WO2018044986A1 - Inhibiteurs chimiques de biogenèse de ribosomes eucaryotes à base de triazinoindole - Google Patents

Inhibiteurs chimiques de biogenèse de ribosomes eucaryotes à base de triazinoindole Download PDF

Info

Publication number
WO2018044986A1
WO2018044986A1 PCT/US2017/049295 US2017049295W WO2018044986A1 WO 2018044986 A1 WO2018044986 A1 WO 2018044986A1 US 2017049295 W US2017049295 W US 2017049295W WO 2018044986 A1 WO2018044986 A1 WO 2018044986A1
Authority
WO
WIPO (PCT)
Prior art keywords
chosen
halogen
alkyl
hydrocarbyl
compound
Prior art date
Application number
PCT/US2017/049295
Other languages
English (en)
Inventor
Tarun M. Kapoor
Original Assignee
The Rockefeller University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Rockefeller University filed Critical The Rockefeller University
Publication of WO2018044986A1 publication Critical patent/WO2018044986A1/fr

Links

Classifications

    • 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

Definitions

  • This invention relates to triazinoindole-based chemical inhibitors of eukaryotic ribosome genesis.
  • the compounds disclosed are useful in treatment of fungal diseases.
  • RNA Every minute a growing cell can generate -2000 ribosomes, ribonucleoprotein- based nanomachines that translate genomes to synthesize proteins (Warner, 1999).
  • the assembly of ribosomes which contain -5500 nucleotides of RNA and -80 proteins that are together organized into the 60S and 40S subunits, starts in the nucleolus with the transcription of ribosomal DNA.
  • Subsequent steps lead to the processing of RNA to generate 25S, 5.8S, and 5S rRNAs (ribosomal RNAs) for the 60S subunit and the 18S rRNA for the 40S subunit.
  • the assembly of the 60S subunit involves multiple intermediates, collectively referred to as pre-60S particles (Nissan et al., 2002).
  • pre-60S particles The rearrangements of these particles have been characterized using imaging and immunoprecipitation-based approaches, and the key intermediates are linked to specific proteins that associate with distinct particles, such as the Nsal -associated complex (named the Nsal -particles) that assemble in the nucleolus and the Rixl -associated complex (named the Rixl -particles) that are processed in the nucleoplasm (Kressler et al., 2012).
  • the fidelity of this dynamic assembly process depends on precise spatial and temporal regulation at each stage.
  • the Nug2 GTPase-dependent checkpoint blocks export of pre-60S particles to the cytoplasm until it is properly assembled (Matsuo et al., 2014).
  • the sequential processing of these ribosome assembly intermediates requires numerous energy-consuming enzymes, including ATPases in the AAA+ (ATPases associated with diverse cellular activities) family (Thomson et al., 2013).
  • the AAA+ family is a large and functionally diverse group of enzymes, that can couple ATP-hydrolysis with mechanical work (e.g. protein unfolding and directional transport).
  • At least three AAA+ ATPases, Drgl, Rix7, and Midasin are involved in the assembly of the 60S subunit (Kressler et al. Biochim. Biophys. Acta 1823, 92-100. (2012)).
  • Drgl and Rix7 are closely related to the well-studied Cdc48 (p97/VCP in mammals) and likely function as homohexamers with six equivalent ATPase sites.
  • Midasin has all six ATPase domains within a single long polypeptide, an organization that is similar to that of the microtubule motor cytoplasmic dynein.
  • Midasin' s ATPase domains form a ring-like domain from which extends an elongated 'tail' with a Metal Ion-Dependent Adhesion Site (MIDAS) domain at its end.
  • MIDAS Metal Ion-Dependent Adhesion Site
  • the invention relates to compounds of general formula I:
  • R 2 is halogen, trifluoromethyl, or hydrogen
  • R 3 is halogen or trifluoromethyl, or when R 1 is other than benzyl, R 3 may additionally be hydrogen, wherein at least one of R 2 and R 3 is halogen or trifluoromethyl;
  • R 4 is chosen from hydrogen and -CH 2 CH 2 C ⁇ CH.
  • the invention relates to methods of inhibiting midasin comprising contacting midasin with a compound of general formula I
  • R 1 is chosen from -(C 3 -Ci 2 )hydrocarbyl and -(CH2)heteroaryl, wherein any one of said -(C 3 -Ci2)hydrocarbyl or -(CH 2 )heteroaryl may be optionally substituted with one or two substituents chosen independently from OH, halogen, nitro, (Ci- C 3 )alkylamino, (Ci-C 3 )dialkylamino, (Ci-C 3 )acylamino, (Ci-C 3 )alkylsulfonyl, (Ci- C 3 )alkylthio, (Ci-C 3 )alkyl, (Ci-C 3 )haloalkyl, (Ci-C 3 )haloalkoxy, (Ci- C 3 )haloalkylthio, and (Ci-C 3 )alkoxy, and wherein said -(C 3 - Ci 2 )hydrocar
