WO2023021498A1 - Analogues de nucléosides à base d'azole et de furane fusionnés et leurs utilisations - Google Patents

Analogues de nucléosides à base d'azole et de furane fusionnés et leurs utilisations Download PDF

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Publication number
WO2023021498A1
WO2023021498A1 PCT/IL2022/050668 IL2022050668W WO2023021498A1 WO 2023021498 A1 WO2023021498 A1 WO 2023021498A1 IL 2022050668 W IL2022050668 W IL 2022050668W WO 2023021498 A1 WO2023021498 A1 WO 2023021498A1
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alkyl
compound
optionally substituted
combination
aryl
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PCT/IL2022/050668
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English (en)
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Eyal BEN AMI
Orli EVEN-OR
Noa BALABAN
Joel VAN GELDER
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Moya Bio Ltd.
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered

Definitions

  • the present invention relates generally to the field of nucleoside analogs comprising fused heterocyclic rings (e.g. azole-based and/or furan-based rings) and is directed to methods of using the same such as for treating a disease responsive to inhibition of one or more El enzyme(s).
  • fused heterocyclic rings e.g. azole-based and/or furan-based rings
  • Ubiquitin is an 8.5-kDa, 76-amino acid polypeptide that serves as a post- translational modifier of cellular substrates.
  • Post-translational modifications of cellular substrates with ubiquitin or a family of closely related proteins known collectively as ubiquitin-like proteins (UBLs) play a key role in regulating many aspects of cell biology (such as cellular signaling cascades, protein degradation, intracellular localization, etc.).
  • UBLs share a common biochemical mechanism by which they covalently attach to a lysine residue on their target protein through formation of an isopeptide bond with the C-terminal glycine residue of the UBL.
  • UBLs In eukaryotes, there are more than a dozen UBLs, including: Ubiquitin, neural precursor cell-expressed developmentally downregulated protein 8 (NEDD8), small ubiquitin-like modifier (SUMO), human leukocyte antigen F-associated transcript 10 (FAT10, also known as ubiquitin D), interferon- stimulated gene (ISG) 15, autophagy-related protein (ATG) 8, ATG12, ubiquitin-fold modifier 1 (UFM1), and ubiquitin-related modifier 1 (URM1).
  • NEDD8 neural precursor cell-expressed developmentally downregulated protein 8
  • SUMO small ubiquitin-like modifier
  • FATG interferon- stimulated gene
  • UMG autophagy-related protein
  • UFM1 ubiquitin-fold modifier 1
  • URM1 ubiquitin-related modifier 1
  • the UBL conjugation cascade is initiated by a family of ATP-dependent enzymes termed El enzymes.
  • El enzymes In the human proteome, eight El enzymes are known to activate UBLs. These include ubiquitin activating enzyme (UAE), NEDD8-activating enzyme (NAE), SUMO-activating enzyme (SAE), Ubiquitin-like modifier-activating enzyme 6 (UBA6), UBA7, UBA4, UBA5 and autophagy-related enzyme 7 (ATG7).
  • UAE ubiquitin activating enzyme
  • NAE NEDD8-activating enzyme
  • SAE SUMO-activating enzyme
  • Ubiquitin-like modifier-activating enzyme 6 UBA7
  • UBA4 UBA5
  • ATG7 autophagy-related enzyme 7
  • the activated UBL is transferred to a catalytic cysteine residue in a separate domain within the enzyme, to form an El -UBL intermediate connected via a thioester bond.
  • the El -UBL intermediate transfers the UBL to an E2 enzyme by transthioesterification resulting in an E2 bound UBL.
  • the UBL is further transferred to the target protein, either directly or in conjugation with an E3 ligase enzyme.
  • Human cells contain more than 35 ubiquitin E2 enzymes and more than 500 E3 enzymes which mediate the selective targeting of specific cellular substrate proteins.
  • UBA6 is also the El for FAT10.
  • the cellular pathways mediated by UAE and UBA6 through ubiquitination and FATlOylation of target proteins are diverse. These include key pathways crucial for cellular physiology such as receptor internalization and lysosomal degradation, autophagy, signal transduction, regulation of transcription, DNA repair and maintenance of protein homeostasis. Many of these processes are important for cancer cell survival and metastasis. Multiple evidence gathered over the years support the notion that tumor cells may have increased sensitivity to UAE inhibition as a result of their rapid growth rate, increased metabolic demands and oncogene-fueled protein stress.
  • Maintaining protein homeostasis is critical for eukaryotic cell survival.
  • disruption of protein homeostasis using a small molecule is a validated therapeutic approach for the treatment of cancer.
  • Bortezomib (Velcade), Ixazomib (Ninlaro) and Carfilzomib (Kyprolis) are all drugs that disrupt protein homeostasis by inhibiting the proteasome, approved for the treatment of multiple myeloma and mantle cell lymphoma.
  • the ubiquitin system of trypanosomatids is essential for cell viability. It is involved in basic cellular processes, but also in trypanosomatid-specific processes such as the remodeling of the flagellum during differentiation and tolerance to ionizing radiation. Ubiquitin-mediated proteasome inhibition has been shown to kill trypanosomes in animal infection models and RNAi knockdown of UAE in the eukaryotic parasite Trypanosoma brucei has been shown to lead to severe growth defects in the parasite.
  • a compound or a salt thereof wherein the compound is represented by Formula 1:
  • each R, Rl, R2, X and Y is the same or different, and wherein at least one X comprises O, NH, or NR.
  • At least one X’ comprises O, NH, or NR.
  • the compound is represented by Formula 2: , wherein at least one X comprises O, N, NH, or
  • the compound is represented by Formula 2B: wherein at least one X and at least one X’ comprises O, NH, or NR; or by Formula 2B’ : wherein at least one X comprises O, NH, N, or
  • R2 is H, and wherein the protecting group is selected from hydroxy protecting group and amine protecting group.
  • the hydroxy protecting group comprises any of acetal (e.g. acetonide), tert-butyl, acyl, benzyl, O-nitrobenzyl, allyl, and silyl ether.
  • Y is H.
  • the compound is represented by Formula 3’ : , wherein each R3 is independently H or comprises an optionally substituted cyclyl; and wherein at least one X comprises O, NH, N, or NR.
  • the cyclyl comprises an aryl, a polycyclyl, a heteroaryl, a cycloalkyl, or heterocyclyl or any combinations thereof.
  • the substituted cyclyl comprises one or more substituents selected from halo, cyano, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, hydroxy, amino, mercapto, thioalkyl, aminoalkyl, haloalkoxy, haloalkyl and alkoxy, an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted polycyclyl, or any combination thereof.
  • the compound is represented by Formula 4’ : , wherein at least one R3 comprises an optionally substituted cyclyl.
  • n is 1, and at least one R is H.
  • the compound is represented by Formula 5’ : , p sents any of cycloalkyl, aryl, and heteroaryl, a fused aryl, a fused cycloalkyl or any combination thereof.
  • each X is independently CH or N, optionally wherein at least one X is N.
  • the compound comprises: , wherein A represents a cycloalkyl, aryl, heteroaryl, or is absent; wherein each R” independently represents a substituent or is absent, and wherein the substituent comprises any one of: alkoxy, haloalkoxy, halo, alkyl, haloalkyl, mercaptoalkyl, mercapto haloalkyl, a haloalkyl, or any combination thereof.
  • A is absent, and R” represents a substituent selected from alkoxy, haloalkoxy, alkyl, haloalkyl, a mercaptoalkyl, mercapto haloalkyl, a haloalkyl, or any combination thereof; optionally wherein the mercaptohaloalkyl comprises -SCF3.
  • composition comprising the compound of the invention, a pharmaceutically acceptable salt thereof or both; and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition comprises a therapeutically effective amount of the compound, of the pharmaceutically acceptable salt thereof or both. [029] In one embodiment, the pharmaceutical composition is for use as a medicament.
  • the pharmaceutical composition is for use in the inhibition of
  • the pharmaceutical composition is for use in the prevention or treatment of a disease responsive to El enzyme inhibition.
  • the disease is a proliferation disease, or any of autoimmune disorders and inflammation, or a eukaryotic parasitic disease.
  • the inhibition comprises selective inhibition of (i) UAE activity, (ii) UBA6 activity or both.
  • the disease or the disorder comprises a proliferative disease, an inflammatory disease, an autoimmune disease, a eukaryotic parasitic disease, or any combination thereof.
  • the proliferative disease is cancer.
  • Figure 1 represents the chemical structures of the tested compounds (abbreviated with i-01, i-02, i-03, i-04).
  • Figures 2A-2C are bar graphs representing upregulation of the expression of ER- stress related genes by the exemplary compounds of the invention: i-03 in A549 (2A) and HCT-116 (2B) cancer cell lines; and of i-02 in HCT-116 cell-line (2C).
