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WO2008114009A1 - Composés de 10h-benzo[g]ptéridine-2,4-dione pour le traitement de troubles prolifératifs - Google Patents

Composés de 10h-benzo[g]ptéridine-2,4-dione pour le traitement de troubles prolifératifs Download PDF

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WO2008114009A1
WO2008114009A1 PCT/GB2008/000962 GB2008000962W WO2008114009A1 WO 2008114009 A1 WO2008114009 A1 WO 2008114009A1 GB 2008000962 W GB2008000962 W GB 2008000962W WO 2008114009 A1 WO2008114009 A1 WO 2008114009A1
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cancer
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Shankar Balasubramanian
Mallesham Bejugam
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Cambridge Enterprise Ltd
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Cambridge Enterprise Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D475/00—Heterocyclic compounds containing pteridine ring systems
    • C07D475/12—Heterocyclic compounds containing pteridine ring systems containing pteridine ring systems condensed with carbocyclic rings or ring systems
    • C07D475/14—Benz [g] pteridines, e.g. riboflavin

Definitions

  • This invention pertains generally to the field of G-quadruplex ligands, and more particularly, to certain 10H-benzo[g]pteridine-2,4-dione compounds ("BPD compounds") which, inter alia, (selectively.) bind (and stabilize) G-quadruplexes.
  • BPD compounds 10H-benzo[g]pteridine-2,4-dione compounds
  • the present invention also pertains to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions, both in vitro and in vivo, to (selectively) bind (and stabilize) G-quadruplexes, to inhibit telomerase, to alter expression of certain genes, to regulate cell proliferation, and in the treatment of proliferative disorders, such as cancer.
  • Ranges are often expressed herein as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent "about,” it will be understood that the particular value forms another embodiment.
  • Nucleic acid sequences rich in guanine are capable of forming four-stranded structures called G-quadruplexes, stabilized by Hoogsteen hydrogen bonding between a tetrad of guanine bases (see, e.g., Gellert et al., 1962; Guschlbauer et al., 1990).
  • Telomeric repeats in a variety of organisms have been shown to form these structures in vitro and high-resolution structures of the human telomeric sequence d(T2AG3)n have been solved by NMR spectroscopy (see, e.g., Wang et al., 1993) and X-ray crystallography (see, e.g., Parkinson et al., 2002).
  • Quadruplexes have also been shown to exist in vivo in Stylonychia lemnae macronuclei (see, e.g., Schaffitzel et al., 2001 ).
  • telomere formation has been shown to decrease the activity of the enzyme telomerase (see, e.g., Fletcher et al., 1998), which is responsible for elongating telomeres. Since elevated telomerase activity has been implicated in -85% of cancers (see, e.g., Mergny et al., 2002), this has become a significant strategy for drug development (see, e.g., Neidle et al., 2002) and molecules that bind to and stabilize G-quadruplexes have been identified (see, e.g., Mergny et al., 2002).
  • quadruplex-forming sequences include the fragile X syndrome repeat d(CGG)n (see, e.g., Fry et al., 1999; Fry et al., 1994; Fojtik et al., 2004), and the Cystatin B promoter (see, e.g., Saha et al., 2001), which has a region with sequence (CGCG 4 CG 4 ) 4 and is involved in epilepsy.
  • CGG fragile X syndrome repeat
  • Cystatin B promoter see, e.g., Saha et al., 2001
  • G-rich strands of the human insulin gene can form quadruplexes (see, e.g., Castati et al., 1996), as can the mouse Ms6-hm hypervariable satellite repeat (see, e.g., Weitzmann et al., 2002), with sequence (CAGGG) n .
  • the promoter regions of the RET protooncogene see, e.g., Sun et al., 2003
  • c-kit Rankin et al., 2005; Fernando et al., 2006
  • Ki-ras see, e.g., Cogoi et al., 2004
  • G-rich RNA can also fold into quadruplex structures, e.g. the insulin-like growth factor Il mRNA (see, e.g., Christansen et al., 1994). Efforts have been made to gauge the prevalence of G- quadruplexes in the human genome (see, e.g., Huppert et al., 2005).
  • quadruplex nucleic acids in nature is also supported by the observation of a number of naturally occurring quadruplex binding proteins, such as the helicases implicated in Bloom's (see, e.g., Sun et al., 1998) and Werner's (see, e.g., Fry et al., 1999) syndromes, the yeast telomere binding protein Rap1 (see, e.g., Giraldo et al., 1994) and the ⁇ subunit of the Oxytrchia nova telomere binding protein (see, e.g., Laporte et al., 1998).
  • the fragile X syndrome mental retardation protein has been shown to target G-quadruplex mRNA with a physiological consequence (see, e.g., Darnell et al., 2001).
  • telomeres The use of a quadruplex-specific small molecule probe (see, e.g., Chang et al., 2004) and also an antibody probe (see, e.g., Schaffitzel et al., 2001) has shown that quadruplex structures can exist at the ends of telomeres.
  • telomeres are those derived from telomeric repeat sequences.
  • Zahler et al. demonstrated that telomeric DNA folded into a quadruplex was not a competent substrate for the enzyme telomerase (see, e.g., Zahler et al., 1991).
  • Hurley and Neidle subsequently demonstrated that a quadruplex stabilising ligand could inhibit the extension of DNA substrate by human telomerase (see, e.g., Sun et al., 1997).
  • Telomerase is critical for immortality in most human cancers (see, e.g., Morin et al.,
  • telomeric DNA has potential as an anti-cancer therapeutic strategy (see, e.g., Neidle et al., 2002).
  • Biophysical studies on the human telomeric quadruplex have provided valuable insights into its structural (see, e.g., Wang et al., 1993; Parkinson et al., 2002) and dynamic properties (see, e.g., Phan et al., 2003; Ying et al., 2003).
  • telomere length a number of first generation of ligands that selectively bind quadruplexes and inhibit the action of human telomerase. It has been shown that quadruplex inhibitors can induce relatively rapid telomere attrition, and cell growth inhibition, usually accompanied by apoptosis (see, e.g., Gowan et al., 2002; Sumi et al., 2004; Shammas et al., 2004(a); Riou et al., 2002; Shammas et al., 2003), although there is one reported case of cell growth effects without changes in telomere length (see, e.g., Leonetti et al., 2004).
