate 92: methyl 2-[(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000091_0001
0.090 g (71 %) of methyl 2-[(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 92) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.74 (m, 1 H), 8.45 - 8.41 (m, 1 H), 7.85 - 7.78 (m, 1 H), 6.96 - 6.80 (m, 4 H), 5.57 - 5.51 (m, 1 H), 5.41 - 5.30 (m, 2 H), 3.98 - 3.86 (m, 4 H), 3.72 - 3.63 (m, 1 H), 3.12 - 2.94 (m, 4 H), 1.88 - 1.56 (m, 12 H); MS (ESI): m/z: 454 [M+H]+.
Intermediate 93: 2-[(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000091_0002
0.045 g (60%) of 2-[(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 93) was prepared according to the procedure described for Intermediate 23, starting from 0.078 g (0.17 mmol) of methyl 2- [(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 92). 1H NMR (CDCI3) δ (ppm): 8.79 - 8.73 (m, 1 H), 8.40 - 8.35 (m, 1 H), 7.88 - 7.81 (m, 1 H), 7.20 - 7.09 (m, 2 H), 6.95 - 6.87 (m, 2 H), 5.56 - 5.51 (m, 1 H), 5.47 - 5.35 (m, 2 H), 3.96 - 3.87 (m, 1 H), 3.72 - 3.64 (m, 1 H), 3.23 - 3.09 (m, 4 H), 1.97 - 1.55 (m, 12 H); MS (ESI): m/z: 440 [M+H]+. Intermediate 94: methyl 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000092_0001
0.090 g (71 %) of methyl 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 94) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.74 (m, 1 H), 8.48 - 8.41 (m, 1 H), 7.87 - 7.80 (m, 1 H), 7.46 - 7.29 (m, 5 H), 6.94 - 6.84 (m, 4 H), 5.57 - 5.51 (m, 1 H), 5.42 - 5.32 (m, 2 H), 5.01 (s, 2 H), 3.97 (s, 3 H), 3.94 - 3.86 (m, 1 H), 3.71 - 3.63 (m, 1 H), 1.89 - 1.80 (m, 3 H), 1.71 - 1.57 (m, 3 H); MS (ESI): m/z: 477 [M+H]+.
Intermediate 95: 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000092_0002
0.085 g (80%) of 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 95) was prepared according to the procedure described for Intermediate 23, starting from 0.1 1 g (0.23 mmol) of methyl 2- [(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylate (Intermediate 94). 1H NMR (CDCI3) δ (ppm): 8.82 - 8.74 (m, 1 H), 8.52 - 8.45 (m, 1 H), 7.88 - 7.80 (m, 1 H), 7.46 - 7.29 (m, 5 H), 6.94 - 6.83 (m, 4 H), 5.64 - 5.56 (m, 1 H), 5.44 - 5.33 (m, 2 H), 5.01 (s, 2 H), 3.99 - 3.88 (m, 1 H), 3.79 - 3.69 (m, 1 H), 1.93 - 1.56 (m, 6 H); MS (ESI): m/z: 463 [M+H]+.
Intermediate 96: methyl 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox late
Figure imgf000093_0001
0.10 g (quantitative) of methyl 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyhdine-4-carboxylate (Intermediate 96) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.79 - 8.75 (m, 1 H), 8.49 - 8.43 (m, 1 H), 7.87 - 7.81 (m, 1 H), 7.21 - 7.14 (m, 1 H), 7.05 - 7.00 (m, 1 H), 6.97 - 6.91 (m, 1 H), 6.88 - 6.82 (m, 1 H), 5.59 - 5.53 (m, 1 H), 5.48 - 5.34 (m, 2 H), 4.01 - 3.88 (m, 4 H), 3.74 - 3.65 (m, 1 H), 1.92 - 1.59 (m, 6 H); MS (ESI): m/z: 405 [M+H]+.
Intermediate 97: 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000093_0002
0.074 g (quantitative) of 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 97) was prepared according to the procedure described for Intermediate 23, starting from 0.075 g (0.19 mmol) of methyl 2- [(Z)-C-[(3-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 96). 1H NMR (CDCI3) δ (ppm): 8.82 - 8.76 (m, 1 H), 8.54 - 8.49 (m, 1 H), 7.89 - 7.82 (m, 1 H), 7.22 - 7.13 (m, 1 H), 7.06 - 7.01 (m, 1 H), 6.98 - 6.91 (m, 1 H), 6.89 - 6.82 (m, 1 H), 5.66 - 5.60 (m, 1 H), 5.48 - 5.37 (m, 2 H), 4.01 - 3.91 (m, 1 H),
3.81 - 3.73 (m, 1 H), 1.95 - 1.60 (m, 6 H); MS (ESI): m/z: 391 [M+H]+.
Intermediate 98: methyl 2-[(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-
2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000094_0001
0.090 g (78%) of methyl 2-[(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 98) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.76 (m, 1 H), 8.50 - 8.43 (m, 1 H), 7.88 - 7.81 (m, 1 H), 7.34 - 7.29 (m, 2 H), 7.08 - 7.00 (m, 1 H), 6.98 - 6.90 (m, 6 H), 5.58 - 5.53 (m, 1 H), 5.47 - 5.35 (m, 2 H), 4.00 - 3.88 (m, 4 H), 3.74 - 3.63 (m, 1 H), 1.91 - 1.80 (m, 3 H), 1.70 - 1.61 (m, 3 H); MS (ESI): m/z: 463 [M+H]+.
Intermediate 99: 2-[(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000094_0002
0.074 g (97%) of 2-[(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 99) was prepared according to the procedure described for Intermediate 23, starting from 0.080 g (0.17 mmol) of methyl 2- [(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 98). 1H NMR (CDCI3) δ (ppm): 8.84 - 8.77 (m, 1 H), 8.54 - 8.48 (m, 1 H), 7.89 - 7.82 (m, 1 H), 7.34 - 7.27 (m, 2 H), 7.09 - 7.02 (m, 1 H), 6.99 - 6.90 (m, 6 H), 5.65 - 5.60 (m, 1 H), 5.47 - 5.37 (m, 2 H), 4.01 - 3.92 (m, 1 H), 3.80 - 3.71 (m, 1 H), 1.93 - 1.64 (m, 6 H); MS (ESI): m/z: 449 [M+H]+.
Intermediate 100: methyl 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000095_0001
0.093 g (85%) of methyl 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 100) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.74 (m, 1 H), 8.50 - 8.45 (m, 1 H), 7.88 - 7.82 (m, 1 H), 7.32 - 7.28 (m, 1 H), 7.16 - 7.1 1 (m, 1 H), 6.86 - 6.79 (m, 1 H), 5.59 - 5.52 (m, 1 H), 5.44 - 5.35 (m, 2 H), 4.01 - 3.88 (m, 4 H), 3.74 - 3.66 (m, 1 H), 1.93 - 1.80 (m, 3 H), 1.72 - 1.60 (m, 3 H); MS (ESI): m/z: 440 [M+H]+.
Intermediate 101 : 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000095_0002
0.074 g (87%) of 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 101 ) was prepared according to the procedure described for Intermediate 23, starting from 0.087 g (0.20 mmol) of methyl 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 100). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.75 (m, 1 H), 8.54 - 8.48 (m, 1 H), 7.88 - 7.83 (m, 1 H), 7.34 - 7.28 (m, 1 H), 7.18 - 7.12 (m, 1 H), 6.86 - 6.80 (m, 1 H), 5.65 - 5.59 (m, 1 H), 5.45 - 5.35 (m, 2 H), 4.00 - 3.90 (m, 1 H), 3.81 - 3.72 (m, 1 H), 1.97 - 1.63 (m, 6 H); MS (ESI): m/z: 426 [M+H]+.
Intermediate 102: methyl 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000096_0001
0.070 g (64%) of methyl 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 102) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.74 (m, 1 H), 8.51 - 8.43 (m, 1 H), 7.88 - 7.81 (m, 1 H), 6.99 - 6.94 (m, 1 H), 6.94 - 6.88 (m, 2 H), 5.59 - 5.54 (m, 1 H), 5.45 - 5.35 (m, 2 H), 4.01 - 3.87 (m, 4 H), 3.75 - 3.66 (m, 1 H), 1.94 - 1.82 (m, 3 H), 1.73 - 1.63 (m, 3 H); MS (ESI): m/z: 440 [M+H]+.
Intermediate 103: 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000096_0002
0.045 g (60%) of 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 103) was prepared according to the procedure described for Intermediate 23, starting from 0.060 g (0.14 mmol) of methyl 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 102). 1H NMR (CDCI3) δ (ppm): 8.82 - 8.77 (m, 1 H), 8.55 - 8.49 (m, 1 H), 7.90 - 7.83 (m, 1 H), 7.00 - 6.96 (m, 1 H), 6.94 - 6.88 (m, 2 H), 5.64 - 5.58 (m, 1 H), 5.47 - 5.37 (m, 2 H), 3.99 - 3.91 (m, 1 H), 3.79 - 3.70 (m, 1 H), 1.96 - 1.87 (m, 3 H), 1.76 - 1.66 (m, 3 H); MS (ESI): m/z: 426 [M+H]+.
Intermediate 104: methyl 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000097_0001
0.10 g (91 %) of methyl 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 104) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.74 (m, 1 H), 8.51 - 8.43 (m, 1 H), 7.87 - 7.81 (m, 1 H), 7.59 - 7.52 (m, 2 H), 7.46 - 7.40 (m, 2 H), 7.37 - 7.30 (m, 2 H), 7.23 - 7.16 (m, 2 H), 6.99 - 6.94 (m, 1 H), 5.58 - 5.55 (m, 1 H), 5.54 - 5.43 (m, 2 H), 4.00 - 3.87 (m, 4 H), 3.71 - 3.62 (m, 1 H), 1 .90 - 1.76 (m, 3 H), 1.72 - 1.59 (m, 3 H); MS (ESI): m/z: 447 [M+H]+.
Intermediate 105: 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000097_0002
0.062 g (90%) of 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 105) was prepared according to the procedure described for Intermediate 23, starting from 0.070 g (0.16 mmol) of methyl 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridin 4-carboxylate (Intermediate 104). 1H NMR (CDCI3) δ (ppm): 8.83 - 8.77 (m, 1 H), 8.54 - 8.49 (m, 1 H), 7.87 - 7.81 (m, 1 H), 7.59 - 7.52 (m, 2 H), 7.46 - 7.39 (m, 2 H), 7.37 - 7.30 (m, 2 H), 7.24 - 7.17 (m, 2 H), 6.99 - 6.95 (m, 1 H), 5.64 - 5.58 (m, 1 H), 5.55 - 5.44 (m, 2 H), 3.99 - 3.90 (m, 1 H), 3.77 - 3.68 (m, 1 H), 1.91 - 1.78 (m, 3 H), 1.71 - 1.59 (m, 3 H); MS (ESI): m/z: 433 [M+H]+.
Intermediate 106: methyl 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000098_0001
0.1 1 g (93%) of methyl 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 106) was prepared according to the procedure described for Intermediate 22, starting from 0.090 g (0.25 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.77 - 8.72 (m, 1 H), 8.46 - 8.41 (m, 1 H), 7.86 - 7.79 (m, 1 H), 7.36 - 7.30 (m, 2 H), 7.23 - 7.17 (m, 1 H), 7.13 - 7.08 (m, 1 H), 7.02 - 6.97 (m, 2 H), 6.74 - 6.69 (m, 1 H), 6.68 - 6.65 (m, 1 H), 6.62 - 6.58 (m, 1 H), 5.52 - 5.47 (m, 1 H), 5.45 - 5.34 (m, 2 H), 3.97 (s, 3 H), 3.94 - 3.87 (m, 1 H), 3.69 - 3.62 (m, 1 H), 1.87 - 1.76 (m, 3 H), 1.72 - 1.59 (m, 3 H); MS (ESI): m/z: 463 [M+H]+.
Intermediate 107: 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000098_0002
0.063 g (83%) of 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 107) was prepared according to the procedure described for Intermediate 23, starting from 0.080 g (0.17 mmol) of methyl 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylate (Intermediate 106). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.71 (m, 1 H), 8.53 - 8.44 (m, 1 H), 7.86 - 7.77 (m, 1 H), 7.37 - 7.29 (m, 2 H), 7.23 - 7.17 (m, 1 H), 7.14 - 7.07 (m, 1 H), 7.03 - 6.96 (m, 2 H), 6.75 - 6.56 (m, 3 H), 5.59 - 5.53 (m, 1 H), 5.46 - 5.34 (m, 2 H), 3.99 - 3.89 (m, 1 H), 3.76 - 3.67 (m, 1 H), 1.85 - 1.53 (m, 6 H); MS (ESI): m/z: 449 [M+H]+.
Intermediate 108: methyl 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox late
Figure imgf000099_0001
0.12 g (quantitative) of methyl 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 108) was prepared according to the procedure described for Intermediate 22, starting from 0.10 g (0.28 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.80 - 8.74 (m, 1 H), 8.49 - 8.43 (m, 1 H), 7.88 - 7.82 (m, 1 H), 7.39 - 7.32 (m, 2 H), 6.90 - 6.83 (m, 2 H), 5.58 - 5.53 (m, 1 H), 5.45 - 5.34 (m, 2 H), 3.98 (s, 3 H), 3.95 - 3.87 (m, 1 H), 3.73 - 3.65 (m, 1 H), 1.91 - 1.64 (m, 6 H); MS (ESI): m/z: 449 [M+H]+.
Intermediate 109: 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000100_0001
0.044 g (quantitative) of 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 109) was prepared according to the procedure described for Intermediate 23, starting from 0.045 g (0.10 mmol) of methyl 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 108). MS (ESI): m/z: 436 [M+H]+.
Intermediate 110: methyl 2-[(Z)-C-[(4-phenylphenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox late
Figure imgf000100_0002
0.065 g (65%) of methyl 2-[(Z)-C-[(4-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 110) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.76 (m, 1 H), 8.50 - 8.46 (m, 1 H), 7.87 - 7.82 (m, 1 H), 7.57 - 7.53 (m, 2 H), 7.52 - 7.48 (m, 2 H), 7.44 - 7.39 (m, 2 H), 7.34 - 7.29 (m, 1 H), 7.08 - 7.03 (m, 2 H), 5.58 (s, 1 H), 5.51 - 5.43 (m, 2 H), 3.99 - 3.92 (m, 4 H), 3.73 - 3.67 (m, 1 H), 1.96 - 1.55 (m, 6 H); MS (ESI): m/z: 447 [M+H]+.
Intermediate 111 : 2-[(Z)-C-[(4-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000101_0001
0.025 g (43%) of 2-[(Z)-C-[(4-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 111 ) was prepared according to the procedure described for Intermediate 23, starting from 0.060 g (0.13 mmol) of methyl 2-[(Z)-C-[(4-phenylphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylate (Intermediate 110). 1H NMR (CDCI3) δ (ppm): 8.82 - 8.76 (m, 1 H), 8.56 - 8.51 (m, 1 H), 7.87 - 7.81 (m, 1 H), 7.57 - 7.48 (m, 4 H), 7.45 - 7.38 (m, 2 H), 7.34 - 7.28 (m, 1 H), 7.08 - 7.02 (m, 2 H), 5.67 - 5.61 (m, 1 H), 5.52 - 5.42 (m, 2 H), 4.02 - 3.93 (m, 1 H), 3.81 - 3.72 (m, 1 H), 1.99 - 1.54 (m, 6 H); MS (ESI): m/z: 433 [M+H]+.
Intermediate 112: methyl 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]- N-tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylate
Figure imgf000101_0002
0.1 1 g (97%) of methyl 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 112) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2-[(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.83 - 8.72 (m, 1 H), 8.48 - 8.38 (m, 1 H), 7.83 (dd, J = 1.7, 5.1 Hz, 1 H), 7.44 - 7.33 (m, 2 H), 7.1 1 - 7.02 (m, 2 H), 6.95 - 6.80 (m, 4 H), 5.58 - 5.49 (m, 1 H), 5.43 - 5.31 (m, 2 H), 4.97 (s, 2 H), 4.00 - 3.58 (m, 5 H), 1.92 - 1.53 (m, 6 H); MS (ESI): m/z: 495 [M+H]+.
Intermediate 113: 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000102_0001
0.095 g (94%) of 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 113) was prepared according to the procedure described for Intermediate 23, starting from 0.10 g (0.21 mmol) of methyl 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 112). 1H NMR (CDCIs) δ (ppm): 8.87 - 8.70 (m, 1 H), 8.55 - 8.35 (m, 1 H), 7.83 (dd, J = 1.6, 4.9 Hz, 1 H), 7.51 - 7.30 (m, 2 H), 7.14 - 6.99 (m, 2 H), 6.95 - 6.73 (m, 4 H), 5.69 - 5.58 (m, 1 H), 5.44 - 5.32 (m, 2 H), 4.97 (s, 2 H), 4.01 - 3.69 (m, 2 H), 1.97 - 1.53 (m, 6 H); MS (ESI): m/z: 481 [M+H]+.
Intermediate 114: methyl 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carbox late
Figure imgf000102_0002
0.087 g (96%) of methyl 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 114) was prepared according to the procedure described for Intermediate 22, starting from 0.080 g (0.22 mmol) of methyl 2- [(Z)-C-(bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 12). 1H NMR (CDCI3) δ (ppm): 8.84 - 8.69 (m, 1 H), 8.51 - 8.42 (m, 1 H), 7.91 - 7.81 (m, 1 H), 7.24 - 7.19 (m, 2 H), 6.93 - 6.88 (m, 2 H), 5.58 - 5.53 (m, 1 H), 5.44 - 5.37 (AB System: VA=5.41 , VB=5.39, JAB= 1 1 Hz), 4.00 - 3.87 (m, 4 H), 3.73 - 3.65 (m, 1 H), 1.96 - 1.51 (m, 6 H); MS (ESI): m/z: 405 [M+H]+. Intermediate 115: 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000103_0001
0.062 g (72%) of 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 115) was prepared according to the procedure described for Intermediate 23, starting from 0.079 g (0.22 mmol) of methyl 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylate (Intermediate 114). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.76 (m, 1 H), 8.53 - 8.49 (m, 1 H), 7.88 - 7.84 (m, 1 H), 7.25 - 7.19 (m, 2 H), 6.94 - 6.87 (m, 2 H), 5.66 - 5.60 (m, 1 H), 5.45 - 5.36 (m, 2 H), 4.00 - 3.91 (m, 1 H), 3.82 - 3.73 (m, 1 H), 1.99 - 1.54 (m, 6 H); MS (ESI): m/z: 389 [M+H]+.
Intermediate 116: 2-[(Z)-C-(cyclopentoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000103_0002
0.010 g (13%) of 2-[(Z)-C-(cyclopentoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 116) was prepared according to the procedure described for Intermediate 19, starting from 0.080 g (0.22 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 24 h. 1H NMR (CDCI3) δ (ppm): 8.82 - 8.77 (m, 1 H), 8.49 - 8.45 (m, 1 H), 7.85 - 7.80 (m, 1 H), 5.63 - 5.56 (m, 1 H), 4.83 (s, 2 H), 4.09 - 4.02 (m, 1 H), 4.01 - 3.93 (m, 1 H), 3.79 - 3.71 (m, 1 H), 1.98 - 1.86 (m, 3 H), 1.79 - 1.60 (m, 9 H), 1.55 - 1.44 (m, 2 H); MS (ESI): m/z: 449 [M+H]+. Intermediate 117: 2-[(Z)-C-(cyclobutoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000104_0001
0.016 g (21 %) of 2-[(Z)-C-(cyclobutoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 117) was prepared according to the procedure described for Intermediate 19, starting from 0.080 g (0.22 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 24 h. 1H NMR (CDCI3) δ (ppm): 8.82 - 8.77 (m, 1 H), 8.49 - 8.45 (m, 1 H), 7.84 - 7.79 (m, 1 H), 5.63 - 5.56 (m, 1 H), 4.79 (s, 2 H), 4.12 - 4.04 (m, 1 H), 4.01 - 3.93 (m, 1 H), 3.79 - 3.72 (m, 1 H), 2.00 - 1.42 (m, 12 H); MS (ESI): m/z: 335 [M+H]+.
Intermediate 118: 2-[(Z)-C-(propoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000104_0002
0.0090 g (13%) of 2-[(Z)-C-(propoxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 118) was prepared according to the procedure described for Intermediate 19, starting from 0.080 g (0.22 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 20 h. 1H NMR (CDCI3) δ (ppm): 8.80 (s, 1 H), 8.49 - 8.45 (m, 1 H), 7.85 - 7.79 (m, 1 H), 5.65 - 5.57 (m, 1 H), 4.93 - 4.82 (m, 2 H), 4.02 - 3.92 (m, 1 H), 3.81 - 3.71 (m, 1 H), 3.50 (t, J = 6.9 Hz, 2 H), 2.00 - 1.61 (m, 6 H), 1.61 - 1.52 (m, 2 H), 0.83 (t, J = 7.4 Hz, 3 H); MS (ESI): m/z: 323 [M+H]+. Intermediate 119: ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate
Figure imgf000105_0001
A solution of 5.00 g (0.028 mol) of ethyl 2-formylpyhdine-4-carboxylate (FLRCHEM Cat No. 220837) in 48 ml of dry isopropanol was treated with 3.3 ml (0.036 mmol) of aniline and 10.5 g (0.045 mol) of diphenyl phosphite, at r.t. under stirring. The reaction was stirred at r.t. for about 2.5 h; the resulting precipitate was filtered, washed with cold 1 :1 EtOH:water, then cold EtOH and dried under vacuum. About 1 1.6 g (83%) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119) was obtained as a colourless solid. 1H NMR (DMSO-d6) δ (ppm): 8.84 - 8.72 (m, 1 H), 8.29 - 8.21 (m, 1 H), 7.81 - 7.73 (m, 1 H), 7.38 - 6.87 (m, 14 H), 6.83 - 6.74 (m, 1 H), 6.65 - 6.55 (m, 1 H), 5.89 - 5.75 (m, 1 H), 4.41 - 4.29 (m, 2 H), 1.32 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 489 [M+H]+.
