EP1844062A2 - Inhibiteurs d'adn-methyltransferase - Google Patents

Inhibiteurs d'adn-methyltransferase

Info

Publication number
EP1844062A2
EP1844062A2 EP06718809A EP06718809A EP1844062A2 EP 1844062 A2 EP1844062 A2 EP 1844062A2 EP 06718809 A EP06718809 A EP 06718809A EP 06718809 A EP06718809 A EP 06718809A EP 1844062 A2 EP1844062 A2 EP 1844062A2
Authority
EP
European Patent Office
Prior art keywords
alkyl
aryl
optionally substituted
heteroaryl
mmol
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP06718809A
Other languages
German (de)
English (en)
Inventor
Amal Wahhab
Jeffrey M. Besterman
Daniel Delorme
Ljubomir Isakovic
David Llewellyn
Jubrail Rahil
Oscar Saavedra
Robert Deziel
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Methylgene Inc
Original Assignee
Methylgene Inc
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 Methylgene Inc filed Critical Methylgene Inc
Publication of EP1844062A2 publication Critical patent/EP1844062A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

  • This invention relates to inhibition of the DNA methyltransferase isoforms DNMT1 and DNMT3b2. More particularly, the invention relates to compounds and methods for the inhibition of DNMT1 and DNMT3b2.
  • DNA methyltransferase 1 (DNMT1) protein is a major contributor to DNA methyltransferase activity in human cells and is required to maintain methylation patterns in differentiated cells (M. F. Robert, S. Morin, N. Beaulieu, F. Gauthier, I.C. Chute, A. Barsalou, A.R. MacLeod, Nat. Genet. 33 (2003) 61-65).
  • the de novo DNA methyltransferases DNMT3A and DNMT3B establish DNA methylation during early embryogenesis (M. Okano, S. Xie, E. Li, Nat. Genet, 1998, 19, 219-220; M. Okano, D.W. Bell, DA Haber, E.
  • SAH can bind to DNA methyltransferases and inhibit their catalytic reaction and is an important molecule in the regulation of biological transmethylation (T. Deguchi and J. Barchas, J. Biol. Chem., 1971 , 246, 3175; A. Oliva, P. Galletti, and V. Zappia, Eurp. J. Biochem., 1980, 104, 595; P.M. Ueland and J. Saebo, Biochemistry, 1979, 18, 4130). [0005] Inhibitors of DNMT1 and DNMT3B are expected to have value as anti-cancer agents (M. Szyf, Fontiers in Bioscience 2001 , 6, 599-609; M.F. Robert, S. Morin, N.
  • SAH is a known nonselective inhibitor of human DNA methyltransferase and many other methyltransferases.
  • a number of simplified and less rigid analogues of SAH have been reported. Such compounds, however, were devoid of activity against human DNA methyltransferase (M. Botta, R. Saladino, G. Pedraly-Noy, and R. Nicoletti, Med. Chem. Res., 1994, 4, 323).
  • SAM and SAH as inhibitors are not good drugs and they are unstable in plasma due to hydrolases and ribonucleases, they have poor absorption due to the Zwitterionic nature, are rapidly excreted, and have a short half life.
  • SAH is an endogenous inhibitor of numerous methyltransferases and as such is non-selective, making it undesirable as a drug.
  • Some more stable nitrogen anlogues of SAM and SAH have been reported. Chi-Deu Chang and J. K. Coward, J. Med.
  • Sinefungin a natural product, is a nitrogen analogue of SAH and has been reported to inhibit human DNA methyltransferase. It is also a non-selective inhibitor with potential for toxicity (C. Barbes, J. Sanchez, M.J. Yebra, M.Robert-Gero, and C. Hardisson, FEMS Microbiology Letters, 1990, 69, 239).
  • DNA methyltransferase inhibitors are known to be DNA binding agents, however, and thus may inhibit the DNMTs indirectly.
  • This invention provides compounds and methods for the inhibition of human C-5
  • the compounds of the invention are inhibitors of the DNA methyltransferase isoforms DNMT1 and DNMT3b2. Accordingly, the invention provides new inhibitors of DNMT1 and DNMT3b2. [0016] In a first aspect, the invention provides compounds of formula (I) (and their pharmaceutically acceptable salts) that are useful as inhibitors of DNMT1 and/or DNMT3b2 and therefore are useful for studying the role of DNA methyltransferase in biological processes.
  • compositions comprising a compound that is an inhibitor of DNMT1 and/or DNMT3b2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient, or diluent.
  • the invention provides a method of inhibiting DNMT1 and/or DNMT3b2 enzymes in a cell, comprising contacting a cell in which inhibition of DNMT1 or DNMT3b2 is desired with a compound of the invention.
  • Figure 1 displays the full length DNMT1 (Swissprot accession number P26358 (SEQ ID NO.1)).
  • Figure 2 displays the DNMT3 splice variant 2 of DNMT3b2 (Swissprot accession number Q9UBC3-2 (SEQ ID NO.2))
  • the invention provides compounds and methods for inhibiting DNMT1 and DNMT3b2.
  • inhibitor of DNMT1 and DNMT3b2 is used to identify a compound having a structure as defined herein, which is capable of interacting with DNMT1 , DNMT3b2 or both DNMT1 and DNMT3b2 and inhibiting the activity of DNMT1 , DNMT3b2, or both DNMT1 and DNMT3b2. In some preferred embodiments, such reduction of activity is at least about 50%, more preferably at least about 75%, and still more preferably at least about 90%.
  • chemical moieties are defined and referred to throughout primarily as univalent chemical moieties (e.g., alkyl, aryl, etc.).
  • alkyl generally refers to a monovalent radical (e.g. CH 3 -CH 2 -)
  • a bivalent linking moiety can be "alkyl,” in which case those skilled in the art will understand the alkyl to be a divalent radical (e.g., -CH 2 - CH 2 -), which is equivalent to the term "alkylene.”
  • aryl refers to the corresponding divalent moiety, arylene.
  • All atoms are understood to have their normal number of valences for bond formation ⁇ i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of the S).
  • a moiety may be defined, for example, as (A) 3 -B-, wherein a is 0 or 1. In such instances, when a is 0 the moiety is B- and when a is 1 the moiety is A-B-. Also, a number of moieties disclosed herein exist in multiple tautomeric forms, all of which are intended to be encompassed by any given tautomeric structure.
  • hydrocarbyl refers to a straight, branched, or cyclic alkyl, alkenyl, or alkynyl, each as defined herein.
  • a “C 0 " hydrocarbyl is used to refer to a covalent bond.