  • R 2 and R 3 are chosen independently from a halogen, trifluoromethyl, and hydrogen;
  • R 4 is chosen from hydrogen and -CH 2 CH 2 C ⁇ CH.
  • the invention relates to methods of treating a disease or disorder in a subject where the disease or disorder involves the dysregulation of the midasin signaling pathway, said method comprising administering to the subject a therapeutically effective amount of at least one compound of general formula I:
  • R 2 and R 3 are chosen independently from a halogen, trifluoromethyl, and hydrogen;
  • R 4 is chosen from hydrogen and -CH2CH 2 C ⁇ CH.
  • compositions comprising at least one compound of general formula I:
  • the invention relates to compounds having general formula
  • R 1 may be chosen from -(C3-Ci 2 )hydrocarbyl and -(CH 2 )heteroaryl, wherein any one of said -(C3-Ci 2 )hydrocarbyl or -(CH 2 )heteroaryl may be optionally substituted with one or two substituents chosen independently from OH, halogen, nitro, (Ci-C3)alkylamino, (Ci-C3)dialkylamino, (Ci-C3)acylamino, (Ci-C3)alkylsulfonyl, (Ci- C 3 )alkylthio, (Ci-C 3 )alkyl, (Ci-C 3 )haloalkyl, (Ci-C 3 )haloalkoxy, (Ci- C 3 )haloalkylthio,
  • R 2 may be halogen or hydrogen.
  • R 3 may be halogen, or when R 1 is other than benzyl, R 3 may additionally be hydrogen, wherein at least one of R 2 and R 3 is halogen.
  • R 4 may be chosen from hydrogen and -CH 2 CH 2 C ⁇ CH.
  • R 3 may be halogen. In some embodiments, one or both of R 2 and R 3 may be independently Br or I. In some embodiments, R 3 may be Br. In some embodiments of formula I, R 3 may be trifluorom ethyl.
  • R 4 may be hydrogen
  • R 1 may be— CH 2 -R 6 and R 6 may be chosen from (C2-C 6 )alkenyl, (C3-C 6 )cycloalkyl, phenyl, substituted phenyl, pyridinyl substituted pyridinyl, pyrazolyl and substituted pyrazolyl.
  • -((CH 2 )i -6 )CH 3 -CH 2 (CH)(CH 2 ) 2 , -CH 2 (fluorophenyl), -CH 2 (naphthyl), -CH 2 (pyridinyl), and - CH 2 (pyrazolyl).
  • the -CH 2 (naphthyl), -CH 2 (pyridinyl), and -CH 2 (pyrazolyl) may be optionally substituted.
  • Preferred substituents on pyridine and pyrazole are (Ci-C 3 )alkyl.
  • R 1 CH 2 ), -(CH 2 ) 2 CH 3 and e
  • R 4 may be H or
  • R 1 may be R 2 may be H
  • R 3 may be Br
  • R 4 may be H.
  • R 1 , R 2 may be H
  • R 3 may be Br
  • R 4 may be H
  • R 5 may be chosen from -H, -halogen, -N0 2 , and -OCH 3 .
  • R 1 may be , R 2 may be H, R 3 may be Br, and R 4 may be H.
  • alkyl (or alkylene) is intended to include linear or branched saturated hydrocarbon structures and combinations thereof.
  • Alkyl refers to alkyl groups of from 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, s- butyl, t-butyl and the like.
  • Cycloalkyl is a subset of hydrocarbon and includes cyclic hydrocarbon groups of from 3 to 8 carbon atoms. Examples of cycloalkyl groups include c-propyl, c-butyl, c-pentyl, norbornyl and the like.
  • Ci to C20 hydrocarbon includes alkyl, cycloalkyl, polycycloalkyl, alkenyl, alkynyl, aryl and combinations thereof. Examples include benzyl, phenethyl, cyclohexylmethyl, adamantyl, camphoryl and naphthyl ethyl.
  • Hydrocarbon or hydrocarbyl refer to any substituent comprised of hydrogen and carbon as the only elemental constituents.
  • Aliphatic hydrocarbons or hydrocarbyls are hydrocarbons or hydrocarbyls that are not aromatic; they may be saturated or unsaturated, cyclic, linear or branched.
  • aliphatic hydrocarbons or hydrocarbyls examples include isopropyl, 2-butenyl, 2-butynyl, cyclopentyl, norbornyl, etc.
  • Aromatic hydrocarbons or hydrocarbyls include benzene (phenyl), naphthalene (naphthyl), anthracene, etc.
  • carbocycle is intended to include ring systems in which the ring atoms are all carbon but of any oxidation state.
  • C3-C12 carbocycle refers to both non-aromatic and aromatic systems, including such systems as cyclopropane, benzene and cyclohexene.
  • Carbocycle if not otherwise limited, refers to monocycles, bicycles and polycycles.
  • Cs-Co Carbopolycycle refers to such systems as norbornane, decalin, indane and naphthalene.
  • Heterocycle means an aliphatic or aromatic carbocycle residue in which from one to four carbons is replaced by a heteroatom selected from the group consisting of N, O, and S.
  • the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized.
  • a heterocycle may be non-aromatic (heteroaliphatic) or aromatic (heteroaryl).