  • Figure 3 is an image representing inhibition of E2-UBL adduct formation by the exemplary compound of the invention (i-03).
  • Figures 4A-4C are images representing the effect of i-03 in inducing accumulation of IkBa in A549 (4A) and HCT-116 (4B) cancer cell lines; and the effect of i-02 in HCT- 116 cell-line (4C).
  • Figure 5 represents the chemical structures of the R group for the tested furan- based compounds of the invention (abbreviated with F-01, F-02, F-03, F-04, F-05, F-06, F- [044]
  • Figure 6 are images representing the accumulation of ubiquitin substrates in HCT- 116 cancer cell line following El inhibition by F-01.
  • the compound of the invention is represented by Formula 1, wherein W, R, Rl, R2, n and Y are as described hereinabove; Xa’ is C or CH; each X’, and Xb’ is independently selected from CH2, CH, C(R)2, O, S, NH, and NR including any combination thereof as allowed by valency, wherein at least one of X’, and Xb’ is selected from O, S, NH, and NR as allowed by valency; and wherein each X is independently selected from CH, CR, C(R)2, O, NH, and NR including any combination thereof as allowed by valency.
  • the compound of the invention is as described hereinabove, wherein at least one X is or comprises O, S, NH, or NR as allowed by valency.
  • each X’ and Xb’ is independently selected from CH2, CH, C(R)2, O, and S, as allowed by valency, optionally, wherein at least one of X’ and Xb’ is or comprises a heteroatom.
  • the compound of the invention is represented by Formula: described herein; and wherein W’ represents O, S, NH, or NR.
  • W’ comprises a heteroatom (e.g. W is O, N, or NH).
  • W’ is N or NH.
  • at least one X and/or at least one X’ is or comprises a heteroatom (such as O, S, NH, or NR).
  • R is or comprises an optionally substituted cyclyl.
  • the compound of the invention is represented by Formula: described herein; and wherein W’ represents O, S, NH, or NR.
  • W’ comprises a heteroatom (e.g. W is O, N, or NH).
  • W’ is N or NH.
  • at least one X and/or at least one X’ is or comprises a heteroatom (such as O, S, NH, or NR).
  • R is or comprises an optionally substituted cyclyl.
  • the compound of the invention is represented by Formula: described herein;
  • the compound is represented by Formula 2B or by Formula 2B’, wherein at least one X comprises O, NH, N, or NR.
  • n is any of 0, 1 and 2.
  • each Y independently is or comprises H.
  • each Y independently represents a protecting group, selected from amine protecting group, and hydroxy protecting group.
  • amine protecting groups include but are not limited to: 9-fluorenylmethyloxycarbonyl (Fmoc), Alloc, Dde, iv-Dde, benzyl, benzyloxycarbonyl, tert-butyloxycarbonyl (Boc) and 2- [biphenylyl-(4)]-propyl-2-oxycarbonyl, dimethyl-3,5dimethoxybenzyloxycarbonyl, 2-(4- Nitrophenylsulfonyl)ethoxycarbonyl, 1,1 -Dioxobenzo [b]thiophene-2- ylmethyloxycarbonyl, 2,7-Di-tert-butyl-Fmoc, 2-Fluoro-Fmoc, Nitrobenzenesulfonyl,
  • Non-limiting examples of hydroxy protecting groups include but are not limited to: a silyl ether (or silyl) such as triisopropylsilyl ether (TIPS), tert-Butyldimethyl silyl ether (TBDMS); an acetal (e.g.
  • the compound is represented by Formula 3 ’ : , wherein R, n, and R2 are as described hereinabove, and wherein each R3 is independently H or comprises an optionally substituted cyclyl; and wherein at least one X comprises O, NH, N, or NR. In some embodiments, R3 comprises an optionally substituted cyclyl or is absent.
  • R3 represents hydrogen. In some embodiments, R3 represents a substituent as defined for R and as described herein. In some embodiments, R3 represents a substituted cyclyl.
  • cyclyl comprises an aryl, a polycyclyl, a heteroaryl, a cycloalkyl, or heterocyclyl or any combinations thereof.
  • polycyclic ring or “polycyclyl” encompasses a plurality (e.g. 2, 3, 4, 5 or 6) of fused or adjacent rings (e.g. biaryl or bicyclohexyl), wherein each ring is independently selected from aryl, heteroaryl, an optionally unsaturated cycloalkyl, an optionally unsaturated heterocyclyl, or any combination thereof.
  • polycyclyl encompasses a polycyclic aromatic ring, a polycyclic aliphatic ring, or a mixed polycyclic ring.
  • the term “mixed polycyclic ring” refers to any plurality of rings covalently bound to each other (e.g. fused rings, dicylyls, spirocyclic rings etc.) comprising at least one aromatic ring (aryl, or heteroaryl) and at least one aliphatic or nonaromatic ring (optionally a heterocyclyl and/or unsaturated cyclyl).
  • R3 represents a substituted cyclyl comprising one or more substituents, wherein each substituent independently is selected from the group as defined for R.
  • R3 is or comprises an optionally substituted aryl (e.g. C6- aryl), an optionally substituted heteroaryl (e.g. C2-C5 heteroaryl), an optionally substituted cycloalkyl (e.g. C3-C10 aliphatic or unsaturated cycloalkyl), an optionally substituted heterocyclyl, an optionally substituted polycyclic ring or a combination thereof.
  • the polycyclic ring comprises a bicyclic ring.
  • the term “bicyclic ring” encompasses a fused ring (fused aromatic and/or heteroaromatic ring), spirocyclic ring, a bridged ring, a dicyclyl (two aromatic and/or aliphatic rings joined by a single carbon-carbon bond).
  • fused ring fused aromatic and/or heteroaromatic ring
  • spirocyclic ring a bridged ring
  • dicyclyl two aromatic and/or aliphatic rings joined by a single carbon-carbon bond
  • R3 represents hydrogen, or a substituent comprising aryl (e.g. phenyl, naphthyl), optionally substituted; or 5- 6-membered heteroaryl (e.g. pyrrole, furan, thiophene, thiazole, pyrazole, isothiazole, imidazole, pyridine, pyrimidine, etc.), or a fused heterocyclic ring (e.g. indole, isoindole, benzofuran, benzothiophene, benzotriazole, quinoline, chromene, chroman, quinazoline), optionally substituted.
  • substituted encompasses substitution by one or more R, wherein R is as described hereinabove.
  • R3 comprises 7-10 bicyclic ring optionally substituted with one or more R, wherein the bicyclic ring optionally comprises 1 or 2 heteroatoms (e.g. O, S or N or a combination thereof).
  • R3 comprises a fused 7-10 aliphatic ring optionally substituted with one or more R.
  • fused 7-10 aliphatic ring comprises norbornane, bicyclooctane, bicyclodecane or a combination thereof.
  • R3 is or comprises a heterocyclyl (e.g. pyrrolidine, oxirane, tetrahydrofuran, aziridine, pyran, dioxane, thiolane, oxathiolane, piperidine, and/or morpholine) optionally substituted with one or more R.
  • R3 is or comprises a fused aryl-heteroaryl, or a fused aryl-heterocyclyl.
  • R3 comprises a cyclyl selected from phenyl, pyridine, pyrazole, pyrimidine, indole, naphthyl, oxazole, thiazole, benzofuran and imidazole or any combination thereof; wherein each cyclyl is optionally substituted by one or more R”.
  • each R is independently selected from halo, cyano, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl (e.g. acetylenyl), hydroxy, amino, mercapto, thioalkyl, aminoalkyl, haloalkoxy, haloalkyl and alkoxy, an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted polycyclyl, or any combination thereof.
  • each R represents one or more substituents each independently selected from halo, cyano, Cl- C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, hydroxy, amino, mercapto, thioalkyl, aminoalkyl, haloalkoxy, haloalkyl and alkoxy, an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted polycyclyl, or any combination thereof.
  • the compound is represented by Formula 4’ : wherein R, n, X, and R2 are as described hereinabove, and wherein each at least one R3 comprises an optionally substituted cyclyl.
  • R represents H or one or more substituents as described herein, and wherein if R represents a plurality of substituents, the substituents are the same or different.
  • at least one X is or comprises a heteroatom, and at least one R is or comprises one or more H.
  • the compound is represented by Formula: , wherein each X is independently N or CR, and wherein R3, n, and R2 are as described hereinabove. In some embodiments, at least one X is N.
  • the compound is represented by Formula 5’ : or by Formula 5 A’ : , wherein A represents a cyclyl or is absent, and wherein
  • X is as described herein (e.g., optionally at least one X is N), and wherein R represents one or more substituents, wherein each substituent independently is as described herein.