  • C 0 -C 3 -hydrocarbyF' includes a covalent bond, methyl, ethyl, ethenyl, ethynyl, propyl, propenyl, propynyl, and cyclopropyl.
  • alkyl refers to straight and branched chain aliphatic groups having from 1 to 12 carbon atoms, preferably 1-8 carbon atoms, and more preferably 1-6 carbon atoms, which is optionally substituted with one, two or three substituents.
  • Preferred alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert- butyl, pentyl, and hexyl.
  • a "C 0 " alkyl (as in "C 0 -C 3- alkyr') is a covalent bond (like "C 0 " hydrocarbyl).
  • alkenyl as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon double bonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms, which is optionally substituted with one, two or three substituents.
  • Preferred alkenyl groups include, without limitation, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.
  • alkynyl as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon triple bonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms, which is optionally substituted with one, two or three substituents.
  • Preferred alkynyl groups include, without limitation, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.
  • alkylene is an alkyl, alkenyl, or alkynyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups.
  • Preferred alkylene groups include, without limitation, methylene, ethylene, propylene, and butylene.
  • Preferred alkenylene groups include, without limitation, ethenylene, propenylene, and butenylene.
  • Preferred alkynylene groups include, without limitation, ethynylene, propynylene, and butynylene.
  • cycloalkyl as employed herein includes saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbons, preferably 3 to 8 carbons, and more preferably 3 to 6 carbons, wherein the cycloalkyl group additionally is optionally substituted.
  • Preferred cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.
  • heteroalkyl refers to an alkyl group, as defined hereinabove, wherein one or more carbon atoms in the chain are replaced by a heteroatom selected from the group consisting of O, S, NH, N-alkyl, SO, SO 2 , SO 2 NH, or NHSO 2 .
  • An "aryl” group is a C 6 -Ci 4 aromatic moiety comprising one to three aromatic rings, which is optionally substituted.
  • the aryl group is a C 6 -Ci 0 aryl group.
  • Preferred aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl.
  • An “aralkyl” or “arylalkyl” group comprises an aryl group covalently linked to an alkyl group, either of which may independently be optionally substituted or unsubstituted.
  • the aralkyl group is (C 1 - C ⁇ aIk(C 6 -C 10 )aryl, including, without limitation, benzyl, phenethyl, and naphthylmethyl.
  • a "heterocyclyl” or “heterocyclic” group is a ring structure having from about 3 to about 12 atoms, wherein one or more atoms are selected from the group consisting of N, O, S, SO, and SO 2 .
  • the heterocyclic group is optionally substituted on carbon at one or more positions.
  • heterocyclic group is also independently optionally substituted on nitrogen with alkyl, aryl, aralkyl, alkylcarbonyl, alkylsulfonyl, arylcarbonyl, arylsulfonyl, alkoxycarbonyl, or aralkoxycarbonyl.
  • Preferred heterocyclic groups include, without limitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl, and morpholino.
  • the heterocyclic group is fused to an aryl, heteroaryl, or cycloalkyl group.
  • fused heterocyles include, without limitation, tetrahydroquinoline and dihydrobenzofuran.
  • tetrahydroquinoline and dihydrobenzofuran.
  • compounds where an annular O or S atom is adjacent to another O or S atom are also included.
  • heteroaryl refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 ⁇ -electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S.
  • heteroaryl is also meant to encompass monocyclic and bicyclic groups.
  • a heteroaryl group may be pyrimidinyl, pyridinyl, benzimidazolyl, thienyl, benzothiazolyl, benzofuranyl and indolinyl.
  • Preferred heteroalkyl groups comprise a C 1 -C 6 alkyl group and a heteroaryl group having 5, 6, 9, or 10 ring atoms.
  • Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms.
  • heteroaralkyl groups examples include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imida ⁇ olylethyl, thiazolylmethyl, and thiazolylethyl.
  • pyridylmethyl pyridylethyl
  • pyrrolylmethyl pyrrolylethyl
  • imidazolylmethyl imida ⁇ olylethyl
  • thiazolylmethyl examples of preferred heteroaralkyl groups.
  • C n -C n heterocyclyl or “C n -C m “ heteroaryl means a heterocyclyl or heteroaryl having from “n” to "m” annular atoms, where "n” and “m” are integers.
  • a C 5 -C 6 -heterocyclyl is a 5- or 6- membered ring having at least one heteroatom, and includes pyrrolidinyl (C 5 ) and piperidinyl (C 6 );
  • C 6 -hetoaryl includes, for example, pyridyl and pyrimidyl.
  • arylene is an aryl, heteroaryl, or heterocyclyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups.
  • Preferred heterocyclyls and heteroaryls include, but are not limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, pyridotriazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H.6H-1 ,5,2- dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1 H-indazolyl, indolenyl, ind
  • a moiety e.g., cycloalkyl, hydrocarbyl, aryl, heteroaryl, heterocyclic, urea, etc.
  • a moiety e.g., cycloalkyl, hydrocarbyl, aryl, heteroaryl, heterocyclic, urea, etc.
  • the group optionally has from one to four, preferably from one to three, more preferably one or two, non- hydrogen substituents.
  • Suitable substituents include, without limitation, halo, hydroxy, oxo (e.g., an annular -CH- substituted with oxo is -C(O)-) nitro, halohydrocarbyl, hydrocarbyl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, acyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido groups.
  • Preferred substituents, which are themselves not further substituted are:
  • R 30 and R 31 are each independently hydrogen, cyano, oxo, carboxamido, amidino, Ci-C 8 hydroxyalkyl, C 1 -C 3 alkylaryl, aryl-C r C 3 alkyl, C 1 -C 8 alkyl, C r C 8 alkenyl, C 1 -C 8 alkoxy, Ci-C 8 alkoxycarbonyl, aryloxycarbonyl, 3IyI-C 1 -C 3 alkoxycarbonyl, C 2 -C 8 acyl, Ci-C 8 alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, aroyl, aryl, cycloalkyl, heterocyclyl, or heteroaryl, wherein