  • heterocycle s examples include pyrrolidine, pyrazole, pyrrole, indole, quinoline, isoquinoline, tetrahydroisoquinoline, benzofuran, benzodioxan, benzodioxole (commonly referred to as methylenedioxyphenyl, when occurring as a substituent), tetrazole, morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene, furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and the like.
  • heterocyclyl residues include piperazinyl, piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, tetrahydrofuryl, tetrahydropyranyl, thienyl (also historically called thiophenyl), benzothienyl, thiamorpholinyl, oxadiazolyl, triazolyl and tetrahydroquinolinyl.
  • Hydrocarbyloxy refers to groups of from 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms attached to the parent structure through an oxygen.
  • Alkoxy is a subset of hydrocarbyloxy and includes groups of a straight or branched configuration. Examples include methoxy, ethoxy, propoxy, isopropoxy and the like.
  • Lower- alkoxy refers to groups containing one to four carbons.
  • halogen means fluorine, chlorine, bromine or iodine atoms.
  • acyl refers to formyl and to groups of 1, 2, 3, 4, 5, 6, 7 and 8 carbon atoms of a straight, branched, cyclic configuration, saturated, unsaturated and aromatic and combinations thereof, attached to the parent structure through a carbonyl functionality. Examples include acetyl, benzoyl, propionyl, isobutyryl and the like. Lower- acyl refers to groups containing one to four carbons.
  • the double bonded oxygen, when referred to as a substituent itself is called "oxo".
  • Alkoxy or alkoxyl refers to groups of from 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms of a straight or branched configuration attached to the parent structure through an oxygen. Examples include methoxy, ethoxy, propoxy, isopropoxy and the like.
  • Oxaalkyl refers to alkyl residues in which one or more carbons (and their associated hydrogens) have been replaced by oxygen. Examples include methoxypropoxy, 3,6,9-trioxadecyl and the like.
  • oxaalkyl is intended as it is understood in the art [see Naming and Indexing of Chemical Substances for Chemical Abstracts, published by the
  • thiaalkyl and azaalkyl refer to alkyl residues in which one or more carbons has been replaced by sulfur or nitrogen, respectively. Examples of azaalkyl include ethylaminoethyl and aminohexyl.
  • substituted refers to the replacement of one or more hydrogen atoms in a specified group with a specified radical. For example, substituted alkyl, aryl, cycloalkyl, heterocyclyl etc.
  • alkyl)(aryl)aminoalkyl alkylaminoalkyl (including cycloalkylaminoalkyl), dialkylaminoalkyl, dialkylaminoalkoxy, heterocyclylalkoxy, mercapto, alkylthio, sulfoxide, sulfone, sulfonylamino, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino, arylsulfonyl, arylsulfonylamino,
  • acylaminoalkyl acylaminoalkoxy, acylamino, amidino, aryl, benzyl, heterocyclyl,
  • heterocyclylalkyl phenoxy, benzyloxy, heteroaryloxy, hydroxyimino, alkoxyimino, oxaalkyl, aminosulfonyl, trityl, amidino, guanidino, ureido, benzyl oxyphenyl, and benzyloxy.
  • Oxo is also included among the substituents referred to in "optionally substituted”; it will be appreciated by persons of skill in the art that, because oxo is a divalent radical, there are circumstances in which it will not be appropriate as a substituent (e.g. on phenyl). In one embodiment, 1, 2 or 3 hydrogen atoms are replaced with a specified radical.
  • haloalkyl or halophenyl refers to an alkyl or phenyl in which at least one, but perhaps more than one, hydrogen is replaced by halogen.
  • substituents are halogen, haloalkyl, alkyl, acyl, hydroxyalkyl, hydroxy, alkoxy, haloalkoxy, oxaalkyl, carboxy, cyano, acetoxy, nitro, amino, alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonylamino arylsulfonyl, arylsulfonylamino and benzyloxy.
  • Substituents R n are generally defined when introduced and retain that definition throughout the specification and in all independent claims.
  • Suitable pharmaceutically acceptable acids for salts of the compounds of the present invention include, for example, acetic, adipic, alginic, ascorbic, aspartic, benzenesulfonic (besylate), benzoic, boric, butyric, camphoric, camphorsulfonic, carbonic, citric, ethanedisulfonic, ethanesulfonic, ethylenediaminetetraacetic, formic, fumaric, glucoheptonic, gluconic, glutamic, hydrobromic, hydrochloric, hydroiodic, hydroxynaphthoic, isethionic, lactic, lactobionic, laurylsulfonic, maleic, malic, mandelic, methanesulfonic, mucic, naphthylenesulfonic, ni