  • telomere elongation by telomerase an enzyme up-regulated in cancer cells
  • telomere elongation by telomerase an enzyme up-regulated in cancer cells
  • a large number of other putative quadruplex forming sequences have been identified in the human genome (see, e.g., Huppert et al., 2005; see, e.g., Todd et al., 2005).
  • one aim of the present invention is the provision of compounds which (selectively) bind G-quadruplexes, which are (selective) G-quadruplex ligands, telomerase inhibitors, antiproliferative agents, anti-cancer agents, etc., and which offer one or more of the above properties and advantages.
  • the present invention pertains to certain 10H-benzo[g]pteridine-2,4-dione compounds ("BPD compounds”), and the discovery of their surprising and unexpected activity as (selective) G-quadruplex ligands, telomerase inhibitors, antiproliferative agents, anti-cancer agents, etc.
  • BPD compounds 10H-benzo[g]pteridine-2,4-dione compounds
  • One aspect of the invention pertains to certain compounds, specifically, certain 10H-benzo[g]pteridine-2,4-dione compounds ("BPD compounds”), and their surprising and unexpected activity as (selective) (stabilizing) ligands of G-quadruplxes.
  • BPD compounds 10H-benzo[g]pteridine-2,4-dione compounds
  • Another aspect of the invention pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a BPD compound, as described herein, and a pharmaceutically acceptable carrier, diluent, or excipient.
  • Another aspect of the present invention pertains to a method of stabilizing a G-quadruplex, comprising contacting said G-quadruplex (e.g., within a living cell) with an effective amount of a BPD compound, as described herein. Such a method may be performed in vitro or in vivo.
  • Another aspect of the present invention pertains to a method of inhibiting telomerase (for example, inhibiting telomerase activity, inhibiting formation of telomerase complexes, inhibiting activity of telomerase complexes, etc.), comprising contacting said telomerase (e.g., within a living cell) with an effective amount of a BPD compound, as described herein.
  • a method of inhibiting telomerase for example, inhibiting telomerase activity, inhibiting formation of telomerase complexes, inhibiting activity of telomerase complexes, etc.
  • Such a method may be performed in vitro or in vivo.
  • Another aspect of the present invention pertains to a method of regulating cell proliferation comprising contacting a living cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • Another aspect of the present invention pertains to a method of treating a proliferative disorder in a subject comprising administering to said subject a therapeutically-effective amount of a BPD compound, as described herein.
  • the proliferative disorder is cancer.
  • Another aspect of the present invention pertains to a BPD compound as described herein for use in a method of treatment of the human or animal body by therapy, for example, in a method of treatment of a proliferative disorder, for example, cancer.
  • Another aspect of the present invention pertains to use of a BPD compound, as described herein, in the manufacture of a medicament for use in the treatment of a proliferative disorder.
  • the proliferative disorder is cancer.
  • Another aspect of the present invention pertains to a method of treatment comprising administering to a patient in need of treatment a therapeutically effective amount of a BPD compound as described herein, preferably in the form of a pharmaceutical composition.
  • the treatment is treatment of a proliferative disorder, for example, cancer.
  • kits comprising (a) a BPD compound as described herein, preferably provided as a pharmaceutical composition and in a suitable container and/or with suitable packaging; and (b) instructions for use, for example, written instructions on how to administer the compound.
  • Figure 1 illustrates the circular dichroism (CD) spectral data (ellipticity (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 (40 ⁇ M) with G-quadruplex DNA of htelo (4 ⁇ M), in K + and Na + containing buffer (50 mM Tris.HCI, pH 7.4, 10O mM KCI, and 10O mM NaCI):
  • Figure 2 illustrates the circular dichroism (CD) spectral data (ellipticity (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 with G-quadruplex DNA of htelo in K + containing buffer (50 mM Tris.HCI, pH 7.4, 100 mM KCI): (a) folded in the presence of 100 mM potassium without ligand (O 1 open hexagon)
  • CD circular dichroism
  • Figure 3 illustrates the circular dichroism (CD) spectral data (ellipticity (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 (40 ⁇ M) with G- quadruplex DNA of c-kit (4 ⁇ M) in K + containing buffer (50 mM Tris.HCI, pH 7.4, 100 mM KCI):
  • Ckit without salt and ligand (o, open circle)
  • Ckit-BPD5 BDP 5 (A, filled triangle)
  • Figure 4 is bar graph showing the percentage gene expression of c-kit in MCF-7 cells treated with BPD5 or BPD8. The figure shows the levels of expression for control cells treated with 10% DMSO in water (100% expression, blank bar), and for cells treated with the isoalloxazines BPD5 or BPD8 at 6 hours at 5 ⁇ M concentration (shaded bars).
  • Figure 5 is a bar graph showing the percentage gene expression of c-kit in HGC-27 cells treated with BPD8. The figure shows the levels of expression for control cells treated with 10% DMSO in water (100% expression, blank bar), and for cells treated with BPD8 at 2, 4 and 8 hours at 5 ⁇ M concentration (shaded bars).
  • Isoalloxazine also known as 10H-benzo[g]pteridine-2,4-dione, has the structure shown below.
  • riboflavin i.e., Vitamin B2
  • the present invention pertains generally to compounds which may be described as "10H-benzo[g]pteridine-2,4-dione compounds" ("BPD compounds”), and their surprising and unexpected ability to (selectively) bind (and stabilize) G-quadruplexes and to inhibit telomerase, and their use to inhibit telomerase, to regulate cell proliferation, and in the treatment of proliferative disorders, such as cancer.