Intermediate 120: ethyl 2-[2-(4-chlorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000105_0002
A flask charged with 0.35 g (2.28 mmol) of cesium carbonate was dried at 125°C under vacuum for 16 h. After cooling at r.t. 0.40 g, (0.82 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119), 0.32 g (2.28 mmol) of 4-chlorobenzaldehyde, 2 ml of dry THF and 0.5 ml of dry isopropanol were sequentially added under Ar atmosphere. The resulting yellow suspension was stirred at r.t. for 20 h. 3 ml of 1 M aqueous HCI was added and the resulting red solution was stirred for 2 h at r.t.. The reaction mixture was neutralized with 0.8 ml of 1 M NaOH and then extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2S04 and concentrated in vacuum. The residue was purified by flash chromatography (eluent hexane:EtOAc from 92:8 to 85:15) to give 0.073 g (29%) of ethyl 2-[2-(4- chlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 120) as a yellow oil. 1H NMR (CDCIs) δ (ppm): 8.82 - 8.92 (m, 1 H), 8.54 - 8.59 (m, 1 H), 8.03 - 8.08 (m, 1 H), 7.25 - 7.32 (m, 4H), 4.54 (s, 2H), 4.44 (q, J = 7.01 Hz, 2H), 1.43 (t, J = 7.09 Hz, 3H); MS (ESI): m/z: 304 [M+H]+.
Intermediate 121 : ethyl 2-[2-(4-benzyloxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000106_0001
0.068 g (29%) of ethyl 2-[2-(4-benzyloxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 121 ) was prepared according to the procedure described for Intermediate 120, starting from 0.31 g (0.63 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.83 - 8.92 (m, 1 H), 8.51 - 8.60 (m, 1 H), 8.01 - 8.08 (m, 1 H), 7.30 - 7.48 (m, 5H), 6.90 - 7.30 (m, 5H), 5.05 (s, 2H), 4.50 (s, 2H), 4.44 (q, J = 6.85 Hz, 2H), 1.41 (t, J = 6.80 Hz, 3H); MS (ESI): m/z: 376 [M+H]+.
Intermediate 122: ethyl 2-[2-(4-phenoxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000106_0002
0.070 g (38%) of ethyl 2-[2-(4-phenoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 122) was prepared according to the procedure described for Intermediate 120, starting from 0.25 g (0.51 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.91 - 8.83 (m, 1 H), 8.61 - 8.52 (m, 1 H), 8.06 (dd, J = 2.0, 4.9 Hz, 1 H), 7.37 - 7.28 (m, 4 H), 7.14 - 7.07 (m, 1 H), 7.04 - 6.92 (m, 4 H), 4.54 (s, 2 H), 4.45 (q, J = 7.3 Hz, 2 H), 1.45 - 1.40 (m, 3 H); MS (ESI): m/z: 362 [M+H]+.
Intermediate 123: ethyl 2-[2-[4-[(4-fluorophenyl)methoxy]phenyl]acetyl]pyridine-4- carboxylate
Figure imgf000107_0001
0.13 g (65%) of ethyl 2-[2-[4-[(4-fluorophenyl)methoxy]phenyl]acetyl]pyridine-4- carboxylate (Intermediate 123) was prepared according to the procedure described for Intermediate 120, starting from 0.25 g (0.51 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (CDCIs) δ (ppm): 8.92 - 8.85 (m, 1 H), 8.57 - 8.54 (m, 1 H), 8.04 (dd, J = 2.0, 4.9 Hz, 1 H), 7.43 - 7.35 (m, 2 H), 7.31 - 7.23 (m, 2 H), 7.1 1 - 7.03 (m, 2 H), 6.96 - 6.88 (m, 2 H), 5.00 (s, 2 H), 4.55 - 4.38 (m, 4 H), 1.47 - 1.36 (m, 3 H); MS (ESI): m/z: 394 [M+H]+.
Intermediate 124: ethyl 2-[2-[3-[(4-fluorophenyl)methoxy]phenyl]acetyl]pyridine-4- carboxylate
Figure imgf000107_0002
0.12 g (64%) of ethyl 2-[2-[3-[(4-fluorophenyl)methoxy]phenyl]acetyl]pyridine-4- carboxylate (Intermediate 124) was prepared according to the procedure described for Intermediate 120, starting from 0.25 g (0.51 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.90 - 8.81 (m, 1 H), 8.60 - 8.51 (m, 1 H), 8.05 (dd, J = 2.0, 4.9 Hz, 1 H), 7.43 - 7.35 (m, 2 H), 7.27 - 7.19 (m, 1 H), 7.1 1 - 7.03 (m, 2 H), 6.99 - 6.93 (m, 2 H), 6.89 - 6.82 (m, 1 H), 5.01 (s, 2 H), 4.54 (s, 2 H), 4.44 (q, J = 7.3 Hz, 2 H), 1.47 - 1.37 (m, 3 H); MS (ESI): m/z: 394 [M+H]+.
Intermediate 125: ethyl 2-[2-(4-phenylphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000108_0001
0.14 g (81 %) of ethyl 2-[2-(4-phenylphenyl)acetyl]pyridine-4-carboxylate (Intermediate 125) was prepared according to the procedure described for Intermediate 120, starting from 0.25 g (0.51 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (DMSO-d6) δ (ppm): 9.06 - 8.97 (m, 1 H), 8.35 - 8.28 (m, 1 H), 8.12 (dd, J = 2.0, 4.9 Hz, 1 H), 7.71 - 7.29 (m, 9 H), 4.61 (s, 2 H), 4.44 - 4.32 (m, 2 H), 1.35 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 346 [M+H]+.
Intermediate 126: ethyl 2-[2-(3,5-dichlorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000108_0002
0.050 g (33%) of ethyl 2-[2-(3,5-dichlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 126) was prepared according to the procedure described for Intermediate 120, starting from 0.22 g (0.45 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). MS (ESI): m/z: 339 [M+H]+.
Intermediate 127: ethyl 2-[2-(3,4-dichlorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000109_0001
0.045 g (30%) of ethyl 2-[2-(3,4-dichlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 127) was prepared according to the procedure described for Intermediate 120, starting from 0.22 g (0.45 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). MS (ESI): m/z: 339 [M+H]+.
Intermediate 128: ethyl 2-[2-(3-benzyloxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000109_0002
0.17 g (55%) of ethyl 2-[2-(3-benzyloxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 128) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). MS (ESI): m/z: 376 [M+H]+.
Intermediate 129: ethyl 2-[2-(3-chlorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000109_0003
0.13 g (43%) of ethyl 2-[2-(3-chlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 129) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). MS (ESI): m/z: 304 [M+H]+.
Intermediate 130: ethyl 2-[2-(3-phenoxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000110_0001
0.13 g (44%) of ethyl 2-[2-(3-phenoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 130) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). MS (ESI): m/z: 362 [M+H]+.
Intermediate 131 : ethyl 2-[2-(3-phenylphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000110_0002
0.070 g (25%) of ethyl 2-[2-(3-phenylphenyl)acetyl]pyridine-4-carboxylate (Intermediate 131 ) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). MS (ESI): m/z: 346 [M+H]+.
Intermediate 132: ethyl 2-[2-(1 -naphthyl)acetyl]pyridine-4-carboxylate
Figure imgf000110_0003
0.135 g (52%) of ethyl 2-[2-(1 -naphthyl)acetyl]pyridine-4-carboxylate (Intermediate 132) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.96 - 8.89 (m, 1 H), 8.59 - 8.52 (m, 1 H), 8.12 - 8.05 (m, 1 H), 8.02 - 7.39 (m, 7 H), 5.04 (s, 2 H), 4.43 (q, J = 7.0 Hz, 2 H), 1.41 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 320 [M+H]+.
Intermediate 133: ethyl 2-[2-(2-naphthyl)acetyl]pyridine-4-carboxylate
Figure imgf000111_0001
0.130 g (50%) of ethyl 2-[2-(2-naphthyl)acetyl]pyridine-4-carboxylate (Intermediate 133) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). MS (ESI): m/z: 320 [M+H]+.
Intermediate 134: ethyl 2-[2-[3-(4-methylpiperazin-1 -yl)phenyl]acetyl]pyridine-4- carboxylate
Figure imgf000111_0002
0.230 g (87%) of ethyl 2-[2-[3-(4-methylpiperazin-1 -yl)phenyl]acetyl]pyridine-4- carboxylate (Intermediate 134) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (DMSO-de) δ (ppm): 9.06 - 8.94 (m, 1 H), 8.33 - 8.25 (m, 1 H), 8.10 (dd, J = 1.5, 4.9 Hz, 1 H), 7.15 - 7.02 (m, 1 H), 6.88 - 6.61 (m, 3 H), 4.53 - 4.32 (m, 4 H), 3.16 - 3.03 (m, 4 H), 2.46 - 2.36 (m, 4 H), 2.20 (s, 3 H), 1.42 - 1.25 (m, 3 H) MS (ESI): m/z: 368 [M+H]+.
Intermediate 135: ethyl 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000111_0003
0.030 g (10%) of ethyl 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 135) was prepared according to the procedure described for Intermediate 12, starting from 0.23 g (0.62 mmol) of ethyl 2-[2-[3-(4-methylpiperazin-1 -yl)phenyl]acetyl]pyridine-4-carboxylate (Intermediate 134). 1H NMR (CDCI3) δ (ppm): 8.82 - 8.64 (m, 1 H), 8.53 - 8.42 (m, 1 H), 7.79 (dd, J = 1.5, 4.9 Hz, 1 H), 7.17 - 7.05 (m, 1 H), 6.99 - 6.90 (m, 1 H), 6.84 - 6.54 (m, 2 H), 5.60 - 5.49 (m, 1 H), 4.49 - 4.30 (m, 4 H), 3.80 - 3.55 (m, 2 H), 3.16 (bs, 4 H), 2.55 (bs, 4 H), 2.35 (s, 3 H), 1.96 - 1.48 (m, 6 H), 1.41 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 467 [M+H]+.
Intermediate 136: 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000112_0001
0.026 g (93%) of 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N-tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 136) was prepared according to the procedure described for Intermediate 23, starting from 0.030 g (0.06 mmol) of ethyl 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 135). 1H NMR (DMSO-d6) δ (ppm): 8.81 - 8.71 (m, 1 H), 8.32 - 8.18 (m, 1 H), 7.79 (dd, J = 1 .5, 4.9 Hz, 1 H), 7.10 - 6.99 (m, 1 H), 6.93 - 6.84 (m, 1 H), 6.77 - 6.58 (m, 2 H), 5.43 (bs, 1 H), 4.35 - 4.19 (m, 2 H), 3.59 - 3.45 (m, 2 H), 3.16 - 2.97 (m, 4 H), 2.60 (bs, 4 H), 2.33 (s, 3 H), 1.84 - 1.35 (m, 6 H); MS (ESI): m/z: 439 [M+H]+.
Intermediate 137: ethyl 2-[2-(4-fluorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000112_0002
0.018 g (10%) of ethyl 2-[2-(4-fluorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 137) was prepared according to the procedure described for Intermediate 120, starting from 0.30 g (0.64 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.93 - 8.80 (m, 1 H), 8.61 - 8.45 (m, 1 H), 8.06 (dd, J = 1.7, 5.1 Hz, 1 H), 7.36 - 6.96 (m, 4 H), 4.61 - 4.32 (m, 4 H), 1.42 (t, J = 7.3 Hz, 3 H); MS (ESI): m/z: 288 [M+H]+.
Intermediate 138: [1 -(2-bromo-4-pyri trifluoro-ethoxy]-trimethyl-silane
Figure imgf000113_0001
To a solution of 1.00 g (5.38 mmol) of 2-bromo-4-pyridincarboxaldehyde (FLRCHEM Cat No. 047751 ) and 1.19 ml (8.1 mmol) of trimethyl(trifluoromethyl)silane in 16 ml of dry DMF, 0.037 g (0.27 mmol) of potassium carbonate was added. The mixture was vigorously stirred at r.t. for about 3 h, poured into brine and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2S04 and concentrated in vacuum. The residue was purified by flash chromatography (eluent hexane: EtOAc from 98:2 to 80:20) to give 1.20 g (68%) of [1 -(2-bromo-4-pyridyl)-2,2,2-trifluoro-ethoxy]- trimethyl-silane (Intermediate 138) as a light yellow oil. 1H NMR (DMSO-d6) δ (ppm): 8.61 - 8.22 (m, 1 H), 7.66 - 7.52 (m, 1 H), 7.43 - 7.30 (m, 1 H), 4.95 - 4.83 (m, 1 H), 0.18 (s, 9 H); MS (ESI): m/z: 328 [M+H]+.
Intermediate 139: [1 -[2-(1 -ethoxyvinyl)-4-pyridyl]-2,2,2-trifluoro-ethoxy]-trimethyl- silane
Figure imgf000113_0002
0.088 g (0.080 mmol) of Tetrakis(triphenylphosphine)palladium(0) was added to a degassed solution of 0.50 g (1 .5 mmol) of [1 -(2-bromo-4-pyridyl)-2,2,2-trifluoro-ethoxy]- trimethyl-silane (Intermediate 138) and 0.62 ml (1.8 mmol) of tributyl(1 - ethoxyvinyl)stannane in 5.5 ml of dry 1 ,4-dioxane and 2.3 ml of dry DMF; argon was bubbled for 5 more min. and the mixture was heted at 1 10°C for about 24 h, then filtered thorough a thin pad of celite, washing with EtOAc. The combined organics were concentrated in vacuum. The residue was purified by flash chromatography (eluent hexane:EtOAc from 98:2 to 80:20) to give 0.30 g (62%) of [1 -[2-(1 -ethoxyvinyl)-4-pyridyl]- 2,2,2-trifluoro-ethoxy]-trimethyl-silane (Intermediate 139) as an orange oil. 1H NMR (DMSO-de) δ (ppm): 8.64 - 8.53 (m, 1 H), 7.81 - 7.71 (m, 1 H), 7.49 - 7.35 (m, 1 H), 5.72 - 5.62 (m, 1 H), 5.42 (d, J = 1.5 Hz, 1 H), 4.46 (d, J = 1 .5 Hz, 1 H), 4.02 - 3.87 (m, 2 H), 1.37 (t, J = 6.8 Hz, 3 H), 0.23 -0.06 (m, 9 H); MS (ESI): m/z: 320 [M+H]+.
Intermediate 140: 1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone hydrochloride
Figure imgf000114_0001
A solution of 0.15 g (0.47 mmol) of [1 -[2-(1 -ethoxyvinyl)-4-pyridyl]-2,2,2-trifluoro-ethoxy]- trimethyl-silane (Intermediate 139) in 3 ml of THF was treated with 0.47 ml (0.94 mmol) of 2N HCI in water at rt for about for about 20h. Solvents were evaporated, the residue treated with Et20 and evaporated (X4); then it was dried under vacuum at rt to yield 0.1 1 mg (93%) of 1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone hydrochloride (Intermediate 140) as a reddish solid.1H NMR (DMSO-d6) δ (ppm): 8.83 - 8.71 (m, 1 H), 8.1 1 - 8.02 (m, 1 H), 7.80 - 7.71 (m, 1 H), 5.52 - 5.40 (m, 1 H), 2.65 (s, 3 H); MS (ESI): m/z: 256 [M+H]+.
Intermediate 141 : 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]-4-pyridyl]ethanol
Figure imgf000114_0002
0.1 1 g (79%) of 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]-4-pyridyl]ethanol (Intermediate 141 ) was prepared according to the procedure described for Intermediate 12, starting from 0.12 g (0.44 mmol) of 1 -[4-(2,2,2- trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone hydrochloride (Intermediate 140) and heating the reaction mixture at reflux for about 4 h. 1H NMR (DMSO-d6) δ (ppm): 8.70 - 8.58 (m, 1 H), 8.06 - 7.96 (m, 1 H), 7.55 - 7.50 (m, 1 H), 7.23 - 7.12 (m, 1 H), 5.46 - 5.32 (m, 2 H), 3.82 - 3.48 (m, 2 H), 2.32 (s, 3 H), 1.86 - 1.45 (m, 6 H); MS (ESI): m/z: 319 [M+H]+.
Intermediate 142: 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy- carbonimidoyl]-4-pyridyl]ethane-1 ,1 -diol
Figure imgf000115_0001
A solution of 0.075 g (0.24 mmol) of 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N-tetrahydropyran- 2-yloxy-carbonimidoyl]-4-pyridyl]ethanol (Intermediate 141 ) in 1.8 ml of EtOAc was treated with 0.13 g (0.48 mmol) of 2-lodoxybenzoic acid. After stirring at reflux for about 4 h. the mixture was allowed to cool to r.t. and filtered, washing the solid residue with EtOAc. The filtrate was concentrated in vacuum and the residue purified by reverse phase chromatography (eluent H2O:ACN from 80:20 to 40:60) to give 0.030 g (40%) of 2,2,2- trifluoro-1 -[2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy-carbonimidoyl]-4-pyridyl]ethane- 1 ,1 -diol (Intermediate 142). 1H NMR (DMSO-d6) δ (ppm): 8.75 - 8.62 (m, 1 H), 8.14 - 7.90 (m, 3 H), 7.63 - 7.47 (m, 1 H), 5.40 (bs, 1 H), 3.82 - 3.49 (m, 2 H), 2.34 (s, 3 H), 1.80 (d, J = 7.3 Hz, 6 H); MS (ESI): m/z: 335 [M+H]+.
Intermediate 143: ethyl 2-[2-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]acetyl]pyridine- 4-carboxylate
Figure imgf000115_0002
0.19 g (58%) of ethyl 2-[2-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]acetyl]pyridine-4- carboxylate (Intermediate 143) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2- [anilino(diphenoxyphosphoryl)methyl]pyridine-4-carboxylate (Intermediate 119). 1H NMR (CDCIs) δ (ppm): 8.90 - 8.84 (m, 1 H), 8.58 - 8.52 (m, 1 H), 8.07 - 7.99 (m, 1 H), 7.21 - 7.1 1 (m, 1 H), 6.96 - 6.88 (m, 1 H), 6.88 - 6.82 (m, 1 H), 6.75 - 6.67 (m, 1 H), 4.50 (s, 2 H), 4.44 (q, J = 7.3 Hz, 2 H), 1.42 (t, J = 7.3 Hz, 3 H), 0.99 - 0.95 (m, 9 H), 0.20 - 0.15 (m, 6 H); MS (ESI): m/z: 400 [M+H]+.
Intermediate 144: ethyl 2-[2-(3-hydroxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000116_0001
0.13 g (0.51 mmol) of tetrabutylammonium fluoride was added dropwise to a solution of 0.17 g (0.43 mmol) of ethyl 2-[2-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]acetyl]pyridine-4- carboxylate (Intermediate 143) in 10 ml of dry THF at 0 °C. The resulting mixture was stirred at 0 °C for about 2 h, washed successively with water and saturated NaCI. The resulting organic layer was dried over anhydrous Na2S04, and concentrated. The residue was purified by column chromatography (eluent Hexane:EtOAc from 95:5 to 80:20) to obtain 0.10 g (82%) of ethyl 2-[2-(3-hydroxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 144) as a beige solid. 1H NMR (CDCI3) δ (ppm): 8.91 - 8.84 (m, 1 H), 8.59 - 8.53 (m, 1 H), 8.08 - 8.02 (m, 1 H), 7.22 - 7.15 (m, 1 H), 6.94 - 6.88 (m, 1 H), 6.86 - 6.80 (m, 1 H), 6.76 - 6.69 (m, 1 H), 4.52 (s, 2 H), 4.44 (q, J = 7.3 Hz, 2 H), 1 .42 (t, J = 7.3 Hz, 3 H); MS (ESI): m/z: 286 [M+H]+.
Intermediate 145: ethyl 2-[(E)-C-[(3-hydroxyphenyl)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000116_0002
A mixture of 0.035 g (0.12 mmol) of ethyl 2-[2-(3-hydroxyphenyl)acetyl]pyridine-4- carboxylate (Intermediate 144) and 0.043 g (0.37 mmol) of O-tetrahydropyran-2- ylhydroxylamine in 3 ml of EtOH was heated at 68°C. Solvent was removed under vacuum and the residue purified by flash chromatography (eluent Hexane:EtOAc 80:20) to obtain 0.030 g (65%) of ethyl 2-[(E)-C-[(3-hydroxyphenyl)methyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 145) as a colourless oil. 1H NMR (CDCIs) δ (ppm): 8.79 - 8.73 (m, 1 H), 8.54 - 8.48 (m, 1 H), 7.85 - 7.78 (m, 1 H), 7.14 - 7.05 (m, 1 H), 6.95 - 6.87 (m, 1 H), 6.83 - 6.77 (m, 1 H), 6.67 - 6.60 (m, 1 H), 5.58 - 5.51 (m, 1 H), 4.64 (bs, 1 H), 4.48 - 4.30 (m, 4 H), 3.76 - 3.58 (m, 2 H), 1.91 - 1.79 (m, 3 H), 1.60 - 1.49 (m, 3 H), 1.45 - 1.35 (m, 3 H); MS (ESI): m/z: 385 [M+H]+.
Intermediate 146: ethyl 2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl- amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-
4-carboxylate
Figure imgf000117_0001
A mixture of 0.030 g (0.08 mmol) of ethyl 2-[(E)-C-[(3-hydroxyphenyl)methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 145) and 0.022 g (0.16 mmol) of K2CO3 in 3 ml of dry DMF was stirred at r.t. for about 15 min., then 0.018 g (0.09 mmol) of 2-chloro-N-[(4-fluorophenyl)methyl]-N-methyl-ethanamine was added. Stirring was continued at r.t. for about 10 min. and at reflux for about 5 h., solvent was removed under vacuum and the residue purified by flash chromatography (eluent Hexane:EtOAc from 95:5 to 80:20) to obtain 0.035 g (80%) of ethyl 2-[(E)-C-[[3-[2-[(4- fluorophenyl)methyl-methyl-amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 146) as a yellow solid. 1H NMR (CDCI3) δ (ppm): 8.79 - 8.72 (m, 1 H), 8.53 - 8.47 (m, 1 H), 7.84 - 7.75 (m, 1 H), 7.33 - 7.28 (m, 1 H), 7.17 - 7.08 (m, 1 H), 7.05 - 6.95 (m, 3 H), 6.93 - 6.88 (m, 2 H), 6.71 - 6.65 (m, 1 H), 5.58 - 5.52 (m, 1 H), 4.46 - 4.31 (m, 4 H), 4.06 - 3.95 (m, 2 H), 3.76 - 3.47 (m, 6 H), 2.33 - 2.27 (m, 3 H), 1 .92 - 1.79 (m, 3 H), 1.69 - 1.59 (m, 3 H), 1.44 - 1.36 (m, 3 H); MS (ESI): m/z: 550 [M+H]+.