  • Suitable pharmaceutically acceptable base addition salts for the compounds of the present invention include, but are not limited to, metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from lysine, arginine, ⁇ , ⁇ '-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium cations and carboxylate, sulfonate and phosphonate anions attached to alkyl having from 1 to 20 carbon atoms.
  • radiolabeled form i.e., the compounds may contain one or more atoms containing an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • a plurality of molecules of a single structure may include at least one atom that occurs in an isotopic ratio that is different from the isotopic ratio found in nature.
  • Radioisotopes of hydrogen, carbon, phosphorous, fluorine, chlorine and iodine include 2 H, 3 H, U C, 13 C, 14 C, 15 N, 35 S, 18 F, 36 C1, 125 I, 124 I and 131 I respectively.
  • Compounds that contain those radioisotopes and/or other radioisotopes of other atoms are within the scope of this invention.
  • Radiolabeled compounds of formula I of this invention and prodrugs thereof can generally be prepared by methods well known to those skilled in the art. Conveniently, such radiolabeled compounds can be prepared by carrying out the procedures disclosed in the Examples and Schemes by substituting a readily available radiolabeled reagent for a non-radiolabeled reagent.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, together with one or more pharmaceutically carriers thereof and optionally one or more other therapeutic ingredients.
  • the carrier(s) must be "acceptable” in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • the compositions may be formulated for oral, topical or parenteral administration. For example, they may be given intravenously, intraarterially, subcutaneously, and directly into the CNS - either intrathecally or intracerebroventricularly.
  • Formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), rectal and topical (including dermal, buccal, sublingual and intraocular) administration.
  • the compounds are preferably administered orally or by injection (intravenous or subcutaneous).
  • the precise amount of compound administered to a patient will be the responsibility of the attendant physician. However, the dose employed will depend on a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity. Also, the route of administration may vary depending on the condition and its severity.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
  • Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
  • the active ingredient may also be presented as a bolus, electuary or paste.
  • formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.
  • treatment or “treating,” or “palliating” or “ameliorating” are used interchangeably herein. These terms refers to an approach for obtaining a therapeutic benefit in the form of eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological systems associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder.
  • the compositions may be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made.
  • a protecting group refers to a group which is used to mask a functionality during a process step in which it would otherwise react, but in which reaction is undesirable.
  • the protecting group prevents reaction at that step, but may be subsequently removed to expose the original functionality.
  • the removal or "deprotection” occurs after the completion of the reaction or reactions in which the functionality would interfere.
  • the compounds of the present invention may be prepared by the methods illustrated in the general reaction scheme as, for example, described below, or by modifications thereof, using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants that are in themselves known, but are not mentioned here.
  • the starting materials are either commercially available, synthesized as described in the examples or may be obtained by the methods well known to persons of skill in the art.
  • Evaporation of solvents was accomplished by rotary evaporation using a Biichi rotary evaporator, equipped with a dry ice-acetone condenser, at 5-75 mm Hg at temperatures between 35°C and 50°C.
  • Experiments were monitored by thin layer chromatography (TLC) or liquid chromatography mass spectrometry (LC-MS). The maintenance of 30 °C to 150 °C reaction temperatures was accomplished by the use of an oil bath. Products obtained as solids or high boiling oils were dried under vacuum ( ⁇ 1 mmHg).