  • A represents any of cycloalkyl, aryl, heteroaryl, a bicyclyl or a polycyclyl, or any combination thereof.
  • the compound is represented by Formula 5A’, wherein A is absent, or comprises a cycloalkyl, an aryl, a heteroaryl, a fused aryl, a fused cycloalkyl or any combination thereof.
  • the compound is represented by Formula:
  • R independently represents one or more substituents or is absent, and wherein the substituent is as described herein.
  • each independently R” is H or represents a substituent comprising any one of -NO2, -CN, -OH, -NH2, carbonyl, -CONH 2 , -CONR’ 2 , -NHCOR’, -SO 2 R’, -SOR’, -SR’, -SO 2 OR”, -SO 2 N(R’) 2 , -
  • A is absent or represents a cycloalkyl, aryl, heteroaryl, or any combination thereof.
  • A is absent or comprises phenyl, indole, benzofuran, pyridine, pyrimidine, naphthyl, oxazole, pyrazole, indazole, thiazole, biphenyl benzimidazole, and imidazole or any combination thereof.
  • A is absent.
  • each R independently represents one or more substituent or is absent, wherein one or more substituent independently comprises any one of: alkoxy, haloalkoxy, halo, alkyl (linear or cyclic), heterocyclyl, haloalkyl, mercaptoalkyl, mercapto haloalkyl, or any combination thereof.
  • each R independently represents one or more substituent or is absent, wherein one or more substituent independently comprises any one of C 1 -C 10 alkyl, C 1 -C 10 alkoxy, C 1 -C 10 haloalkoxy, Ci- C10 haloalkyl, C 1 -C 10 mercaptoalkyl, C 1 -C 10 mercapto haloalkyl, 5-6 membered cycloalkyl, 5-6 membered heterocyclyl, or any combination thereof.
  • A is absent, and R” represents a substituent selected from C 1 -C 10 alkyl, C 1 -C 10 alkoxy (e.g. -OMe), C 1 -C 10 haloalkoxy (e.g., -OCF2, -OCF3), C 1 -C 10 haloalkyl (e.g., -CF3), C 1 -C 10 mercaptoalkyl, C 1 -C 10 mercapto haloalkyl, 5-6 membered heterocyclyl (e.g. morpholine) or any combination thereof.
  • R represents a substituent selected from C 1 -C 10 alkyl, C 1 -C 10 alkoxy (e.g. -OMe), C 1 -C 10 haloalkoxy (e.g., -OCF2, -OCF3), C 1 -C 10 haloalkyl (e.g., -CF3), C 1 -C 10 mercaptoalkyl
  • C 1 -C 10 haloalkoxy comprises mono-, di-, or tri-halogenated (e.g. fluorinated ) haloalkoxy.
  • C 1 -C 10 haloalkyl comprises mono-, di-, or tri-halogenated (e.g. fluorinated ) haloalkyl.
  • C 1 -C 10 mercaptohaloalkyl comprises mono-, di-, or tri- halogenated (e.g. fluorinated ) mercaptohaloalkyl.
  • C 1 -C 10 mercaptohaloalkyl comprises -SCF3.
  • the compound is or comprises: , wherein R” represents a substituent selected from Ci-
  • R is at at ortho-, and/or at meta-position.
  • the compound of the invention is represented by Formula: are as described hereinabove; wherein each X’, Xa’ and Xb’ is independently selected from CH2, CH, C(R)2, NH, and NR including any combination thereof, wherein at least one of X’, Xa’ and Xb’ is selected from NH, and NR; and if Xa’ and Xb’ are N, then X’ is NH, or NR; and wherein each X is independently selected from CH, CR, C(R)2, O, NH, and NR including any combination thereof.
  • one or more X independently comprises O, NH, or NR, as allowed by valency.
  • At least one of X’, Xa’ and Xb’ is N.
  • (i) Xa’ and X’ are N, and Xb’ is selected from CH, and C as allowed by valency; or
  • (ii) Xb’ and X’ are N or NH, and Xa’ is C.
  • X’ is N or NH as allowed by valency; and Xa’ and Xb’ are selected from CH, and C as allowed by valency.
  • n is any of 0, 1 and 2.
  • the compound of the invention is represented by Formula 1A: described herein, XI is selected from O and N-Y; and wherein W’ represents any of CFh, CH, C(R) 2 , O, NH, and NR.
  • W’ and W are the same group.
  • W’ comprises a heteroatom (e.g. O, N, or NH).
  • W’ is
  • the compound of the invention is represented by Formula
  • the compound of the invention is represented by any one of Formulae 1-2 A, wherein R2 is H.
  • each Y is or comprises H.
  • Y is a protecting group, selected from amine protecting group, and hydroxy protecting group.
  • amine protecting groups include but are not limited to: 9- fluorenylmethyloxycarbonyl (Fmoc), Alloc, Dde, iv-Dde, benzyl, benzyloxycarbonyl, tertbutyloxycarbonyl (Boc) and 2-[biphenylyl-(4)]-propyl-2-oxycarbonyl, dimethyl- 3 ,5dimethoxybenzyloxycarbonyl, 2-(4-Nitrophenylsulfonyl)ethoxycarbonyl, 1,1-
  • Non-limiting examples of hydroxy protecting groups include but are not limited to: a silyl ether (or silyl) such as triisopropylsilyl ether (TIPS), tert-Butyldimethyl silyl ether (TBDMS); an acetal (e.g.
  • the compound is represented by Formula 3: , wherein W, R, Rl, R2, X, n and Y are as described herein.
  • R3 comprises an optionally substituted cyclyl or is absent.
  • R3 represents hydrogen.
  • R3 is a substituent as defined for R and as described herein.
  • cyclyl comprises an aryl, a polycyclyl, a heteroaryl, a cycloalkyl, or heterocyclyl or any combinations thereof.
  • polycyclic ring or “polycyclyl” encompasses a plurality (e.g. 2, 3, 4, 5 or 6) of fused or adjacent rings (e.g. biaryl or bicyclohexyl), wherein each ring is independently selected from aryl, heteroaryl, an optionally unsaturated cycloalkyl, an optionally unsaturated heterocyclyl, or any combination thereof.
  • polycyclyl encompasses a polycyclic aromatic ring, a polycyclic aliphatic ring, or a mixed polycyclic ring.
  • the term “mixed polycyclic ring” refers to any plurality of rings covalently bound to each other (e.g. fused rings, dicylyls, spirocyclic rings etc.) comprising at least one aromatic ring (aryl, or heteroaryl) and at least one aliphatic or nonaromatic ring (optionally a heterocyclyl and/or unsaturated cyclyl).
  • R3 represents a substituted cyclyl comprising one or more substituents, wherein each substituent independently is selected from the group as defined for R.
  • R3 is or comprises an optionally substituted aryl (e.g. C6- aryl), an optionally substituted heteroaryl (e.g. C2-C5 heteroaryl), an optionally substituted cycloalkyl (e.g. C3-C10 aliphatic or unsaturated cycloalkyl), an optionally substituted heterocyclyl, an optionally substituted polycyclic ring or a combination thereof.
  • aryl e.g. C6- aryl
  • heteroaryl e.g. C2-C5 heteroaryl
  • cycloalkyl e.g. C3-C10 aliphatic or unsaturated cycloalkyl
  • heterocyclyl e.g. C3-C10 aliphatic or unsaturated cycloalkyl
  • the polycyclic ring comprises a bicyclic ring.
  • the term “bicyclic ring” encompasses a fused ring (fused aromatic and/or heteroaromatic ring), spirocyclic ring, a bridged ring, a dicylyl (two aromatic and/or aliphatic rings joined by a single carbon-carbon bond).
  • fused ring fused aromatic and/or heteroaromatic ring
  • spirocyclic ring a bridged ring
  • a dicylyl two aromatic and/or aliphatic rings joined by a single carbon-carbon bond
  • R3 represents hydrogen, or a substituent comprising aryl (e.g. phenyl, naphthyl), optionally substituted; or 5- 6-membered heteroaryl (e.g. pyrrole, furan, thiophene, thiazole, pyrazole, isothiazole, imidazole, pyridine, pyrimidine, etc.), or a fused heterocyclic ring (e.g. indole, isoindole, benzofuran, benzothiophene, benzotriazole, quinoline, chromene, chroman, quinazoline), optionally substituted.
  • aryl e.g. phenyl, naphthyl
  • 5- 6-membered heteroaryl e.g. pyrrole, furan, thiophene, thiazole, pyrazole, isothiazole, imidazole, pyridine, pyrimidine, etc.