  • BPD compounds 10H-benzo[g]pteridine-2,4-dione compounds
  • One aspect of the present invention pertains to compounds of the following formula, and pharmaceutically acceptable salts, solvates, and hydrates thereof (collectively referred to herein as "10H-benzo[g]pteridine-2,4-dione" or "BPD” compounds):
  • R C6 is independently -H, -G 1 , or -G 2 ;
  • R C7 is independently -H, -G 1 , or -G 2 ;
  • R C9 is independently -H, -G 1 , or -G 2 ;
  • each -G 1 if present, is independently -Ph, -NHPh, -NR A1 Ph, or -OPh;
  • -R -,A1 is independently saturated aliphatic C ⁇ alkyl; and each Ph is independently unsubstituted phenyl or phenyl substituted with one or more groups G 2 ;
  • each -G 2 if present, is independently:
  • each -X 1 - is independently -O-, -NH-, or -NR ⁇ -; each -R* 2 is independently saturated aliphatic C 1-4 alkyl; each -NR A3 R M is independently -Q 1 ; each -L A1 - is independently saturated aliphatic C ⁇ alkylene;
  • R N8B is independently -H or -G 3 ;
  • each G 3 is independently saturated aliphatic C ⁇ alkyl
  • R N1 ° is independently saturated aliphatic C ⁇ alkyl, -G 4 , -
  • G 4 is phenyl or Cs- ⁇ heteroaryl, and is independently unsubstituted or substituted with one or more substituents -G 6 ;
  • G 5 is phenyl or C 5-6 heteroaryl, and is independently unsubstituted or substituted with one or more substituents -G 6 ;
  • -L 10 - is independently saturated aliphatic C ⁇ alkylene; and -
  • each -G 6 is independently: -F, -Cl, -Br, -I, -R A5 , -CF 3 , -OH, -OR A5 , -OCF 3 , -SR A5 ,
  • each -X 2 - is independently -O-, -NH-, or -NR A5 -; each -R A5 is independently saturated aliphatic C 1-4 alkyl; each -NR A6 R A7 is independently -Q 5 ; each -I_ A2 - is independently saturated aliphatic C 2-8 alkylene;
  • each of -L 3 - and -L 8 - is independently saturated aliphatic C 2 -ealkylene;
  • -NR 3PA R 3PB is independently -Q 6 ;
  • -NR 8PA R 8PB is independently -Q 7 ;
  • each of -Q 1 , -Q 2 , -Q 3 , -Q 4 , -Q 5 , -Q 6 , and -Q 7 is independently: -NH 2 , -NHR B1 , -NR B1 2 , or -NR 82 R 83 ;
  • each R B1 is independently saturated aliphatic C ⁇ alkyl; and in each group -NR 82 R 83 , R 82 and R 83 , taken together with the nitrogen atom to which they are attached, form a 5-, 6-, or 7-membered non-aromatic ring having exactly 1 ring heteroatom or exactly 2 ring heteroatoms, wherein one of said exactly 2 ring heteroatoms is N, and the other of said exactly 2 ring heteratoms is independently selected from N and O.
  • R C6 is independently -H or -G 2 .
  • R C6 is independently -G 1 or -G 2 .
  • R C6 is independently -H.
  • R C6 is independently -G 1 . In one embodiment, R C6 is independently -G 2 .
  • R C7 is independently -H or -G 2 . In one embodiment, R C7 is independently -G 1 or -G 2 . In one embodiment, R C7 is independently -H. In one embodiment, R C7 is independently -G 1 . In one embodiment, R C7 is independently -G 2 . In one embodiment, R C9 is independently -H or -G 2 . In one embodiment, R C9 is independently -G 1 or -G 2 . In one embodiment, R C9 is independently -H. 5 In one embodiment, R C9 is independently -G 1 . In one embodiment, R C9 is independently -G 2 .
  • R A1 if present, is independently -Me, -Et, -nPr, or -iPr. In one embodiment, R A1 , if present, is independently -Me. 10
  • each -G 2 is independently -F, -Cl, -Br, -I 1 -R* 2 , -CF 3 , -OR A2 , or -OCF 3 .
  • each R* 2 if present, is independently -Me, -Et, -nPr, or -iPr. 15 In one embodiment, each R A2 , if present, is independently -Me.
  • each -L A1 - is independently -(CH 2 ) n i-, wherein n1 is independently 2, 3, 4, 5, 6, 7, or 8. In one embodiment, n1 is independently 2, 3, 4, 5, or 6. In one embodiment, n1 is independently 2, 3, or 4. 20
  • R N8B is independently -H. In one embodiment, R N8B is independently -G 3 .
  • G 3 if present, is independently -Me, -Et, -nPr, or -iPr. 25 In one embodiment, G 3 , if present, is independently -Me.
  • R N1 ° is independently saturated aliphatic C 1-6 alkyl, -G 4 , or -
  • R N1 ° is independently -L 10 -Q 3 or -L 10P -Q 4 .
  • R N1 ° is independently -G 4 or -L 10G -G 5 . 30
  • R N1 ° is independently saturated aliphatic C h alky!.
  • R N1 ° is independently -G 4 .
  • R N1 ° is independently -L 10G -G 5 .
  • R N1 ° is independently -Q 2 . 35 In one embodiment, R N1 ° is independently -L 10 -Q 3 .
  • R N1 ° is independently -L 10P -Q 4 .
  • R N1 ° is independently -Me, -Et, -nPr, or -iPr. In one embodiment, R N1 ° is independently -Me. W In one embodiment, -L 10 -, if present, is independently -(CH 2 ) n 3-, wherein n3 is independently 2, 3, 4, 5, 6, 7, or 8. In one embodiment, n3 is independently 2, 3, 4, 5, or 6. In one embodiment, n3 is independently 2, 3, or 4.
  • -L 10G - is independently -(CH 2 ) n2 -, wherein n2 is independently 1 , 2, 3, or 4. In one embodiment, n2 is independently 1 or 2. In one embodiment, n2 is independently 1.
  • -L 10P - is independently phenylene, oxazol-di-yl, thiazol-di-yl, isoxazol-di-yl, isothiazol-di-yl, pyrazol-di-yl, pyridin-di-yl, pyrimidin-di-yl, or pyrazin-di-yl; and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -L 10P - is independently phenylene (e.g., 1 ,4-phenylene, 1 ,3-phenylene, 1 ,2-phenylene); and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 4 is independently phenyl or C 5-6 heteroaryl, and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 4 is independently phenyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridinyl, pyrimidinyl, or pyrazinyl; and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 4 if present, is independently phenyl; and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 5 is independently phenyl or Cs-eheteroaryl, and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 5 is independently phenyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridinyl, pyrimidinyl, or pyrazinyl; and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • -G 5 if present, is independently phenyl; and is independently unsubstituted or substituted with one or more substituents -G 6 .