  • tetramethylsilane itself: dimethylsulfoxide-i/6 ( ⁇ 2.50, quintet) and deuterium oxide-i/2 ( ⁇ 4.80, singlet). Multiplicities are given as: s (singlet), d (doublet), t (triplet), or m (multiplet). Coupling constants are reported as a J value in Hertz (Hz). The number of protons (n) for a given resonance is indicated by nH.
  • Liquid chromatography mass spectral analyses were obtained using a Waters MicroMassZQ mass spectrometer, with an electron spray ionization (ESI) probe, connected to a Waters 2795 HT Separation Module Alliance HT HPLC system running MassLynx (V4.0).
  • the system used a Waters 996 Photodiode Array Detector set to 254 nm for peak detection, and a Symmetry® C18 (3.5 micron) 2.1 x 50 mm column for separation (mobile phase for positive mode: solvent A: water with 0.1% formic acid, solvent B: acetonitrile; mobile phase for negative mode: solvent A: water with 0.1% morpholine, solvent B: acetonitrile). Values are reported in units of mass to charge (m/z).
  • Cell-permeable chemical inhibitors can be powerful tools for examining dynamic cellular processes, such as ribosome biogenesis, as the functions of target proteins can be blocked within minutes.
  • a chemical inhibitor that directly targets eukaryotic ribosome assembly factors is diazaborine, an antibacterial compound active at -0.4 mM in S. cerevisiae, a concentration at which selective target inhibition can be difficult to achieve.
  • diazaborine blocks cytoplasmic steps (i.e. pre-60S maturation) of ribosome biogenesis, chemical probes for the several distinct assembly steps that occur in the nucleolus and nucleus may be lacking.
  • Another chemical inhibitor of ribosome assembly factors is lamotrigine, a compound that blocks this process in bacteria.
  • RBins for ribosome biogenesis inhibitors
  • RBins are identified as potent, reversible and specific inhibitors of Midasin.
  • Live cell imaging, biochemical approaches, and the use of RBins are combined to inhibit or activate Midasin on the timescale of minutes to analyze ribosome assembly dynamics in cells.
  • the present disclosure uncovers a previously uncharacterized function of Midasin in assembling nucleolar Nsal -particles.
  • fission yeast As a model system has been developed that allows for efficient combining of genetic and chemical approaches.
  • fission yeast strains (named 'MDR-sup' strains) lacking critical factors for multi-drug resistance have been generated and they have been used for chemical screens that mimic synthetic lethal genetic screens.
  • a compound, example 1, hereinafter called RBin-1 was identified.
  • Example 15 As a potent analog of RBin-1 that is likely to have a similar mechanism of action.
  • the fact that compound sensitivity varies significantly by mutations in mdnl indicates that compounds of the invention could directly target Mdnl .
  • ATP -binding in Mdnl is at the interface of two AAA domains and involves Walker A and B motifs from one AAA domain while the arginine finger motif extends from the following AAA domain (e.g. ATP binding at the AAA1 site depends on Walker Al, Walker Bl, and the arginine finger R2 motifs).
  • Mdnl-F1093L Full-length recombinant Mdnl-F1093L, containing a mutation that confers resistance to the RBins in cell-based assays was generated.
  • Mdnl-F1093L is an active enzyme whose ATPase activity (1.8 ATP s "1 ) is comparable to that of wild-type Mdnl .
  • the F1093L mutation in Mdnl also confers resistance to example 15 in the biochemical assay.
  • the pre-rRNAs such as 35S, 27S and 7S pre-rRNA, accumulated only after 60min of RBin-1 treatment, reaching a maximum level in 90-120 min after treatment.
  • RBins inhibit or activate Midasin function on the minutes timescale in fission yeast cells.
  • Eukaryotic ribosome biogenesis involves complex multi-step RNA processing, the recruitment of ribosomal proteins and dynamic associations of multiple non-ribosomal proteins. Briefly, two ribosomal RNA precursors, the 35S and the pre-5S, are transcribed in the nucleolus. The 35S RNA is processed to generate 27S RNA, which is subsequently cleaved to generate 25.5S and 7S RNAs, which contain sequences corresponding to the 25S and 5.8S in the 60S subunit of the mature ribosome. Findings from studies in budding yeast suggest that Mdnl/Real is likely involved in two steps of ribosome biogenesis. The data indicate that many aspects of Mdnl/Real -dependent ribosome biogenesis are likely to be conserved across fungi.
  • ESI-MS (m/z)
  • ESI-MS (m/z) [M+H]+ Calculated: 245.32, found: 245.57.
  • ESI-MS (m/z)
  • ESI-MS (m/z) [M+H]+ Calculated: 293.36, found: 293.62.