  • R3 comprises 7-10 bicyclic ring optionally substituted with one or more R, wherein the bicyclic ring optionally comprises 1 or 2 heteroatoms (e.g. O, S or N or a combination thereof).
  • R3 comprises a fused 7-10 aliphatic ring optionally substituted with one or more R.
  • fused 7-10 aliphatic ring comprises norbornane, bicyclooctane, bicyclodecane or a combination thereof.
  • R3 is or comprises a heterocyclyl (e.g. pyrrolidine, oxirane, tetrahydrofuran, aziridine, pyran, dioxane, thiolane, oxathiolane, piperidine, and/or morpholine) optionally substituted with one or more R.
  • R3 is or comprises a fused aryl-heteroaryl, or a fused aryl-heterocyclyl.
  • R3 comprises a cyclyl optionally substituted by one or more R”.
  • R3 comprises a cyclyl selected from phenyl, pyridine, pyrazole, pyrimidine, indole, naphthyl, oxazole, thiazole, benzofuran and imidazole or any combination thereof; wherein each cyclyl is optionally substituted by one or more R”.
  • each R” is independently selected from halo (e.g. Br, Cl, etc), cyano, C1-C6 alkyl (linear or branched), C1-C6 alkenyl (linear or branched), C1-C6 alkynyl (linear or branched e.g. acetylenyl), C1-C6 haloalkyl (e.g.
  • the compound is represented by Formula 4: , wherein R, R2, X, n and R3 are as described herein.
  • n is 1; and R2 and at least one R are both H.
  • at least one X is N. In some embodiments, all X are CH or CH2.
  • the compound of Formula 4 includes any stereoisomer thereof (e.g. enantiomer or diastereomer).
  • the compound of the invention is represented by Formula any of cycloalkyl, aryl, heteroaryl, a bicyclyl or a polycyclyl, or any combination thereof.
  • the compound of the invention is represented by Formula 5A: , wherein A, R”, and X are as described herein, optionally wherein one X is N and another X is CH.
  • A is or comprises phenyl, indole, benzofuran, pyridine, pyrimidine, naphthyl, oxazole, pyrazole, indazole, thiazole, biphenyl benzimidazole, and imidazole or any combination thereof.
  • A is or comprises phenyl, indole, benzofuran, pyridine, pyrimidine, naphthyl, or oxazole.
  • A is or comprises phenyl pyridine, phenyl pyrimidine, phenyl oxazole; optionally having a variable attachment to triazole via phenyl or via a heteroaryl.
  • R is H.
  • each R” comprises one or more substituents each independently selected from halo, haloalkyl, C1-C6 alkyl, C1-C6 alkynyl (e.g. acetylenyl), thioalkyl, thiohaloalkyl (e.g. -SCF3), haloalkoxy (e.g. -OCF2H, - OCFH2, -OCF3), alkoxy (e.g.
  • -OMe, -OEt cyano, hydroxy, amino, mercapto, aminoalkyl, an optionally substituted aryl, an optionally substituted heterocyclyl (e.g. morpholine, pyrrolidine, etc.) or any combination thereof.
  • the compound is selected from the group consisting of:
  • the compound of the invention is represented by Formula
  • each X is N or CR, and one of X is N;
  • A represents an aryl, or a heteroaryl (e.g. a 5-6 membered heteroaryl or a fused heteroaryl such as indole, isoindole, benzofuran, benzothiophene, benzotriazole, quinoline, chromene, chroman, quinazoline, etc.), and wherein R” represents one or more substituents, each substituent is indepedntly H or is selected from -NO2, -CN, -OH, -NH2, carbonyl, -CONH2, -CONR’ 2 , -NHCOR’, -SO 2 R’, -SOR’, -SR’, -SO2OR”, -SO 2 N(R’) 2 , -
  • the compound of the invention is represented by Formula , wherein each R” and R1 is independently H or a substituent as described herein.
  • at least one R’ ’ comprises halo, a mercaptoalkyl (e.g. C 1 -C 6 mercaptoalkyl), a mercaptohaloalkyl (e.g. Ci-Ce mercaptohaloalkyl), a haloalkyl (e.g. C 1 -C 6 haloalkyl), alkoxy(e.g. C 1 -C 6 alkoxy), haloalkoxy (e.g. C 1 -C 6 haloalkoxy), or a combination thereof.
  • R1 is or comprises halo.
  • the compound of the invention is as presented above, wherein R’ ’ comprises a mercaptoalkyl, a haloalkyl, optionally wherein the mercaptoalkyl comprises -SCF3.
  • the term “one or more” refers to any numerical value selected form of 1, 2, 3, 4, 5, or 6.
  • the heteroatom comprises any of N, O, NH, or S.
  • the compounds described herein are chiral compounds (i.e. possess an asymmetric carbon atom). In some embodiments, diastereomers, geometric isomers and individual isomers are encompassed within the scope of the present invention. In some embodiments, a chiral compound described herein is in form of a racemic mixture. In some embodiments, a chiral compound is in form of a single enantiomer, with an asymmetric carbon atom having the R configuration. In some embodiments, a chiral compound is in form of a single enantiomer, with an asymmetric carbon atom having the S configuration as described hereinabove.
  • a chiral compound is in form of a single enantiomer with enantiomeric purity of more than 70%. In some embodiments, a chiral compound is in form of a single enantiomer with enantiomeric purity of more than 80%. In some embodiments, a chiral compound is in form of a single enantiomer with enantiomeric purity of more than 90%. In some embodiments, a chiral compound is in form of a single enantiomer with enantiomeric purity of more than 95%.
  • the compound of the invention comprising an unsaturated bond is in a form of a trans-, or cis-isomer.
  • the composition of the invention comprises a mixture of cis- and trans-isomers, as described hereinabove.
  • the compounds described herein can exist in unsolvated form as well as in solvated form, including hydrated form.
  • the solvated form is equivalent to the unsolvated form and is encompassed within the scope of the present invention.
  • Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.
  • solvate refers to a complex of variable stoichiometry (e.g., di-, tri-, tetra-, penta-, hexa-, and so on), which is formed by a solute (the conjugate described herein) and a solvent, whereby the solvent does not interfere with the biological activity of the solute.
  • Suitable solvents include, for example, ethanol, acetic acid and the like.
  • hydrate refers to a solvate, as defined hereinabove, where the solvent is water.
  • structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, geometric, conformational, and rotational) forms of the structure.
  • isomeric e.g., enantiomeric, diastereomeric, geometric, conformational, and rotational
  • the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers are included in this invention.
  • a substituent can freely rotate around any rotatable bonds. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, geometric, conformational, and rotational mixtures of the present compounds are within the scope of the invention.
  • structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a hydrogen by 18 F, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as imaging probes.
  • a pharmaceutical composition comprising the compound of the present invention (including any salt or derivative thereof) and a pharmaceutically acceptable carrier.
  • the salt of the compound is a pharmaceutically acceptable salt.
  • the pharmaceutical composition comprises a therapeutically effective amount of the compound of the invention, and/or of the pharmaceutically acceptable salt thereof.
  • the pharmaceutical composition is for use in the inhibition of El enzyme activity.
  • the pharmaceutical composition is for use in the prevention or treatment of a disease, a disorder, and/or a condition responsive to El enzyme inhibition.
  • the El enzyme comprises UAE and/or UBA6.
  • UBA6 As used herein, the terms “UBA1” and “UAE” are used herein interchangeably.
  • the pharmaceutical composition is for use in the prevention or treatment of a disease, a disorder, and/or a condition responsive to inhibition of enzymatic activity of UAE and/or UBA6.
  • inhibition comprises selective inhibition of UAE and/or UBA6 enzymatic activity.
  • the disease or the disorder comprises a proliferation disease. In some embodiments, the disease or the disorder comprises inflammation. In some embodiments, the disease or the disorder comprises an autoimmune disorder. In some embodiments, the disease or the disorder comprises an infectious disease caused by one or more eukaryotic parasite.
  • Non-limiting examples of parasitic diseases include but are not limited to: trypanosomiasis (also known as African sleeping sickness), leishmaniasis, toxoplasmosis, Chagas disease, toxocariasis, cysticercosis, giardiasis and trichomoniasis, or any combination thereof.
  • the composition is a pharmaceutical composition or a kit (e.g. a kit of parts), comprising the compound of the invention (and/or any pharmaceutically acceptable salt, or any derivative thereof) and a pharmaceutically acceptable carrier.
  • the compound or the composition of the present invention selectively inhibits UAE and/or UBA6 enzymatic activity.
  • the compound or the composition of the present invention has at least 2 times, at least 5 times, at least 10 times, at least 15 times, at least 20 times, at least 30 times, at least 30 times, at least 50 times, at least 80 times, at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 700 times, at least 1000 times, at least 10,000 times, at least 50,000 times, at least 100,000 times lower IC50 for the UAE and/or UBA6 as compared to other El enzyme (e.g. NAE, or SAE).