  • each -L* 2 - is independently -(CH 2 ),*-, wherein n4 is independently 2, 3, 4, 5, 6, 7, or 8. In one embodiment, n4 is independently 2, 3, 4, 5, or 6. In one embodiment, n4 is independently 2, 3, or 4. In one embodiment, each -G 6 , if present, is independently -F 1 -Cl, -Br 1 -I, -R A5 , -CF 3 , -OR A5 , or -OCF 3 .
  • each R A5 if present, is independently -Me, -Et, -nPr, or -iPr. In one embodiment, each R A5 , if present, is independently -Me.
  • -L 3 - is independently -(CH 2 ) n5 -, wherein n5 is independently 2, 3, 4, 5, 6, 7, or 8. In one embodiment, n5 is independently 2, 3, 4, 5, or 6. In one embodiment, n5 is independently 2, 3, or 4.
  • -L 8 - is independently -(CH 2 W. wherein n6 is independently 2, 3, 4, 5, 6, 7, or 8. In one embodiment, n6 is independently 2, 3, 4, 5, or 6. In one embodiment, n6 is independently 2, 3, or 4.
  • -L 3 - and -L 8 - are the same.
  • each of -Q 1 , -Q 2 , -Q 3 , -Q 4 , -Q 5 , -Q 6 , and -Q 7 is independently: -NH 2 , -NHR B1 , or -NR 62 R 63 .
  • each -Q 1 if present, is independently -NH 2 , -NHR 61 , -NR B1 2l or -NR 62 R 63 . In one embodiment, each -Q 1 , if present, is independently -NH 2 , -NHR 61 , or -NR 62 R 63 .
  • -Q 2 if present, is independently -NH 2 , -NHR 61 , -NR B1 2 , or -NR 62 R 63 . In one embodiment, -Q 2 , if present, is independently -NH 2 , -NHR 61 , or -NR 62 R 63 .
  • -Q 3 if present, is independently -NH 2 , -NHR 61 , -NR B1 2 , or -NR 62 R 63 . In one embodiment, -Q 3 , if present, is independently -NH 2 , -NHR 61 , or -NR 62 R 63 .
  • -Q 4 if present, is independently -NH 2 , -NHR 61 , -NR B1 2 , or -NR 62 R 63 . In one embodiment, -Q 4 , if present, is independently -NH 2 , -NHR 61 , or -NR 62 R 63 .
  • each -Q 5 is independently -NH 2 , -NHR 61 , -NR 61 2 , or -NR 82 R 63 . In one embodiment, each -Q 5 , if present, is independently -NH 2 , -NHR 61 , or -NR 62 R 83 .
  • -Q 6 is independently -NH 2 , -NHR 61 , -NR 61 2 , or -NR 62 R 63 . In one embodiment, -Q 6 is independently -NH 2 , -NHR 61 , or -NR 82 R 63 .
  • -Q 7 is independently -NH 2 , -NHR 61 , -NR B1 2 , or -NR 62 R 63 . In one embodiment, -Q 7 is independently -NH 2 , -NHR 61 , or -NR 82 R 63 . In one embodiment, each -Q 1 , if present, is independently -NH 2 , -NR B1 2l or -NHR B1 . In one embodiment, each -Q 1 , if present, is independently -NH 2 or -NHR B1 .
  • each -Q 5 if present, is independently -NH 2 , -NR B1 2 , or -NHR B1 . In one embodiment, each -Q 5 , if present, is independently -NH 2 or -NHR B1 .
  • -Q 2 if present, is independently -NR B2 R B3 .
  • -Q 3 if present, is independently -NR B2 R B3 .
  • -Q 4 if present, is independently -NR B2 R B3 .
  • -Q 6 is independently -NR 82 R 63 .
  • -Q 7 is independently -NR 82 R 63 .
  • -Q 6 and -Q 7 are the same.
  • each R B1 if present, is independently -Me, -Et, -nPr, or -iPr. In one embodiment, each R 81 , if present, is independently -Me.
  • each -NR 82 R 83 is independently pyrrolidino, imidazolidino, N-CCvsalkyO-imidazolidino, pyrazolidino, N-(C 1-3 alkyl)-pyrazolidino, piperidino, N-Cd ⁇ alkyl ⁇ piperidino, piperizino, morpholino, azepino, diazepino, or N ⁇ CvaalkyO-diazepino.
  • -Q 1 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(JPr) 2 , pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 1 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2l or -N(JPr) 2 .
  • -Q 5 is independently -NH 2 , -NHMe 1 -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(JPr) 2 , pyrrolidino, imidazolidino,
  • N-(methyl)-imidazolidino pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 5 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , or -N(JPr) 2 .
  • -Q 2 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(JPr) 2 , pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 2 is independently pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 3 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(JPr) 2 , pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 3 is independently pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 4 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(JPr) 2 , pyrrolidino, imidazolidino,
  • N-(methyl)-imidazolidino pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 4 is independently pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 6 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr),
  • -Q 6 is independently pyrrolidino, imidazolidino,
  • N-(methyl)-imidazolidino pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 7 is independently -NH 2 , -NHMe, -NHEt, -NH(nPr), -NH(iPr), -N(Me) 2 , -N(Et) 2 , -N(nPr) 2 , -N(iPr) 2 , pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • -Q 7 is independently pyrrolidino, imidazolidino, N-(methyl)-imidazolidino, pyrazolidino, N-(methyl)-pyrazolidino, piperidino, N-(methyl)-piperidino, piperizino, morpholino, azepino, diazepino, or N-(methyl)-diazepino.