Intermediate 147: 2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl- amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine- 4-carboxylic acid
Figure imgf000118_0001
0.015 g (58%) of 2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl- amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy-carbo
carboxylic acid (Intermediate 147) was prepared according to the procedure described for Intermediate 23, starting from 0.030 g (0.05 mmol) of ethyl 2-[(E)-C-[[3-[2-[(4- fluorophenyl)methyl-methyl-amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 146). 1H NMR (CDCI3) δ (ppm): 8.69 - 8.61 (m, 1 H), 8.50 - 8.40 (m, 1 H), 7.75 - 7.65 (m, 1 H), 7.47 - 7.38 (m, 2 H), 7.15 - 7.03 (m, 3 H), 7.00 - 6.91 (m, 2 H), 6.65 (d, J = 8.3 Hz, 1 H), 5.52 - 5.41 (m, 1 H), 4.47 - 4.33 (m, 2 H), 4.27 - 4.16 (m, 2 H), 4.08 - 3.97 (m, 2 H), 3.82 - 3.70 (m, 2 H), 3.21 - 3.07 (m, 2 H), 2.55 (s, 3 H), 1.89 - 1.79 (m, 3 H), 1.65 - 1.47 (m, 3 H); MS (ESI): m/z: 522 [M+H]+. Intermediate 148: methyl 2-(4-fluorobenzoyl)pyridine-4-carboxylate
Figure imgf000118_0002
0.10 g (26%) of methyl 2-(4-fluorobenzoyl)pyridine-4-carboxylate (Intermediate 147) was prepared according to the procedure described for Intermediate 1 , starting from 0.20 g (1.46 mmol) of methyl isonicotinate and 0.24 g (1.94 mmol) of 4-fluorobenzaldehyde. 1 H NMR (CDCI3) δ (ppm): 8.92 - 8.86 (m, 1 H), 8.63 - 8.56 (m, 1 H), 8.22 - 8.15 (m, 2 H), 8.10 - 8.03 (m, 1 H), 7.24 - 7.14 (m, 2 H), 4.02 (s, 3 H); MS (ESI): m/z: 260 [M+H]+. Intermediate 149: ethyl 2-[2-(3-fluorophenyl)acetyl]pyridine-4-carboxylate
Figure imgf000119_0001
0.10 g (42%) of ethyl 2-[2-(3-fluorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 149) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.91 - 8.85 (m, 1 H), 8.60 - 8.54 (m, 1 H), 8.09 - 8.03 (m, 1 H), 7.32 - 7.28 (m, 1 H), 7.15 - 7.04 (m, 2 H), 7.00 - 6.90 (m, 1 H), 4.56 (s, 2 H), 4.45 (q, J = 7.3 Hz, 2 H), 1.47 - 1.37 (m, 3 H); MS (ESI): m/z: 288 [M+H]+. Intermediate 150: ethyl 2-[2-(3-methoxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000119_0002
0.16 g (67%) of ethyl 2-[2-(3-methoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 150) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.91 - 8.84 (m, 1 H), 8.60 - 8.54 (m, 1 H), 8.07 - 8.02 (m, 1 H), 7.27 - 7.20 (m, 1 H), 6.97 - 6.87 (m, 2 H), 6.83 - 6.76 (m, 1 H), 4.54 (s, 2 H), 4.44 (q, J = 7.2 Hz, 2 H), 3.80 (s, 3 H), 1.42 (t, J = 7.2 Hz, 3 H); MS (ESI): m/z: 300 [M+H]+.
Intermediate 151 : ethyl 2-[2-(4-methoxyphenyl)acetyl]pyridine-4-carboxylate
Figure imgf000119_0003
0.080 g (33%) of ethyl 2-[2-(4-methoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 151 ) was prepared according to the procedure described for Intermediate 120, starting from 0.40 g (0.82 mmol) of ethyl 2-[anilino(diphenoxyphosphoryl)methyl]pyridine-4- carboxylate (Intermediate 119). 1H NMR (CDCI3) δ (ppm): 8.90 - 8.83 (m, 1 H), 8.58 - 8.53 (m, 1 H), 8.06 - 8.00 (m, 1 H), 7.30 - 7.23 (m, 2 H), 6.90 - 6.82 (m, 2 H), 4.50 (s, 2 H), 4.44 (q, J = 7.3 Hz, 2 H), 3.79 (s, 3 H), 1 .45 - 1.37 (m, 3 H); MS (ESI): m/z: 300 [M+H]+. Intermediate 152: 2-[(Z)-C-[2-(4-phenyl-1 -piperidyl)ethoxymethyl]-N- tetrahydropyran-2-yloxy-carbonimido l]pyridine-4-carboxylic
Figure imgf000120_0001
0.025 g (13%) of 2-[(Z)-C-[2-(4-phenyl-1 -piperidyl)ethoxymethyl]-N-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic (Intermediate 152) was prepared according to the procedure described for Intermediate 19, starting from 0.15 g (0.42 mmol) of 2-Z-(C- (bromomethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl)pyridine-4-carboxylate
(Intermediate 12). The reaction was carried out at r.t. for about 20 h. 1H NMR (DMSO-d6) δ (ppm): 12.68 (bs, 1 H), 8.84 - 8.62 (m, 1 H), 8.29 - 8.09 (m, 1 H), 7.79 (dd, J = 1.5, 4.9 Hz, 1 H), 7.35 - 7.02 (m, 5 H), 5.43 (bs, 1 H), 4.84 - 4.62 (m, 2 H), 4.10 (bs, 2 H), 3.86 - 3.51 (m, 4 H), 3.09 - 2.54 (m, 4 H), 2.50 - 2.39 (m, 1 H), 2.29 - 1.31 (m, 10 H); MS (ESI): m/z: 468 [M+H]+.
Example 1 : methyl 2-[(E)-A -hydroxy-C-methyl-carbonimidoyl]pyridine-4- carboxylate
Figure imgf000120_0002
A mixture of 0.10 g (0.56 mmol) of methyl 2-acetylpyridine-4-carboxylate (Intermediate 1 ), 0.045 g (0.64 mmol) of hydroxylamine hydrochloride and 0.053 g (0.64 mmol) of sodium acetate in 0.90 mL of ethanol and 0.45 mL of water was stirred at 55°C for about 14 h. Further 0.0060 g (0.08 mmol) of hydroxylamine hydrochloride and 0.0070 g (0.08 mmol) of sodium acetate were added and heating was continued for another 8 h. The reaction mixture was allowed to cool down to r.t., 1 mL of water was added while cooling in an ice/water bath. The resulting solid was filtered, washed with water and dried to give 0.080 g (74%) of methyl 2-[(E)-N-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylate (Example 1 ) as a colourless solid. 1H NMR (DMSO-d6) δ (ppm): 1 1.72 (s, 1 H), 8.84-8.64 (m, 1 H), 8.38-8.23 (m, 1 H), 7.79 (dd, J=1 .7, 5.1 Hz, 1 H), 3.90 (s, 3 H), 2.23 (s, 3 H); MS (ESI): m/z: 195 [M+H]+.
Example 2: methyl 2-[(E)-/V-hydroxy-C-phenyl-carbonimidoyl]pyridine-4- carboxylate and Example 3: methyl 2-[(Z)-/V-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000121_0001
0.018 g (28%) of methyl 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylate (Example 2) and 0.010 g (16%) of methyl 2-[(Z)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 3) were prepared according to the procedure described for Example 1 , starting from 0.060 g (0.25 mmol) of methyl 2- benzoylpyridine-4-carboxylate (Intermediate 4). The reaction gave two products, the £- isomer and the Z-isomer of the oxime. The two isomers were separated by flash chromatography (eluent DCM:EtOAc from 98:2 to 80:20). Example 2 (E-isomer): 1H NMR (ACN-ds) δ (ppm): 9.47 (s, 1 H), 8.66 (dd, J=1 .0, 4.9 Hz, 1 H), 8.41-8.31 (m, J=1 .0 Hz, 1 H), 7.83 (dd, J=1 .5, 4.9 Hz, 1 H), 7.53-7.30 (m, 5 H), 3.98 (s, 3 H); MS (ESI): m/z: 257 [M+H]+; Example 3 (Z-isomer): 1H NMR (ACN-d3) δ (ppm): 9.89 (s, 1 H), 8.94-8.81 (m, 1 H), 8.07-7.85 (m, 2 H), 7.50-7.30 (m, 5 H), 3.97 (s, 3 H); MS (ESI): m/z: 257 [M+H]+. Example 4: methyl 2-[(E)-/V-hydroxy-C-cyclopropyl-carbonimidoyl]pyridine-4- carboxylate and Example 5: methyl 2-[(Z)-A -hydroxy-C-cyclopropyl- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000122_0001
0.035 g (54%) of methyl 2-[(E)-A/-hydroxy-C-cyclopropyl-carbonimidoyl]pyridine-4- carboxylate (Example 4) and 0.012 g (19%) of methyl 2-[(Z)-N-hydroxy-C-cyclopropyl- carbonimidoyl]pyridine-4-carboxylate (Example 5) were prepared according to the procedure described for Example 1 , starting from 0.060 g (0.29 mmol) of methyl 2- (cyclopropanecarbonyl)pyridine-4-carboxylate (Intermediate 5). The reaction gave two products, the E-isomer and the Z-isomer of the oxime. The two isomers were separated by flash chromatography (eluent DCM:EtOAc from 85:15 to 50:50). Example 4 (E- isomer): 1 H NMR (ACN-d3) δ (ppm): 9.12 (bs, 1 H), 8.79-8.61 (m, 1 H), 8.12-7.98 (m, 1 H), 7.86-7.69 (m, 1 H), 3.95 (s, 3 H), 2.59-2.41 (m, 1 H), 1 .31-0.80 (m, 4 H); MS (ESI): m/z: 221 [M+H]+; Example 5 (Z-isomer): 1 H NMR (ACN-d3) δ (ppm): 1 1 .35 (s, 1 H), 8.90- 8.74 (m, J=4.4 Hz, 1 H), 8.28-8.10 (m, 1 H), 7.91 (dd, J=1 .5, 4.9 Hz, 1 H), 3.97 (s, 3 H), 2.07-1 .96 (m, 1 H), 0.91 -0.71 (m, 4 H); MS (ESI): m/z: 221 [M+H]+.
Example 6: methyl 2-[(E)-C-ethyl-/V-hydroxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000122_0002
0.046 g (70.5%) of methyl 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 6) was prepared according to the procedure described for Example 1 , starting from 0.060 g (0.10 mmol) of methyl 2-propanoylpyridine-4-carboxylate (Intermediate 6). 1 H NMR (DMSO-de) δ (ppm): 8.86-8.76 (m, 1 H), 8.49-8.31 (m, 1 H), 7.86 (dd, J=1 .5, 4.9 Hz, 1 H), 4.00 (s, 3 H), 3.01 (q, J=7.4 Hz, 2 H), 1 .19 (t, J=7.4 Hz, 3 H); MS (ESI): m/z: 209 [M+H]+.
Example 7: methyl 2-[(E)-/V-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine- 4-carboxylate and Example 8: methyl 2-[(Z)-/V-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate
Figure imgf000123_0001
0.041 g (43%) of methyl 2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate (Example 7) and 0.035 g (37%) of methyl 2-[(Z)-N-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyhdine-4-carboxylate (Example 8) were prepared according to the procedure described for Example 1 , starting from 0.090 g (0.33 mmol) of methyl 2-(2-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 7) and using MeOH as solvent. The reaction gave two products, the E-isomer and the Z-isomer of the oxime. The two isomers were separated by flash chromatography (eluent DCM:EtOAc from 95:5 to 70:30). Example 7 (E-isomer): 1 H NMR (DMSO-d6) δ (ppm): 1 1 .69 (s, 1 H), 8.64-8.60 (m, 1 H), 8.32-8.28 (m, 1 H), 7.74 (dd, J=1 .5, 4.9 Hz, 1 H), 7.41 -6.93 (m, 4 H), 3.92 (s, 3 H), 3.58 (s, 3 H); MS (ESI): m/z: 287 [M+H]+; Example 8 (Z-isomer): 1H NMR (DMSO-d6) δ (ppm): 1 1 .83 (s, 1 H), 8.76-8.61 (m, 1 H), 8.34-8.21 (m, 1 H), 7.83-7.70 (m, 1 H), 7.48- 7.31 (m, 2 H), 7.07-6.89 (m, 2 H), 3.93 (s, 3 H), 3.39 (s, 3 H); MS (ESI): m/z: 287 [M+H]+. Example 9: methyl 2-[(E)-/V-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine- 4-carboxylate and Example 10: methyl 2-[(Z)-/V-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate
Figure imgf000123_0002
0.022 g (42%) of methyl 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate (Example 9) and 0.027 g (51 %) of methyl 2-[(Z)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 10) were prepared according to the procedure described for Example 1 , starting from 0.050 g (0.18 mmol) of methyl 2-(3-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 9) and using 2 ml_ of MeOH as solvent. The reaction gave two products, the E-isomer and the Z-isomer of the oxime. The two isomers were separated by flash chromatography (DCM:EtOAc from 95:5 to 70:30). Example 9 (E-isomer): 1H NMR (DMSO-d6) δ (ppm): 1 1.84 (s, 1 H), 8.70- 8.67 (m, 1 H), 8.31-8.28 (m, 1 H), 7.79 (dd, J=1.7, 5.1 Hz, 1 H), 7.36-7.31 (m, 1 H), 6.98- 6.82 (m, 3 H), 3.93 (s, 3 H), 3.74 (s, 3 H); MS (ESI): m/z: 287 [M+H]+; Example 10 (Z- isomer): 1H NMR (DMSO-d6) δ (ppm): 1 1.68 (s, 1 H), 8.89 (dd, J=1.0, 4.9 Hz, 1 H), 7.95- 7.92 (m, 1 H), 7.88 (dd, J=1.5, 4.9 Hz, 1 H), 7.30-7.23 (m, 1 H), 6.99-6.80 (m, 3 H), 3.92 (s, 2 H), 3.73 (s, 3 H); MS (ESI): m/z: 287 [M+H]+.
Example 11 : ferf-butyl 2-[(E)-C-benzyl-/V-hydroxy-carbonimidoyl]pyridine-4- carboxylate
Figure imgf000124_0001
0.055 g (70%) of te/t-butyl 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylate (Example 11 ) was prepared according to the procedure described for Example 1 , starting from 0.075 g (0.25 mmol) of te/t-butyl 2-(2-phenylacetyl)pyridine-4- carboxylate (Intermediate 10). 1H NMR (DMSO-d6) 5 (ppm): 1 1.93 (s, 1 H), 8.85-8.67 (m, 1 H), 8.30-8.19 (m, 1 H), 7.73 (dd, J=1.5, 4.9 Hz, 1 H), 7.26-7.06 (m, 5 H), 4.26 (s, 2 H), 1.69-1.37 (m, 9 H); MS (ESI): m/z: 313 [M+H]+.
Example 12: 2-[(E)-/V-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000124_0002
A solution of 0.065 g (0.34 mmol) of methyl 2-[(E)-A/-hydroxy-C-methyl- carbonimidoyl]pyridine-4-carboxylate (Example 1 ) and 0.020 g (0.84 mmol) of LiOH in 9 mL of THF and 1.8 mL of water was stirred at r.t. for about 1.5 h. THF was removed under vacuum, the residue diluted with water and washed with DCM. The aqueous phase was brought to pH 3 with AcOH, while cooling in an ice/water bath, and the product was extracted with DCM: n-BuOH 8:2 and 1 :1 . The combined organic layers were evaporated. The residue was treated with water at r.t, filtered, washed with little cold water and dried under vacuum at 40°C. About 0.036 g (60%) of 2-[(E)-N-hydroxy-C-methyl- carbonimidoyl]pyridine-4-carboxylic acid (Example 12) were obtained as a beige solid. 1 H NMR (DMSO-de) δ (ppm): 13.73 (bs, 1 H), 1 1 .68 (s, 1 H), 8.82-8.66 (m, 1 H), 8.35-8.21 (m, 1 H), 7.77 (dd, J=1 .5, 5.4 Hz, 1 H), 2.23 (s, 3 H. MS (ESI): m/z: 181 [M+H]+.
Example 13: 2-[(E)-/V-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000125_0001
0.014 g (82%) of 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid (Example 13) was prepared according to the procedure described for Example 12, starting from 0.018 g (0.070 mmol) of methyl 2-[(E)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 2). 1H NMR (DMSO-d6) δ (ppm): 13.81 (bs, 1 H), 1 1 .80 (s, 1 H), 8.72-8.57 (m, 1 H), 8.38-8.1 1 (m, 1 H), 7.77 (dd, J=1 .5, 4.9 Hz, 1 H), 7.48-7.17 (m, 5 H); MS (ESI): m/z: 243 [M+H]+.
Example 14: 2-[(Z)-/V-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000125_0002
0.0090 g (95%) of 2-[(Z)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid (Example 14) was prepared according to the procedure described for Example 12, starting from 0.010 g (0.040 mmol) of methyl 2-[(Z)-A/-hydroxy-C-phenyl- carbonimidoyl]pyridine-4-carboxylate (Example 3). 1H NMR (DMSO-d6) δ (ppm): 13.84 (bs, 1 H), 1 1 .64 (s, 1 H), 8.91-8.74 (m, 1 H), 8.00-7.78 (m, 2 H), 7.52-7.10 (m, 5 H); MS (ESI): m/z: 243 [M+H]+.
Example 15: 2-[(E)-C-cyclopropyl-/V-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000126_0001
0.010 g (31 %) of 2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 15) was prepared according to the procedure described for Example 12, starting from 0.035 g (0.59 mmol) of methyl 2-[(E)-C-cyclopropyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 4). 1H NMR (ACN-d3) δ (ppm): 8.77-8.63 (m, 1 H), 8.14-7.97 (m, 1 H), 7.80 (dd, J=1.5, 4.9 Hz, 1 H), 2.56-2.43 (m, 1 H), 1.31-0.81 (m, 4 H); MS (ESI): m/z: 207 [M+H]+.
Example 16: 2-[(Z)-C-cyclopropyl-/V-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000126_0002
0.008 g (68%) of 2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 16) was prepared according to the procedure described for Example 12, starting from 0.012 g (0.054 mmol) of methyl 2-[(Z)-C-cyclopropyl-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 5). 1H NMR (ACN-d3) δ (ppm): 8.92-8.70 (m, 1 H), 8.28-8.13 (m, 1 H), 7.92 (dd, J=1.5, 5.4 Hz, 1 H), 2.09-1.96 (m, 1 H), 0.94-0.72 (m, 4 H); MS (ESI): m/z: 207 [M+H]+.
Example 17: 2-[(E)-C-ethyl-/V-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000126_0003
0.023 g (65%) of 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 17) was prepared according to the procedure described for Example 12, starting from 0.039 g (0.19 mmol) of methyl 2-[(E)-C-ethyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 6). 1H NMR (DMSO-d6) δ (ppm): 13.70 (bs, 1 H), 1 1.60 (s, 1 H), 8.85-8.68 (m, 1 H), 8.37-8.07 (m, 1 H), 7.76 (dd, J=1.7, 5.1 Hz, 1 H), 2.85 (q, J=7.5 Hz, 2 H), 1.15-0.84 (m, 3 H); MS (ESI): m/z: 195 [M+H]+.
Example 18: 2-[(E)-/V-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000127_0001
0.0080 g (42%) of 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 18) was prepared according to the procedure described for Example 12, starting from 0.020 g (0.070 mmol) of methyl 2-[(E)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 9). 1H NMR (DMSO-de) δ (ppm): 13.59 (bs, 1 H), 1 1.79 (s, 1 H), 8.73-8.57 (m, 1 H), 8.36-8.17 (m, 1 H), 7.76 (dd, J=1.5, 5.4 Hz, 1 H), 7.39-7.25 (m, 1 H), 6.99-6.91 (m, 1 H), 6.89-6.81 (m, 2 H), 3.78- 3.69 (m, 3 H); MS (ESI): m/z: 273 [M+H]+.
Example 19: 2-[(Z)-/V-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000127_0002
0.020 g (84%) of 2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 19) was prepared according to the procedure described for Example 12, starting from 0.025 g (0.090 mmol) of methyl 2-[(Z)-A/-hydroxy-C-(3- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 10). 1H NMR (DMSO-de) δ (ppm): 11.97 (bs, 1 H), 1 1.64 (s, 1 H), 8.93-8.74 (m, 1 H), 7.92-7.87 (m, 1 H), 7.84 (dd, J=1.5, 4.9 Hz, 1 H), 7.31-7.22 (m, 1 H), 7.00-6.93 (m, 2 H), 6.85-6.78 (m, 1 H), 3.73 (s, 3 H); MS (ESI): m/z: 273 [M+H]+.
Example 20: 2-[(E)-/V-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000128_0001
0.010 g (23%) of 2-[(E)-N-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 20) was prepared according to the procedure described for Example 12, starting from 0.040 g (0.14 mmol) of methyl 2-[(E)-N-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 7). 1H NMR (DMSO-d6) δ (ppm):13.69 (bs, 1 H), 1 1 .65 (s, 1 H), 8.66-8.49 (m, 1 H), 8.35-8.21 (m, 1 H), 7.72 (dd, J=1 .5, 4.9 Hz, 1 H), 7.40-7.31 (m, 1 H), 7.15-7.10 (m, 1 H), 7.08-7.02 (m, 1 H), 7.02- 6.95 (m, 1 H), 3.59 (s, 3 H); MS (ESI): m/z: 273 [M+H]+.
Example 21 : 2-[(Z)-/V-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000128_0002
0.012 g (37%) of 2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 21 ) was prepared according to the procedure described for Example 12, starting from 0.034 g (0.12 mmol) of methyl 2-[(Z)-A/-hydroxy-C-(2- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 8). 1H NMR (DMSO-de) δ (ppm): 13.88-13.60 (m, 1 H), 1 1 .85 (s, 1 H), 8.74-8.54 (m, 1 H), 8.26 (s, 1 H), 7.82-7.62 (m, 1 H), 7.49-7.29 (m, 2 H), 7.10-6.87 (m, 2 H), 3.40 (s, 3 H); MS (ESI): m/z: 271 [M- H].
Example 22: 2-[(E)-C-benzyl-/V-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000129_0001
0.023 g (62%) of 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 22) was prepared according to the procedure described for Example 12, starting from 0.045 g (0.14 mmol) of te/t-butyl 2-[(E)-C-benzyl-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 11 ). 1H NMR (DMSO-d6) δ (ppm): 13.66 (bs, 1 H), 1 1.88 (s, 1 H), 8.82-8.66 (m, 1 H), 8.40-8.21 (m, 1 H), 7.76 (dd, J=1.2, 5.1 Hz, 1 H), 7.28-6.99 (m, 5 H), 4.26 (s, 2 H); MS (ESI): m/z: 257 [M+H]+.