  • the compound is substantially devoid of inhibitory activity with respect to El enzymes which are not UAE or UBA6 (e.g. NAE, or SAE).
  • compounds of the invention inhibit 50 % of UAE and/or UBA6 activity at a concentration (e.g. IC50) of less than 10000 nM, less than 5000 nM, less than 2000 nM, less than 1000 nM, less than 200 nM, less than 500 nM, less than 50 nM, less than 10 nM, including any range between.
  • the present invention provides a compound or a composition (e.g. a pharmaceutical composition) as described herein, for use in the treatment of disease, a disorder, and/or a condition responsive to inhibition of UAE and/or UBA6 activity.
  • the disease, the disorder, and/or the condition responsive to inhibition of UAE and/or UBA6 is selected from a proliferation disease, an autoimmune disorder and inflammation, a parasitic disease, or any combination thereof.
  • the compound of the invention includes any salt, any solvate, any hydrate, any stereoisomer, any isotope (e.g. a deuterated compound), and/or any derivative (e.g. a biologically active derivative) of any of the compounds or of the Formulae disclosed herein.
  • n is any of 0, 1 and 2.
  • each W, R, Rl, R2, X and Y is the same or different.
  • each of R and R2 is represents one or more substituents or hydrogen(s), as described herein.
  • the term “one or more” and the term “a” encompasses a singular or a plural (e.g. 1, 2, 3, 4, 5 or 6) substituents.
  • any one of the variables disclosed herein (such as X, R, Y, W, including any variants or combinations thereof) optionally represent one or more hydrogens.
  • the present invention provides a method for preventing or treating a disease or a disorder responsive to inhibition of: (i) UAE activity, (ii) UBA6 activity or both in a subject, comprising administering to the subject a therapeutically effective amount of the compound (e.g. an inhibitor) or the pharmaceutical composition described herein.
  • administering is by an oral administration, a systemic administration, or a combination thereof.
  • administration routes are as described hereinbelow.
  • the method is for preventing or treating a disease or disorder responsive to inhibition of UAE activity. In some embodiments, the method is for preventing or treating a disease or disorder responsive to inhibition of UBA6 activity.
  • UAE activity as used herein, refers to ubiquitination of a target protein, catalyzed by UAE. In some embodiments, UBA6 activity refers to ubiquitination and/or FATlOylation of the target protein, catalyzed by UBA6.
  • inhibition refers to an inhibition of UAE and/or UBA6 enzymatic activity within at least one cell and/or tissue of the subject, wherein inhibition is by at least 10%, at least 20%, at least 50%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, compared to an initial enzymatic activity (e.g. enzymatic activity within a cell/tissue without administration of the compound of the invention).
  • an initial enzymatic activity e.g. enzymatic activity within a cell/tissue without administration of the compound of the invention.
  • the disease or the disorder is selected from a proliferation disease, an autoimmune disorder, inflammation, and a parasitic disease, or any combination thereof.
  • administering comprises any of oral administration, topical administration, systemic administration, or any combination thereof.
  • the present invention provides a method for treating or ameliorating the effects of cancer, inflammation and/or of an autoimmune disease, a parasitic disease, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound and/or the pharmaceutical composition disclosed herein.
  • the subject is a mammal.
  • the mammal is selected from the group consisting of humans, veterinary animals, and agricultural animals.
  • the subject is a human.
  • the subject is afflicted with a disease or a disorder responsive to inhibition of: (i) UAE activity, (ii) UBA6 activity or both.
  • the subject is afflicted with a proliferative disease (e.g. cancer). In some embodiments, the subject is afflicted with a UAE and/or UBA6-related proliferative disease.
  • a proliferative disease e.g. cancer
  • the subject is afflicted with a UAE and/or UBA6-related proliferative disease.
  • Non-limiting examples of cancer comprise but are not limited to: colon cancer, glioblastoma, breast cancer, ovarian cancer, lung cancer, pancreatic cancer, Melanoma, Prostate cancer, Neuroblastoma, Mantle Cell Lymphoma, Diffuse large B-cell lymphoma, multiple myeloma, T Cell Lymphoma or any combination thereof.
  • the method of the invention in some embodiments thereof, may also encompass ameliorating or treating of additional cancer types which are well-known in the art.
  • the subject is afflicted with an inflammatory disease. In some embodiments, the subject is afflicted with a UAE and/or UBA6-related inflammatory disease.
  • the subject is afflicted with an autoimmune disease. In some embodiments, the subject is afflicted with a UAE and/or UBA6-related autoimmune disease.
  • the subject is afflicted with a parasitic disease. In some embodiments, the subject is afflicted with a UAE and/or UBA6-related parasitic disease. [0138] In some embodiments, the method further comprises co-administering to the subject an effective amount of one or more additional therapeutic agents.
  • the pharmaceutical composition of the present invention is administered in a therapeutically safe and effective amount.
  • safe and effective amount refers to the quantity of a component which is sufficient to yield a desired therapeutic response without undue adverse side effects, including but not limited to toxicity, such as calcemic toxicity, irritation, or allergic response, commensurate with a reasonable benefit/risk ratio when used in the presently described manner.
  • the actual amount administered, and the rate and time-course of administration, will depend on the nature and severity of the condition being treated. Prescription of treatment, e.g.
  • toxicity and therapeutic efficacy of the active ingredients described herein can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals.
  • the data obtained from these in vitro and cell culture assays and animal studies can be used in formulating a range of dosage for use in human.
  • the dosages may vary depending on the dosage form employed and the route of administration utilized.
  • the exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. [See e.g., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 13th Ed., McGraw-Hill/Education, New York, NY (2017)].
  • the effective amount or dose of the active ingredient can be estimated initially from in vitro assays.
  • a dose can be formulated in animal models and such information can be used to more accurately determine useful doses in humans.
  • the effective amount or dose of the active ingredient can be estimated by performing a diagnostic method (e.g. a detectable probe-based imaging).
  • a pharmaceutical composition comprising the compound of the invention, a pharmaceutically acceptable salt thereof or both.
  • the pharmaceutical composition of the invention comprises a therapeutically effective amount of the compound of the invention and/or any pharmaceutically acceptable salt and/or derivative thereof.
  • therapeutically effective amount is sufficient for reduction of at least one symptom, or for substantial reduction in the severity and/or inhibition of the progression of a disease, disorder, or condition as described hereinabove.
  • the therapeutically effective amount can be determined as described hereinabove.
  • compositions comprising one or more compounds of the invention, including any salt (e.g. a pharmaceutically acceptable salt), any tautomer, and/or any stereoisomer thereof.
  • a composition comprising one or more of the compounds disclosed herein individually or in combination, and a pharmaceutically acceptable carrier, adjuvant or vehicle.
  • the compound as described hereinabove is the only active ingredient within the composition of the invention (e.g. pharmaceutical composition).
  • the composition of the invention is a pharmaceutical composition comprising at least one compound of the invention and a pharmaceutically acceptable carrier. In some embodiments, the composition of the invention is a pharmaceutical composition comprising at least one compound of the invention as a first active ingredient and an additional active ingredient.
  • the pharmaceutical composition comprises one or more compounds of the invention (including any pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier, wherein the compound of the invention is a pharmaceutically pure compound.
  • the pharmaceutical composition comprises a therapeutically effective amount of the compound of the invention and the pharmaceutically acceptable carrier.
  • the pharmaceutical composition is in a form of a combination or of a kit of parts.
  • the pharmaceutical composition of the invention is for use as a medicament.
  • the term "pharmaceutically acceptable” can mean approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
  • the compound of the invention is referred to herein as an active ingredient of a pharmaceutical composition.
  • the pharmaceutical composition as described herein is a topical composition.
  • the pharmaceutical composition is an oral composition.
  • the pharmaceutical composition is an injectable composition.
  • the pharmaceutical composition is for a systemic use.
  • the term "carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the active ingredient is administered.
  • Such carriers can be sterile liquids, such as water-based and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents.
  • carriers include, but are not limited to: terpenes derived from Cannabis, or total terpene extract from Cannabis plants, terpenes from coffee or cocoa, mint-extract, eucalyptus-extract, citrus-extract, tobacco-extract, anis-extract, any vegetable oil, peppermint oil, d-limonene, P-myrcene, a-pinene, linalool, anethole, a- bisabolol, camphor, P-caryophyllene and caryophyllene oxide, 1,8-cineole, citral, citronella, delta-3-carene, farnesol, geraniol, indomethacin, isopulegol, linalool, unalyl acetate, b-myrcene, myrcenol, 1-menthol, menthone, menthol and neomenthol,
  • the carrier improves the stability of the active ingredient in a living organism. In some embodiments, the carrier improves the stability of the active ingredient within the pharmaceutical composition. In some embodiments, the carrier enhances the bioavailability of the active ingredient.