  • the group -NR N8B -L 8 -NR 8PA R 8PB is independently selected from:
  • the group -L -NR R is independently selected from:
  • the group -L -G is independently selected from:
  • the group -G 4 is independently selected from:
  • the group -L -Q is independently selected from: In one embodiment, the group -L -Q is independently selected from Molecular Weight
  • the compound has a molecular weight of from 329 to 1200.
  • the bottom of range is 330; 350; 375; 400; 425; 450.
  • the top of range is 1100; 1000, 900, 800, 700.
  • the range is 329 to 700.
  • Examples of some preferred compounds include the following compounds, and pharmaceutically acceptable salts, solvates, and hydrates thereof:
  • One aspect of the present invention pertains to BPD compounds, as described herein, in substantially purified form and/or in a form substantially free from contaminants.
  • the substantially purified form is at least 50% by weight, e.g., at least 60% by weight, e.g., at least 70% by weight, e.g., at least 80% by weight, e.g., at least 90% by weight, e.g., at least 95% by weight, e.g., at least 97% by weight, e.g., at least 98% by weight, e.g., at least 99% by weight.
  • the substantially purified form refers to the compound in any stereoisomeric or enantiomeric form.
  • the substantially purified form refers to a mixture of stereoisomers, i.e., purified with respect to other compounds.
  • the substantially purified form refers to one stereoisomer, e.g., optically pure stereoisomer.
  • the substantially purified form refers to a mixture of enantiomers.
  • the substantially purified form refers to a equimolar mixture of enantiomers (i.e., a racemic mixture, a racemate).
  • the substantially purified form refers to one enantiomer, e.g., optically pure enantiomer.
  • the contaminants represent no more than 50% by weight, e.g., no more than 40% by weight, e.g., no more than 30% by weight, e.g., no more than 20% by weight, e.g., no more than 10% by weight, e.g., no more than 5% by weight, e.g., no more than 3% by weight, e.g., no more than 2% by weight, e.g., no more than 1% by weight.
  • the contaminants refer to other compounds, that is, other than stereoisomers or enantiomers. In one embodiment, the contaminants refer to other compounds and other stereoisomers. In one embodiment, the contaminants refer to other compounds and the other enantiomer.
  • the substantially purified form is at least 60% optically pure (i.e., 60% of the compound, on a molar basis, is the desired stereoisomer or enantiomer, and 40% is the undesired stereoisomer or enantiomer), e.g., at least 70% optically pure, e.g., at least 80% optically pure, e.g., at least 90% optically pure, e.g., at least 95% optically pure, e.g., at least 97% optically pure, e.g., at least 98% optically pure, e.g., at least 99% optically pure.
  • 60% optically pure i.e., 60% of the compound, on a molar basis, is the desired stereoisomer or enantiomer, and 40% is the undesired stereoisomer or enantiomer
  • at least 70% optically pure e.g., at least 80% optically pure, e.g., at least 90% optically pure, e
  • Certain compounds may exist in one or more particular geometric, optical, enantiomeric, diasteriomeric, epimeric, atropic, stereoisomeric, tautomeric, conformational, or anomeric forms, including but not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, and r- forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d- and l-forms; (+) and (-) forms; keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal- and anticlinal-forms; ⁇ - and ⁇ -forms; axial and equatorial forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and combinations thereof, hereinafter collectively referred to as "isomers” (or "isomeric forms").
  • isomers are structural (or constitutional) isomers (i.e., isomers which differ in the connections between atoms rather than merely by the position of atoms in space).
  • a reference to a methoxy group, -OCH 3 is not to be construed as a reference to its structural isomer, a hydroxymethyl group, -CH 2 OH.
  • a reference to ortho-chlorophenyl is not to be construed as a reference to its structural isomer, meta-chlorophenyl.
  • a reference to a class of structures may well include structurally isomeric forms falling within that class (e.g., C 1-7 alkyl includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).
  • C 1-7 alkyl includes n-propyl and iso-propyl
  • butyl includes n-, iso-, sec-, and tert-butyl
  • methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl
  • keto-, enol-, and enolate-forms as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro.
  • keto enol enolate as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro.
  • H may be in any isotopic form, including 1 H, 2 H (D), and 3 H (T); C may be in any isotopic form, including 12 C, 13 C, and 14 C; O may be in any isotopic form, including 16 O and 18 O; and the like.
  • a corresponding salt of the compound for example, a pharmaceutically-acceptable salt.
  • pharmaceutically acceptable salts are discussed in Berge et al., 1977, "Pharmaceutically Acceptable Salts," J. Pharrn. ScL. Vol. 66, pp. 1-19.
  • a salt may be formed with a suitable cation.
  • suitable inorganic cations include, but are not limited to, alkali metal ions such as Na + and K + , alkaline earth cations such as Ca 2+ and Mg 2+ , and other cations such as Al +3 .
  • Suitable organic cations include, but are not limited to, ammonium ion (i.e., NH 4 + ) and substituted ammonium ions (e.g., NH 3 R + , NH 2 R 2 + , NHR 3 + , NR 4 + ).
  • suitable substituted ammonium ions are those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids, such as lysine and arginine.
  • An example of a common quaternary ammonium ion is N(CH 3 ) 4 + .
  • a salt may be formed with a suitable anion.
  • suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous.
  • Suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, and valeric.
  • solvate is used herein in the conventional sense to refer to a complex of solute (e.g., compound, salt of compound) and solvent. If the solvent is water, the solvate may be conveniently referred to as a hydrate, for example, a mono-hydrate, a di-hydrate, a tri-hydrate, etc.
  • a reference to a particular compound also includes solvate and hydrate forms thereof.
  • the 10H-benzo[g]pteridine-2,4-dione (“BPD”) compounds described herein may be prepared, for example, by methods in which, first, 2,4-dichloro-i-nitro-benzene (1) is reacted with a suitable amine to form the corresponding (5-chloro-2-nitro-phenyl) amine (2). This amine (2) is then reduced, for example, with zinc dust, and then reacted with alloxan and boric acid to give the corresponding 8-chloro-10-substituted- 10H-benzo[g]pteridine-2,4-dione (3).
  • This compound is then reacted with a suitable chloride to form the corresponding 3,10-disubstituted-8-chloro-10H-benzo[g]pteridine-2,4- dione (4).