Example 23: 2-[(E)-/V-hydroxy-(C-[(4-methylpiperazin-1 - yl)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000129_0002
0.0040 g (0.17 mmol) of LiOH was added at 0°C to a solution of 0.032 g (0.085 mmol) of methyl 2-[(E)-C-[(4-methylpiperazin-1 -yl)methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 16) in a mixture of 1 mL of THF and 0.3 mL of water. After stirring 1 h at r.t, THF was removed under vacuum, the residue diluted with water and washed with DCM. The aqueous phase was acidified with 5% citric acid and the product was extracted with DCM: n-BuOH 8:2. The combined organic layers were evaporated, the residue was dissolved in water, passed through a RP-SPE cartridge and eluted with water followed by MeOH to afford 0.020 g (69%) of 2-[(E)-C- (phenoxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid. 0.070 mL (0.28 mmol) of 4 M HCI in 1 ,4-dioxane was added at 0°C to a suspension of 0.020 g (0.055 mmol) of 2-[(E)-C-(phenoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid in 1 mL of 1 ,4-dioxane. The mixture was stirred at r.t. for about 1 h, then diluted with water and brought to pH 4-5 with AcONa and finally concentrated to dryness. The residue was dissolved in water and passed onto a RP-SPE cartridge (eluent: water, then MeOH) to afford 0.004 g (17%) of 2-[(E)-N-hydroxy-(C-[(4- methylpiperazin-1 -yl)methyl]carbonimidoyl]pyridine-4-carboxylic acid (Example 23). 1 H NMR (DMSO-de) δ (ppm): 12.27-1 1 .33 (bs, 2 H), 8.62-8.49 (m, 1 H), 8.14-8.03 (m, 1 H), 7.72-7.58 (m, 1 H), 3.90-3.76 (m, 2H), 3.49-3.20 (m, 4 H), 2.57-2.23 (m, 7 H); MS (ESI): m/z: 279 [M+H]+.
Example 24: 2-[(Z)-/V-hydroxy-C-(methoxymethyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000130_0001
0.072 mL (0.29 mmol) of 4 M HCI in 1 ,4-dioxane was added at 0°C to a solution of 0.017 g (0.058 mmol) of 2-[(Z)-C-(methoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 23). The mixture was stirred at r.t. for about 1 h, then the solvents were decanted, the residue treated successively with
DCM and Et20 and decanted, then dried under vacuum to afford about 0.01 1 g (91 %) of
2-[(Z)-A/-hydroxy-C-(methoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid (Example 24). 1 H NMR (DMSO-d6) δ (ppm): 14.66-12.66 (m, 1 H), 12.06 (s, 1 H), 8.85-8.69 (m, 1
H), 8.25-8.16 (m, 1 H), 7.84-7.74 (dd, J=1 .5, 5.2 Hz, 1 H), 4.62 (s, 2 H), 3.29-3.16 (m, 3
H); MS (ESI): m/z: 21 1 [M+H]+.
Example 25: 2-[(Z)-A -hydroxy-C-(2- methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid trifluoroacetate
Figure imgf000130_0002
0.070 g (69%) of 2-[(Z)-A/-hydroxy-C-(2- methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid trifluoroacetate (Example 25) was prepared according to the procedure described for Example 24, starting from 0.020 9 (0.046 mmol) 2-[(Z)-C-[2-(fert- butoxycarbonyl(methyl)amino)ethoxymethyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 19). The crude product was purified by preparative HPLC to afford 2-[(Z)-N-hydroxy-C-(2- methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid as trifluoroacetate salt. 1 H NMR (DMSO-d6) δ (ppm): 14.10-13.52 (m, 1 H), 12.23 (s, 1 H), 8.78 (m, 1 H), 8.43-8.18 (m, 2 H), 7.87-7.71 (m, 1 H), 4.76 (s, 2 H), 3.75-3.58 (m, 2 H), 3.14-2.95 (m, 2 H), 2.58-2.41 (m, 3 H); MS (ESI): m/z: 254 [M+H]+.
Example 26: 2-[(Z)-C-[2-(benzylamino)ethoxymethyl]-/V-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000131_0001
0.0070 g (76%) of 2-[(Z)-C-[2-(benzylamino)ethoxymethyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 26) was prepared according to the procedure described for Example 24, starting from 0.013 g (0.025 mmol) of 2-[(Z)-C-[2-(benzyl(te/t-butoxycarbonyl)amino)ethoxymethyl]-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 21 ) and using 2 M HCI in Et20 and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 14.14-13.43 (bs, 1 H), 12.24 (s, 1 H), 9.01 (bs, 2 H), 8.80-8.65 (m, 1 H), 8.32-8.21 (m, 1 H), 7.85-7.73 (dd, J=1 .4, 5.2 Hz, 1 H), 7.48-7.29 (m, 5 H), 4.77 (s, 2 H), 4.17-3.98 (m, 2 H), 3.80-3.67 (m, 2 H), 3.1 1 - 2.97 (m, 2 H); MS (ESI): m/z: 330 [M+H]+.
Example 27: 2-[(Z)-/V-hydroxy-C-(phenoxymethyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000131_0002
0.0060 g (46%) of 2-[(Z)-A/-hydroxy-C-(phenoxymethyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 27) was prepared according to the procedure described for Example 24, starting from 0.017 g (0.048 mmol) of 2-[(Z)-C-(phenoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 24) and using DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 12.36 (s, 1 H), 8.78-8.75 (m, 1 H), 8.25-8.23 (m, 1 H), 7.80 (dd, J=1 .5, 4.9 Hz, 1 H), 7.30-7.24 (m, 2 H), 6.98-6.90 (m, 3 H), 5.23 (s, 2 H); MS (ESI): m/z: 273 [M+H]+.
Example 28: 2-[(Z)-C-(ethoxymethyl)-/V-hydroxy-carbonimidoyl)pyridine-4- carboxylic acid
Figure imgf000132_0001
0.0050 g (46%) of 2-[(Z)-C-(ethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4- carboxylic acid (Example 28) was prepared according to the procedure described for Example 24, starting from 0.015 g (0.049 mmol) of 2-[(Z)-C-(ethoxymethyl)-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 18) and using 2 M HCI in Et20 and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 13.76 (bs, 1 H), 12.02 (s, 1 H), 8.78 (m, 1 H), 8.20 (m, 1 H), 7.78 (dd, J=1 .6, 4.9 Hz, 1 H), 4.65 (s, 2 H), 3.45 (q, J=6.8 Hz, 2 H), 1 .03 (t, J=6.9 Hz, 3 H); MS (ESI): m/z: 225[M+H]+.
Example 29: 2-[(E)-/V-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide hydrochloride
Figure imgf000132_0002
0.015 g (71 %) of 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide hydrochloride (Example 29) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.095 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2- yloxy-carbonimidoyl]pyridine-4-carboxamide (Intermediate 25) and 25 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 6 days. 1H NMR (DMSO-de) δ (ppm): 1 1 .63 (bs, 1 H), 8.74-8.58 (m, 1 H), 8.39-8.09 (m, 2 H), 7.80-7.63 (m, 2 H), 4.56 (bs, 1 H), 2.28 (s, 3 H); MS (ESI): m/z: 180 [M+H]+.
Example 30: A -ethyl-2-[(E)-A -hydroxy-C-methyl-carbonimidoyl]pyridine-4- carboxamide hydrochloride
Figure imgf000133_0001
0.018 g (85%) of A/-ethyl-2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyndine-4- carboxamide hydrochloride (Example 30) was prepared according to the procedure described for Example 24, starting from 0.026 g (0.089 mmol) of N-ethyl-2-[(E)-C-methyl- N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxamide (Intermediate 26) and 20 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 24 h. 1 H NMR (DMSO-de) δ (ppm): 1 1 .63 (bs, 1 H), 8.86 (t, J=5.1 Hz, 1 H), 8.72-8.58 (m, 1 H), 8.28-8.13 (m, 1 H), 7.76-7.58 (m, 1 H), 5.21 (bs, 1 H), 3.37-3.17 (m, 2 H), 2.22 (s, 3 H), 1.12 (t, J=7.3 Hz, 3 H); MS (ESI): m/z: 208 [M+H]+.
Example 31 : 2-[(E)-/V-hydroxy-C-methyl-carbonimidoyl]pyridine-4- carbohydroxamic acid
Figure imgf000133_0002
0.018 g (84%) of 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbohydroxamic acid (Example 31 ) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.1 1 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]-A/-tetrahydropyran-2-yloxy-pyridine-4-carboxamide (Intermediate 27) and 20 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 18 h. 1 H NMR (DMSO-de) δ (ppm): 1 1.60 (bs, 2 H), 8.76-8.55 (m, 1 H), 8.22-8.05 (m, 1 H), 7.70- 7.57 (m, 1 H), 2.22 (s, 3 H); MS (ESI): m/z: 196 [M+H]+.
Example 32: (E)-1 -[4-(1 H-tetrazol-5-yl)-2-pyridyl]ethanone oxime
Figure imgf000134_0001
0.016 g (90%) of (E)-1 -[-4-(1 H-tetrazol-5-yl)-2-pyridyl]ethanone oxime (Example 32) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.087 mmol) of (E)-A/-tetrahydropyran-2-yloxy-1 -[4-(1 H-tetrazol-5-yl)-2- pyridyl]ethanimine (Intermediate 28) and 10 equiv. of 4 M HCI in 1 ,4-dioxane and using 1 ,4-dioxane/MeOH as solvent. The reaction was carried out at r.t. for about 20 h. 1H NMR (DMSO-de) δ (ppm): 1 1.73 (bs, 1 H), 8.88-8.75 (m, 1 H), 8.56-8.41 (m, 1 H), 7.98 (dd, J=1.5, 4.9 Hz, 1 H), 2.26 (s, 3 H); MS (ESI): m/z: 205 [M+H]+.
Example 33: 2-[(E)-/V-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbaldehyde
Figure imgf000134_0002
0.012 g (52%) 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbaldehyde (Example 33) was prepared according to the procedure described for Example 24, starting from 0.035 g (0.14 mmol) of 2-[(E)-C-methyl-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carbaldehyde (Intermediate 29) and 10 equiv. of 4 M HCI in 1 ,4- dioxane. The reaction was carried out at r.t. for about 20 h. 1H NMR (ACN-d3) δ (ppm): 10.1 1 (s, 1 H), 9.46 (bs, 1 H), 8.91-8.75 (m, 1 H), 8.35-8.17 (m, 1 H), 7.73 (dd, J=1.5, 4.9 Hz, 1 H), 2.38-2.28 (m, 3 H); MS (ESI): m/z: 165 [M+H]+.
Example 34: (E)-1 -[4-(hydroxymethyl)-2-pyridyl]ethanone oxime
Figure imgf000135_0001
0.010 g (84%) (E)-1 -[4-(hydroxymethyl)-2-pyridyl]ethanone oxime (Example 34) was prepared according to the procedure described for Example 24, starting from 0.018 g (0.072 mmol) of [2-[(E)-C-methyl-N-tetrahydropyran-2-yloxy-carbonimidoyl]-4- pyridyl]methanol (Intermediate 30) and 10 equiv. of 4 M HCI in 1 ,4-dioxane. The reaction was carried out at r.t. for about 20 h. 1H NMR (DMSO-d6) δ (ppm): 12.08 (bs, 1 H), 8.70- 8.40 (m, 1 H), 8.08-7.88 (m, 1 H), 7.60 (bs, 1 H), 4.68 (s, 2 H), 2.24 (s, 3 H); MS (ESI): m/z: 167 [M+H]+.
Example 35: 2-[(E)-/V-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbonitrile
Figure imgf000135_0002
A mixture of 0.10 g (0.68 mmol) of 2-acetylpyridine-4-carbonitrile (Intermediate 3), 0.055 g (0.78 mmol) of hydroxylamine hydrochloride and 0.065 g (0.78 mmol) of AcONa in 1.0 mL of EtOH and 0.5 mL of water was stirred at 55°C for about 1.5 h. The reaction mixture was allowed to cool down to r.t. and about 1 mL of water was added while cooling in an ice/water bath. The resulting solid was filtered off, washed with little water and dried to give 0.073 g (66%) of 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbonitrile (Example 35) as a colourless solid. 1H NMR (DMSO-d6) δ (ppm): 1 1.87 (s, 1 H), 8.92- 8.73 (m, 1 H), 8.19-8.08 (m, 1 H), 7.83 (dd, J=1.2, 5.1 Hz, 1 H), 2.21 (s, 3 H); MS (ESI): m/z: 162 [M+H]+.
Example 36: 2-[(Z)-C-(benzyloxymethyl)-/V-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000136_0001
0.0050 g (43%) of 2-[(Z)-C-(benzyloxymethyl)-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 36) was prepared according to the procedure described for Example 24, starting from 0.015 g (0.041 mmol) of 2-[(Z)-C-(benzyloxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 37) and using 0.10 mL (0.20 mmol) of 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) 5 (ppm): 13.72 (bs, 1 H), 12.10 (bs, 1 H), 1 1 .83-1 1 .82 (m, 1 H), 8.81-8.77 (m, 1 H), 8.22- 8.19 (m, 1 H), 7.82-7.78 (m, 1 H), 7.33-7.21 (m, 5 H), 4.74 (s, 2 H), 4.49 (s, 2 H); MS (ESI): m/z: 287 [M+H]+.
Example 37: methyl 2-[(E)-A -hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine- 4-carboxylate and Example 38: methyl 2-[(Z)-/V-hydroxy-C-(4- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate
Figure imgf000136_0002
0.032 g (20%) of methyl 2-[(E)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate (Example 37) and 0.032 g (20%) of methyl 2-[(Z)-A/-hydroxy-C-(4- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 38) were prepared according to the procedure described for Example 1 , starting from 0.15 g (0.55 mmol) of methyl 2-(4-methoxybenzoyl)pyridine-4-carboxylate (Intermediate 31 ). The reaction gave two products, the E-isomer and the Z-isomer of the oxime. The two isomers were separated by flash chromatography (eluent DCM:EtOAc from 95:5 to 70:30). Example 37 (E-isomer): 1 H NMR (DMSO-d6) δ (ppm): 1 1 .77 (s, 1 H), 8.70-8.68 (m, 1 H), 8.27-8.25 (m, 1 H), 7.80 (dd, J=1 .5, 4.9 Hz, 1 H), 7.33-7.29 (m, 2 H), 6.98-6.94 (m, 2 H), 3.92 (s, 3 H), 3.79 (s, 3 H); MS (ESI): m/z: 287 [M+H]+; Example 38 (Z-isomer): 1 H NMR (DMSO- d6) δ (ppm): 1 1 .42 (s, 1 H), 8.88 (dd, J=1 .0, 5.1 Hz, 1 H), 7.91-7.90 (m, 1 H), 7.87 (dd, J=1 .7, 5.1 Hz, 1 H), 7.29-7.25 (m, 2 H), 6.93-6.90 (m, 2 H), 3.91 (s, 3 H), 3.75 (s, 3 H); MS (ESI): m/z: 287 [M+H]+.
Example 39: 2-[(E)-/V-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000137_0001
0.020 g (78%) of 2-[(E)-N-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 39) was prepared according to the procedure described for Example 12, starting from 0.027 g (0.094 mmol) of methyl 2-[(E)-N-hydroxy-C-(4- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 37). 1H NMR (DMSO-de) 5 (ppm): 1 1 .72 (s, 1 H), 8.72-8.57 (m, 1 H), 8.31-8.16 (m, 1 H), 7.87-7.68 (m, 1 H), 7.43- 7.20 (m, 2 H), 7.06-6.86 (m, 2 H), 3.79 (s, 3 H); MS (ESI): m/z: 273 [M+H]+.
Example 40: 2-[(Z)-/V-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000137_0002
0.01 1 g (41 %) of 2-[(Z)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 40) was prepared according to the procedure described for Example 12, starting from 0.028 g (0.098 mmol) of methyl 2-[(Z)-A/-hydroxy-C-(4- methoxyphenyl)carbonimidoyl]pyridine-4-carboxylate (Example 38). 1H NMR (DMSO-de) δ (ppm): 1 1 .38 (s, 1 H), 8.92-8.76 (m, 1 H), 7.89-7.85 (m, 1 H), 7.85-7.82 (m, 1 H), 7.37- 7.18 (m, 2 H), 6.99-6.82 (m, 2 H), 3.75 (s, 3 H); MS (ESI): m/z: 273 [M+H]+.
Example 41 : methyl 2-[(Z)-/V-hydroxy-C-(phenoxymethyl)carbonimidoyl]pyridine-4- carboxylate
Figure imgf000138_0001
Methyl 2-[(Z)-N-hydroxy-C-(phenoxymethyl)carbonimidoyl]pyridine-4-carboxylate example 41 ) was obtained by reacting 2-[(Z)-C-(benzyloxymethyl)-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 36) with diazomethane in a DCM/MeOH mixture (4; 1 , v:v). 1H NMR (DMSO-d6) δ (ppm): 12.40 (s, 1 H), 8.80 (m, 1 H), 8.30-8.18 (m, 1 H), 7.82 (dd, J=1 .6, 4.9 Hz, 1 H), 7.32-7.19 (m, 2 H), 7.03-6.84 (m, 3 H), 5.23 (s, 2 H), 3.91 (s, 3 H); MS (ESI): m/z: 287 [M+H]+.
Example 42: 2-[(Z)-C-[[3-(3-aminopropanoylamino)phenoxy]methyl]-/V-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000138_0002
0.033 g (91 %) of 2-[(Z)-C-[[3-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 42) was prepared according to the procedure described for Example 24, starting from 0.050 g (0.092 mmol) of 2-[(Z)-C-(2-acetamidoethoxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 36) and using 0.12 mL (0.48 mmol) of 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 12.38 (s, 1 H), 10.27-10.18 (m, 1 H), 8.79-8.73 (m, 1 H), 8.27-8.23 (m, 1 H), 7.99-7.77 (m, 4 H), 7.35-7.30 (m, 1 H), 7.24-7.09 (m, 2 H), 6.73-6.64 (m, 1 H), 5.20 (s, 2 H), 3.1 1-2.98 (m, 2 H), 2.75-2.66 (m, 2 H); MS (ESI): m/z: 359 [M+H]+.
Example 43: 2-[(Z)-C-(2-acetamidoethoxymethyl)-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000139_0001
0.015 g (85%) of 2-[(Z)-C-(2-acetamidoethoxymethyl)-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid (Example 43) was prepared according to the procedure described for Example 24, starting from 0.023 g (0.060 mmol) of 2-[(Z)-C-(2-acetamidoethoxymethyl)- N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 35) and using 0.16 ml_ (0.32 mmol) of 2 M HCI in Et20 and DCM as solvent. 1 H NMR (ACN- d3) δ (ppm): 9.78 (bs, 1 H), 8.85-8.71 (m, 1 H), 8.36-8.23 (m, 1 H), 7.90-7.79 (m, 1 H), 6.45 (bs, 1 H), 4.80 (s, 2 H), 3.54 (m, 2 H), 3.31 (s, 3 H), 3.26 (m, 2 H), 1 .80 (s, 3 H); MS (ESI): m/z: 282 [M+H]+.
Example 44: 2-[(Z)-A -hydroxy-C-(2- methoxyphenyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000139_0002
0.035 g (60%) of 2-[(Z)-A/-hydroxy-C-(2-methoxy- phenyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid (Example 44) was prepared according to the procedure described for Example 24, starting from 0.075 g (0.19 mmol) of 2-[(Z)-C-(2-methoxyphenyloxymethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 40), using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 12.32 (s, 1 H), 8.80-8.71 (m, 1 H), 8.29- 8.23 (m, 1 H), 7.83-7.76 (m, 1 H), 7.12-7.02 (m, 1 H), 6.95-6.80 (m, 3 H), 5.19 (s, 2 H), 3.64 (s, 3 H); MS (ESI): m/z: 301 [M+H]+.
Example 45: 2-[(Z)-A -hydroxy-C-(3- methoxyphenyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000140_0001
0.034 g (50%) of 2-[(Z)-N-hyd roxy-C-(3-methoxy- phenyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid (Example 45) was prepared according to the procedure described for Example 24, starting from 2-[(Z)-C-(3- methoxyphenyloxymethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylic acid (Intermediate 41 ), using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-de) δ (ppm): 12.38 (s, 1 H), 8.81-8.72 (m, 1 H), 8.27-8.19 (m, 1 H), 7.84-7.73 (m, 1 H), 7.22-7.08 (m, 1 H), 6.61-6.41 (m, 3 H), 5.21 (s, 2 H), 3.70 (s, 3 H); MS (ESI): m/z: 301 [M+H]+.
Example 46: 2-[(E)-C-(anilinomethyl)-/V-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000140_0002
0.010 g (66%) of 2-[(E)-C-(anilinomethyl)-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid (Example 46) was prepared according to the procedure described for Example 24, starting from 0.022 g (0.062 mmol) of 2-[(E)-C-(anilinomethyl)-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 43) using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 12.27 (bs, 1 H), 8.84-8.71 (m, 1 H), 8.24-8.10 (m, 1 H), 7.84-7.74 (m, 1 H), 7.12-7.06 (m, 2 H), 6.79 (m, 2 H), 6.71 -6.64 (m, 1 H), 4.51 (m, 2 H); MS (ESI): m/z: 272 [M+H]+.
Example 47: 2-[(E)-A -hydroxy-C-(methylaminomethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride
Figure imgf000141_0001
0.0050 g (50%) of 2-[(E)-N-hydroxy-C-(methylaminomethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride (Example 47) was prepared according to the procedure described for Example 24, starting from 0.022 g (0.062 mmol) of 2-[(E)-C- (methylaminomethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 44), using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.71 (bs, 1 H), 12.06 (s, 1 H), 8.79-8.75 (m, 1 H), 8.21-8.18 (m, 1 H), 7.80- 7.77 (m, 1 H), 4.62 (s, 2 H), 3.23 (s, 3 H); MS (ESI): m/z: 210 [M+H]+.