  • Water may be used as a carrier such as when the active ingredient has a sufficient aqueous solubility, so as to be administered intravenously.
  • Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions.
  • the carrier is a liquid carrier. In some embodiments, the carrier is an aqueous carrier.
  • Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like.
  • the composition if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates.
  • Antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; and agents for the adjustment of tonicity such as sodium chloride or dextrose are also envisioned.
  • the carrier may comprise, in total, from 0.1% to 99.99999% by weight of the composition/s or the pharmaceutical composition/s presented herein.
  • the pharmaceutical composition includes incorporation of any one of the active ingredients into or onto particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, etc., or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts.
  • polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, etc.
  • liposomes such as polylactic acid, polyglycolic acid, hydrogels, etc.
  • microemulsions such as polylactic acid, polyglycolic acid, hydrogels, etc.
  • Such compositions may influence the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance.
  • the pharmaceutical composition comprising the compound of the invention is in a unit dosage form.
  • the pharmaceutical composition is prepared by any of the methods well known in the art of pharmacy.
  • the unit dosage form is in the form of a tablet, capsule, lozenge, wafer, patch, ampoule, vial or pre-filled syringe.
  • in vitro assays may optionally be employed to help identify optimal dosage ranges.
  • the precise dose to be employed in the formulation will also depend on the route of administration, and the nature of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. Effective doses can be extrapolated from dose-response curves derived from in-vitro or in-vivo animal model test bioassays or systems. In some embodiments, the effective dose is determined as described hereinabove.
  • the pharmaceutical composition of the invention is administered in any conventional oral, parenteral or transdermal dosage form.
  • administering refers to any method which, in sound medical practice, delivers a composition containing an active agent to a subject in such a manner as to provide a therapeutic effect.
  • administering is by an oral administration, a systemic administration or a combination thereof.
  • the pharmaceutical composition is administered via oral (i.e., enteral), rectal, vaginal, topical, nasal, ophthalmic, transdermal, subcutaneous, intramuscular, intraperitoneal or intravenous routes of administration.
  • oral i.e., enteral
  • vaginal topical
  • nasal ophthalmic
  • transdermal subcutaneous
  • intramuscular intraperitoneal
  • intravenous routes of administration i.e., transdermal, subcutaneous, intramuscular, intraperitoneal or intravenous routes of administration.
  • routes of administration of the pharmaceutical composition will depend on the disease or condition to be treated. Suitable routes of administration include, but are not limited to, parenteral injections, e.g., intradermal, intravenous, intramuscular, intralesional, subcutaneous, intrathecal, and any other mode of injection as known in the art.
  • intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir.
  • Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer.
  • the pharmaceutical composition or is in the form of a tablets or a capsule, which can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; or a glidant such as colloidal silicon dioxide.
  • a liquid carrier such as fatty oil.
  • dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or other enteric agents.
  • the tablet of the invention is further film coated.
  • oral application of the pharmaceutical composition or of the kit is in a form of a drinkable liquid.
  • oral application of the pharmaceutical composition or of the kit is in a form of an edible product.
  • aqueous solutions may be suitably buffered, if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose.
  • aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal injection purposes.
  • the compounds of the present invention can exist in free form for treatment, or as a pharmaceutically acceptable salt.
  • the term "pharmaceutically acceptable salt” refers to any non-toxic salt of a compound of the present invention that, upon administration to a subject, e.g., a human, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof.
  • pharmaceutically acceptable can mean approved by a regulatory agency of the Federal or a state government or listed in the U. S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
  • Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19.
  • Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. These salts can be prepared in situ during the final isolation and purification of the compounds. Acid addition salts can be prepared by 1) reacting the purified compound in its free -based form with a suitable organic or inorganic acid and 2) isolating the salt thus formed.
  • Non-limiting examples of pharmaceutically acceptable salts include but are not limited to: acetate, aspartate, benzenesulfonate, benzoate, bicarbonate, carbonate, halide (such as bromide, chloride, iodide, fluoride), bitartrate, citrate, salicylate, stearate, succinate, sulfate, tartrate, decanoate, edetate, fumarate, gluconate, and lactate or any combination thereof.
  • halide such as bromide, chloride, iodide, fluoride
  • Additional examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate
  • Base addition salts can be prepared by 1) reacting the purified compound in its acid form with a suitable organic or inorganic base and 2) isolating the salt thus formed.
  • Salts derived from appropriate bases include alkali metal (e.g., sodium, lithium, and potassium), alkaline earth metal (e.g., magnesium and calcium), ammonium and N+(C1- 4alkyl)4 salts.
  • alkali metal e.g., sodium, lithium, and potassium
  • alkaline earth metal e.g., magnesium and calcium
  • ammonium and N+(C1- 4alkyl)4 salts e.g., sodium, lithium, and potassium
  • alkaline earth metal e.g., magnesium and calcium
  • ammonium and N+(C1- 4alkyl)4 salts e.g., sodium, lithium, and potassium
  • alkaline earth metal e.g., magnesium and calcium
  • ammonium e.g., sodium, lithium, and calcium
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
  • Other acids and bases while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid or base addition salts.
  • alkyl describes an aliphatic hydrocarbon including straight chain and branched chain groups.
  • alkyl also encompasses saturated or unsaturated hydrocarbon, hence this term further encompasses alkenyl and alkynyl.
  • alkenyl describes an unsaturated alkyl, as defined herein, having at least two carbon atoms and at least one carbon-carbon double bond.
  • the alkenyl may be substituted or unsubstituted by one or more substituents, as described hereinabove.
  • alkynyl as defined herein, is an unsaturated alkyl having at least two carbon atoms and at least one carbon-carbon triple bond.
  • the alkynyl may be substituted or unsubstituted by one or more substituents, as described hereinabove.
  • cycloalkyl describes an all-carbon monocyclic or fused ring (i.e. rings which share an adjacent pair of carbon atoms) group where one or more of the rings does not have a completely conjugated pi-electron system.
  • the cycloalkyl group may be substituted or unsubstituted, as indicated herein.
  • aryl describes an all-carbon monocyclic or fused-ring polycyclic (i.e. rings which share adjacent pairs of carbon atoms) groups having a completely conjugated pi-electron system.
  • the aryl group may be substituted or unsubstituted, as indicated herein.
  • alkoxy describes both an O-alkyl and an -O-cycloalkyl group, as defined herein.
  • aryloxy describes an -O-aryl, as defined herein.
  • Each of the alkyl, cycloalkyl and aryl groups in the general formulas herein may be substituted by one or more substituents, whereby each substituent group can independently be, for example, halide, alkyl, alkoxy, cycloalkyl, nitro, amino, hydroxyl, thiol, thioalkoxy, carboxy, amide, aryl and aryloxy, depending on the substituted group and its position in the molecule. Additional substituents are also contemplated.
  • halide describes fluorine, chlorine, bromine or iodine.
  • haloalkyl describes an alkyl group as defined herein, further substituted by one or more halide(s).
  • haloalkoxy describes an alkoxy group as defined herein, further substituted by one or more halide(s).
  • hydroxyl or “hydroxy” describes a -OH group.
  • mercapto or “thiol” describes a -SH group.
  • thioalkoxy describes both an -S-alkyl group, and a -S-cycloalkyl group, as defined herein.
  • thioaryloxy describes both an -S-aryl and a -S-heteroaryl group, as defined herein.
  • amino describes a -NR’R” group, or a salt thereof, with R’ and R” as described herein.
  • heterocyclyl describes a monocyclic or fused ring group having in the ring(s) one or more atoms such as nitrogen, oxygen and sulfur.
  • the rings may also have one or more double bonds. However, the rings do not have a completely conjugated pi- electron system.
  • Representative examples are piperidine, piperazine, tetrahydrofuran, tetrahydropyran, morpholino and the like.
  • carboxy describes a -C(O)OR' group, or a carboxylate salt thereof, where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl (bonded through a ring carbon) or heterocyclyl (bonded through a ring carbon) as defined herein, or "carboxylate”
  • carbonyl describes a -C(O)R' group, where R' is as defined hereinabove. The above-terms also encompass thio-derivatives thereof (thiocarboxy and thiocarbonyl).
  • thiocarbonyl describes a -C(S)R' group, where R' is as defined hereinabove.
  • a "thiocarboxy” group describes a -C(S)OR' group, where R' is as defined herein.
  • a "sulfinyl” group describes an -S(O)R' group, where R' is as defined herein.