  • halohydrocarbyl is a hydrocarbyl moiety in which from one to all hydrogens have been replaced with one or more halo.
  • halogen refers to chlorine, bromine, fluorine, or iodine.
  • acyl refers to an alkylcarbonyl or arylcarbonyl substituent.
  • acylamino refers to an amide group attached at the nitrogen atom (Ae., R-CO-NH-).
  • carbamoyl refers to an amide group attached at the carbonyl carbon atom (i.e., NH 2 -CO-). The nitrogen atom of an acylamino or carbamoyl substituent is additionally substituted.
  • sulfonamido refers to a sulfonamide substituent attached by either the sulfur or the nitrogen atom.
  • amino is meant to include NH 2 , alkylamino, arylamino, and cyclic amino groups.
  • ureido refers to a substituted or unsubstituted urea moiety.
  • radical means a chemical moiety comprising one or more unpaired electrons.
  • a moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent.
  • substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluoro- 3-propylphenyl.
  • substituted n-octyls include 2,4-dimethyI-5- ethyl-octyl and 3-cyclopentyl-octyl. Included within this definition are methylenes (-CH 2 -) substituted with oxygen to form carbonyl -CO-).
  • an "unsubstituted" moiety as defined above e.g., unsubstituted cycloalkyl, unsubstituted heteroaryl, etc. means that moiety as defined above that does not have any of the optional substituents for which the definition of the moiety (above) otherwise provides.
  • an "aryl” includes phenyl and phenyl substituted with a halo
  • "unsubstituted aryl” does not include phenyl substituted with a halo.
  • the invention comprises compounds of formula (I), that are inhibitors DNMT1 and DNMT3b2:
  • R 3 and R 4 independently represent H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C r C 6 -alkyl-cycloalkyl, -Ci-C 6 -alkyl- heterocyclyl, -C r C 6 -alkyl-aryl, -CrC 6 -alkyl-heteroaryl, -d-Cealkoxy-aryl or - (CH 2 ) 1-6 -T, wherein C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, at each occurrence, are optionally substituted; or R 3 and R 4 taken together with the nitrogen to which they are attached form a C 5 -C 9 heterocyclyl
  • R 2 is H, halo, CF 3 , C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 alkoxy, -NH-C 1 -C 6 alkyl, or -S- C 1 -C 6 alkyl, wherein C 1 -C 6 alkyl, C 2 -C 6 alkenyl, and C 2 -C 6 alkynyl are each optionally substituted;
  • a and B independently are F, Cl, -OH, H 1 -NHR, or -OR;
  • R at each occurrence is independently benzyl or C 1 -C 4 alkyl, wherein benzyl and C 1 -C 4 alkyl are optionally substituted;
  • W is CH, N, CR, or C-halogen;
  • X is CH, N, C-C r C 6 -alkyl, or C-halogen;
  • D is CH, or N;
  • Z is -L-C(H)(NH 2 )-COOR 7 , -L-NR 19 R 20 , or heterocyclyl, wherein heterocyclyl is optionally substituted;
  • L is a bond or is -(CR 17 Ri 8 )I -6 -;
  • each R 17 and R 18 independently is H or C r C 6 -alkyl, wherein Ci-C 6 -alkyl is optionally substituted;
  • C-rC 6 -alkyl and heteroaryl are optionally substituted; and R 7 is H or Ci-C 6 -alkyl.
  • R 2 is H, halo, C 1 -C 6 alkyl, C 2 -C 6 alkenyl or -S- Ci-C 6 alkyl, wherein C 1 -C 6 alkyl and C 2 -C 6 alkenyl, at each occurrence, are optionally substituted;
  • R 3 and R 4 independently represent H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C r C 6 -alkyl-cycloalkyl, -C-i-Ce-alkyl-heterocyclyl, -C r C 6 -a!ky!-aryl, -C 1 -C 6 - alkyl-heteroaryl, -C r C 6 alkoxy-aryl or -(CH 2 )i
  • Ri 4 is C 1 -C 6 alkyl, aryl or heteroaryl and R 15 is aryl, wherein C 1 -C 6 alkyl, aryl, and heteroaryl, at each occurrence, are optionally substituted;
  • W and X are independently CH or N;
  • R 16 is H, C 1 -C 6 alkyl, -C-pCe-alkyl-aryl, -CrCe-alkyl-heteroaryl, or -C 2 -C 6 alkenyl-aryl, wherein C 1 -C 6 alkyl, aryl, and heteroaryl, at each occurrence, are optionally substituted;
  • Z is -L-C(H)(NH 2 )-COOR 7 , -L-NR 19 R 20 , or heterocyclyl, wherein heterocyclyl is optionally substituted;
  • L is a bond or is -(CR 17 Ris)i-6-;
  • R 7 is H or CrCe-alkyl.
  • H-A and pharmaceutically acceptable salts and complexes thereof, wherein A is H, halogen, or OH;
  • R 2 is H, halo, Ci-C 6 alkyl, C 2 -C 6 alkenyl or -S- C 1 -C 6 alkyl, wherein Ci-C 6 alkyl and C 2 -C 6 alkenyl, at each occurrence, are optionally substituted;
  • R 3 and R 4 independently represent H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C-i-Ce-alkyl-cycloalkyl, -Ci-Ce-alkyl-heterocyclyl, -Ci-C 6 -alkyl-aryl, -C 1 -C 6 - alkyl-heteroaryl, -Ci-C 6 alkoxy-aryl or -(CH 2 )i
  • R 14 is C 1 -C 6 alkyl, aryl or heteroaryl and R 15 is aryl, wherein C 1 -C 6 alkyl, aryl, and heteroaryl, at each occurrence, are optionally substituted
  • R 7 is H or Ci-C 6 -alky ⁇
  • Y is S, O, or -N(R 16 )-
  • R 16 is H, C 1 -C 6 alkyl, -C r C 6 -alkyl-aryl, -CrCe-alkyl-heteroaryl, Or -C 2 -C 6 alkenyl-aryl, wherein C 1 -C 6 alkyl, aryl, and heteroaryl, at each occurrence, are optionally substituted
  • R 7 is H or Ci-C 6 -alky
  • L 1 is -(CR 17 Ri 8 )i-6-;
  • R 17 and R 18 independently are H or C r C 6 -alkyl, wherein C r C 6 -alkyl is optionally substituted.
  • R 7 is H.
  • L 1 is -CH 2 CH 2 -.
  • R 2 is H, halogen, C 1 -C 3 alkyl, -S-C 1 -C 2 alkyl, or C 2 -C 3 alkenyl.
  • R 2 is H or halogen.
  • Y is S.
  • R 3 and R 4 are independently H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CrCe-alkyl-cycloalkyl, -CrC 6 -alkyl-aryl, -CpCe-alkyl-heteroaryl, -CrCealkoxy-aryl or
  • R 4 are both H.
  • R 3 is H and R 4 is C 1 -C 6 alkyl optionally substituted with 1, 2, or 3 groups independently selected from
  • both R 3 and R 4 are C 1 -C 6 alkyl, wherein Ci-C 6 alkyl is independently optionally substituted with 1 , 2, or 3 groups independently selected from OH, CO 2 H, NH 2 , N(C 1 -C 3 alkyl) 2 , C 1 -C 3 alkoxy, and phenyl.
  • R 3 is -
  • C- ⁇ -C 3 -all ⁇ yl-aryl and R 4 is C 1 -C 6 alkyl, wherein C 1 -C 6 alkyl is optionally substituted with 1 , 2, or 3 groups independently selected from OH, CO 2 H 1 NH 2 , N(C 1 -C 3 alkyl) 2 , C 1 -C 3 alkoxy, and phenyl.
  • R 3 is H and R 4 is C 3 -C 8 -cycloalkyl.
  • cycloalkyl is cyclopropyl, cyclohexyl, or cyclooctanyl.