Example 48: 2-[(Z)-C-[(2-fluorophenoxy)methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000141_0002
0.029 g (94%) of 2-[(Z)-C-[(2-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 48) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.1 1 mmol) of 2-[(Z)-C-[(2-fluorophenoxy)methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 46), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 13.73 (bs, 1 H), 12.43 (s, 1 H), 8.77 (s, 1 H), 8.28-8.23 (m, 1 H), 7.82-7.78 (m, 1 H), 7.35-7.28 (m, 1 H), 7.19-7.10 (m, 2 H), 6.98-6.90 (m, 1 H), 5.30 (s, 2 H); MS (ESI): m/z: 291 [M+H]+. Example 49: 2-[(Z)-C-[[4-(3-aminopropanoylamino)phenoxy]methyl]-/V-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000141_0003
0.016 g (41 %) of 2-[(Z)-C-[[4-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 49) was prepared according to the procedure described for Example 24, starting from 0.030 g (0.055 mmol) of 2-[(Z)-C-[[4-[3-(te/t-butoxycarbonylamino)propanoylamino]phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 48), using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.71 (bs, 1 H), 12.36 (s, 1 H), 10.08 (s, 1 H), 8.79-8.73 (m, 1 H), 8.26-8.22 (m, 1 H), 7.94-7.77 (m, 4 H), 7.52-7.46 (m, 2 H), 6.96-6.88 (m, 2 H), 5.19 (s, 2 H), 3.12-3.01 (m, 2 H), 2.71-2.64 (m, 2 H); MS (ESI): m/z: 359 [M+H]+.
Example 50: 2-[(Z)-C-[(3-aminophenoxy)methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000142_0001
0.020 g (65%) of 2-[(Z)-C-[(3-aminophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid hydrochloride (Example 50) was prepared according to the procedure described for Example 24, starting from 0.045 g (0.095 mmol) of 2-[(Z)-C-[[3-(fert- butoxycarbonylamino)phenoxy]methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 50), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.73 (bs, 1 H), 12.46 (s, 1 H), 10.05 (bs, 3 H), 8.83-8.69 (m, 1 H), 8.29-8.25 (m, 1 H), 7.82-7.79 (m, 1 H), 7.38 (m, 1 H), 7.05-6.99 (m, 1 H), 6.94-6.87 (m, 2 H), 5.20 (s, 2 H); MS (ESI): m/z: 288 [M+H]+.
Example 51 : 2-[(Z)-C-[(4-aminophenoxy)methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000142_0002
0.034 g (99%) of 2-[(Z)-C-[(4-aminophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid hydrochloride (Example 51 ) was prepared according to the procedure described for Example 24, starting from 0.050 g (0.1 1 mmol) of 2-[(Z)-C-[[4-(fert- butoxycarbonylamino)phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 50), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.81 (bs, 1 H), 12.45 (s, 1 H), 10.13 (bs, 1 H), 8.81-8.68 (m, 1 H), 8.32-8.18 (m, 1 H), 7.86-7.73 (m, 1 H), 7.36-7.23 (m, 2 H), 7.15-7.03 (m, 2 H), 5.25 (s, 2 H); MS (ESI): m/z: 288 [M+H]+.
Example 52: 2-[(Z)-C-[(4-fluorophenoxy)methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000143_0001
0.013 g (48%) of 2-[(Z)-C-[(4-fluorophenoxy)methyl]-A/-hydroxy-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 52) was prepared according to the procedure described for Example 24, starting from 0.035 g (0.094 mmol) of 2-[(Z)-C-[(4- fluorophenoxy)methyl]-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 54), using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.84 (bs, 1 H), 12.39 (s, 1 H), 8.86-8.70 (m, 1 H), 8.30-8.18 (m, 1 H), 7.85- 7.73 (m, 1 H), 7.13-7.07 (m, 2 H), 7.01-6.95 (m, 2 H), 5.21 (s, 2 H); MS (ESI): m/z: 291 [M+H]+.
Example 53: 2-[(Z)-C-[(3-fluorophenoxy)methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000143_0002
0.015 g (77%) of 2-[(Z)-C-[(3-fluorophenoxy)methyl]-A/-hydroxy-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 53) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.067 mmol) of 2-[(Z)-C-[(3- fluorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 56), using 4 M HCI in dioxane and DCM as solvent. 1H NMR (DMSO- d6) δ (ppm): 13.68 (bs, 1 H), 12.45 (s, 1 H), 8.84-8.68 (m, 1 H), 8.31-8.16 (m, 1 H), 7.87- 7.70 (m, 1 H), 7.36-7.22 (m, 1 H), 6.98-6.63 (m, 3 H), 5.25 (s, 2 H); MS (ESI): m/z: 291 [M+H]+.
Example 54: 2-[(Z)-/V-hydroxy-C-(2-pyridyloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000144_0001
A solution of 0.10 g (0.27 mmol) of methyl 2-[(Z)-C-(2-pyridyloxymethyl)-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 57) in 4 mL of a water/THF mixture (3:1 , v:v) was treated with 0.013 mg (0.54 mmol) of LiOH at r.t. for about 1 h. The mixture was diluted with water and EtOAc; the aqueous phase was extracted with EtOAc, acidified with 5% citric acid and extracted with DCM. The combined organic layers were dried over Na2SO4 and concentrated to dryness. The crude product was dissolved in 3 mL of dry DCM and treated with 0.37 mL (1.4 mmol) of 2 M HCI in Et2O, at 0°C. The resulting suspension was stirred at r.t. for about 3 h and decanted. Purification by preparative HPLC provided 0.0050 g (5%) of 2-[(Z)-A/-hydroxy-C-(2- pyridyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid as its trifluoroacetate salt (Example 54) and as a colourless solid. 1H NMR (DMSO-d6) δ (ppm): 13.77 (bs, 1 H), 12.1 1 (s, 1 H), 8.74-8.62 (m, 1 H), 8.19-8.09 (m, 1 H), 7.80-7.70 (m, 1 H), 7.67-7.57 (m, 1 H), 7.39-7.24 (m, 1 H), 6.29-6.08 (m, 2 H), 5.16 (s, 2 H); MS (ESI): m/z: 274[M+H]+. Example 55: 2-[(Z)-/V-hydroxy-C-(pyrimidin-2-yloxymethyl)carbonimidoyl]pyridine- 4-carboxylic acid
Figure imgf000145_0001
0.025 g (24%) of 2-[(Z)-N-hydroxy-C-(pyrimidin-2-yloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid was prepared according to the procedure described for Example 54, starting from 0.10 g (0.27 mmol) of methyl 2-[(Z)-C-(pyrimidin-2-yloxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylate (Intermediate 58) as its trifluoroacetate salt. 1H NMR (DMSO-d6) δ (ppm): 13.74 (bs, 1 H), 12.27 (s, 1 H), 8.71- 8.66 (m, 1 H), 8.56-8.50 (m, 1 H), 8.45-8.38 (m, 1 H), 8.20-8.14 (m, 1 H), 7.82-7.74 (m, 1 H), 6.54-6.46 (m, 1 H), 5.23 (s, 2 H); MS (ESI): m/z: 275[M+H]+.
Example 56: 2-[(Z)-/V-hydroxy-C-(1 -naphthyloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride
Figure imgf000145_0002
0.013 g (74%) of 2-[(Z)-A/-hydroxy-C-(1 -naphthyloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride (Example 56) was prepared according to the procedure described for Example 24, starting from 0.020 g (0.049 mmol) of 2-[(Z)-C-(1 - naphthyloxymethyl)-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 60), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO- d6) δ (ppm): 13.74 (bs, 1 H), 12.41 (s, 1 H), 8.85-8.63 (m, 1 H), 8.35-8.18 (m, 1 H), 7.96- 7.70 (m, 3 H), 7.54-7.30 (m, 4 H), 7.20-7.08 (m, 1 H), 5.45 (s, 2 H); MS (ESI): m/z: 323[M+H]+.
Example 57: 2-[(Z)-A -hydroxy-C-(2-naphthyloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride
Figure imgf000146_0001
0.048 g (91 %) of 2-[(Z)-N-hydroxy-C-(2-naphthyloxymethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride (Example 57) was prepared according to the procedure described for Example 24, starting from 0.060 g (0.15 mmol) of 2-[(Z)-C-(2- naphthyloxymethyl)-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 62), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO- d6) δ (ppm): 13.74 (bs, 1 H), 12.44 (s, 1 H), 8.85-8.65 (m, 1 H), 8.38-8.17 (m, 1 H), 7.88- 7.66 (m, 4 H), 7.55-7.28 (m, 3 H), 7.16-7.00 (m, 1 H), 5.36 (s, 2 H); MS (ESI): m/z: 323[M+H]+.
Example 58: 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000146_0002
0.015 g (67%) of 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 58) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.055 mmol) of 2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 67), using 4 M HCI in 1 ,4-dioxane and DCM-1 ,4-dioxane as solvent. 1 H NMR (DMSO-d6) δ (ppm): 1 1 .75 (bs, 1 H), 8.62- 8.71 (m, 1 H), 8.23 (s, 1 H), 7.73-7.84 (m, 1 H,) 7.46-7.59 (m, 2 H), 7.28-7.41 (m, 2 H), 7.17-7.27 (m, 2 H), 6.98-7.10 (m, 2 H), 5.13 (s, 2 H); MS (ESI): m/z: 367[M+H]+.
Example 59: 2-[(Z)-A -Hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000147_0001
0.020 g (51 %) of 2-[(Z)-N-hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 59) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.088 mmol) of 2-[(Z)-C-[[3-(4-methylpiperazin-1 -yl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 69), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 12.39 (s, 1 H), 10.41 (bs, 1 H), 8.80-8.70 (m, 1 H), 8.29-8.12 (m, 1 H), 7.80 (dd, J=1 .5, 4.9 Hz, 1 H), 7.19-7.04 (m, 1 H), 6.61 -6.38 (m, 3 H), 5.22 (s, 2 H), 3.89-2.88 (m, 8 H), 2.84-2.69 (m, 3 H); MS (ESI): m/z: 371 [M+H]+.
Example 60: 2-[(Z)-/V-Hydroxy-C-[[2-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000147_0002
0.012 g (31 %) of 2-[(Z)-A/-hydroxy-C-[[2-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 60) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.088 mmol) of 2-[(Z)-C-[[2-(4-methylpiperazin-1 -yl)phenoxy]methyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 71 ), using 4 M HCI in 1 ,4-dioxane and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 12.41 (s, 1 H), 10.1 1 (bs, 1 H), 8.82-8.72 (m, 1 H), 8.30-8.17 (m, 1 H), 7.82 (dd, J=1 .5, 4.9 Hz, 1 H), 7.19-7.07 (m, 1 H), 7.05-6.83 (m, 3 H), 5.26 (s, 2 H), 3.42-3.17 (m, 4 H), 2.99-2.66 (m, 7 H); MS (ESI): m/z: 371 [M+H]+. Example 61 : 2-[(Z)-A -hydroxy-C-[[3-
(trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000148_0001
0.020 g (52%) of 2-[(Z)-N-hydroxy-C-[[3- (trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyndine-4-carboxylic acid was prepared according to the procedure described for Example 54, starting from 0.050 g (0.1 1 mmol) of methyl 2-[(Z)-N-tetrahydropyran-2-yloxy-C-[[3-
(trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylate (Intermediate 72). 1 H NMR (DMSO-de) δ (ppm): 12.49 (s, 1 H), 8.81-8.73 (m, 1 H), 8.28-8.20 (m, 1 H), 7.85-7.75 (m, 1 H), 7.55-7.45 (m, 1 H), 7.38-7.21 (m, 3 H), 5.33 (s, 2 H); MS (ESI): m/z: 341 [M+H]+.
Example 62: 2-[(Z)-A -hydroxy-C-[(4- methoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000148_0002
0.025 g (66%) of 2-[(Z)-A/-hydroxy-C-[(4- methoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid was prepared according to the procedure described for Example 54, starting from 0.050 g (0.12 mmol) of methyl 2-[(Z)-C-[(4-methoxyphenoxy)methyl]-A/-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylate (Intermediate 73). 1 H NMR (DMSO-de) δ (ppm): 12.33 (s, 1 H), 8.82-8.69 (m, 1 H), 8.26-8.18 (m, 1 H), 7.84-7.73 (m, 1 H), 6.95-6.75 (m, 4 H), 5.17 (s, 2 H), 3.68 (s, 3 H); MS (ESI): m/z: 303[M+H]+.
Example 63: methyl 2-[(E)-/V-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4- carboxylate
Figure imgf000149_0001
0.10 g (79%) of methyl 2-[(E)-N-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4- carboxylate (Example 63) was prepared according to the procedure described for Example 1 , starting from 0.12 g (0.45 mmol) of methyl 2-(3-phenylpropanoyl)pyridine-4- carboxylate (Intermediate 75). 1 H NMR (DMSO-d6) δ (ppm): 1 1 .77 (s, 1 H), 8.73-8.87 (m, 1 H), 8.18-8.34 (m, 1 H), 7.72-7.84 (m, 1 H), 7.05-7.33 (m, 5 H), 3.91 (s, 3 H), 3.03-3.23 (m, 2 H), 2.69-2.87 (m, 2 H); MS (ESI): m/z: 285 [M+H]+.
Example 64: 2-[(E)-/V-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000149_0002
0.028 g (39%) of 2-[(E)-A/-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylic acid (Example 64) was prepared according to the procedure described for Example 12, starting from 0.075 g (0.26 mmol) of methyl 2-[(E)-A/-hydroxy-C-phenethyl- carbonimidoyl]pyridine-4-carboxylate (Example 63). 1 H NMR (DMSO-d6) δ (ppm): 13.72 (bs, 1 H), 1 1 .73 (s, 1 H), 8.67-8.87 (m, 1 H), 8.09-8.36 (m, 1 H), 7.64-7.83 (m, 1 H), 7.05-7.34 (m, 5 H), 3.06-3.23 (m, 2 H), 2.69-2.87 (m, 2 H); MS (ESI): m/z: 271 [M+H]+. Example 65: 2-[/V-hydroxy-C-[[3-
(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000149_0003
0.020 g (64%) of 2-[N-hydroxy-C-[[3-
(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 65) was prepared according to the procedure described for Example 54, starting from 0.035 g (0.079 mmol) of 2-[(Z)-N-tetrahydropyran-2-yloxy-C-[[3- (trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 77). 1 H NMR (DMSO-de) δ (ppm): 13.73 (bs, 1 H), 12.47 (s, 1 H), 8.82-8.69 (m, 1 H), 8.29-8.14 (m, 1 H), 7.80 (dd, J=1 .5, 4.9 Hz, 1 H), 7.45-7.33 (m, 1 H), 7.07-6.83 (m, 3 H), 5.28 (s, 2 H); MS (ESI): m/z: 357[M+H]+.
Example 66: 2-[(Z)-C-[2-[(4-fluorophenyl)methylamino]ethoxymethyl]-/V-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000150_0001
0.010 g (63%) of 2-[(Z)-C-[2-[(4-fluorophenyl)methylamino]ethoxymethyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 66) was prepared according to the procedure described for Example 24, starting from 0.026 g (0.042 mmol) of 2-[(Z)-C-[2-[tert-butoxycarbonyl-[(4-fluorophenyl)methyl]amino]ethoxymethyl]-/V- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 78) and using 2 M HCI in Et20 and DCM as solvent. 1 H NMR (DMSO-d6) δ (ppm): 13.78 (bs, 1 H), 12.24 (s, 1 H), 9.00 (bs, 2 H), 8.77-8.68 (m, 1 H), 8.29-8.21 (m, 1 H), 7.85-7.73 (m, 1 H), 7.55-7.41 (m, 2 H), 7.31-7.18 (m, 2 H), 4.76 (s, 2 H), 4.14-4.01 (m, 2 H), 3.77-3.67 (m, 2 H), 3.08-2.99 (m, 2 H); MS (ESI): m/z: 348 [M+H]+.
Example 67: 2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-A -hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000150_0002
0.0080 g (55%) of 2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 67) was prepared according to the procedure described for Example 24, starting from 0.019 g (0.05 mmol) of 2-[(Z)-C-[[2- (dimethylamino)-2-oxo-ethoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 79) and using 2 M HCI in Et.20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.64 (bs, 1 H), 12.15 (s, 1 H), 8.81-8.76 (m, 1 H), 8.25-8.23 (m, 1 H), 7.81 (s, 1 H), 4.78-4.60 (m, 2 H), 4.14 (s, 2 H), 2.84-2.76 (m, 6 H); MS (ESI): m/z: 282 [M+H]+.
Example 68: 2-[(Z)-A -hydroxy-C-(3^yridylmethoxymethyl)carbonimidoyl]pyridine- 4-carboxylic acid hydrochloride
Figure imgf000151_0001
0.0055 g (6%) of 2-[(Z)-A/-hydroxy-C-(3-pyridylmethoxymethyl)carbonimidoyl]pyridine-4- carboxylic acid hydrochloride (Example 68) was prepared according to the procedure described for Example 24, starting from 2-[(Z)-C-[[2-(dimethylamino)-2-oxo- ethoxy]methyl]-A/-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 80) and using 2 M HCI in Et20 and DCM as solvent. 1H NMR (DMSO-d6) δ (ppm): 13.78 (bs, 1 H), 12.24 (s, 1 H), 8.83-8.74 (m, 3 H), 8.38-8.33 (m, 1 H), 8.23 (s, 1 H), 7.97-7.90 (m, 1 H), 7.83-7.78 (m, 1 H), 4.82-4.71 (m, 4 H); MS (ESI): m/z: 288 [M+H]+.
Example 69: 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000151_0002
0.013 g (50%) of 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 69) was prepared according to the procedure described for Example 24, starting from 0.033 g (0.094 mmol) of 2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 81 ). 1H NMR (DMSO-de) δ (ppm): 13.86 (bs, 1 H), 12.26 (s, 1 H), 9.77 (bs, 1 H), 8.83 - 8.72 (m, 1 H), 8.31 - 8.19 (m, 1 H), 7.81 (s, 1 H), 4.75 (s, 2 H), 3.80 - 3.75 (m, 2 H), 3.22 (s, 2 H), 2.71 - 2.67 (m, 6 H); MS (ESI): m/z: 268 [M+H]+.
Example 70: 2-[(E)-C-[(4-fluoroanilino)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid hydrochloride
Figure imgf000152_0001
0.0070 g (57%) of 2-[(E)-C-[(4-fluoroanilino)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid hydrochloride (Example 70) was prepared according to the procedure described for Example 24, starting from 0.014 g (0.037 mmol) of 2-[(E)-C-[(4- fluoroanilino)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 83). 1H NMR (DMSO-d6) δ (ppm): 13.70 (bs, 1 H), 12.26 (bs, 1 H), 8.82 - 8.74 (m, 1 H), 8.19 - 8.12 (m, 1 H), 7.81 - 7.76 (m, 1 H), 6.97 - 6.89 (m, 2 H), 6.82 - 6.73 (m, 2 H), 4.52 - 4.29 (m, 2 H); MS (ESI): m/z: 289 [M+H]+.
Example 71 : 2-[(Z)-N-hydroxy-C-[(4- morpholinophenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000152_0002
0.060 g (97%) of 2-[(Z)-N-hydroxy-C-[(4- morpholinophenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 71 ) was prepared according to the procedure described for Example 24, starting from 0.070 g (0.16 mmol) of 2-[(Z)-C-[(4-morpholinophenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 85). 1H NMR (DMSO-de) δ (ppm): 13.73 (bs, 1 H), 12.40 (s, 1 H), 8.83 - 8.69 (m, 1 H), 8.29 - 8.20 (m, 1 H), 7.80 (dd, J = 1.5, 4.9 Hz, 1 H), 7.17 (bs, 4 H), 5.24 (s, 2 H), 3.91 (bs, 4 H), 3.36 (bs, 4 H); MS (ESI): m/z: 358 [M+H]+.
Example 72: 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000153_0001
0.040 g (72%) of 2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 72) was prepared according to the procedure described for Example 24, starting from 0.068 g (0.14 mmol) of 2-[(Z)-C-[[3- [(4-fluorophenyl)methoxy]phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 87). 1H NMR (DMSO-de) δ (ppm): 13.76 (bs, 1 H), 12.39 (s, 1 H), 8.82 - 8.71 (m, 1 H), 8.29 - 8.15 (m, 1 H), 7.80 (dd, J = 1.5, 5.4 Hz, 1 H), 7.53 - 7.42 (m, 2 H), 7.26 - 7.10 (m, 3 H), 6.68 - 6.48 (m, 3 H), 5.22 (s, 2 H), 5.03 (s, 2 H); MS (ESI): m/z: 397 [M+H]+.
Example 73: 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000153_0002
0.034 g (91 %) of 2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 73) was prepared according to the procedure described for Example 24, starting from 0.045 g (0.10 mmol) of 2-[(Z)-C-[[3- [(phenyl)methoxy]phenoxy]methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylic acid (Intermediate 89). 1H NMR (DMSO-d6) δ (ppm): 13.71 (bs, 1 H), 12.39 (s, 1 H), 8.84 - 8.65 (m, 1 H), 8.32 - 8.14 (m, 1 H), 7.80 (dd, J = 1.5, 4.9 Hz, 1 H), 7.50 - 7.05 (m, 6 H), 6.70 - 6.48 (m, 3 H), 5.22 (s, 2 H), 5.07 (s, 2 H); MS (ESI): m/z: 379 [M+H]+. Example 74: 2-[(Z)-N-hydroxy-C-[[4-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimido l]pyridine-4-carboxylic acid 2,2,2-trifluoroacetate
Figure imgf000154_0001
0.0060 g (15%) of 2-[(Z)-N-hydroxy-C-[[4-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid 2,2,2-trifluoroacetate (Example 74) was prepared according to the procedure described for Example 24, starting from 0.035 g (0.080 mmol) of 2-[(Z)-C-[[4-(4-methylpiperazin-1 - yl)phenoxy]methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 91 ). The crude product was purified by preparative HPLC, affording 2-[(Z)- N-hydroxy-C-[[4-(4-methylpiperazin-1 -yl)phenoxy]methyl]carbonimidoyl]pyridine-4- carboxylic acid as trifluoro acetate salt. 1H NMR (DMSO-d6) δ (ppm): 12.33 (s, 1 H), 9.58 (bs, 1 H), 8.79 - 8.73 (m, 1 H), 8.26 - 8.19 (m, 1 H), 7.82 - 7.77 (m, 1 H), 6.97 - 6.86 (m, 4 H), 5.18 (s, 2 H), 3.69 - 3.61 (m, 2 H), 3.53 - 3.44 (m, 2 H), 3.20 - 3.09 (m, 2 H), 2.90 - 2.77 (m, 5 H); MS (ESI): m/z: 371 [M+H]+.