  • This compound is then reacted with suitable amine to give the corresponding 3,8,10-trisubstituted-8-chloro-10H-benzo[g]pteridine-2,4-dione (5).
  • R i -NH 2 anhydrous K 2 CO 3 , DMF, 9O 0 C, 24-36 hours or: R 1 -NH 2 , neat, 100 0 C, 2-3 hours
  • Zinc dust AcOH, room temperature, 20-30 minutes
  • Alloxan H 2 O, boric acid, AcOH, room temperature, 0.5 to 2 hours or 100°C, 0.5 to 5 hours
  • Another aspect of the invention pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a BPD compound as described herein.
  • Another aspect of the invention pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a BPD compound as described herein, and a pharmaceutically acceptable carrier, diluent, or excipient.
  • Another aspect of the invention pertains to a method of preparing a pharmaceutical composition
  • a method of preparing a pharmaceutical composition comprising admixing a BPD compound as described herein and a pharmaceutically acceptable carrier, diluent, or excipient.
  • Another aspect of the present invention pertains to a method of selectively binding a G-quadruplex, comprising contacting said G-quadruplex with an effective amount of a BPD compound, as described herein.
  • said G-quadruplex is within a living cell. In one embodiment, said G-quadruplex is within a living cell that is not an integral part of a living human or multicellular animal body. In one embodiment, said G-quadruplex is within a living cell that is an integral part of a living human or multicellular animal body.
  • the method is a method of selectively binding a G-quadruplex within a living cell, comprising contacting said cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • a BPD compound as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • Such a method may be performed in vitro or in vivo.
  • Another aspect of the present invention pertains to a method of stabilizing a
  • G-quadruplex comprising contacting said G-quadruplex with an effective amount of a BPD compound, as described herein.
  • said G-quadruplex is within a living cell. In one embodiment, said G-quadruplex is within a living cell that is not an integral part of a living human or multicellular animal body. In one embodiment, said G-quadruplex is within a living cell that is an integral part of a living human or multicellular animal body.
  • the method is a method of stabilizing a G-quadruplex within a living cell, comprising contacting said cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • a BPD compound as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • Such a method may be performed in vitro or in vivo.
  • Another aspect of the present invention pertains to a method of inhibiting telomerase (for example, inhibiting telomerase activity, inhibiting formation of telomerase complexes, inhibiting activity of telomerase complexes, etc.), comprising contacting said telomerase with an effective amount of a BPD compound, as described herein.
  • telomerase is within a living cell.
  • said telomerase is within a living cell that is not an integral part of a living human or multicellular animal body.
  • said telomerase js within a living cell that is an integral part of a living human or multicellular animal body.
  • the method is a method of inhibiting telomerase within a living cell, comprising contacting said cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • a BPD compound as described herein, preferably in the form of a pharmaceutically acceptable composition.
  • Such a method may be performed in vitro or in vivo.
  • telomerase activity One of ordinary skill in the art is readily able to determine whether or not a candidate compound inhibits telomerase activity.
  • Another aspect of the present invention pertains to a method reducing, repressing, or inhibiting gene expression in a cell, comprising contacting a living cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition. Such a method may be performed in vitro or in vivo.
  • the gene expression is c-kit gene expression (e.g., expression of the gene that encodes the c-kit receptor).
  • the gene expression is vascular endothelial growth factor (VEGF) gene expression (e.g., expression of the gene that encodes the VEGF receptor).
  • VEGF vascular endothelial growth factor
  • the gene expression is c-myc gene expression (e.g., expression of the gene that encodes the c-myc receptor).
  • the gene expression is BCL-2 gene expression (e.g., expression of the gene that encodes the BCL-2 receptor). In one embodiment, the gene expression is B-raf gene expression (e.g., expression of the gene that encodes the B-raf receptor).
  • the gene expression is k-ras gene expression (e.g., expression of the gene that encodes the f k-ras receptor).
  • said cell is a living cell that is not an integral part of a living human or multicellular animal body. In one embodiment, said cell is a living cell that is an integral part of a living human or multicellular animal body.
  • One of ordinary skill in the art is readily able to determine whether or not a candidate compound regulates cell proliferation for any particular cell line.
  • Another aspect of the present invention pertains to a method of regulating (e.g., inhibiting) cell proliferation comprising contacting a living cell with an effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutically acceptable composition. Such a method may be performed in vitro or in vivo.
  • said cell is a living cell that is not an integral part of a living human or multicellular animal body. In one embodiment, said cell is a living cell that ⁇ s an integral part of a living human or multicellular animal body.
  • One of ordinary skill in the art is readily able to determine whether or not a candidate compound regulates cell proliferation for any particular cell line.
  • a sample of cells e.g., from a tumour
  • a candidate compound brought into contact with the cells, and the effect of the compound on those cells observed.
  • effect the morphological status of the cells may be determined (e.g., alive or dead), or the expression levels of genes associated with cell cycle regulation determined.
  • this may be used as a prognostic or diagnostic marker of the efficacy of the compound in methods of treating a patient with cells of the same type (e.g., the tumour or a tumour of the same cellular type).
  • Another aspect of the present invention pertains to a method of treatment comprising administering to a patient in need of treatment a therapeutically-effective amount of a BPD compound, as described herein, preferably in the form of a pharmaceutical composition.
  • Another aspect of the present invention pertains to a BPD compound, as described herein, for use in a method of treatment, for example, a method of treatment of the human or animal body by therapy.
  • Another aspect of the present invention pertains to use of a BPD compound, as described herein, in the manufacture of a medicament for use in treatment.
  • the medicament comprises the BPD compound.
  • the BPD compounds described herein are useful in the treatment of disorders that are mediated by a G-quadruplex.
  • the treatment is treatment of a disorder that is mediated by a G-quadruplex.
  • a disorder that is mediated by a G-quadruplex is a disorder that, for initiation or maintenance or progress of the disorder, requires the formation or destruction of a
  • G-quadruplex For such disorders, treatment with a G-quadruplex ligand, e.g., a selective G-quadruplex ligand, e.g., a selective stabilising G-quadruplex ligand, is beneficial.