  • R 3 is H and R 4 is aryl, wherein aryl is optionally substituted.
  • aryl is phenyl, naphthyl, or fluorenyl.
  • aryl is unsubstituted or is substituted with 1 , 2, or 3 groups independently selected from OCH 3 , NH 2 , NHCH 3 , NO 2 , C 1 -C 3 alkyl, halogen, CF 3 , CN, OH, NH 2 SO 2 -, and phenyl.
  • R 3 is H and R 4 is heterocyclyl.
  • heterocyclyl is pyrrolidinonyl.
  • R 3 is H and R 4 is -C r C 6 -alkyl-heteroaryl, wherein heteroaryl and C 1 -C 6 alkyl are optionally substituted.
  • heteroaryl is imidazolyl, indolyl, thiophenyl, pyridinyl, or dihydroindenyl.
  • heteroaryl is unsubstituted or is substituted with 1, 2, or 3 groups independently selected from methoxy and phenyl-CrC 3 -alkoxy-.
  • R 3 is H and R 4 is -C r C 6 -alkyl-aryl, wherein aryl and C 1 -C 6 alkyl are optionally substituted.
  • aryl is phenyl, naphthyl, or fluorenyl.
  • aryl is unsubstituted or is substituted with 1 , 2, or 3 groups independently selected from OCH 3 , NH 2 , NHCH 3 , NO 2 , C 1 -C 3 alkyl, halogen, CF 3 ,
  • aryl is phenyl, optionally substituted with 1 , 2, or 3 groups independently selected from OCH 3 , NH 2 ,
  • T is selected from -NH-CO-phenyl, NH-SO 2 -naphthyl, -S-CH 2 -phenyl, -NH-CO-methyl, and -NH-CO- furanyl.
  • R 3 and R 4 are selected from -NH-CO-phenyl, NH-SO 2 -naphthyl, -S-CH 2 -phenyl, -NH-CO-methyl, and -NH-CO- furanyl.
  • R 4 taken together with the nitrogen to which they are attached form a C 5 -C 9 heterocyclyl ring or a heteroaryl ring, wherein said ring is optionally substituted.
  • R 4 taken together with the nitrogen to which they are attached form a C 4 -C 8 heterocyclyl ring, wherein said ring is optionally substituted.
  • R 3 and R 4 taken together with the nitrogen to which they are attached form a pyrrolidinyl, azetidinyl, or piperidinyl ring.
  • Y is -N(R 16 )-.
  • R 16 is
  • R 16 is
  • CrC 6 -alkyl is unsubstituted or is substituted with NO 2 .
  • R 16 is aryl, wherein aryl is optionally substituted.
  • R 16 is -
  • Ri 6 is -
  • aryl is phenyl, wherein phenyl is unsubstituted or is substituted with 1 , 2, or 3 groups independently selected from NO 2 , Ci-C 3 -alkoxy, CN, or CF 3 .
  • R 16 is -
  • heteroaryl is pyridinyl.
  • R 3 and R 4 are both H.
  • R 3 is H, and R 4 is CrC 6 -alkyl.
  • L 2 is a bond or is -CH 2 -;
  • R 2 is H or halogen;
  • R 3 is H, C 1 -C 6 alkyl, or -Ci-C 6 -alkyl-aryl, wherein C 1 -C 6 alkyl and aryl, at each occurrence, are optionally substituted;
  • R 4 is H or Ci-C 6 alkyl, wherein C 1 -C 6 alkyl is optionally substituted;
  • R 8 is H, -CO 2 H, or CO 2 CH 3 ;
  • R 9 is absent, H or C 1 -C 6 alkyl, wherein C 1 -C 6 alkyl is optionally substituted; W is N or CH; Y is S or O; and
  • Q is N, CH or O, provided that when Q is O, R 9 is absent.
  • R 3 and R 4 are both H.
  • R 3 is - C-i-Ce-alkyl-aryl and R 4 is H, wherein aryl is optionally substituted.
  • aryl is phenyl.
  • phenyl is unsubstituted or is substituted with phenyl.
  • R 2 is H, or halogen
  • R 3 and R 4 independently represent H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, cycloalkyl, aryl or (-Ci-C 6 -alkyl)-aryl, wherein C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, cycloalkyl, aryl and (-Ci-C 6 -alkyl)-aryl are each optionally substituted; or R 3 and R 4 taken together with the nitrogen to which they are attached form a C 5 -C 9 - heterocyclyl ring or a heteroaryl ring, wherein said ring is optionally substituted;
  • L 3 is a bond or is -(CRi 7 Ri8)i-e-;
  • Ri 7 and R 18 at each occurrence are independently H or Ci-C 6 -alkyl, wherein C 1 -
  • C 6 -alkyl is optionally substituted;
  • L 3 is - CHRi 7 CHR 18 -, wherein R 17 and Ri 8 independently are H, or CrC 6 alkyl, and where C 1 -C 6 alkyl at each occurrence is optionally substituted.
  • C 1 -C 6 alkyl is unsubstituted or is substituted with NH 2 .
  • L 3 is - CH 2 CH 2 CH 2 -.
  • R 19 is C 1 -C 6 alkyl, wherein C 1 -C 6 alkyl is optionally substituted.
  • C 1 -C 6 alkyl is unsubstituted or is substituted with NH 2 .
  • R 19 is heteroaryl.
  • heteroaryl is pyrimidin-2(1 H)-one.
  • pyrimidin-2(1H)-one is unsubstituted or is substituted with amine.
  • the invention provides a composition comprising a compound according to any one of paragraphs [0047]-[0096] or as depicted in any of the examples and tables herein together with a pharmaceutically acceptable excipient, diluent, or carrier.
  • a pharmaceutically acceptable excipient diluent, or carrier.
  • Compounds of the invention may be formulated by any method well known in the art.
  • pharmaceutically acceptable means a non-toxic material that is compatible with a biological system in a cell, cell culture, or tissue sample and that does not interfere with the effectiveness of the biological activity of the active ingredient(s).
  • compositions according to the invention may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art.
  • diluents fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art.
  • the preparation of pharmaceutically acceptable formulations is described in, e.g., Remington's Pharmaceutical Sciences, 18th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, PA, 1990.
  • pharmaceutically acceptable salts refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects.
  • salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric add, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid.
  • inorganic acids for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric add, and the like
  • organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid
  • the compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula -NR + Z-, wherein R is hydrogen, alkyl, or benzyl, and Z is a counterion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).
  • R is hydrogen, alkyl, or benzyl
  • Z is a counterion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulf
  • salt is also meant to encompass complexes, such as with an alkaline metal or an alkaline earth metal.
  • the active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to an enzyme-inhibiting effective amount without causing serious toxic effects to the cell.