Example 75: 2-[(Z)-N-hydroxy-C-[[4-(1 - piperidyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000154_0002
0.031 g (quantitative) of 2-[(Z)-N-hydroxy-C-[[4-(1 - piperidyl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 75) was prepared according to the procedure described for Example 24, starting from 0.035 g (0.080 mmol) of 2-[(Z)-C-[[4-(1 -piperidyl)phenoxy]methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 93). 1H NMR (DMSO-de) δ (ppm): 12.47 (s, 1 H), 12.06 (bs, 1 H), 8.80 - 8.72 (m, 1 H), 8.30 - 8.21 (m, 1 H), 7.84 - 7.67 (m, 3 H), 7.20 - 7.12 (m, 2 H), 5.28 (s, 2 H), 3.57 - 3.39 (m, 4 H), 2.1 1 - 1.44 (m, 6 H); MS (ESI): m/z: 356 [M+H]+.
Example 76: 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000155_0001
0.027 g (quantitative) of 2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 76) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.090 mmol) of 2-[(Z)-C-[(4- benzyloxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4- carboxylic acid (Intermediate 95). 1 H NMR (DMSO-d6) δ (ppm): 12.32 (s, 1 H), 8.79 - 8.73 (m, 1 H), 8.25 - 8.18 (m, 1 H), 7.83 - 7.76 (m, 1 H), 7.46 - 7.27 (m, 5 H), 6.94 - 6.83 (m, 4 H), 5.17 (s, 2 H), 5.02 (s, 2 H); MS (ESI): m/z: 379 [M+H]+.
Example 77: 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000155_0002
0.046 g (quantitative) of 2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 77) was prepared according to the procedure described for Example 24, starting from 0.060 g (0.15 mmol) of 2-[(Z)-C-[(3- chlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 97). 1H NMR (DMSO-d6) δ (ppm): 12.45 (s, 1 H), 8.80 - 8.72 (m, 1 H), 8.28 - 8.21 (m, 1 H), 7.84 - 7.76 (m, 1 H), 7.33 - 7.23 (m, 1 H), 7.13 - 7.06 (m, 1 H), 7.03 - 6.89 (m, 2 H), 5.26 (s, 2 H); MS (ESI): m/z: 307 [M+H]+.
Example 78: 2-[(Z)-N-hydroxy-C-[(4- phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000156_0001
0.046 g (97%) of 2-[(Z)-N-hydroxy-C-[(4- phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid (Example 78) was prepared according to the procedure described for Example 24, starting from 0.060 g (0.13 mmol) of 2-[(Z)-C-[(4-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 99). 1H NMR (DMSO-de) δ (ppm): 12.37 (s, 1 H), 8.81 - 8.74 (m, 1 H), 8.27 - 8.20 (m, 1 H), 7.84 - 7.75 (m, 1 H), 7.39 - 7.28 (m, 2 H), 7.09 - 7.04 (m, 1 H), 7.02 - 6.86 (m, 6 H), 5.23 (s, 2 H); MS (ESI): m/z: 365 [M+H]+.
Example 79: 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000156_0002
0.030 g (63%) of 2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 79) was prepared according to the procedure described for Example 24, starting from 0.060 g (0.14 mmol) of 2-[(Z)-C-[(3,4- dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 101 ). 1H NMR (DMSO-d6) δ (ppm): 12.49 (s, 1 H), 8.79 - 8.74 (m, 1 H), 8.27 - 8.22 (m, 1 H), 7.83 - 7.77 (m, 1 H), 7.53 - 7.47 (m, 1 H), 7.37 - 7.31 (m, 1 H), 7.03 - 6.96 (m, 1 H), 5.27 (s, 2 H); MS (ESI): m/z: 342 [M+H]+.
Example 80: 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000156_0003
0.028 g (91 %) of 2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 79) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.090 mmol) of 2-[(Z)-C-[(3,5- dichlorophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 103). 1H NMR (DMSO-d6) δ (ppm): 12.52 (s, 1 H), 8.80 - 8.73 (m, 1 H), 8.28 - 8.23 (m, 1 H), 7.83 - 7.76 (m, 1 H), 7.19 - 7.09 (m, 3 H), 5.28 (s, 2 H); MS (ESI): m/z: 342 [M+H]+.
Example 81 : 2-[(Z)-N-hydroxy-C-[(3- phenylphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000157_0001
0.035 g (84%) of 2-[(Z)-N-hydroxy-C-[(3-phenylphenoxy)methyl]carbonimidoyl]pyridine-
4-carboxylic acid (Example 81 ) was prepared according to the procedure described for
Example 24, starting from 0.050 g (0.12 mmol) of 2-[(Z)-C-[(3-phenylphenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 105). 1H NMR (DMSO-de) δ (ppm): 12.41 (s, 1 H), 8.84 - 8.72 (m, 1 H), 8.29 - 8.20 (m, 1 H), 7.85
- 7.76 (m, 1 H), 7.67 - 7.59 (m, 2 H), 7.49 - 7.16 (m, 6 H), 7.01 - 6.90 (m, 1 H), 5.33 (s, 2
H); MS (ESI): m/z: 349 [M+H]+.
Example 82: 2-[(Z)-N-hydroxy-C-[(3- phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000157_0002
0.025 g (63%) of 2-[(Z)-N-hydroxy-C-[(3- phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid (Example 82) was prepared according to the procedure described for Example 24, starting from 0.050 g (0.1 1 mmol) of 2-[(Z)-C-[(3-phenoxyphenoxy)methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 107). 1H NMR (DMSO-de) δ (ppm): 12.37 (s, 1 H), 8.79 - 8.71 (m, 1 H), 8.25 - 8.20 (m, 1 H), 7.81 - 7.76 (m, 1 H), 7.42 - 7.33 (m, 2 H), 7.29 - 7.21 (m, 1 H), 7.17 - 7.10 (m, 1 H), 7.03 - 6.96 (m, 2 H), 6.78 - 6.71 (m, 1 H), 6.62 - 6.57 (m, 1 H), 6.56 - 6.50 (m, 1 H), 5.22 (s, 2 H); MS (ESI): m/z: 365 [M+H]+. Example 83: 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000158_0001
0.031 g (quantitative) of 2-[(Z)-C-[(4-bromophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 83) was prepared according to the procedure described for Example 24, starting from 0.040 g (0.090 mmol) of 2-[(Z)-C-[(4- bromophenoxy)methyl]-N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 109). 1H NMR (DMSO-d6) δ (ppm): 12.42 (s, 1 H), 8.80 - 8.72 (m, 1 H), 8.27 - 8.20 (m, 1 H), 7.84 - 7.76 (m, 1 H), 7.49 - 7.37 (m, 2 H), 7.00 - 6.90 (m, 2 H), 5.23 (s, 2 H); MS (ESI): m/z: 352 [M+H]+.
Example 84: 2-[(Z)-N-hydroxy-C-[(4- phenylphenoxy)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000158_0002
0.01 1 g (63%) of 2-[(Z)-N-hydroxy-C-[(4-phenylphenoxy)methyl]carbonimidoyl]pyridine- 4-carboxylic acid (Example 84) was prepared according to the procedure described for Example 24, starting from 0.022 g (0.050 mmol) of 2-[(Z)-C-[(4-phenylphenoxy)methyl]- N-tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 111 ). 1H NMR (DMSO-de) δ (ppm): 13.77 (bs, 1 H), 12.41 (s, 1 H), 8.85 - 8.63 (m, 1 H), 8.32 - 8.19 (m, 1 H), 7.81 (dd, J = 1 .4, 5.2 Hz, 1 H), 7.63 - 7.24 (m, 7 H), 7.10 - 6.95 (m, 2 H), 5.29 (s, 2 H); MS (ESI): m/z: 349 [M+H]+.
Example 85: 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000159_0001
0.073 g (93%) of 2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 85) was prepared according to the procedure described for Example 24, starting from 0.095 g (0.20 mmol) of 2-[(Z)-C-[[4- [(4-fluorophenyl)methoxy]phenoxy]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 113). 1H NMR (DMSO-d6) δ (ppm): 13.79 (bs, 1 H), 12.33 (s, 1 H), 8.81 - 8.70 (m, 1 H), 8.24 - 8.20 (m, 1 H), 7.81 - 7.77 (m, 1 H), 7.49 - 7.44 (m, 2 H), 7.23 - 7.16 (m, 2 H), 6.93 - 6.86 (m, 4 H), 5.17 (s, 2 H), 5.06 - 4.98 (m, 2 H); MS (ESI): m/z: 397 [M+H]+.
Example 86: 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000159_0002
0.040 g (98%) of 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid (Example 86) was prepared according to the procedure described for Example 24, starting from 0.052 g (0.13 mmol) of 2-[(Z)-C-[(4-chlorophenoxy)methyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 115). 1H NMR (DMSO-de) δ (ppm): 13.75 (bs, 1 H), 12.42 (s, 1 H), 8.78 - 8.74 (m, 1 H), 8.26 - 8.22 (m, 1 H), 7.81 - 7.78 (m, 1 H), 7.33 - 7.28 (m, 1 H), 7.03 - 6.96 (m, 1 H), 5.23 (s, 2 H), 4.96 - 4.93 (m, 1 H); MS (ESI): m/z: 307 [M+H]+.
Example 87: 2-[(Z)-C-(cyclopentoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000160_0001
0.0050 g (82%) of 2-[(Z)-C-(cyclopentoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 87) was prepared according to the procedure described for Example 24, starting from 0.0080 g (0.020 mmol) of 2-[(Z)-C-(cyclopentoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 116). 1H NMR (DMSO-d6) 5 (ppm): 13.64 (bs, 1 H), 11.98 (s, 1 H), 8.81 - 8.70 (m, 1 H), 8.25 - 8.13 (m, 1 H), 7.84 - 7.72 (m, 1 H), 4.61 (s, 2 H), 4.04 - 3.87 (m, 1 H), 1.68 - 1.33 (m, 8 H); MS (ESI): m/z: 265 [M+H]+.
Example 88: 2-[(Z)-C-(cyclobutoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000160_0002
0.0030 g (27%) of 2-[(Z)-C-(cyclobutoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 88) was prepared according to the procedure described for Example 24, starting from 0.015 g (0.040 mmol) of 2-[(Z)-C-(cyclobutoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 117). 1H NMR (DMSO-de) δ (ppm): 14.27 - 13.15 (m, 1 H), 12.02 (s, 1 H), 8.80 - 8.75 (m, 1 H), 8.21 - 8.17 (m, 1 H), 7.79 - 7.76 (m, 1 H), 4.58 (s, 2 H), 3.99 - 3.90 (m, 1 H), 2.13 - 2.03 (m, 2 H), 1.82 - 1.71 (m, 2 H), 1 .61 - 1.51 (m, 1 H), 1.47 - 1.34 (m, 1 H); MS (ESI): m/z: 251 [M+H]+
Example 89: 2-[(Z)-C-(propoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000160_0003
0.0050 g (21 %) of 2-[(Z)-C-(propoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 89) was prepared according to the procedure described for Example 24, starting from 0.010 g (0.031 mmol) of 2-[(Z)-C-(propoxymethyl)-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 118). 1H NMR (DMSO-de) δ (ppm): 13.71 (bs, 1 H), 12.01 (s, 1 H), 8.83 - 8.71 (m, 1 H), 8.25 - 8.13 (m, 1 H), 7.85 - 7.75 (m, 1 H), 4.65 (s, 2 H), 3.35 (t, J = 7.0 Hz, 2 H), 1.47 - 1.36 (m, 2 H), 0.74 (t, J = 7.0 Hz, 3 H); MS (ESI): m/z: 239 [M+H]+
Example 90: ethyl 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox late
Figure imgf000161_0001
0.043 g (60%) of ethyl 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 90) was prepared according to the procedure described for Example 1 , starting from 0.068 g (0.22 mmol) of ethyl 2-[2-(4- chlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 120).1H NMR (CDCI3) δ (ppm): 8.75 - 8.82 (m, 1 H), 8.36 - 8.45 (m, 1 H), 7.77 - 7.87 (m, 1 H), 7.13 - 7.35 (m, 4H), 4.34 - 4.48 (m, 4H), 1.41 (t, J = 7.09 Hz, 3H); MS (ESI): m/z: 319 [M+H]+.
Example 91 : 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid
Figure imgf000161_0002
0.025 g (69%) of 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 91 ) was prepared according to the procedure described for Example 12, starting from 0.040 g (0.13 mmol) of ethyl 2-[(E)-C-[(4-chlorophenyl)methyl]- N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 90). 1H NMR (DMSO-d6) δ (ppm): 13.75 (bs, 1 H), 1 1.94 (s, 1 H), 8.67 - 8.87 (m, 1 H), 8.23 - 8.41 (m, 1 H), 7.70 - 7.83 (m, 1 H), 7.18 - 7.39 (m, 4H), 4.23 (s, 2H); MS (ESI): m/z: 291 [M+H]+. Example 92: ethyl 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox late
Figure imgf000162_0001
0.048 g (72%) of ethyl 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 92) was prepared according to the procedure described for Example 1 , starting from 0.064 g (0.17 mmol) of ethyl 2-[2-(4- benzyloxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 121 ).1H NMR (DMSO-de) δ (ppm): 1 1.88 (s, 1 H), 8.69 - 8.89 (m, 1 H), 8.21 - 8.40 (m, 1 H), 7.68 - 7.86 (m, 1 H), 7.24 - 7.46 (m, 5H), 7.03 - 7.23 (m, 2H), 6.73 - 6.94 (m, 2H), 5.00 (s, 2H), 4.35 (q, J = 6.85 Hz, 2H), 4.18 (s, 2H), 1.32 (t, J = 7.10 Hz, 3H); MS (ESI): m/z: 391 [M+H]+.
Example 93: 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000162_0002
0.021 g (58%) of 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 93) was prepared according to the procedure described for Example 12, starting from 0.039 g (0.10 mmol) of 2-[(E)-C-[(4- benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 92). 1H NMR (DMSO-de) δ (ppm): 13.70 (bs, 1 H), 1 1.82 (s, 1 H), 8.70 - 8.84 (m, 1 H), 8.22 - 8.35 (m, 1 H), 7.70 - 7.82 (m, 1 H), 7.23 - 7.45 (m, 5H), 7.07 - 7.22 (m, 2H), 6.74 - 6.94 (m, 2H), 5.00 (s, 2H), 4.18 (s, 2H); MS (ESI): m/z: 363 [M+H]+.
Example 94: ethyl 2-[(E)-N-hydroxy-C-[(4- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate
Figure imgf000163_0001
0.038 g (61 %) of ethyl 2-[(E)-N-hydroxy-C-[(4- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate (Example 94) was prepared according to the procedure described for Example 1 , starting from 0.060 g (0.17 mmol) of ethyl 2-[2-(4-phenoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 122). 1H NMR (DMSO-de) δ (ppm): 1 1.95 (s, 1 H), 8.86 - 8.74 (m, 1 H), 8.40 - 8.28 (m, 1 H), 7.79 (dd, J = 1.7, 5.1 Hz, 1 H), 7.40 - 7.20 (m, 4 H), 7.14 - 7.05 (m, 1 H), 6.98 - 6.79 (m, 4 H), 4.35 (q, J = 6.8 Hz, 2 H), 4.24 (s, 2 H), 1.33 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 377 [M+H]+. Example 95: 2-[(E)-N-hydroxy-C-[(4- phenoxyphenyl)methyl]carbonimido l]pyridine-4-carboxylic acid
Figure imgf000163_0002
0.027 g (78%) of 2-[(E)-N-hydroxy-C-[(4-phenoxyphenyl)methyl]carbonimidoyl]pyridine- 4-carboxylic acid (Example 95) was prepared according to the procedure described for Example 12, starting from 0.038 g (0.10 mmol) of ethyl 2-[(E)-N-hydroxy-C-[(4- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate (Example 94). 1H NMR (DMSO-de) δ (ppm): 13.70 (bs, 1 H), 1 1.89 (s, 1 H), 8.87 - 8.65 (m, 1 H), 8.39 - 8.24 (m, 1 H), 7.82 - 7.71 (m, 1 H), 7.43 - 7.16 (m, 4 H), 7.13 - 7.03 (m, 1 H), 6.99 - 6.79 (m, 4 H), 4.24 (s, 2 H); MS (ESI): m/z: 349 [M+H]+.
Example 96: ethyl 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N- hydroxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000163_0003
0.068 g (65%) of ethyl 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 96) was prepared according to the procedure described for Example 1 , starting from 0.10 g (0.25 mmol) of ethyl 2-[2-[4-[(4- fluorophenyl)methoxy]phenyl]acetyl]pyridine-4-carboxylate (Intermediate 123). 1H NMR (DMSO-de) δ (ppm): 1 1.88 (s, 1 H), 8.84 - 8.71 (m, 1 H), 8.35 - 8.22 (m, 1 H), 7.77 (dd, J = 1.7, 5.1 Hz, 1 H), 7.49 - 7.39 (m, 2 H), 7.24 - 7.07 (m, 4 H), 6.91 - 6.77 (m, 2 H), 4.98 (s, 2 H), 4.35 (q, J = 7.3 Hz, 2 H), 4.18 (s, 2 H), 1 .32 (t, J = 7.3 Hz, 3 H); MS (ESI): m/z: 409 [M+H]+.
Example 97: 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000164_0001
0.050 g (79%) of 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 97) was prepared according to the procedure described for Example 12, starting from 0.068 g (0.17 mmol) of ethyl 2-[(E)-C- [[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate (Example 96). 1H NMR (DMSO-d6) δ (ppm): 13.69 (bs, 1 H), 1 1.82 (s, 1 H), 8.82 - 8.62 (m, 1 H), 8.35 - 8.19 (m, 1 H), 7.75 (dd, J = 1.7, 5.1 Hz, 1 H), 7.52 - 7.32 (m, 2 H), 7.27 - 7.07 (m, 4 H), 6.91 - 6.66 (m, 2 H), 4.98 (s, 2 H), 4.18 (s, 2 H); MS (ESI): m/z: 381 [M+H]+.
Example 98: ethyl 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N- hydroxy-carbonimidoyl]pyridine-4-carboxylate
Figure imgf000165_0001
0.081 g (78%) of ethyl 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 98) was prepared according to the procedure described for Example 1 , starting from 0.10 g (0.25 mmol) of ethyl 2-[2-[3-[(4- fluorophenyl)methoxy]phenyl]acetyl]pyridine-4-carboxylate (Intermediate 124). 1H NMR (DMSO-de) δ (ppm): 1 1.94 (s, 1 H), 8.83 - 8.73 (m, 1 H), 8.35 - 8.27 (m, 1 H), 7.78 (dd, J = 1.7, 5.1 Hz, 1 H), 7.48 - 7.38 (m, 2 H), 7.22 - 6.70 (m, 6 H), 4.98 (s, 2 H), 4.35 (q, J = 6.8 Hz, 2 H), 4.22 (s, 2 H), 1.33 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 409 [M+H]+.
Example 99: 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000165_0002
0.048 g (86%) of 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 99) was prepared according to the procedure described for Example 12, starting from 0.060 g (0.15 mmol) of ethyl 2-[(E)-C- [[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate (Example 98). 1H NMR (DMSO-de) δ (ppm): 13.81 - 13.63 (bs, 1 H), 11.87 (s, 1 H), 8.81 - 8.69 (m, 1 H), 8.33 - 8.25 (m, 1 H), 7.79 - 7.68 (m, 1 H), 7.49 - 7.38 (m, 2 H), 7.24 - 6.68 (m, 6 H), 4.98 (s, 2 H), 4.22 (s, 2 H; MS (ESI): m/z: 381 [M+H]+. Example 100: ethyl 2-[(E)-N-hydroxy-C-[(4- phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate
Figure imgf000166_0001
0.085 g (82%) of ethyl 2-[(E)-N-hydroxy-C-[(4- phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate (Example 100) was prepared according to the procedure described for Example 1 , starting from 0.10 g (0.29 mmol) of ethyl 2-[2-(4-phenylphenyl)acetyl]pyridine-4-carboxylate (Intermediate 125). 1H NMR (DMSO-de) δ (ppm): 1 1.97 (s, 1 H), 8.85 - 8.76 (m, 1 H), 8.38 - 8.32 (m, 1 H), 7.83 - 7.76 (m, 1 H), 7.64 - 7.24 (m, 9 H), 4.42 - 4.25 (m, 4 H), 1.33 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 361 [M+H]+.
Example 101 : 2-[(E)-N-hydroxy-C-[(4- phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000166_0002
0.060 g (81 %) of 2-[(E)-N-hydroxy-C-[(4-phenylphenyl)methyl]carbonimidoyl]pyridine-4- carboxylic acid (Example 101 ) was prepared according to the procedure described for Example 12, starting from 0.080 g (0.22 mmol) of ethyl 2-[(E)-N-hydroxy-C-[(4- phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate (Example 100). 1 H NMR (DMSO-d6) δ (ppm): 13.74 (bs, 1 H), 1 1.92 (s, 1 H), 8.88 - 8.68 (m, 1 H), 8.41 - 8.21 (m, 1 H), 7.77 (dd, J = 1 .5, 4.9 Hz, 1 H), 7.63 - 7.26 (m, 9 H), 4.30 (s, 2 H); MS (ESI): m/z: 333 [M+H]+.
Example 102: ethyl 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000167_0001
0.015 g (35%) of ethyl 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 102) was prepared according to the procedure described for Example 1 , starting from 0.040 g (0.12 mmol) of ethyl 2-[2-(3,5- dichlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 126). 1H NMR (CDCI3) δ (ppm): 8.84 - 8.74 (m, 1 H), 8.48 - 8.42 (m, 1 H), 7.89 - 7.83 (m, 1 H), 7.30 - 7.25 (m, J = 2.0 Hz, 2 H), 7.20 - 7.14 (m, 1 H), 4.43 (q, J = 7.2 Hz, 2 H), 4.36 (s, 2 H), 1.46 - 1.37 (m, 3 H); MS (ESI): m/z: 354 [M+H]+.
Example 103: 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000167_0002
0.0070 g (54%) of 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 103) was prepared according to the procedure described for Example 12, starting from 0.015 g (0.040 mmol) of 2-[(E)-C-[(3,5- dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 102). 1 H NMR (DMSO-d6) δ (ppm): 13.77 (bs, 1 H), 12.08 (s, 1 H), 8.83 - 8.74 (m, 1 H), 8.36 - 8.29 (m, 1 H), 7.82 - 7.74 (m, 1 H), 7.45 - 7.37 (m, 1 H), 7.31 - 7.22 (m, 2 H), 4.25 (s, 2 H); MS (ESI): m/z: 326 [M+H]+.