  • ESI-MS (m/z) M+H]+ Calculated: 294.35, found: 294.59.
  • ESI-MS (m/z) M+H]+ Calculated: 243.42, found: 243.65.
  • ESI-MS (m/z) [M+H]+ Calculated: 323.39, found: 323.67.
  • ESI-MS (m/z) [M+H]+ Calculated: 338.36, found: 338.61.
  • ESI-MS (m/z) M+H]+ Calculated: 338.36, found:
  • ESI-MS (m/z) [M+H]+ Calculated: 311.35, found: 311.57.
  • ESI-MS (m/z) [M+H]+ Calculated: 419.26, found: 419.48.
  • ESI-MS (m/z) [M+H]+ Calculated: 419.26, found: 419.55.
  • ESI-MS (m/z) [M]+ Calculated: 335.22, found: 335.45 and [M+2H]+ Calculated: 337.22, found: 337.48 (bromine pattern).
  • ESI-MS (m/z) [M]+ Calculated: 371.26, found: 371.49 and [M+2H]+ Calculated: 373.26, found: 373.48 (bromine pattern).
  • ESI-MS (m/z) [M]+ Calculated: 335.22, found: 335.52 and [M+2H]+ Calculated: 337.22, found: 337.55 (bromine pattern).
  • ESI-MS (m/z) [M]+ Calculated: 497.15, found: 497.43 and [M+2H]+ Calculated: 499.15, found: 499.46 (bromine pattern).
  • ESI-MS (m/z) [M]+ Calculated: 497.15, found: 497.44 and [M+2H]+ Calculated: 499.15, found: 499.45 (bromine pattern).
  • ESI-MS (m/z) [M]+ Calculated: 319.18, found: 319.49 and [M+2H]+ Calculated: 321.18, found: 321.51 (bromine pattern).
  • ESI-MS (m/z) [M+H]+ Calculated: 309.40, found: 309.65.
  • Schizosaccharomyces pombe S.pombe strains were grown to exponential phase in 5g of yeast extract (Fisher BioReagents), 30 g of Dextrose Anhydrous Powder (J.T. Baker), 250 mg of L- Histidine (Sigma- Aldrich), 250 mg of L-Leucine (Sigma- Aldrich), 250 mg of adenine
  • hemisulfate salt (Sigma-Aldrich) and 250 mg of uridine per liter of solution.
  • the samples were diluted to ODeoo-0.01, and 1 ⁇ L ⁇ DMSO or 1 ⁇ > ⁇ a DMSO solution of test compound (lOOOx) were mixed with 1 ⁇ _, diluted cell suspensions.
  • the cells were placed in shaker (220 rpm) for 18 hr at 29°C till ODeoo of DMSO control reached ⁇ 1. Relative growth was calculated by dividing the measured OD at a specific concentration by the OD for the DMSO control.
  • Half maximum growth inhibition (GI50) was determined by fitting relative growth to a four-parameter sigmoidal dose-response curve in PRISM.
  • the cells were lysed by sonication in equal volume of lysis buffer (50mMTris, 400mM NaCl, 20 mM imidazole, 1 mM MgCb, 5 ⁇ 2-mercaptoethanol, 200 ⁇ ATP, 3 U/mL benzonase, IX Roche complete protease inhibitor without EDTA, 10% glycerol [pH 7.5]).
  • lysis buffer 50mMTris, 400mM NaCl, 20 mM imidazole, 1 mM MgCb, 5 ⁇ 2-mercaptoethanol, 200 ⁇ ATP, 3 U/mL benzonase, IX Roche complete protease inhibitor without EDTA, 10% glycerol [pH 7.5].
  • the homogenized lysate was then centrifuged at 55,000 rpm for 1 hr.
  • the supernatant was incubated with Ni-NTA beads (QIAGEN) for 40 min.
  • the beads were extensively washed using Washing buffer (50 mM Tris, 400 mM NaCl, 20 mM imidazole, 1 mM MgCb, 5 ⁇ 2-mercaptoethanol, 10% glycerol [pH 7.5]).
  • Washing buffer 50 mM Tris, 400 mM NaCl, 20 mM imidazole, 1 mM MgCb, 5 ⁇ 2-mercaptoethanol, 10% glycerol [pH 7.5]
  • the protein was then eluted by high imidazole buffer (20 mM Tris [pH 7.5], 120 mM NaCl, 300 mM imidazole, 1 mM MgCb, 5 ⁇ 2-mercaptoethanol, 200 ⁇ ATP (A2383)).
  • the eluted fraction was filtered and loaded onto a Mono Q column 5/50 GL (GE Healthcare Life Sciences).
  • the reactions were then incubated at room temperature for 30 or 60 min before quenching with 12 ⁇ 0.2 M EDTA. 1 ⁇ from each reaction mixture was spotted on to TLC PEI cellulose F plates (Millipore, 105579).
  • the TLC buffer contained 0.15 M formic acid and 0.15 M lithium chloride.
  • the TLC plates were then imaged using the Typhoon Scanner 9400 (GE Healthcare Life Sciences). ImageJ was used to calculate the densitometric ratio of the spots corresponding to radioactive free phosphate and ATP to determine the percent of ATP hydrolyzed.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des inhibiteurs chimiques de la genèse des ribosomes eucaryotes à base de Triazinoindole. Les composés ont la structure suivante : les composés décrits sont utiles dans le traitement d'une maladie fongique et de maladies similaires associées à la dysrégulation de la voie de signalisation de de midasin.
PCT/US2017/049295 2016-08-30 2017-08-30 Inhibiteurs chimiques de biogenèse de ribosomes eucaryotes à base de triazinoindole WO2018044986A1 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US201662381355P 2016-08-30 2016-08-30
US62/381,355 2016-08-30
US201662382597P 2016-09-01 2016-09-01
US62/382,597 2016-09-01
US201662410605P 2016-10-20 2016-10-20
US62/410,605 2016-10-20