  • a G-quadruplex ligand e.g., a selective G-quadruplex ligand, e.g., a selective stabilising G-quadruplex ligand
  • C-kit is a tyrosine kinase receptor for the growth-promoting cytokine SCF (stem cell factor) which plays an important biological role in the control of differentiation.
  • SCF stem cell factor
  • Two quadruplex forming sequence stretches have been identified within the c-kit promoter. Expression of mutant c-kit/SCF has been implicated in the genesis of several solid tumour types (see, e.g., Rankin et al., 2005).
  • c-kit activity can be inhibited with the new therapeutic agent Gleevec®, and in Ewing's sarcoma and gastrointestinal stromal tumours, there is at least in vitro evidence that c-kit inhibition can retard tumor growth (see, e.g., DeMatteo, 2002; Druker, 2002).
  • Other targets on the basis of c-kit expression include: prostate cancer (see, e.g., Simak et al., 2000), and adenocarcinoma lung cancers (see, e.g., Micke et al., 2004).
  • c-kit is a receptor tyrosine kinase expressed in a number of different normal cell types, whose dysregulated expression or activity can drive the proliferation of cancer cells.
  • c-kit dysregulation is a common feature of gastro-intestinal stromal tumours (GIST), a leiomyosarcomatous neoplasm with an estimated incidence of 1200 cases per year in the UK and 5000 cases per year in the USA.
  • GIST gastro-intestinal stromal tumours
  • tumour mass does not usually regress completely, and the usual clinical course involves a period of quiescence followed by the outgrowth of cells from the persisting tumour mass.
  • Clinical recrudescence of this kind is often accompanied by refractoriness to c-kit inhibition by Gleevec®, through acquired mutations in the kinase activation loop.
  • activation loop mutations in c-kit found in certain cases of acute myeloid leukaemia (incidence -2000 cases/year in the UK) or testicular seminomas (incidence -2000 cases/year in the UK) are also typically resistant to Gleevec® therapy.
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of c-kit.
  • a disorder e.g., cancer
  • c-kit e.g., cancer
  • the treatment is treatment of a solid tumour, gastro-intestinal stromal tumour (GIST), small cell lung cancer, adenocarcinoma lung cancer, Ewing's sarcoma, acute myeloid leukaemia, prostate cancer, or testicular seminoma; for example, that is characterised by expression (or overexpression) of c-kit.
  • GIST gastro-intestinal stromal tumour
  • small cell lung cancer adenocarcinoma lung cancer
  • Ewing's sarcoma acute myeloid leukaemia
  • prostate cancer prostate cancer
  • testicular seminoma for example, that is characterised by expression (or overexpression) of c-kit.
  • VEGF Human Vascular Endothelial Growth Factor
  • VEGF Vascular endothelial growth factor receptor
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of VEGF.
  • a disorder e.g., cancer
  • VEGF vascular endothelial growth factor
  • the treatment is treatment of a solid tumour; for example, that is characterised by expression (or overexpression) of VEGF.
  • a quadruplex forming sequence has been identified within the c-myc promoter.
  • c-MYC oncogene Overexpression of the c-MYC oncogene is linked with a wide range of cancers that include colon cancer, breast cancer, small-cell lung cancer, osteosarcomas, glioblastomas, and myeloid leukaemia.
  • cancers that include colon cancer, breast cancer, small-cell lung cancer, osteosarcomas, glioblastomas, and myeloid leukaemia.
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of c-myc.
  • a disorder e.g., cancer
  • c-myc e.g., cancer
  • the treatment is treatment of colon cancer, breast cancer, small-cell lung cancer, osteosarcoma, glioblastoma, or myeloid leukaemia; for example, that is characterised by expression (or overexpression) of c-myc.
  • BCL-2 promoter Quadruplex forming sequences have been identified within the BCL-2 promoter.
  • BCL-2 promoter quadruplexes are important for B-cell and T-cell lymphomas, breast, prostate, cervical, colorectal and non-small cell lung carcinomas.
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of BCL-2.
  • the treatment is treatment of B-cell lymphoma, T-cell lymphoma, breast cancer, prostate cancer, cervical cancer, colorectal cancer, and non-small cell lung cancer; for example, that is characterised by expression (or overexpression) of BCL-2.
  • B-raf encodes an intracellular serine-threonine kinase that is critical for growth signalling through the ras pathway.
  • B-raf activity is dysregulated by mutations in up to 70% of malignant melanoma, an aggressive and hard-to-treat neoplasm of the skin, whose annual incidence is estimated to be about 7500 cases/year in the UK or 30000 cases/year in the USA.
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of B-raf.
  • a disorder e.g., cancer
  • B-raf e.g., cancer
  • the treatment is treatment of skin cancer or melanoma; for example, that is characterised by expression (or overexpression) of B-raf.
  • the treatment is treatment of a disorder (e.g., cancer) that is characterised by expression (or overexpression) of k-ras.
  • a disorder e.g., cancer
  • k-ras e.g., cancer
  • the treatment is treatment of a proliferative disorder.
  • proliferative condition refers to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells that is undesired, such as, neoplastic or hyperplastic growth.
  • the compounds are anti-proliferative agents.
  • antiproliferative agent refers to a compound that treats a proliferative disorder (i.e., a compound which is useful in the treatment of a proliferative disorder).
  • the treatment is treatment of a proliferative disorder characterised by benign, pre-malignant, or malignant cellular proliferation, including but not limited to, neoplasms, hyperplasias, and tumours (e.g., histocytoma, glioma, astrocyoma, osteoma), cancers (see below), psoriasis, bone diseases, fibroproliferative disorders (e.g., of connective tissues), pulmonary fibrosis, atherosclerosis, smooth muscle cell proliferation in the blood vessels, such as stenosis or restenosis following angioplasty.
  • a proliferative disorder characterised by benign, pre-malignant, or malignant cellular proliferation
  • neoplasms e.g., hyperplasias, and tumours (e.g., histocytoma, glioma, astrocyoma, osteoma), cancers (see below), psoriasis, bone diseases, fibroprolifer
  • the treatment is treatment of cancer.