Example 104: ethyl 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000167_0003
0.012 g (28%) of ethyl 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 104) was prepared according to the procedure described for Example 1 , starting from 0.040 g (0.12 mmol) of ethyl 2-[2-(3,4- dichlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 127). 1H NMR (CDCI3) δ (ppm): 8.79 - 8.73 (m, 1 H), 8.47 - 8.41 (m, 1 H), 7.86 - 7.80 (m, 1 H), 7.50 - 7.45 (m, 1 H), 7.32 - 7.26 (m, 1 H), 7.24 - 7.18 (m, 1 H), 4.42 (q, J = 7.0 Hz, 2 H), 4.36 (s, 2 H), 1.44 - 1.39 (m, 3 H); MS (ESI): m/z: 354 [M+H]+.
Example 105: 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000168_0001
0.0050 g (51 %) of 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 105) was prepared according to the procedure described for Example 12, starting from 0.010 g (0.030 mmol) of ethyl 2-[(E)- C-[(3,4-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate
(Example 104). 1 H NMR (DMSO-d6) δ (ppm): 13.78 (bs, 1 H), 12.03 (s, 1 H), 8.80 - 8.74 (m, 1 H), 8.36 - 8.28 (m, 1 H), 7.80 - 7.74 (m, 1 H), 7.53 - 7.45 (m, 2 H), 7.26 - 7.18 (m, 1 H), 4.24 (s, 2 H); MS (ESI): m/z: 326 [M+H]+.
Example 106: ethyl 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox late
Figure imgf000168_0002
0.070 g (40%) of ethyl 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 106) was prepared according to the procedure described for Example 1 , starting from 0.17 g (0.45 mmol) of ethyl 2-[2-(3- benzyloxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 128). 1H NMR (CDCI3) δ (ppm): 8.78 - 8.73 (m, 1 H), 8.43 - 8.38 (m, 1 H), 7.83 - 7.78 (m, 1 H), 7.57 (bs, 1 H), 7.43 - 7.28 (m, 5 H), 7.19 - 7.12 (m, 1 H), 7.02 - 6.92 (m, 2 H), 6.81 - 6.75 (m, 1 H), 5.01 (s, 2 H), 4.46 - 4.34 (m, 4 H), 1.44 - 1.37 (m, 3 H); MS (ESI): m/z: 391 [M+H]+.
Example 107: 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000169_0001
0.0050 g (51 %) of 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid (Example 107) was prepared according to the procedure described for Example 12, starting from 0.060 g (0.15 mmol) of ethyl 2-[(E)-C- [(3-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 106). 1H NMR (DMSO-de) δ (ppm): 13.78 (bs, 1 H), 12.03 (s, 1 H), 8.80 - 8.74 (m, 1 H), 8.36 - 8.28 (m, 1 H), 7.80 - 7.74 (m, 1 H), 7.53 - 7.45 (m, 2 H), 7.26 - 7.18 (m, 1 H), 4.24 (s, 2 H); MS (ESI): m/z: 363 [M+H]+.
Example 108: ethyl 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000169_0002
0.012 g (9%) of ethyl 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 108) was prepared according to the procedure described for Example 1 , starting from 0.13 g (0.43 mmol) of ethyl 2-[2-(3- chlorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 129). 1H NMR (CDCI3) δ (ppm): 8.79 - 8.73 (m, 1 H), 8.47 - 8.41 (m, 1 H), 7.86 - 7.80 (m, 1 H), 7.50 - 7.45 (m, 1 H), 7.32 - 7.26 (m, 1 H), 7.24 - 7.18 (m, 1 H), 4.42 (q, J = 7.0 Hz, 2 H), 4.36 (s, 2 H), 1 .44 - 1.39 (m, 3 H); MS (ESI): m/z: 319 [M+H]+.
Example 109: 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000170_0001
0.038 g (82%) of 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyndine-4- carboxylic acid (Example 109) was prepared according to the procedure described for Example 12, starting from 0.050 g (0.16 mmol) of ethyl 2-[(E)-C-[(3-chlorophenyl)methyl]- N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 108). 1H NMR (DMSO-d6) δ (ppm): 13.76 (bs, 1 H), 1 1.98 (s, 1 H), 8.80-8.71 (m, 1 H), 8.36-8.28 (m, 1 H), 7.80-7.73 (m, 1 H), 7.31 -7.14 (m, 4 H), 4.26 (s, 2H) ; MS (ESI): m/z: 291 [M+H]+.
Example 110: ethyl 2-[(E)-N-hydroxy-C-[(3- phenoxyphenyl)methyl]carbonimido l]pyridine-4-carboxylate
Figure imgf000170_0002
0.070 g (40%) of ethyl 2-[(E)-N-hydroxy-C-[(3- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylate (Example 110) was prepared according to the procedure described for Example 1 , starting from 0.17 g (0.45 mmol) of ethyl 2-[2-(3-phenoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 130). 1H NMR (CDCIs) δ (ppm): 8.78 - 8.73 (m, 1 H), 8.43 - 8.38 (m, 1 H), 7.83 - 7.78 (m, 1 H), 7.57 (bs, 1 H), 7.43 - 7.28 (m, 5 H), 7.19 - 7.12 (m, 1 H), 7.02 - 6.92 (m, 2 H), 6.81 - 6.75 (m, 1 H), 5.01 (s, 2 H), 4.46 - 4.34 (m, 4 H), 1.44 - 1.37 (m, 3 H); MS (ESI): m/z: 391 [M+H]+. Example 111 : ethyl 2-[(E)-N-hydroxy-C-[(3- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000171_0001
0.007 g (29%) of ethyl 2-[(E)-N-hydroxy-C-[(3- phenoxyphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid (Example 111 ) was prepared according to the procedure described for Example 12, starting from 0.025 g (0.07 mmol) of ethyl 2-[(E)-C-[(3-phenoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 110). 1H NMR (DMSO-d6) δ (ppm): 13.76 (bs, 1 H), 1 1.88 (s, 1 H), 8.75 - 8.67 (m, 1 H), 8.32 - 8.22 (m, 1 H), 7.79 - 7.71 (m, 1 H), 7.38 - 7.31 (m, 2 H), 7.25 - 7.19 (m, 1 H), 7.14 - 7.09 (m, 1 H), 7.02 - 6.98 (m, 1 H), 6.95 - 6.90 (m, 2 H), 6.89 - 6.84 (m, 1 H), 6.78 - 6.73 (m, 1 H), 4.23 (s, 2 H); MS (ESI): m/z: 349 [M+H]+
Example 112: ethyl 2-[(E)-C-[(3-phenylphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox late
Figure imgf000171_0002
0.036 g (50%) of ethyl 2-[(E)-C-[(3-phenylphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 112) was prepared according to the procedure described for Example 1 , starting from 0.070 g (0.20 mmol) of ethyl 2-[2-(3- phenylphenyl)acetyl]pyridine-4-carboxylate (Intermediate 131 ). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.74 (m, 1 H), 8.46 - 8.40 (m, 1 H), 7.85 - 7.78 (m, 1 H), 7.66 - 7.28 (m, 10 H), 4.49 (s, 2 H), 4.45 - 4.36 (m, 2 H), 1.40 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 361 [M+H]+. Example 113: 2-[(E)-N-hydroxy-C-[(3- phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000172_0001
0.015 g (56%) of 2-[(E)-N-hydroxy-C-[(3-phenylphenyl)methyl]carbonimidoyl]pyridine-4- carboxylic acid (Example 113) was prepared according to the procedure described for Example 12, starting from 0.030 g (0.080 mmol) of ethyl 2-[(E)-C-[(3- phenylphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 112). 1H NMR (DMSO-de) δ (ppm): 13.72 (bs, 1 H), 1 1.92 (s, 1 H), 8.81 - 8.75 (m, 1 H), 8.35 - 8.30 (m, 1 H), 7.79 - 7.73 (m, 1 H), 7.57 - 7.18 (m, 9 H), 4.34 (s, 2 H); MS (ESI): m/z: 333 [M+H]+.
Example 114: ethyl 2-[N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4- carboxylate
Figure imgf000172_0002
0.10 g (79%) of ethyl 2-[N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4- carboxylate (Example 114) was prepared according to the procedure described for Example 1 , starting from 0.12 g (0.38 mmol) of ethyl 2-[2-(1 -naphthyl)acetyl]pyridine-4- carboxylate (Intermediate 132). It was obtained as a 2:1 mixture of the two diastereoisomers and used in the next step without any further purification. MS (ESI): m/z: 335 [M+H]+.
Example 115: 2-[(E)-N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000173_0001
0.022 g (27%) of 2-[(E)-N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 115) was prepared according to the procedure described for Example 12, starting from 0.090 g (0.27 mmol) of ethyl 2-[N-hydroxy-C-(1 - naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate (Example 114). The crude product was purified by preparative HPLC to afford 2-[(E)-N-hydroxy-C-(1 - naphthylmethyl)carbonimidoyl]pyridine-4-carboxylic acid (Example 115) as a pale rose solid. 1H NMR (DMSO-de) δ (ppm): 13.75 (bs, 1 H), 1 1.99 (s, 1 H), 8.72 - 8.62 (m, 1 H), 8.42 - 8.34 (m, 1 H), 8.31 - 8.22 (m, 1 H), 7.96 - 7.86 (m, 1 H), 7.77 - 7.67 (m, 2 H), 7.62 - 7.46 (m, 2 H), 7.36 - 7.26 (m, 1 H), 7.12 - 7.03 (m, 1 H), 4.68 (s, 2 H) ; MS (ESI): m/z: 307 [M+H]+.
Example 116: ethyl 2-[N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4- carboxylate
Figure imgf000173_0002
0.040 g (32%) of ethyl 2-[N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4- carboxylate (Example 116) was prepared according to the procedure described for Example 1 , starting from 0.12 g (0.38 mmol) of ethyl 2-[2-(2-naphthyl)acetyl]pyridine-4- carboxylate (Intermediate 133).1H NMR (CDCI3) δ (ppm): 8.82 - 8.75 (m, 1 H), 8.47 - 8.39 (m, 1 H), 7.86 - 7.32 (m, 9 H), 4.59 (s, 2 H), 4.41 (q, J = 7.3 Hz, 2 H), 1.40 (t, J = 7.3 Hz, 3 H); MS (ESI): m/z: 335 [M+H]+.
Example 117: 2-[(E)-N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000174_0001
0.030 g (99%) of 2-[(E)-N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4- carboxylic acid (Example 117) was prepared according to the procedure described for Example 12, starting from 0.035 g (0.10 mmol) of ethyl 2-[N-hydroxy-C-(2- naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate (Example 116). 1H NMR (DMSO- d6) δ (ppm): 13.73 (bs, 1 H), 1 1.95 (s, 1 H), 8.82 - 8.72 (m, 1 H), 8.39 - 8.30 (m, 1 H), 7.84 - 7.73 (m, 4 H), 7.71 - 7.64 (m, 1 H), 7.47 - 7.36 (m, 3 H), 4.43 (s, 2 H) ; MS (ESI): m/z: 307 [M+H]+.
Example 118: 2-[(E)-N-hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenyl]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000174_0002
CI H
0.023 g (quantitative) of 2-[(E)-N-hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenyl]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 118) was prepared according to the procedure described for Example 24, starting from 0.026 g (0.060 mmol) of 2-[(E)-C-[[3-(4-methylpiperazin-1 -yl)phenyl]methyl]-N-tetrahydropyran- 2-yloxy-carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 136). 1H NMR (DMSO-de) 5 (ppm): 1 1.90 (bs, 1 H), 10.83 (bs, 1 H), 8.87 - 8.53 (m, 1 H), 8.35 - 8.19 (m, 1 H), 7.76 (dd, J = 1.5, 4.9 Hz, 1 H), 7.16 - 7.00 (m, 1 H), 6.94 - 6.86 (m, 1 H), 6.81 - 6.64 (m, 2 H), 4.22 (s, 2 H), 3.75 - 3.37 (m, 4 H), 3.20 - 2.90 (m, 4 H), 2.78 (d, J = 4.4 Hz, 3 H); MS (ESI): m/z: 391 [M+H]+
Example 119: ethyl 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000175_0001
0.019 g (60%) of ethyl 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 119) was prepared according to the procedure described for Example 1 , starting from 0.030 g (0.10 mmol) of ethyl 2-[2-(4- fluorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 137). 1H NMR (DMSO-d6) δ (ppm): 1 1.97 (s, 1 H), 8.83 - 8.71 (m, 1 H), 8.38 - 8.28 (m, 1 H), 7.86 - 7.73 (m, 1 H), 7.32 - 6.95 (m, 4 H), 4.41 - 4.17 (m, 4 H), 1.32 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 303 [M+H]+. Example 120: 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid
Figure imgf000175_0002
0.010 g (58%) of 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 120) was prepared according to the procedure described for Example 12, starting from 0.019 g (0.063 mmol) of ethyl 2-[(E)-C-[(4-fluorophenyl)methyl]- N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 119). 1H NMR (DMSO-d6) δ (ppm): 13.74 (bs, 1 H), 1 1.91 (s, 1 H), 8.81 - 8.71 (m, 1 H), 8.34 - 8.26 (m, 1 H), 7.76 (dd, J = 1 .5, 4.9 Hz, 1 H), 7.31 - 7.21 (m, 2 H), 7.09 - 6.99 (m, 2 H), 4.23 (s, 2 H); MS (ESI): m/z: 275 [M+H]+.
Example 121 : 1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride
Figure imgf000175_0003
0.022 g (73%) of 1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride (Example 121 ) was prepared according to the procedure described for Example 24, starting from 0.035 g (0.1 1 mmol) of 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N- tetrahydropyran-2-yloxy-carbonimidoyl]-4-pyridyl]ethanol (Intermediate 141 ). 1H NMR (DMSO-de) δ (ppm): 1 1.69 (bs, 1 H), 8.70 - 8.53 (m, 1 H), 8.09 - 7.93 (m, 1 H), 7.59 - 7.46 (m, 1 H), 7.14 (bs, 1 H), 5.49 - 5.15 (m, 1 H), 2.22 (s, 3 H); MS (ESI): m/z: 271 [M+H]+ Example 122: 1 -[4-(2,2,2-trifluoro-1 ,1 -dihydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride
Figure imgf000176_0001
0.018 g (72%) of 1 -[4-(2,2,2-trifluoro-1 ,1 -dihydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride (Example 122) was prepared according to the procedure described for Example 24, starting from 0.030 g (0.090 mmol) of 2,2,2-trifluoro-1 -[2-[(E)-C-methyl-N- tetrahydropyran-2-yloxy-carbonimidoyl]-4-pyridyl]ethane-1 ,1 -diol (Intermediate 142). 1H NMR (DMSO-de) δ (ppm): 1 1.63 (bs, 1 H), 8.70 - 8.54 (m, 1 H), 8.21 - 8.07 (m, 1 H), 7.86 (bs, 2 H), 7.60 - 7.32 (m, 1 H), 6.75 (bs, 1 H), 2.22 (s, 3 H); MS (ESI): m/z: 287 [M+H]+ Example 123: 2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl- amino]ethoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000176_0002
0.0070 g (49%) of 2-[(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl- amino]ethoxy]phenyl]methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 123) was prepared according to the procedure described for Example 24, starting from 0.015 g (0.030 mmol) of 2-[(E)-C-[[3-[2-[(4- fluorophenyl)methyl-methyl-amino]ethoxy]phenyl]methyl]-N-tetrahydropyran-2-yloxy- carbonimidoyl]pyridine-4-carboxylic acid (Intermediate 147). 1H NMR (DMSO-de) δ (ppm): 1 1.93 (bs, 1 H), 10.81 (bs, 1 H), 8.84 - 8.68 (m, 1 H), 8.37 - 8.20 (m, 1 H), 7.85 - 7.57 (m, 3 H), 7.36 - 7.09 (m, 3 H), 6.93 - 6.69 (m, 3 H), 4.44 - 4.10 (m, 6 H), 3.51 - 3.30 (m, 2 H), 2.71 (bs, 3 H); MS (ESI): m/z: 438 [M+H]+.
Example 124: methyl 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylate
Figure imgf000177_0001
0.0060 g (6%) of methyl 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate (Example 124) were prepared according to the procedure described for Example 1 , starting from 0.090 g (0.35 mmol) of methyl 2-(4-fluorobenzoyl)pyridine-4- carboxylate (Intermediate 147). 1H NMR (DMSO-d6) δ (ppm): 1 1.94 (s, 1 H), 8.73 - 8.65 (m, 1 H), 8.36 - 8.28 (m, 1 H), 7.84 - 7.76 (m, 1 H), 7.43 - 7.35 (m, 2 H), 7.30 - 7.20 (m, 2 H), 3.93 (s, 3 H); MS (ESI): m/z: 275 [M+H.
Example 125: 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid
Figure imgf000177_0002
0.0030 g (63%) of 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 125) was prepared according to the procedure described for Example 12, starting from 0.0050 g (0.020 mmol) of methyl 2-[(E)-C-(4-fluorophenyl)-N- hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 124). 1H NMR (DMSO-de) δ (ppm): 13.79 (bs, 1 H), 1 1.92 (s, 1 H), 8.69 - 8.61 (m, 1 H), 8.33 - 8.28 (m, 1 H), 7.82 - 7.73 (m, 1 H), 7.44 - 7.34 (m, 2 H), 7.30 - 7.19 (m, 2 H); MS (ESI): m/z: 261 [M+H]+. Example 126: ethyl 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000178_0001
0.030 g (32%) of ethyl 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 126) was prepared according to the procedure described for Example 1 , starting from 0.090 g (0.31 mmol) of ethyl 2-[2-(3- fluorophenyl)acetyl]pyridine-4-carboxylate (Intermediate 149). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.72 (m, 1 H), 8.48 - 8.39 (m, 1 H), 7.88 - 7.79 (m, 1 H), 7.60 (bs, 1 H), 7.24 - 7.04 (m, 3 H), 6.90 - 6.81 (m, 1 H), 4.46 - 4.36 (m, 4 H), 1.45 - 1.37 (m, 3 H); MS (ESI): m/z: 303 [M+H]+.
Example 127: 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid
Figure imgf000178_0002
0.015 g (66%) of 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid (Example 127) was prepared according to the procedure described for Example 12, starting from 0.025 g (0.08 mmol) of ethyl 2-[(E)-C-[(3-fluorophenyl)methyl]- N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 126). 1H NMR (DMSO-d6) δ (ppm): 13.74 (bs, 1 H), 1 1.97 (s, 1 H), 8.82 - 8.74 (m, 1 H), 8.37 - 8.27 (m, 1 H), 7.82 - 7.70 (m, 1 H), 7.34 - 7.20 (m, 1 H), 7.12 - 6.88 (m, 3 H), 4.27 (s, 2 H); MS (ESI): m/z: 275 [M+H]+.
Example 128: Ethyl 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000178_0003
0.14 g (89%) of Ethyl 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 128) was prepared according to the procedure described for Example 1 , starting from 0.15 g (0.50 mmol) of ethyl 2-[2-(3- methoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 150). 1H NMR (CDCI3) δ (ppm): 8.81 - 8.74 (m, 1 H), 8.44 - 8.38 (m, 1 H), 7.97 - 7.65 (m, 2 H), 7.20 - 7.1 1 (m, 1 H), 6.97 - 6.88 (m, 2 H), 6.74 - 6.67 (m, 1 H), 4.46 - 4.36 (m, 4 H), 3.76 (s, 3 H), 1.41 (t, J = 7.1 Hz, 3 H); MS (ESI): m/z: 315 [M+H]+.
Example 129: 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid
Figure imgf000179_0001
0.091 g (83%) of 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid (Example 129) was prepared according to the procedure described for Example 12, starting from 0.12 g (0.38 mmol) of ethyl 2-[(E)-C-[(3- methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 128). 1H NMR (DMSO-de) δ (ppm): 13.73 (bs, 1 H), 1 1.87 (s, 1 H), 8.81 - 8.72 (m, 1 H), 8.33 - 8.26 (m, 1 H), 7.79 - 7.72 (m, 1 H), 7.16 - 7.06 (m, 1 H), 6.82 - 6.75 (m, 2 H), 6.73 - 6.66 (m, 1 H), 4.23 (s, 2 H), 3.66 (s, 3 H); MS (ESI): m/z: 287 [M+H]+.
Example 130: Ethyl 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate
Figure imgf000179_0002
0.030 g (41 %) of Ethyl 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate (Example 130) was prepared according to the procedure described for Example 1 , starting from 0.070 g (0.23 mmol) of ethyl 2-[2-(4- methoxyphenyl)acetyl]pyridine-4-carboxylate (Intermediate 151 ). 1H NMR (CDCI3) δ (ppm): 8.78 (m, 1 H), 8.43 - 8.36 (m, 1 H), 7.90 - 7.84 (m, 1 H), 7.31 - 7.22 (m, 2 H), 6.82 - 6.74 (m, 2 H), 4.42 (q, J = 6.8 Hz, 2 H), 4.36 (s, 2 H), 3.76 (s, 3 H), 1.45 - 1.38 (m, 3 H); MS (ESI): m/z: 315 [M+H]+.
Example 131 : 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carbox lic acid
Figure imgf000180_0001
0.025 g (92%) of 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid (Example 131 ) was prepared according to the procedure described for Example 12, starting from 0.030 g (0.10 mmol) of ethyl 2-[(E)-C-[(4- methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate (Example 130). 1H NMR (DMSO-de) δ (ppm): 13.71 (bs, 1 H), 1 1.82 (s, 1 H), 8.82 - 8.71 (m, 1 H), 8.32 - 8.23 (m, 1 H), 7.79 - 7.68 (m, 1 H), 7.20 - 7.08 (m, 2 H), 6.83 - 6.69 (m, 2 H), 4.18 (s, 2 H), 3.66 (s, 3 H); MS (ESI): m/z: 287 [M+H]+.
Example 132: 2-[(Z)-N-hydroxy-C-[2-(4-phenyl-1 - piperidyl)ethoxymethyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride
Figure imgf000180_0002
0.017 g (76%) of 2-[(Z)-N-hydroxy-C-[2-(4-phenyl-1 - piperidyl)ethoxymethyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride (Example 132) was prepared according to the procedure described for Example 24, starting from 0.025 g (0.053 mmol) of 2-[(Z)-C-[2-(4-phenyl-1 -piperidyl)ethoxymethyl]-N- tetrahydropyran-2-yloxy-carbonimidoyl]pyridine-4-carboxylic (Intermediate 152). 1H NMR (DMSO-de) δ (ppm): 13.75 (bs, 1 H), 12.26 (s, 1 H), 9.66 (bs, 1 H), 8.84 - 8.69 (m, 1 H), 8.32 - 8.20 (m, 1 H), 7.80 (dd, J = 1 .5, 4.9 Hz, 1 H), 7.39 - 7.14 (m, 5 H), 4.77 (s, 2 H), 3.88 - 3.76 (m, 2 H), 3.48 - 2.87 (m, 6 H), 2.75 - 2.62 (m, 1 H), 1.97 - 1.74 (m, 4 H); MS (ESI): m/z: 384 [M+H]+.