Publications (1)

Publication Number Publication Date
WO2018044986A1 true WO2018044986A1 (fr) 2018-03-08

Family

ID=61309360

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2017/049295 WO2018044986A1 (fr) 2016-08-30 2017-08-30 Inhibiteurs chimiques de biogenèse de ribosomes eucaryotes à base de triazinoindole

Country Status (1)

Country Link
WO (1) WO2018044986A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080188500A1 (en) * 2006-12-22 2008-08-07 Incyte Corporation Substituted heterocycles as janus kinase inhibitors
US8557848B2 (en) * 2006-12-22 2013-10-15 Merck Sharp & Dohme Corp. 4,5-ring annulated indole derivatives for treating or preventing of HCV and related viral infections
US20140378451A1 (en) * 2011-09-19 2014-12-25 Isis Innovation Limited Cannabinoid-2-Receptor Agonists

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080188500A1 (en) * 2006-12-22 2008-08-07 Incyte Corporation Substituted heterocycles as janus kinase inhibitors
US8557848B2 (en) * 2006-12-22 2013-10-15 Merck Sharp & Dohme Corp. 4,5-ring annulated indole derivatives for treating or preventing of HCV and related viral infections
US20140378451A1 (en) * 2011-09-19 2014-12-25 Isis Innovation Limited Cannabinoid-2-Receptor Agonists