  • the third aspect of the invention provides a method of inhibiting DNMT1 and/or DNMT3b2 enzymes, the method comprising contacting the enzyme(s) with a compound according to any one of paragraphs [0047]-[0097], or as depicted in any of the tables herein, or with a composition according to paragraph [0098]-[0102].
  • l ⁇ h ⁇ bitiori of DNMT1 ahd/or DNMT3b2 " enzymes can be in a cell or a multicellular organism.
  • the method according to this aspect of the invention comprises administering to the organism an effective DNMT1- and/or DNMT3b2-inhibiting amount of a compound according to any one of paragraphs [0047]-[0097] or as depicted in any of the tables herein, or a composition according to paragraph [0098]-[0102].
  • the organism is a mammal, more preferably a primate, most preferably a human.
  • Preferred compounds according to the invention include those described in the examples below. Compounds were named using Chemdraw Ultra version 6.0.2 or version 8.0.3, which are available through Cambridgesoft.com, 100 Cambridge Park Drive, Cambridge, MA 02140, Namepro version 5.09, which is available from ACD labs, 90 Adelaide Street West, Toronto, Ontario, M5H, 3V9, Canada, or were derived therefrom.
  • the compounds of the invention can be prepared according to the reaction schemes or the examples illustrated below utilizing methods known to one of ordinary skill in the art. These schemes serve to exemplify some procedures that can be used to make the compounds of the invention. One skilled in the art will recognize that other general synthetic procedures may be used.
  • the compounds of the invention can be prepared from starting components that are commercially available. Any kind of substitutions can be made to the starting components to obtain the compounds of the invention according to procedures that are well known to those skilled in the art.
  • Step 3 methyl ⁇ -ftert-butoxycarbonyla ' minoy ⁇ -C ⁇ SaS ⁇ S.eR ⁇ aR ⁇ .Z-dimethyl-e-r ⁇ - (phenethylamino)-9H-purin-9-yl)-tetrahvdrofurof3,4-di ⁇ ,31dioxol-4-v ⁇ methylthio')butanoate 5a
  • NaH (30 mg 60% mineral oil suspension, 0.72 mmol) was added to a solution of ((3aR,4R,6R,6aR)-2,2-dimethyl-6-(6-(phenethylamino)-9H-purin-9-yl)-tetrahydrofuro[3,4- d][1,3]dioxol-4-yl)methanol 3a (149 mg, 0.36 mmol) in THF (4 ml_) at O 0 C and stirred for 15 min.
  • pTsCl (76 mg, 0.4 mmol) was then added and the reaction mixture was allowed to stir for 1 hour at O 0 C. It was diluted in EtOAc (15 ml_) and washed sequentially with water (10 mL) and NaCI sat solution (10 mL). The organic layer was dried with Na 2 SO 4 , filtered and concentrated in vacuo. The crude product was dissolved in dry MeOH and treated with a solution prepared by mixing tert-butyl 2-oxo-tetrahydrothiophen-3-ylcarbamate 4 (114 mg, 0.52 mmol) and NaOMe (1.05 mL 0,5M solution, 0.52 mmol) for 15 min at room temperature.
  • Step 4 2-amino-4-f((2S,3S,4R.5R)-3,4-dihvdroxy-5-(6-(phenethylaminoV9H-purin-9-yl)- tetrahvdrofuran-2-yl)methylthio)butanoic acid 6a.
  • Examples 2-9, compounds 5b-5i, Table 2, were prepared in a manner similar to example 1 , scheme 1a, utilizing the appropriate amine.
  • Step 1 methyl 4-(((3aS,4S,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2.2-dimethyl-tetrahvdrofurol ' 3.4- d1f1 ,31dioxol-4-yl)methylthio)-2-(tert-butoxycarbonylamino)butanoate 8
  • NaH 310 mg 60% mineral oil suspension, 7.74 mmol
  • ((3aR,4R,6R,6aR)-6-(6-amino-9H-purin-9-yl)-2,2-dimethyl-tetrahydrofuro[3,4-d][1 ,3]dioxol-4- yOmethanol 7 (1.19 g, 3.87 mmol) in THF (4 ml_) at O 0 C and stirred for 15 min.
  • pTsCI (810 mg, 4.25 mmol) was then added and the reaction mixture was allowed to stir for 1 hour at O 0 C. It was diluted in EtOAc (15 mL) and washed sequentially with water (10 ml_) and NaCI sat solution (10 mL).
  • Step 2 methyl 4-(((3aS,4S.6R.6aR)-6-(6-(4H-t2,4-triazol-4-yl)-9H-purin-9-yl)-2,2-dimethyl- tetrahydrofuro[3,4-di ⁇ ,31dioxol-4-yl)methylthio)-2-(tert-butoxycarbonylamino)butanoate 1
  • the dihydrochloride salt 9 (398 mg, 1.85, 5 equiv.) was added to a solution of 8 (134 mg, 0.37 mmol) in dry pyridine (4 mL) and the reaction mixture was refluxed for 16h.
  • Step 3 methyl 2-(tert-butoxycarbonylamino)-4-(((3aS,4S.6R,6aRV2,2-dimethyl-6-(6-(3- morpholinopropylamino)-9H-purin-9-yl)-tetrahvdrofuror3,4-d1[1 ,31dioxol-4- yl)methylthio)butanoate 11a
  • Step 4 2-amino-4-(((2S.3S.4R,5R)-3,4-dihvdroxy-5-(6-(3-morpholinopropylamino)-9H-purin-9- yl)-tetrahvdrofuran-2-yl)methylthio)butanoic acid 12a.
  • Step 1 methyl 2-(tert-butoxycarbonylamino)-4-(((3aS,4S.6R,6aR)-6-methoxy-2.2-dimethyl- tetrahvdrofurof3,4-di ⁇ ,31dioxol-4-yl)methylthio)butanoate 14
  • Step 2 methyl 2-(((9H-fluoren-9-vnmethoxy ⁇ carbonylaminoV4-(((3aS,4S,6R,6aRV6- methoxy-2,2-dimethyl-tetrahvdrofurof3,4-cn ⁇ ,31dioxol-4-yl)methylthio)butanoate 15
  • a solution of 14 (11.0 g, 25.3 mmol) in dry 1 ,2-dichloroethane (50 ml) was placed under a nitrogen atmosphere and cooled to O 0 C.
  • TMSOTf (4.6 ml, 25.4 mmol) was then added and the reaction was stirred for 1.5 hours at O 0 C.
  • Acetyl chloride (8.9 ml, 0.125 mol) was added slowly to this solution via syringe over a period of one hour and the reaction mixture was stirred overnight as it warmed to room temperature. The reaction was then quenched slowly with saturated NaHCO 3 .
  • Step 5 2-amino-4-(((2S,3S,4R.5R)-5-(6-(2-(dimethylamino)ethylaminoV9H-purin-9-v ⁇ -3,4- dihvdroxy-tetrahvdrofuran-2-yl)methylthio)butanoic acid 19a.
  • Step 1 Tert-butyl 2-(1-(dimethylamino)naphthalene-5-su]fonamido ⁇ ethylcarbamate 20
  • Step 3 2-amino-4-f((2S,3S,4R,5R)-5-(6-(2-(5-(dimethylamino)naphthalene-1- sulfonamido)ethylamino)-9H-purin-9-v ⁇ -3.4-dihvdroxy-tetrahvdrofuran-2- yl)methylthio)butanoic acid 19d.
  • Examples 26-42, compounds 24c-24s, Table 3, were prepared according to scheme 2, utilizing either method A or method B.
  • Step 1 (2R,3R,4S,5R)-2-(6-(2-(biphenyl-4-yl)ethylamino)-2-chloro-9H-purin-9-yl)-5- (hydroxymethyl)-tetrahydrofuran-3,4-diol 26
  • Step 2 ((3aR,4R,6R.6aRV6-(6-(2-(biphenyl-4-vnethylamino)-2-chloro-9H-purin-9-yl)-2,2- dimethyl-tetrahvdrofuror3,4-d1H ,31dioxol-4-v0methanol 27
  • Step 3 ((3aR.4R,6R.6aR)-6-(6-(2-(biphenyl-4-yl)ethylamino)-2-chloro-9H-purin-9-yl)-2,2- dimethyl-tetrahvdrofurof3,4-diri ,31dioxol-4-yl)methyl 4-methylbenzenesulfonate 28