2. BIOLOGICAL TESTING
2.1 Assay of enzyme inhibition of KDM4C (JMJD2C) The KDM4C inhibiting activity was determined using a TR-FRET assay (time resolved fluorescence resonance energy transfer, Lance® Ultra Demethylase technology (Perkin Elmer, Waltham, MA, USA), which comprises a Europium chelate donor dye (TRF0403, Perkin Elmer, Waltham, MA, USA) together with \ Light™ (TR0102, Perkin Elmer, Waltham, MA, USA), a small molecular weight acceptor dye with a red-shifted fluorescent emission, and a biotinylated histone H3 (1 -21 ) peptide tri-methylated at lysine 9 (H3K9me3) [Lys(Me3)9]-histone H3 (1 -21 )-GGK(biotin), (cat. no. 64360, Anaspec, Fremont, CA, USA) as substrate. The intensity of the light emission is proportional to the level of biotinylated reaction product. Human KDM4C (JMJD2C (GenBank Accession No. BC143571 ) (a. a. 2-372) with A/-terminal GST-tag was purchased from BPS Bioscience (50105, BPS Bioscience, Inc., San Diego).
Demethylase assay conditions: The assays were carried out at the final concentrations of 7.5 nM KDM4C, 150 nM H3K9me3, 10 μΜ 2-oxoglutarate (2-OG), 10 μΜ FeSO4, 100 μΜ ascorbate, and 4% DMSO in 96 well half-area flat bottom white plates (3693 Costar, Sigma-Aldrich, St. Louis, MO, USA). The reaction was started by addition of the substrate (H3K9me3) and the mixtures were incubated at r.t. for 30 min.
Detection step conditions: 10 μί of the assay mixture were transferred from the original plate to a 384 well white plate (6007290 OptiPlate™, Perkin Elmer, Waltham, MA, USA) containing 10 μL· of a pre-spotted detection mix (3 nM Eu-antibody, 25 nM \ -Light- Streptavidin, 2 mM EDTA in 1 X Lance Detection Buffer (TRF0403, TR0102, CR97100, Perkin Elmer, Waltham, MA, USA)). The resulting mixture was incubated in the dark for 1 h atr.t. Then, the TR-FRET signal was read by a fluorimeter (Infinite® F200, Tecan, Mannedorf, Swirzerland) (excitation 320nm, emission 665 nm and 620 nm, delay time 50 [is, window time 100 με).
ICso determination: The inhibitor concentrations ranged from 0.025 to 100 μΜ (serial 1 :3 dilutions). The IC50 was calculated using GraphPad Software.
Compounds 12-28, 32, 39-44, 46-57, 59-62, 64-89, 91 , 93, 95, 97, 99, 101 , 103, 105, 107, 109, 111 , 113, 115, 117, 118, 120, 123, 125, 127, 129, 131 , 132 exhibited IC50 values of less than 10 μΜ, compounds 1 , 29-30, and 33-35 exhibited IC50 values of less than 50 μΜ.
2.2 Cell-based Immunofluorescence Assay This procedure was used to demonstrate the ability of the compounds of the disclosure to inhibit specific histone lysine demethylases such as KDM4C exogenously and inducibly expressed in the human osteosarcoma cell line U20S.
The U20S cell line (ATCC® HTB-96™) infected with a Tet-On inducible lentiviral pLIX vector (pl_IX_403 from Addgene) encoding KDM4C was used for these experiments. U20S cells (2x103) were seeded into 96 well black plates with clear bottom (Falcon 353219) in 50 μΙ_ of DMEM medium containing 10% FBS TET-free, 1 mM L-glutamine and 100 U/mL antibiotics (penicillin and streptomycin) and were incubated in a humified chamber. After 3-4 h, 50 μΙ_ of medium containing doxycycline (final concentration 4 μg/mL) were added to the cells to induce over-expression of KDM4C. After 24 h of induction the cells were treated with the compounds at different concentrations (1 :2 serial dilutions from 100 μΜ to 0.39 μΜ). The compounds were added to the cells in 50 μΙ_ of medium comprising doxycycline 4ug/ml_, 0.5% final DMSO. After 24 h of incubation with the compounds, the cells were fixed with paraformaldehyde (4% in PBS) for 15 min. After washing with PBS, the cells were permeabilized for 15 min with PBS containing 0.1 % Triton X-100. The cells were washed twice with PBS containing 0.05% Tween-20 and then incubated with the blocking solution (PBS 2% BSA) for 30 min. Staining with primary antibodies (goat anti JMJD2C S-15, S. Cruz Sc-104949; rabbit anti H3K9 tri-methyl (H3K9me3), AbCAM ab8898; dilution 1 :500) was performed for 45 min in blocking solution. The cells were washed 3 times with PBS and then incubated with the secondary antibodies (donkey anti goat Alexa-488, Thermo Fisher A-1 1055; donkey anti rabbit Alexa-647, Thermo Fisher A-31573; 1 :400) for 45 min. After having stained the nuclei with the DAPI (4',6-diamidino-2-phenylindole), a blue-emitting fluorescent compound widely utilized for nuclear staining, PBS was added and high throughput imaging and analysis were performed by ArrayScan VTI (Thermo Scientific).
The IC50 values were determined based on an average measure of the staining of the H3K9me3 in cells, normalized to basal conditions (not induced cells) and to induced cells over-expressing KDM4C (not treated cells). The IC50 value was calculated using GraphPad Prism version 5.0 (GraphPad Software, San Diego, CA). Compounds 12, 17, 22, 24, 27, 28, 48, 50-53, 62 and 109 exhibited IC50 values of less than 20 μΜ.

Claims

1. A compound of formula (I)
Figure imgf000183_0001
(I)
wherein:
Ci-C6 alkyl;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; Ci-C6 alkyl substituted by X-R* C3-C7-cycloalkyl; heterocyclyl, aryl or heteroaryl;
R2 is CH2OH, (CO)R4, CN, tetrazole, CH(OH)CF3 or C(OH)2CF3; R3 is Ci-C6 alkyl; C3-C7 cycloalkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl;
X O or NH;
R4 is hydrogen, OH, O-Ci-C6 alkyl, NR7R8, or CF3;
P is 0 or 1 ; R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl, said aryl being optionally substituted by one ore more substituents independently selected from the group consisting of halogen, CN, or NH2; Ci-C6 alkyl substituted by C3-C7- cycloalkyl optionally substituted by halogen; or Ci-C6 acyl; R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl or heteroaryl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxy; halogen; C-i- C6 alkyl; Ci-C6 alkyl substituted by NR 6R17; Ci-C6 alkoxy; Ci-C6 haloalkyl; Ci-C6 haloalkoxy; NR9R10; CONR20R21 ; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C6 alkyl; aryl; Ci-C6 alkyl substituted by aryl or heteroaryl; Ci-C6 alkoxy substituted by aryl or heteroaryl; phenyloxy, wherein the phenyl may be optionally substituted by halogen; C2-C6 alkoxy substituted by NR 8R19; Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; or C1 -C6 alkyl substituted by aryloxy or heteroaryloxy, wherein the aryl or heteroaryl on the C1 -C6 alkyl substituted by aryl or heteroaryl, or on the C1 -C6 alkoxy substituted by aryl or heteroaryl, or on the C1 -C6 alkyl substituted by aryloxy or heteroaryloxy may indipendently and optionally be substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of aryl or heteroaryl, wherein the aryl or the heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl or heteroaryl, wherein the aryl or the heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; or Ci-C6 alkyl substituted by OR11 or NR 2R13; or (CO)R 4 ; and
R9, R 0 are, independently, hydrogen; d-C6 alkyl; or COR15; are, independently, hydrogen or Ci-C6 alkyl;
R14 is Ci-C6 alkyl or aryl; R 5 is Ci-C6 alkyl; or Ci-C6 alkyl substituted by NH2;
R 6, R 7, R 8, R 9, R20, R2 are, independently, hydrogen, Ci-C6 alkyl, Ci-C6 alkyl substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or Nh ; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by aryl or heteroaryl, wherein the aryl or heteroaryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, CN, or NH2; or stereoisomers or pharmaceutically acceptable salts thereof and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyridine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-isonicotinic acid.
2. A compound of formula (I) according to claim 1 , or stereoisomers or pharmaceutically acceptable salts thereof wherein
R Ci-C6 alkyl;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl;
Ci-C6 alkyl substituted by X-R3 ;
C3-C7-cycloalkyl; heterocyclyl, aryl or heteroaryl; R2 is CH2OH, (CO)R4, CN, or tetrazole;
R3 is Ci-C6 alkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, or NR7R8; p is O or l ;
R5, R6 are, independently, hydrogen; C1-C6 alkyl; C1-C6 alkyl substituted by aryl; Ci- C-6 alkyl substituted by C3-C7-cycloalkyl optionally substituted by halogen; or Ci-C6 acyl;
R7, R8 are, independently, hydrogen, OH, or C-i-C-6 alkyl; wherein the aryl or heteroaryl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, C1-C6 alkyl, C-i-Ce alkoxy, d-Ce haloalkyl, d-C-e haloalkoxy, NR9R10, unsubstituted heterocyclyl, heterocyclyl substituted by Ci-C6 alkyl, Ci-C6 alkyl substituted by aryl; or Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of C1-C6 alkyl; C1-C6 alkyl substituted by aryl; Ci-C6 alkyl substituted by OR11 or NR 2R13; or (CO)R 4 ; and
R9, R 0 are, independently, hydrogen; Ci-C6 alkyl; or COR15;
R , R 2, R 3 are, independently, hydrogen or Ci-C6 alkyl
R 4 is Ci-C6 alkyl or aryl;
R 5 is Ci-C6 alkyl; or Ci-C6 alkyl substituted by NH2; and with the exclusion of 2-(A/-hydroxy-C-methyl-carbonimidoyl)pyridine-4-carbaldehyde and 2-(alpha-hydroxyimino-3,4-dimethoxy-benzyl)-isonicotinic acid.
3. A compound of formula (I) according to claim 1 , or stereoisomers or pharmaceutically acceptable salts thereof wherein
R is Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl; C1 -C6 alkyl substituted by heterocyclyl, wherein the heterocyclyl may be optionally substituted by C1-C6 alkyl;
Ci-C6 alkyl substituted by X-R3;
C3-C7-cycloalkyl; aryl, wherein the aryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, Ci- Ce alkyl, C1-C6 alkoxy; or C1-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen;
R2 is CH2OH, COR4, CN, tetrazole, CH(OH)CF3 or C(OH)2CF3; R3 is Ci-C6 alkyl;
C3-C7 cycloalkyl
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by heterocyclyl, aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, NR7R8, or CF3; p is O or l ;
R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl, said aryl being optionally substituted by halogen; or Ci-C6 acyl; R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl in R or R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of hydroxyl; halogen; C1 -C6 alkyl; Ci- Ce alkyl substituted by NR 6R17; Ci-C6 alkoxy; Ci-C6 haloalkyl; Ci-C6 haloalkoxy; NR9R10; unsubstituted heterocyclyl; heterocyclyl substituted by Ci-C6 alkyl; phenyl; Ci-C6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; C-i- C-6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; phenyloxy, wherein the phenyl may be optionally substituted by halogen; or C-2- C6 alkoxy substituted by NR18R19 and the heterocyclyl in R and R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of d-C-6 alkyl or aryl, wherein the aryl is optionally substituted by halogen;
R9, R 0 are, independently, hydrogen or COR15;
R 5 is Ci-C6 alkyl substituted by NH2;
R 6, R 7, R 8, R 9 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by phenyl, wherein the phenyl may be optionally substituted by halogen; or taken together and with the nitrogen to which they are attached form a 5-6 member heterocycle, wherein the heterocycle may be optionally substituted by phenyl, wherein the phenyl may be optionally substituted by halogen.
4. A compound of formula (I) according to claim 1 or 2, or stereoisomers or pharmaceutically acceptable salts thereof wherein
R is Ci-C6 alkyl;
Ci-C6 alkyl substituted by aryl;
C-I -C6 alkyl substituted by heterocyclyl, wherein the heterocyclyl may be optionally substituted by C1-C6 alkyl;
Ci-C6 alkyl substituted by X-R3;
C3-C7-cycloalkyl; aryl, wherein the aryl may be optionally substituted by one or more substituents independently selected from the group consisting of halogen, Ci- C6 alkyl, Ci-C6 alkoxy; or Ci-C6 alkoxy substituted by phenyl, wherein the phenyl may be optionally substituted by halogen;
R2 is CH2OH, COR4, CN, or tetrazole; R3 is Ci-C6 alkyl;
Ci-C6 alkyl substituted by (CO)pNR5R6;
Ci-C6 alkyl substituted by aryl or heteroaryl; aryl or heteroaryl; X O or NH;
R4 is hydrogen, OH, 0-Ci-C6 alkyl, or NR7R8; p is O or l ;
R5, R6 are, independently, hydrogen; Ci-C6 alkyl; Ci-C6 alkyl substituted by aryl; or
Ci-Ce acyl; R7, R8 are, independently, hydrogen, OH, or Ci-C6 alkyl; wherein the aryl in R3 is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, d-C-6 alkyl, C-i-C-6 alkoxy, Ci-C6 haloalkyl, Ci-C6 haloalkoxy, NR9R10 or heterocyclyl substituted by Ci-C6 alkyl; and
R9, R 0 are, independently, hydrogen or COR15; R 5 is Ci-C6 alkyl substituted by NH2.
5. A compound of formula (I) according to anyone of claims 1 to 4, or stereoisomers or pharmaceutically acceptable salts thereof, wherein aryl is phenyl or naphthyl.
6. A compound according to claim 1 selected from:
methyl 2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(Z)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate; methyl 2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
methyl 2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate;
methyl 2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyndine-4- carboxylate;
methyl 2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate;
methyl 2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4- carboxylate;
te/t-butyl 2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-phenyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-cyclopropyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-ethyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(3-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-benzyl-A/-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[A/-hydroxy-C-[(4-methylpiperazin-1 -yl)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-(A/-hydroxy-C-(methoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-(A/-hydroxy-C-(2-methylaminoethoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-[C-[2-(benzylamino)ethoxymethyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-(A/-hydroxy-C-(phenoxymethyl)carbonimidoyl)pyridine-4-carboxylic acid;
2-(C-(ethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide;
A/-ethyl-2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carboxamide;
2-[(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyridine-4-carbohydroxamic acid; 1 - [4-(1 H-tetrazol-5-yl)-2-pyridyl]ethanone oxime;
1 -[4-(hydroxymethyl)-2-pyridyl]ethanone oxime;
2- [(E)-A/-hydroxy-C-methyl-carbonimidoyl]pyndine-4-carbonitrile;
methyl 2-(A/-hydroxy-C-(phenoxymethyl)carbonimidoyl)pyridine-4-carboxylate; methyl 2-[(E)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4 carboxylate;
methyl 2-[(Z)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4 carboxylate;
2-[(E)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-A/-hydroxy-C-(4-methoxyphenyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-(C-(benzyloxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid; 2-[C-[[3-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-(C-(2-acetamidoethoxymethyl)-A/-hydroxy-carbonimidoyl)pyridine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-[(2-methoxyphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-A/-hydroxy-C-[(3-methoxyphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-[(E)-C-(anilinomethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(E)-N-hydroxy-C-(methylaminomethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(2-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[[4-(3-aminopropanoylamino)phenoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(3-aminophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(4-aminophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid; 2-[(Z)-C-[(4-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(3-fluorophenoxy)methyl]-A/-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-pyndyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-(pyrimidin-2-yloxymethyl)carbonimidoyl]pyndine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(1 -naphthyloxymethyl)carbonimidoyl]pyndine-4-carboxylic acid;
2-[(Z)-A/-hydroxy-C-(2-naphthyloxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(E)-C-[4-[(4-fluorophenyl)methoxy]phenyl]-A/-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[2-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[3-(trifluoromethyl)phenoxy]methyl]carbonimidoyl]pyndine-4- carboxylic acid;
2-[(Z)-A/-hydroxy-C-[(4-methoxyphenoxy)methyl]carbonimidoyl]pyndine-4- carboxylic acid;
methyl 2-[(E)-A/-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-A/-hydroxy-C-phenethyl-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[3-(trifluoromethoxy)phenoxy]methyl]carbonimidoyl]pyndine- 4-carboxylic acid;
2-[(Z)-C-[2-[(4-fluorophenyl)methylamino]ethoxymethyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[[2-(dimethylamino)-2-oxo-ethoxy]methyl]-A/-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-N-hydroxy-C-(3-pyndylmethoxymethyl)carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-(2-dimethylaminoethyloxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid hydrochloride;
2-[(E)-C-[(4-fluoroanilino)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
2-[(Z)-N-hydroxy-C-[(4-morpholinophenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid hydrochloride;
2-[(Z)-C-[[3-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[[3-[(phenyl)methoxy]phenoxy]methyl]-N-hydroxy-carbonimidoyl]pyridine- 4-carboxylic acid;
2-[(Z)-N-hydroxy-C-[[4-(4-methylpiperazin-1 - yl)phenoxy]methyl]carbonimidoyl]pyridine-4-carboxylic acid 2,2,2-trifluoroacetic acid;
2-[(Z)-N-hydroxy-C-[[4-(1 -piperidyl)phenoxy]methyl]carbonimidoyl]pyridine-4- carboxylic acid hydrochloride;
2-[(Z)-C-[(4-benzyloxyphenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(3-chlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[(4-phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(3,4-dichlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(3,5-dichlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[(3-phenylphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid; 2-[(Z)-N-hydroxy-C-[(3-phenoxyphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[(4-bromophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[(4-phenylphenoxy)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-[[4-[(4-fluorophenyl)methoxy]phenoxy]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-[(4-chlorophenoxy)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-C-(cyclopentoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
2-[(Z)-C-(cyclobutoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; 2-[(Z)-C-(propoxymethyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-N-hydroxy-C-[(4-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-N-hydroxy-C-[(4-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-[[4-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylate;
2-[(E)-C-[[3-[(4-fluorophenyl)methoxy]phenyl]methyl]-N-hydroxy- carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-N-hydroxy-C-[(4-phenylphenyl)methyl]carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-N-hydroxy-C-[(4-phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3,5-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3,4-dichlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-benzyloxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-chlorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
ethyl 2-[(E)-N-hydroxy-C-[(3-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylate;
ethyl 2-[(E)-N-hydroxy-C-[(3-phenoxyphenyl)methyl]carbonimidoyl]pyridine-4- carboxylic acid;
ethyl 2-[(E)-C-[(3-phenylphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-N-hydroxy-C-[(3-phenylphenyl)methyl]carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-N-hydroxy-C-(1 -naphthylmethyl)carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-N-hydroxy-C-(2-naphthylmethyl)carbonimidoyl]pyridine-4-carboxylic acid; 2-[(E)-N-hydroxy-C-[[3-(4-methylpiperazin-1 - yl)phenyl]methyl]carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
ethyl 2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
1 -[4-(2,2,2-trifluoro-1 -hydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride;
1 - [4-(2,2,2-trifluoro-1 ,1 -dihydroxy-ethyl)-2-pyridyl]ethanone oxime hydrochloride;
2- [(E)-C-[[3-[2-[(4-fluorophenyl)methyl-methyl-amino]ethoxy]phenyl]methyl]-N- hydroxy-carbonimidoyl]pyridine-4-carboxylic acid hydrochloride;
methyl 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylate; 2-[(E)-C-(4-fluorophenyl)-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid; ethyl 2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-fluorophenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4-carboxylic acid;
Ethyl 2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(3-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
Ethyl 2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylate;
2-[(E)-C-[(4-methoxyphenyl)methyl]-N-hydroxy-carbonimidoyl]pyridine-4- carboxylic acid;
2-[(Z)-N-hydroxy-C-[2-(4-phenyl-1 -piperidyl)ethoxymethyl]carbonimidoyl]pyridine- 4-carboxylic acid hydrochloride
or stereoisomers or pharmaceutically acceptable salts thereof.
7. A compound of formula (I) as defined in any one of preceding claims for use as a medicament.
8. The compound as defined in any one of claims 1 to 6 being an inhibitor of KDM4C for medical use.
9. The compound for use according to claim 7 or 8, wherein said use is in the treatment and/or prevention of cancer.
10. The compound according to claim 9 for use in the treatment and/or prevention of leukemia, lymphoma, esophageal squamous cell carcinoma, breast cancer, medulloblastoma, prostate cancer, colon cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreas cancer or glioblastoma.
1 1 . The compound for use according to claim 10, wherein the leukemia is acute myeloid leukemia and the lymphoma is selected from the group consisting of: mucosa- associated lymphoid tissue lymphoma, primary mediastinal B cell lymphomas (PMBL), and Hodgkin lymphomas (HL).
12. The compound for use according to claim 10, wherein the glioblastoma is giant cell glioblastoma or gliosarcoma.
13. A pharmaceutical composition comprising the compound as defined in claim 1 to 6 together with a pharmaceutically acceptable excipient and/or diluent.
14. The pharmaceutical composition according to claim 13 further comprising at least one further therapeutic agent selected from the group consisting of histone deacetylase inhibitors, retinoid receptor modulators, anti-proliferative/antineoplastic agents, cytostatic agents, agents which inhibit cancer cell invasion, inhibitors of growth factor function, anti- angiogenic agents, cell cycle inhibitors, proteasome inhibitors, HSP90 inhibitors, selective COX-2 inhibitors or a chemotherapeutic agent.
15. The pharmaceutical composition according to any one of claim 13 or 14 for use in the treatment and/or prevention of cancer.
16. A process for obtaining a compound of formula (I) according to claim 1 or 2, wherein R2 is COOH, the process comprising the preparation of a compound of formula A3 by reaction of a compound of formula A1 with a compound of formula A2 in presence of an oxidant, a transition metal salt, and an acid, the preparation of a compound of formula A4 by reaction of a compound of formula A3 with hydroxylamine hydrochloride and a suitable base or hydroxylamine (NH2OH), and the final deprotection of a compound of formula A4 to obtain a compound of formula (I), as represented in Scheme A-IV:
Figure imgf000198_0001
Scheme A-IV
wherein R is as defined in claim 1 or 2 and PG is a protecting group, preferably PG is methyl or ethyl.
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