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DATABASE Pubchem [O] 9 August 2005 (2005-08-09), "2-propynyl 5H-[1,2,4]triazino[5,6-b]indol-3-yl sulfide", XP055472386, Database accession no. CID 6411022 *
DATABASE PUBCHEM [O] 9 July 2005 (2005-07-09), "Ribozinoindole-1", XP055472376, Database accession no. CID 5731061 *
DATABASE PUBCHEM. 15 July 2005 (2005-07-15), "Compound Summary for CID 5739653", XP055472364, Database accession no. STK509469 *
GARBARINO, JE ET AL.: "Expression and genomic analysis of midasin, a novel and highly conserved AAA protein distantly related to dynein", BMC GENOMICS, vol. 3, no. 18, 8 July 2002 (2002-07-08), pages 1 - 16, XP021014415 *
MATHEIS, N ET AL.: "Proteomics Differentiate Between Thyroid-Associated Orbitopathy and Dry Eye Syndrome", INVESTIGATIVE OPHTHALMOLOGY AND VISUAL SCIENCE, vol. 56, April 2015 (2015-04-01), pages 2649 - 2656, XP055472381 *

Similar Documents

Publication Publication Date Title
KR102630069B1 (ko) 아폽토시스-유도제
RU2704445C2 (ru) Ингибиторы ezh2 человека и способы их применения
SA517380651B1 (ar) مثبطات ديميثيلاز-1 خاص بلايسين
CA3124952A1 (fr) Inhibiteurs aza-heterobicycliques de mat2a et procedes d'utilisation pour le traitement du cancer
KR20150119201A (ko) 메틸 변형 효소 조절제, 이의 조성물 및 용도
CA2817460A1 (fr) Composes heterocycliques azotes 6,6-condenses substitues et leurs utilisations
JP7360745B2 (ja) Ｃ５ａ受容体の調節因子としての縮合ピペリジニル二環式化合物及び関連化合物
WO2020188467A1 (fr) Composé tricyclique condensé utilisé en tant qu'inhibiteur de kinase
JP7125495B2 (ja) テトラヒドロイソキノリン化合物、その調製方法、そのような化合物を含む医薬組成物およびその使用
JP2020520982A (ja) がん治療のためのイオンチャネル阻害剤化合物
AU2011310078B2 (en) Chromene derivatives
Lu et al. Harmine-based dual inhibitors targeting histone deacetylase (HDAC) and DNA as a promising strategy for cancer therapy
KR20160003196A (ko) ＂니코틴아미드 포스포리보실트랜스퍼라제의 억제제, 조성물, 제품 및 그의 용도＂
Yong et al. Synthesis and biological evaluation of quinazoline derivatives as potential anticancer agents (II)
CN103073455A (zh) 一类新型的kcnq钾通道激动剂、其制备方法和用途
WO2017070464A1 (fr) Analogues de cofacteurs utilisés en tant qu'inhibiteurs de méthyltransférase pour le traitement du cancer
Bo-Ru et al. Diversity-oriented synthesis of marine sponge derived hyrtioreticulins and their anti-inflammatory activities
WO2018044986A1 (fr) Inhibiteurs chimiques de biogenèse de ribosomes eucaryotes à base de triazinoindole
WO2023030335A1 (fr) Composé utilisé en tant qu'inhibiteur du domaine de pseudokinase tyk2/jak1, et procédés de synthèse et d'utilisation
CN117321064A (zh) 吉西他滨抗癌衍生物及抗癌医药用途
EP3632912B1 (fr) Inhibiteurs de protéine kinase
AU2009301797A1 (en) Indazole derivative
WO2018053313A1 (fr) Composés analogues d'adénosine utilisés en tant qu'inhibiteurs de méthyltransférase pour le traitement du cancer
RU2792034C2 (ru) Тетрагидроизохинолиновое соединение, способ его получения, фармацевтическая композиция, содержащая такое соединение, и его применение
Montel de la Roche Innovative approaches to cancer therapy

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17847435

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17847435

Country of ref document: EP

Kind code of ref document: A1