  • Step 4 Methyl 4-(((3aS.4S.6R,6aR)-6-(6-(2-( ' biphenyl-4-vnethylamino)-2-chloro-9H-purin-9-vn- 2.2-dimethyl-tetrahvdrofurof3,4-dlf1.31dioxol-4-vnmethylthio)-2-(tert- butoxycarbonylamino)butanoate 29
  • Step 1 (2S.3S.4R.5RV2-((3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-4-methoxy-4- oxobutylth) ⁇ )methyl)-5-(6-(2-(tert-butoxycarbonylamino)ethylamino)-2-chloro-9H-purin-9-v ⁇ - tetrahydrofuran-3,4-diyl diacetate 31
  • Step 2 (2S,3S.4R,5RV2-((3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-4-methoxy-4- oxobutylthio)methyl)-5-(6-(2-arninoethylamino)-2-chloro-9H-pur ' m-9-v ⁇ -tetrahvdrofuran- 3,4-diyl diacetate 32
  • Step 3 2-amino-4-(((2S,3S.4R.5RV5-(2-chloro-6-(2-( r 4-cvanobenzamido)ethylamino)-9H-Durin- 9-yl)-3,4-dihvdroxy-tetrahvdrofuran-2-yl)methylthio)butanoic acid 33a [0143] A solution of 32 (76mg, 0.075 mmol), triethylamine (62.6 ⁇ l, 0.45 mmol) and 4- cyanobenzoyl chloride (18.5 mg, 0.112 mmol) in 0.75 -1.0 ml THF was stirred for 16 hours at room temperature.
  • Examples 51-61 compounds 33b-33l, Table 4 were prepared from compound 32 and the appropriate acid chloride, scheme 3, as described for example 50, step 3. Table 4
  • the title compound 35a was prepared in 35% yield (610 mg) using the general procedure and employing 3.92 g of di-ferf-butyl dicarbonate instead of the indicated amount and the product was obtained as white solid after recrystalized from hexanes.
  • Methane sulfonyl chloride (103 mg, 0.9 mmol) was added to a solution of di-tert-butyl 2-hydroxypropane-1,3-diyldicarbamate 35a (174.2 mg, 0.6 mmol) in 1.0 ml pyridine. The reaction was mixed and left to stand for 30 minutes. Methylene chloride (10 ml) was then added and the reaction was washed with H 2 O (3 x 10 ml). The organic phase was dried with MgSO 4 , filtered and evaporated to give the crude mesylate.
  • Examples 63-66, Table 5, were prepared in a manner similar to example 62, scheme 3, using compounds 35b-e in place of 35a The compounds were isolated as the formate salts after prep-HPLC purification in 20.6%; 33.8%; 18.9; and 47% yields respectively.
  • Step 1 (2S,4S)-methyl 4-(acetylthio)-1-methylpyrrolidine-2-carbo ⁇ ylate 44
  • Mesyl chloride (170 ⁇ L, 251 mg, 2.25 mmol) was added to a solution of (2S.4R)- methyl 4-hydroxy-1-methylpyrrolidine-2-carboxylate 43 (239 mg, 1.5 mmol, prepared according to the procedure of R.L. Elliott et al., Synthesis, 1995, 772) in pyridine (3 mL) and stirred at room temperature overnight.
  • the reaction mixture was diluted in DCM (20 mL) and washed with water (15 mL).
  • Step 2 (2S.4SVmethyl 4-(((3aS,4S.6R.6aRV6-(6-amino-9H-Purin-9-vn-2.2-dimethyl- tetrahvdrofuro[3,4-di ⁇ ,31dioxol-4-yl)methylthio)-1 -methylpyrrolidine-2-carboxylate 46
  • a solution of thioacetate 44 (95 mg, 0.43 mmol) in MeOH (2mL) was treated with 0.5 M solution of NaOMe (876 ⁇ l_).
  • Step 3 (2S.4SV4-(((2S.3S.4R.5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihvdroxy-tetrahvdrofuran-2- yl)methylthio)-1 -methylpyrrolidine-2-carboxylic 47
  • Example 72 compound 48a, was prepared in a manner similar to example 71, were step 1 , was conducted according to the procedure in J. Org. Chem. 1196, 61 , 2226, replacing
  • Example 73, compound 48b, was prepared in a manner similar to example 71, scheme 7. Step 1 was carried out according to the procedure for example 72 replacing (2S.4R)- methyl 4-hydroxy-1-methylpyrrolidine-2-carboxylate 43, with (2S,4R)-1-tert-butyl 2-methyl 4- hydroxypiperidine-1 ,2-dicarboxylate.
  • Example 74 compound 48c, was prepared in a manner similar to example 71. Step
  • Example 75 compound 48d, was prepared in a manner similar to example 71, replacing (2S,4R)-methyl 4-hydroxy-1-methylpyrrolidine-2-carboxylate 43, with (R)-tert-butyl 2-
  • Example 76 compound 48e, was prepared in a manner similar to example 71, replacing (2S,4R)-methyl 4-hydroxy-1-methylpyrrolidine-2-carboxylate 43, with (SJ-tert-butyl 2-
  • Step 1 (2S,4S)-1-tert-butyl 2-methyl 4-(((3aS,4S.6R,6aR)-6-(6-(4H-1 ,2,4-triazol-4-vn-9H-purin-
  • Step 2 (2S,4S)-1-tert-butyl 2-methyl 4-( «3aS,4S,6R,6aRV2,2-dimethyl-6-(6-(3- phenylpropylamino)-9H-purin-9-yl)-tetrahvdrofuror3,4-dlf1 ,31dioxol-4-yl)methylthio)pyrrolidine-
  • Step 3 (2S ⁇ S)-4-(((2S,3S.4R,5R)-3.4-dihvdroxy-5-(6-(3-phenylpropylaminoV9H-purin-9-vO- tetrahvdrofuran-2-v ⁇ methylthio)pyrrolidine-2-carboxylic acid 52a
  • Example 78, compound 52b was prepared according to the method shown in scheme 2a, example 43, replacing 2-(biphenyl-4-yl)ethanamine with 3-phenylpropan-1 -amine in stepi, and utilizing (2S,4S)-1-tert-butyl 2-methyl 4-(acetylthio)pyrrolidine-1,2-dicarboxylate in place of tert-butyl 2-oxo-tetrahydrothiophen-3-ylcarbamate in step 4.
  • Example 79, 52c was prepared as shown in scheme 1a, example 1, in step 3 replacing tert-butyl 2-oxo-tetrahydrothiophen-3-ylcarbamate with (2S,4S)-1 -tert-butyl 2-methyl 4-
  • Example 80, 52d was prepared according to scheme 2a, example 43, step 4 reacting 28 with (2S,4S)-1 -tert-butyl 2-methyl 4-(acetylthio)pyrrolidine-1 ,2-dicarboxylate in place of tert-butyl 2-oxo-tetrahydrothiophen-3-ylcarbamate.
  • Example 81 , 52e was prepared as shown in scheme 1a, example 1, in step 3 replacing tert-butyl 2-oxo-tetrahydrothiophen-3-ylcarbamate with (S)-tert-butyl 3-
  • Example 82, 52f, was prepared accordinglo " scheme 2a ⁇ example 43, " step 4 reacting "
  • Example 83, 52g was prepared according to the method shown in scheme 2a, example 43, replacing 2-(biphenyl-4-yl)ethanamine with 3-phenylpropan-1 -amine in stepi , and utilizing (S)-tert-butyl 3-(acetylthio)pyrrolidine-1-carboxylate in place of tert-butyl 2-oxo- tetrahydrothiophen-3-ylcarbamate in step 4.
  • Step 1 9- ⁇ 3aR,4R,6R.6aR)-6-((tert-butyldimethylsilyloxy)methyl)-2,2-dimethyl- tetrahvdrofurof3,4-d1f1,3ldioxol-4-yl)-6-chloro-2-iodo-9H-purine 55a
  • Step 2 9-((3aR,4R,6R,6aR)-6-((tert-butyldimethylsilyloxy ' )methylV2,2-dimethyl- tetrahvdrofurof3,4-d1f1 ,31dioxol-4-yl)-2-iodo-9H-purin-6-ami ⁇ e 56a
  • Step 3 ( r (3aR.4R,6R,6aR)-6-(6-amino-2-iodo-9H-purin-9-yl)-2.2-dimethyl-tetrahvdrofuror3,4- d1[1 ,31dioxol-4-yl)methanol 57a