  • a "sulfonyl” or “sulfonate” group describes an -S(O)2R' group, where R' is as defined herein.
  • a "carbamyl” or “carbamate” group describes an -OC(O)NR'R" group, where R' is as defined herein and R" is as defined for R'.
  • a “nitro” group refers to a -NO2 group.
  • amide as used herein encompasses C-amide and N-amide.
  • C-amide describes a -C(O)NR'R" end group or a -C(O)NR'-linking group, as these phrases are defined hereinabove, where R' and R" are as defined herein.
  • N-amide describes a -NR"C(O)R' end group or a -NR'C(O)- linking group, as these phrases are defined hereinabove, where R' and R" are as defined herein.
  • a "cyano" or "nitrile” group refers to a -CN group.
  • guanidine describes a - R'NC(N)NR"R"' end group or a -R'NC(N) NR"- linking group, as these phrases are defined hereinabove, where R', R" and R'” are as defined herein.
  • the term “azide” refers to a -N3 group.
  • sulfonamide refers to a -S(O)2NR'R” group, with R' and R" as defined herein.
  • phosphonyl or “phosphonate” describes an -OP(O)-(OR')2 group, with R' as defined hereinabove.
  • phosphinyl describes a -PR'R" group, with R' and R" as defined hereinabove.
  • alkylaryl describes an alkyl, as defined herein, which substituted by an aryl, as described herein.
  • An exemplary alkylaryl is benzyl.
  • heteroaryl describes a monocyclic or fused ring (i.e. rings which share an adjacent pair of atoms) group having in the ring(s) one or more atoms, such as, for example, nitrogen, oxygen and sulfur and, in addition, having a completely conjugated pi- electron system.
  • heteroaryl refers to an aromatic ring in which at least one atom forming the aromatic ring is a heteroatom. Heteroaryl rings can be foamed by three, four, five, six, seven, eight, nine and more than nine atoms. Heteroaryl groups can be optionally substituted.
  • heteroaryl groups include, but are not limited to, aromatic C3-8 heterocyclic groups containing one oxygen or sulfur atom, or two oxygen atoms, or two sulfur atoms or up to four nitrogen atoms, or a combination of one oxygen or sulfur atom and up to two nitrogen atoms, and their substituted as well as benzo- and pyrido- fused derivatives, for example, connected via one of the ring-forming carbon atoms.
  • heteroaryl is selected from among oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinal, pyrazinyl, indolyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinazolinyl or quinoxalinyl.
  • a heteroaryl group is selected from among pyrrolyl, furanyl (furyl), thiophenyl (thienyl), imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3- oxazolyl (oxazolyl), 1,2-oxazolyl (isoxazolyl), oxadiazolyl, 1,3-thiazolyl (thiazolyl), 1,2- thiazolyl (isothiazolyl), tetrazolyl, pyridinyl (pyridyl)pyridazinyl, pyrimidinyl, pyrazinyl,
  • each additional ring is the saturated form (perhydro form) or the partially unsaturated form (e.g., the dihydro form or tetrahydro form) or the maximally unsaturated (nonaromatic) form.
  • heteroaryl thus includes bicyclic radicals in which the two rings are aromatic and bicyclic radicals in which only one ring is aromatic.
  • heteroaryl examples include 3H-indolinyl, 2(lH)-quinolinonyl, 4- oxo-l,4-dihydroquinolinyl, 2H-1 -oxoisoquinolyl, 1,2-dihydroquinolinyl, (2H)quinolinyl N-oxide, 3,4-dihydroquinolinyl, 1,2-dihydroisoquinolinyl, 3,4-dihydro-isoquinolinyl, chromonyl, 3,4-dihydroiso-quinoxalinyl, 4-(3H)quinazolinonyl, 4H-chromenyl, 4- chromanonyl, oxindolyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,2,3,4-tetrahydro-quinolinyl, lH-2,3-dihydroisoindolyl, 2,3-dihydrobenzo[
  • heteroaryl groups are optionally substituted.
  • the one or more substituents are each independently selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-6-alkyl, Ci-6-haloalkyl, Ci-6-hydroxyalkyl, Ci-6- aminoalkyl, Ci -6- alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl, or trifluoromethyl.
  • heteroaryl groups include, but are not limited to, unsubstituted and mono- or di-substituted derivatives of furan, benzofuran, thiophene, benzothiophene, pyrrole, pyridine, indole, oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole, isothiazole, imidazole, benzimidazole, pyrazole, indazole, tetrazole, quinoline, isoquinoline, pyridazine, pyrimidine, purine and pyrazine, furazan, 1,2,3- oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, triazole, benzotriazole, pteridine, phenoxazole, oxadiazole, benzopyrazole, quinolizine, cinnoline
  • compositions, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.
  • method refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.
  • the term “treating” includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.
  • the terms “treatment” or “treating” of a disease, disorder, or condition encompasses substantial alleviation of at least one symptom thereof, substantial reduction in the severity thereof, or inhibition of the progression thereof, wherein substantial is as described herein. Treatment need not mean that the disease, disorder, or condition is totally cured.
  • a useful composition herein needs only to reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide improvement to a patient or subject’s quality of life.
  • prevention of a disease, disorder, or condition encompasses the delay, prevention, suppression, or inhibition of the onset of a disease, disorder, or condition.
  • prevention relates to a process of prophylaxis in which a subject is exposed to the presently described active ingredients prior to the induction or onset of the disease/disorder process. This could be done where an individual has a genetic pedigree indicating a predisposition toward occurrence of the disease/disorder to be prevented.
  • the term “reducing”, including any grammatical from thereof’ relates to at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99% reduction of the UAEand/or UBA6 activity, including any range between.
  • N'-(2-amino-6-chloro-pyrimidin-4-yl)-3-(trifluoromethylsulfanyl) benzo hydrazide (2.7 g, 7.42 mmol, 1 eq) was dissolved in BSA (15 mL) and the mixture was stirred at 100 °C for 12 h under N2. The reaction mixture was added MeOH (30 mL) slowly. Then it was concentrated under reduced pressure to remove solvent. The residue was dissolved into water (60 mL). The aqueous phase was extracted with DCM (30 mL*3). The combined organic phase was washed with brine (60 mL*l), dried with anhydrous Na2SO4, filtered and the filtrate was concentrated in vacuum.
  • Furan -based compounds (F-01-F-07) have been synthesized as follows:
  • the UAE-Ubiquitin conjugation inhibitory activity was measured by quantifying the AMP generated during the UAE-Ubiquitin complex formation.
  • the enzymatic reactions totaled lOpl containing 50mM HEPES (pH 7.5), ImM DTT, 5mM MgC12, 0.1% BSA, lOpM ATP, lOOnM UAE-GST and lOOOnM Ubiquitin- His.
  • the reactions were incubated in the presence of escalating doses of the inhibitory compounds for 2hr at 24°C. Next, detection of the AMP generated during the incubation was performed using the commercial AMP-Glo kit by Promega, according to the manufacturer’s instructions.
  • UBA6-Ubiquitin assay The UBA6-Ubiquitin conjugation inhibitory activity was measured by quantifying the AMP generated during the UBA6 -Ubiquitin complex formation.
  • the enzymatic reactions totaled lOpl containing 50mM HEPES (pH 7.5), ImM DTT, 5mM MgC12, 0.1% BSA, lOpM ATP, 50nM UBA6-GST and lOOOnM Ubiquitin- His.
  • the reactions were incubated in the presence of escalating doses of the inhibitory compounds for 2hr at 24°C. Next, detection of the AMP generated during the incubation was performed using the commercial AMP-Glo kit by Promega, according to the manufacturer’s instructions.
  • UBA6-FAT10 assay The UBA6-FAT10 conjugation inhibitory activity was measured by quantifying the AMP generated during the UBA6-FAT10 complex formation.
  • the enzymatic reactions totaled lOpl containing 50mM HEPES (pH 7.5), ImM DTT, 5mM MgC12, 0.1% BSA, lOpM ATP, 25nM UBA6-GST and lOOOnM FATIO-His.
  • the reactions were incubated in the presence of escalating doses of the inhibitory compounds for 2hr at 24°C. Next, detection of the AMP generated during the incubation was performed using the commercial AMP-Glo kit by Promega, according to the manufacturer’s instructions.
  • NAE-NEDD8 assay The NAE-NEDD8 conjugation inhibitory activity was measured by quantifying the AMP generated during the NAE-NEDD8 complex formation.
  • the enzymatic reactions totaled lOpl containing 50mM HEPES (pH 7.5), ImM DTT, 5mM MgCh, 0.1% BSA, lOpM ATP, lOOnM NAE and 500nM NEDD8-His.