  • Step 4 methyl 4-(((3aS,4S,6R,6aR)-6-(6-amino-2-iodo-9H-purin-9-yl)-2,2-dimethyl- tetrahvdrofurof3,4-din ,31dioxol-4-yl)methylthio)-2-(tert-butoxycarbonylamino)butanoate 58a
  • Step 5 2-amino-4-(((2S,3S,4R,5R)-5-(6-amino-2-iodo-9H-purin-9-yl)-3.4-dihvdroxy- tetrahvdrofuran-2-v0methylthio)butanoic acid 59a
  • Step 1 methyl 4-(((3aS.4S.6R.6aR)-6-(6-amino-2-methyl-9H-purin-9-yl)-2,2-dimethyl- tetrahvdrofuror3,4-diri ,31dioxol-4-yl)methylthio)-2-(tert-butoxycarbonylamino)butanoate 60
  • Step 2 2-Amino-4-(((2S.3S,4R,5R)-5-(6-amino-2-methyl-9H-purin-9-v ⁇ -3,4-dihvdroxy- tetrahydrofuran-2-yl)methylthio)butanoic acid 61
  • Stepi methyl 4-(((3aS,4S,6R,6aR)-6-(6-amino-2-chloro-9H-purin-9-ylV2,2-dimethyl- tetrahvdrofuror3,4-diri ,31dioxol-4-yl)methylthio)-2-(tert-butoxycarbonylamino)butanoate 64
  • Step 2 2-amino-4-(((2S.3S.4R,5R)-5-(6-amino-2-chloro-9H-purin-9-vn-3,4-dihvdroxy- tetrahvdrofuran-2-vDrnethylthio)butanoic acid 65
  • Step 1 (2S.4R)-methyl 4-(((2S,3S.4R.5R)-5-(6-amino-2-chloro-9H-purin-9-vO-3,4-dihvdroxy- tetrahvdrofuran-2-yl)methylthio)pyrrolidine-2-carboxylate 67a
  • Step2 (2S.4R)-4- «(2S,3S ⁇ R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3.4-dihvdroxy- tetrahvdrofuran-2-yl)methylthio)pyrrolidine-2-carboxylic acid 68a
  • Stepi (R)-tert-butyl 3-(((3aS,4S.6R,6aR)-6-(6-amino-2-chloro-9H-purin-9-ylV2.2-dimethyl- tetrahvdrofuror3,4-dlf 1 ,3ldioxol-4-vDmethylthio)pyrrolidine-1 -carboxylate 67b
  • Compound 67b was prepared in 33% yield as a clear syrup following the procedure detailed in step 4 for 58a, example 84, scheme 9, replacing 57a with (S)-tert-butyl 3- (acetylthio)pyrrolidine-i -carboxylate 66b.
  • Step 1 benzyl (S)-3-(((3aR,4R,6R,6aR)-4-(6-amino-9H-purin-9-v ⁇ -tetrahvdro-2.2- dimethylfuro[3,4-cnf 1 ,3ldioxol-6-vDmethylamino)-1 -(tert-butoxycarbonyl)propylcarbamate 71 [0194] To a solution of (S)-tert-butyl 2-(benzyloxycarbonylamino)-4-oxobutanoate 69 (4.0 ml, 1.41 mmol, prepared according to I. Weits et al, J. Org. Chem.
  • Step 3 (S)-4-(N-(2-nitrocinnamv ⁇ -N-(((2R.3S.4R,5RV5-(6-amino-9H-purin-9-yl)-tetrahvdro-3.4- dihvdroxyfuran-2-v0methv0amino)-2-aminobutanoic acid (74a)
  • Example 94 compound 74d, Table 8, was prepared according to the procedure described for example 91 , in step 2 using 72d, 4-nitrobutanal (previously reported by R. Kimura et al. Bull. Chem. Soc. Jpn., 2002, 75(11), 2517-2526, however it was prepared in 94% yield by DIBAL-H reduction of methyl 4-nitrobutanoate in DCM at -78 0 C) in place of aldehyde 72a.
  • Step 1 (S)-tert-butyl 4-((((3aR,4R,6R.6aRV6-(6-amino-9H-purin-9-vn-2.2-dimethyl- tetrahvdrofuror3,4-cn ⁇ ,31dioxol-4-vnmethyl)(methv ⁇ amino)-2-
  • Step 2 benzyl (S)-1-(tert-butoxycarbonv ⁇ -3-(N-(((3aR.4R.6R.6aRV4-(6-(ethvIaminoV9H-purin-)
  • Step 3 (S)-4-(N-(((2R,3S.4R,5R)-5-(6-(ethylamino)-9H-purin-9-vn-tetrahvdro-3.4- dihydroxyfuran-2-yl)methyl)-N-methylamino)-2-aminobutanoic acid 77
  • P26358 (SEQ ID NO.1) was cloned into the BamHt/Sall site of the pBlueBac4.5 vector
  • Hi-5 cells grown in suspension and maintained in serum-free medium (Sf900 Il supplemented with gentamycin) were infected with the abovementioned viruses at multiplicity of infection (MOI) varying from 1 to 3 during 84 hours at 27 0 C with agitation at 120 rpm on a rotary shaker. Infected cells were harvested by centrifugation at 398g for 15 min. after which a nuclear and cytosolic fractionation was performed. Fractions were frozen at -8O 0 C until purifications were performed.
  • MOI multiplicity of infection
  • DTT 0.5% NP-40, 25% glycerol, 1 ⁇ g/ml pepstatin, 2 ⁇ g/ml Aprotinin and leupeptin, 50 ⁇ g/ml
  • Buffer B (2OmM Tris pH 8.0, 1.5mM MgCI 2 , 5mM KCI, 5mM DTT, 0.5% NP-40, 25% glycerol,
  • Buffer B was 10%.
  • DNMT-1 enriched fractions were pooled according to SDS-PAGE analysis (coomassie staining) Final DNMT-1 protein preparations concentration were about 7mg/ml and purity above 95%.
  • DNMT3b2 was purified using only the nuclear extract as starting material.
  • the NaCI concentration of the latter was adjusted to 0.2M by diluting with Buffer D (5OmM NaPO 4 pH 7.8, 10% glycerol and 1mM EDTA) supplemented with the protease inhibitors described in Buffer A followed by centrifugation at 3000Og for 10 min..
  • the supernatant was loaded onto a Hitrap SPsepharose column (Amersham Biosciences) equilibrated with Buffer D with 0.05M NaCI. Column was washed with 8 CV of equilibration buffer and proteins were eluted with an 8 CV linear gradient of NaCI (from 0.05 to 1 M) in Buffer D.
  • DNMT3b2 containing fractions were pooled based on SDS-PAGE analysis. Selected fractions from this elution had conductivity varying from 15.1 to 28.2mS/cm. Finally, this SPsepharose eluate underwent buffer exchange, using PD-10 column (Amersham Biosciences) against Buffer D + 0.3M NaCI. Typical DNMT3b2 enzyme preparation had concentration of about 2.5mg/ml and approximately 70% purity. [0211] Purified DNMT-1 and DNMT3b2 protein stocks were aliquoted and frozen at -8O 0 C prior to use in enzymatic assay. DNA Methyltransferase Assay Reagents : -Enzyme: Cloned human DNA Methyltransferase (DNMT1 and DNMT3b2)
  • MYG 167 ATC GCA TCG ATC GCG ATT CGC GCA TCG GCG ATC MYG 166: GAT XCG XGA TGX GXG AAT XGX GAT XGA TGX GAT (X: 5-methylcytosine) -Substrate2: "" “ " “ ⁇ ' " " "
  • the mixture is pre-incubated at 37 0 C for 10min and then the SAM mix is added (2.25uL of 4OuM of S- Adenosyl-L-Methionine to make 3uM final, 1.65uL of 10X Buffer [5OmM Tris-HCI pH7.6, 5%Glycerol, 1mM EDTA, 100ug/mL BSA, 1 mM DTT]).
  • the final reaction is in 3OuL.
  • the reaction is incubated at 37°C for 15min and then transferred onto DEAE filtermat (Wallac cat# 1450-522) using Tomtec cell harvester.
  • the DEAE filtermat is washed with cold 2OmM NH4HCO3. When the filter is dry, it is covered with MeltiLex TM scintillant (Wallac cat#1450- 441 ). Counting is done in a Wallac beta-counter.
  • DNMT3b2 assay follows identical protocol, at enzyme concentration of 188nM. [0214] The activities of a number of compounds according to the invention measured by various assays are displayed in the table below.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Oncology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Hematology (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