  • the reactions were incubated in the presence of escalating doses of the inhibitory compounds for 2hr at 24°C. Next, detection of the AMP generated during the incubation was performed using the commercial AMP-Glo kit by Promega, according to the manufacturer’s instructions.
  • SAE-SUMQ assay The SAE-SUMO conjugation inhibitory activity was measured by quantifying the AMP generated during the SAE-SUMO complex formation.
  • the enzymatic reactions totaled lOpl containing 50mM HEPES (pH 7.5), ImM DTT, 5mM MgCh, 0.1% BSA, lOpM ATP, lOOnM SAE-CF and 500nM SUMO-His.
  • the reactions were incubated in the presence of escalating doses of the inhibitory compounds for 2hr at 24°C. Next, detection of the AMP generated during the incubation was performed using the commercial AMP-Glo kit by Promega, according to the manufacturer’s instructions.
  • Cell viability assays Cells were plated at a cell density of 8 xlO 4 cells/ml in 96- well culture plates a day before the commencement of the study. Increasing concentrations of the compounds were added to the wells and incubated at 37 °C for 72 hours. Cell viability was assessed using the CellTiter Gio assay kit (Promega) according to the manufacturer’s protocol. Luminescence was measured using a Spark M20 Plate Reader (TECAN).
  • Apoptosis assay Cells were plated at a density of IxlO 6 cells/ml in 6 well plates a day before the commencement of the study. The compounds were added to the wells at the indicated concentrations and the plates were incubated at 37°C for 24 hours. The cells were collected, and the apoptotic cells were labeled using the FITC Annexin V Apoptosis Detection Kit with PI (Biolegend) according to the manufacturer’ s instructions. The labeled cells were quantified by flow cytometry using the MACSQuant Analyzer 10 (Miltenyi Biotec) cytometer.
  • Exemplary compounds of the invention were screened using internally developed cell-free biochemical assays, as described hereinabove.
  • Each assay measures the AMP generated during the conjugation of a specific El-UBL protein pair (e.g., UAE-Ubiquitin; UBA6-Ubiquitin; UBA6-FAT10; NAE-NEDD8; SAE-SUMO).
  • a specific El-UBL protein pair e.g., UAE-Ubiquitin; UBA6-Ubiquitin; UBA6-FAT10; NAE-NEDD8; SAE-SUMO.
  • inhibition of this conjugation attenuates all subsequent El-UBL pathway cascade.
  • Each compound tested in these assays presented a specific set of inhibitory properties against the different El enzymes (Table 1).
  • A ⁇ 10 nM
  • B 10 ⁇ IC50 ⁇ 100 nM
  • C 100 ⁇ IC50 ⁇ l,000 nM
  • D l,000 ⁇ IC50 ⁇ 10,000 nM
  • E >10,000 nM.
  • the structures of the tested compounds are represented in Figure 1.
  • Table 1 IC50 values (nanomolar) for inhibition of conjugation between the El enzyme and its putative UBL obtained for exemplary triazole-based compounds of the invention.
  • A ⁇ 10 nM
  • B 10 ⁇ IC50 ⁇ 100 nM
  • C 100 ⁇ IC50 ⁇ l,000 nM
  • D l,000 ⁇ IC50 ⁇ 10,000 nM
  • E >10,000 nM.
  • Table 1A IC50 values (nanomolar) for inhibition of conjugation between the El enzyme and its putative UBL, obtained for exemplary triazole -based compounds of the invention.
  • A ⁇ 10 nM
  • B 10 ⁇ IC50 ⁇ 100 nM
  • C 100 ⁇ IC50 ⁇ l,000 nM
  • D l,000 ⁇ IC50 ⁇ 10,000 nM
  • E >10,000 nM.
  • Table IB IC50 values (nanomolar) for inhibition of conjugation between the El enzyme and its putative UBL, obtained for exemplary triazole -based compounds of the invention.
  • IC50 values nanomolar for inhibition of conjugation between the El enzyme and its putative UBL, obtained for exemplary triazole -based compounds of the invention.
  • Some of the compounds represented in Tables 1A and IB showed significant in- vitro cytotoxicity in MM1S cancer cell line, exhibiting low nanomolar IC50 values.
  • A ⁇ 10 nM
  • B 10 ⁇ IC50 ⁇ 100 nM
  • C 100 ⁇ IC50 ⁇ l,000 nM
  • D l,000 ⁇ IC50 ⁇ 10,000 nM
  • E >10,000 nM.
  • the structures of the tested compounds are represented in Figure 5.
  • Table 1C IC50 values (nanomolar) for inhibition of conjugation between the El enzyme and its putative UBL, obtained for exemplary furan-based compounds of the invention.
  • MB compounds induce cell cytotoxicity in cancer cells lines
  • the anti-cancer effects of the exemplary compounds of the invention were tested in different solid and liquid cancer cell lines. All the tested compounds showed anti-cancer effects at different potency levels (Table 2). Accordingly, it is postulated that the compounds disclosed herein can be used as anti-cancer agents, i-03 show low nM IC50 values in all cell lines tested. Some of the compounds were found to be effective only against specific types of cancers. For example, i-02 was found to possess nanomolar cytotoxic potency against HCT116, U2932 and MM1S. l,000 ⁇ IC50 ⁇ 10,000 nM; E: >10,000 nM.
  • Table 2 exemplary triazole-based compounds of the invention induced cytotoxicity in different cancer cell lines
  • furan-based compounds of the invention were tested in cell based studies.
  • Exemplary compounds F-01 induced significant cytotoxicity in different cancer cell lines, as represented in Table 2 A, below.
  • A ⁇ 10 nM
  • B 10 ⁇ IC50 ⁇ 100 nM
  • C 100 ⁇ IC50 ⁇ l,000 nM
  • D l,000 ⁇ IC50 ⁇ 10,000 nM
  • E >10,000 nM.
  • Table 2A exemplary furan-based compounds of the invention induced cytotoxicity in different cancer cell lines
  • the tested compounds induce ER-stress in cancer cell lines:
  • the tested compounds induce caspase activation in cancer cell lines:
  • Prolonged ER-stress induction in cells can lead to the activation of caspase proteases which in turn lead to cell apoptosis.
  • the activation of caspases was quantified following treatment of the cells for 24 hours. The obtained results (Table 3) showed that i-02 and i-03 induced up to 27% increase in activated caspases in all tested cell lines.
  • Table 3 i compounds induce caspase activation in cancer cell lines.
  • the tested compounds induce apoptosis in cancer cell lines
  • Table 4 i compounds induced apoptosis in cancer cell lines.
  • Substrates accumulation in cancer cells following treatment with i-03 and F-01 [0245] Inhibition of the Ubiquitin pathway should attenuate substrates degradation (such as Ikba, c-Jun, c-Myc, and p21) and/or activation (such as H2B), leading to the accumulation of these proteins in the cells, preventing the activation of several cellular pathways.
  • substrates degradation such as Ikba, c-Jun, c-Myc, and p21
  • activation such as H2B

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Abstract

La présente invention concerne des composés et des compositions les comprenant. L'invention concerne en outre des procédés d'utilisation, notamment pour le traitement et la prévention du cancer et d'autres maladies sensibles à l'inhibition de UAE et/ou de UBA6 chez un sujet en ayant besoin.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008019124A1 (fr) * 2006-08-08 2008-02-14 Millennium Pharmaceuticals, Inc. Composés hétéroaryles utiles comme inhibiteurs des enzymes d'activation e1
WO2013123169A1 (fr) * 2012-02-17 2013-08-22 Millennium Pharmaceuticals, Inc. Pyrazolopyrimidinyle inhibiteurs de l'enzyme d'activation de l'ubiquitine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008019124A1 (fr) * 2006-08-08 2008-02-14 Millennium Pharmaceuticals, Inc. Composés hétéroaryles utiles comme inhibiteurs des enzymes d'activation e1
WO2013123169A1 (fr) * 2012-02-17 2013-08-22 Millennium Pharmaceuticals, Inc. Pyrazolopyrimidinyle inhibiteurs de l'enzyme d'activation de l'ubiquitine

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MISRA MOHIT; KUHN MAXIMILIAN; LöBEL MARK; AN HEESEON; STATSYUK ALEXANDER V.; SOTRIFFER CHRISTOPH; SCHINDELIN HERMANN: "Dissecting the Specificity of Adenosyl Sulfamate Inhibitors Targeting the Ubiquitin-Activating Enzyme", STRUCTURE, ELSEVIER, AMSTERDAM, NL, vol. 25, no. 7, 1 June 2017 (2017-06-01), AMSTERDAM, NL , pages 1120, XP085115760, ISSN: 0969-2126, DOI: 10.1016/j.str.2017.05.001 *

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