L'invention concerne l'inhibition d'isoformes d'ADN-méthyltransférase DNMT1 et DNMT3b2. L'invention concerne des composés et des procédés permettant d'inhiber les isoformes DNMT1 et DNMT3b2.
EP06718809A 2005-01-21 2006-01-18 Inhibiteurs d'adn-methyltransferase Withdrawn EP1844062A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US64615605P 2005-01-21 2005-01-21
PCT/US2006/001791 WO2006078752A2 (fr) 2005-01-21 2006-01-18 Inhibiteurs d'adn-methyltransferase

Publications (1)

Publication Number Publication Date
EP1844062A2 true EP1844062A2 (fr) 2007-10-17

Family

ID=36692836

Family Applications (1)

Application Number Title Priority Date Filing Date
EP06718809A Withdrawn EP1844062A2 (fr) 2005-01-21 2006-01-18 Inhibiteurs d'adn-methyltransferase

Country Status (2)

Country Link
EP (1) EP1844062A2 (fr)
WO (1) WO2006078752A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011018435A1 (fr) 2009-08-10 2011-02-17 Institut Curie Procédé pour la prévision de la sensibilité d'une tumeur à un traitement épigénétique

Families Citing this family (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2623221T3 (es) 2005-05-20 2017-07-10 Methylgene Inc Inhibidores de la señalización del receptor de VEGF y del receptor de HGF
CA2742869A1 (fr) 2008-11-20 2010-05-27 President And Fellows Of Harvard College Fluoration de composes organiques
EP2646454B1 (fr) 2010-12-03 2015-07-08 Epizyme, Inc. Modulateurs à base de 7-déazapurine de l'histone méthyltransférase et procédés d'utilisation de ceux-ci
CN103391939B (zh) 2010-12-03 2016-03-09 Epizyme股份有限公司 作为表观遗传酶调节剂的经取代的嘌呤和7-氮杂嘌呤化合物
WO2012142162A2 (fr) 2011-04-12 2012-10-18 President And Fellows Of Harvard College Fluoration de composés organiques
WO2013062943A1 (fr) * 2011-10-24 2013-05-02 Glaxosmithkline Intellectual Property Development Limited Nouveaux composés
US9174984B2 (en) 2011-10-24 2015-11-03 Glaxosmithkline Intellectual Property (No.2) Limited Chemical compounds
WO2014026198A1 (fr) 2012-08-10 2014-02-13 Epizyme, Inc. Inhibiteurs de protéine méthyltransférase de dot1l et leurs procédés d'utilisation
US9597348B2 (en) 2012-09-06 2017-03-21 Epizyme, Inc. Method of treating leukemia
US9737493B2 (en) 2012-09-07 2017-08-22 University Of Louisville Research Foundation, Inc. Compositions and methods for modulating DNMT1 inhibitor activity
WO2014052622A1 (fr) 2012-09-26 2014-04-03 President And Fellows Of Harvard College Complexes de fluoration de nickel et leurs utilisations
CA2903303A1 (fr) 2013-03-15 2014-09-25 Epizyme, Inc. Procedes de synthese de composes de purine substitues
BR112016005317A2 (pt) * 2013-09-19 2017-09-12 Centre Nat Rech Scient derivados de quinazolina e seu uso como inibidores de dna metiltransferase
US10759764B2 (en) 2013-10-18 2020-09-01 President And Fellows Of Harvard College Fluorination of organic compounds
TWI791251B (zh) 2015-08-26 2023-02-01 比利時商健生藥品公司 使用作為ｐｒｍｔ５抑制劑之新穎經６-６雙環芳香環取代之核苷類似物
EP3207932A1 (fr) 2016-02-19 2017-08-23 Universität Stuttgart Inhibiteurs de méthyltransférase d'adn pour la thérapie du syndrome de rett
JP6909799B2 (ja) 2016-03-10 2021-07-28 ヤンセン ファーマシューティカ エヌ．ベー． Ｐｒｍｔ５阻害剤として使用するための置換ヌクレオシドアナログ
WO2017216726A1 (fr) 2016-06-13 2017-12-21 Glaxosmithkline Intellectual Property Development Limited Pyridines substituées en tant qu'inhibiteurs de dnmt1
MX2019003843A (es) 2016-10-03 2019-06-24 Janssen Pharmaceutica Nv Análogos novedosos de carbanucleósidos sustituidos de sistema anular monocíclico y bicíclico para su uso como inhibidores de prmt5.
ES2971035T3 (es) 2017-02-27 2024-06-03 Janssen Pharmaceutica Nv Uso de biomarcadores en la identificación de pacientes con cáncer que mostrarán respuesta al tratamiento con un inhibidor de prmt5
JP7352544B2 (ja) 2017-12-08 2023-09-28 ヤンセン ファーマシューティカ エヌ．ベー． 新規なスピロ二環式類似体
WO2020013712A1 (fr) * 2018-07-09 2020-01-16 Victoria Link Limited Inhibiteurs de dnmt1 utilisés en tant qu'agents thérapeutiques anticancéreux
TW202112375A (zh) 2019-06-06 2021-04-01 比利時商健生藥品公司 使用ｐｒｍｔ５抑制劑治療癌症之方法
CN116284188A (zh) * 2023-02-07 2023-06-23 南开大学 一种dna甲基转移酶dnmt1的双底物抑制剂及其应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2006078752A2 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011018435A1 (fr) 2009-08-10 2011-02-17 Institut Curie Procédé pour la prévision de la sensibilité d'une tumeur à un traitement épigénétique

Also Published As

Publication number Publication date
WO2006078752A3 (fr) 2007-02-15
WO2006078752A2 (fr) 2006-07-27

Similar Documents

Publication Publication Date Title
EP1844062A2 (fr) Inhibiteurs d'adn-methyltransferase
US20080132525A1 (en) Inhibitors of DNA Methyltransferase
US5736549A (en) Hypoxanthine and guanine compounds
Mansuri et al. Preparation of 1-(2, 3-dideoxy-. beta.-D-glycero-pent-2-enofuranosyl) thymine (d4T) and 2', 3'-dideoxyadenosine (ddA): general methods for the synthesis of 2', 3'-olefinic and 2', 3'-dideoxy nucleoside analogs active against HIV
FI93217B (fi) Menetelmä terapeuttisesti aktiivisten nukleosidijohdannaisten estereiden ja amidien valmistamiseksi
MXPA97002488A (en) Compounds of purine and guanine as inhibitors of
US20030144501A1 (en) Conformationally constrained L-nucleosides
MXPA92005623A (es) Derivados de nucleosido de pirimidina que tienen actividad antitumoral su preparacion y su uso.
WO2011005595A2 (fr) Analogues de 2-5a et leurs procédés d'utilisation
EP1827460A1 (fr) Nucleosides presentant une activite antivirale et anticancereuse
EP0684953A1 (fr) Inhibiteurs d'adenosine-kinase comprenant des derives de lyxofuranosyle
EP2227481A2 (fr) Composés nucléosidiques antiviraux
DK148745B (da) Ribofuranosylderivater til anvendelse som mellemprodukter ved fremstilling af 5'-deoxy-5-fluorcytidin eller 5'-deoxy-5-fluoruridin eller fysiologisk tolerable syreadditionssalte deraf
JP2022531899A (ja) Ｓｔｉｎｇモジュレータとしての修飾環状ジヌクレオシド化合物
WO2003053989A1 (fr) Derives nucleosides contenant un phosphate masque, et leur utilisation comme agents antiviraux
Zhang et al. Synthesis of neplanocin F analogues as potential antiviral agents
EP1198458B1 (fr) Analogues de ceramide, leur procede de preparation et leur utilisation comme agents antitumoraux
US20040122223A1 (en) Non-natural nucleotides and dinucleotides
EP1968971B1 (fr) Dérivés de dioxolanne pour le traitement du cancer
Silamkoti et al. Synthesis and biological activity of 2-fluoro adenine and 6-methyl purine nucleoside analogs as prodrugs for suicide gene therapy of cancer
CA2290364A1 (fr) Procede de production de tiazofurine et d'autres c-nucleosides
AU626600B2 (en) Nucleoside derivatives
JPH05148261A (ja) 置換２’，３’−ジデオキシ−５−トリフルオロメチルウリジン類
Bhattacharya et al. Studies on the synthesis of furo [3, 2-d] pyrimidine C-nucleosides: new inosine analogues with antiprotozoan activity
WO2006122207A1 (fr) 6-hydrazinopurine 2'-methyl ribonucleosides et nucleotides pour le traitement de hcv

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20070809

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL BA HR MK YU

17Q First examination report despatched

Effective date: 20080227

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20080909