  • the compounds are anti-cancer agents.
  • anti-cancer agent as used herein, pertains to a compound that treats a cancer (i.e., a compound which is useful in the treatment of a cancer).
  • the anti-cancer effect may arise through one or more mechanisms, including but not limited to, the regulation of cell proliferation, the inhibition of cell cycle progression, the inhibition of angiogenesis (the formation of new blood vessels), the inhibition of metastasis (the spread of a tumour from its origin), the inhibition of invasion (the spread of tumour cells into neighbouring normal structures), or the promotion of apoptosis (programmed cell death).
  • One of ordinary skill in the art is readily able to determine whether or not a candidate compound treats a proliferative disorder, or treats cancer, for any particular cell type.
  • the treatment is treatment of: solid tumour cancer, gastrointestinal stromal cancer (GIST), stomach cancer, bowel cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer, gastrointestinal cancer, thyroid cancer, breast cancer, ovarian cancer, epithelial ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, testicular cancer, testicular seminoma, liver cancer, kidney cancer, renal cell carcinoma, bladder cancer, pancreatic cancer, oesophageal cancer, brain cancer, glioblastoma, glioma, sarcoma, osteosarcoma, Ewing's sarcoma, bone cancer, skin cancer (e.g., head and neck cancer), squamous cancer, Kaposi's sarcoma, melanoma, malignant melanoma, lymphoma, B-cell lymphoma, T-cell lymphoma, leuka
  • a pathologist may determine the histological subtype of a cancer based upon the cell morphology, for example, mucinous, adenocarcinoma, serous, papillary, etc.
  • the BPD compounds described herein may be used in the treatment of the cancers described herein, independent of the mechanisms discussed herein. Screeninq
  • a patient Prior to treatment, a patient may be screened in order to determine whether a disease or disorder from which the patient is or may be suffering is one which would be susceptible to treatment with a BPD compound as described herein.
  • treatment pertains generally to treatment and therapy, whether of a human or an animal (e.g., in veterinary applications), in which some desired therapeutic effect is achieved, for example, the inhibition of the progress of the disorder, and includes a reduction in the rate of progress, a halt in the rate of progress, alleviation of symptoms of the disorder, amelioration of the disorder, and cure of the disorder.
  • Treatment as a prophylactic measure i.e., prophylaxis
  • prophylaxis is also included.
  • treatment includes the prophylaxis of cancer, reducing the risk of cancer, alleviating the symptoms of cancer, etc.
  • terapéuticaally-effective amount refers to that amount of a compound, or a material, composition or dosage form comprising a compound, which is effective for producing some desired therapeutic effect, commensurate with a reasonable benefit/risk ratio, when administered in accordance with a desired treatment regimen.
  • treatment includes combination treatments and therapies, in which two or more treatments or therapies are combined, for example, sequentially or simultaneously.
  • the BPD compounds described herein may also be used in combination therapies, e.g., in conjunction with other agents, for example, cytotoxic agents, anticancer agents, etc.
  • treatments and therapies include, but are not limited to, chemotherapy (the administration of active agents, including, e.g., drugs, antibodies (e.g., as in immunotherapy), prodrugs (e.g., as in photodynamic therapy, GDEPT, ADEPT, etc.); surgery; radiation therapy; photodynamic therapy; gene therapy; and controlled diets.
  • a BPD compound as described herein may be beneficial to combine treatment with a BPD compound as described herein with one or more other (e.g., 1, 2, 3, 4) agents or therapies that regulates cell growth or survival or differentiation via a different mechanism, thus treating several characteristic features of cancer development.
  • a BPD compound as described herein is combined with one or more (e.g., 1 , 2, 3, 4) additional therapeutic agents, as described below.
  • One aspect of the present invention pertains to a BPD compound as described herein, in combination with one or more (e.g., 1, 2, 3, 4) additional therapeutic agents, as described below.
  • the agents may be administered simultaneously or sequentially, and may be administered in individually varying dose schedules and via different routes.
  • the agents can be administered at closely spaced intervals (e.g., over a period of 5-10 minutes) or at longer intervals (e.g., 1 , 2, 3, 4 or more hours apart, or even longer periods apart where required), the precise dosage regimen being commensurate with the properties of the therapeutic agent(s).
  • agents i.e., a BPD compound as described here, plus one or more other agents
  • the agents may be formulated together in a single dosage form, or alternatively, the individual agents may be formulated separately and presented together in the form of a kit, optionally with instructions for their use, as described below.
  • the BPD compounds described herein may also be used as cell culture additives to inhibit telomerase, inhibit cell proliferation, etc.
  • the BPD compounds described herein may also be used as part of an in vitro assay, for example, in order to determine whether a candidate host is likely to benefit from treatment with the compound in question.
  • the BPD compounds described herein may also be used as a standard, for example, in an assay, in order to identify other active compounds, other anti-proliferative agents, other anti-cancer agents, etc.
  • kits comprising (a) a BPD compound as described herein, or a composition comprising a compound as described herein, e.g., preferably provided in a suitable container and/or with suitable packaging; and
  • instructions for use e.g., written instructions on how to administer the compound or composition.
  • the written instructions may also include a list of indications for which the active ingredient is a suitable treatment.
  • the BPD compound or pharmaceutical composition comprising the BPD compound may be administered to a subject by any convenient route of administration, whether systemically/peripherally or topically (i.e., at the site of desired action).
  • Routes of administration include, but are not limited to, oral (e.g., by ingestion); buccal; sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal (including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eyedrops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., using an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular
  • the subject/patient may be an animal, a mammal, a placental mammal, a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), murine (e.g., a mouse), a lagomorph (e.g., a rabbit), avian (e.g., a bird), canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a horse), porcine (e.g., a pig), ovine (e.g., a sheep), bovine (e.g., a cow), a primate, simian (e.g., a monkey or ape), a monkey (e.g., marmoset, baboon), an ape (e.g., gorilla, chimpanzee, orangutang, gibbon), or a human.
  • a rodent
  • the subject/patient may be any of its forms of development, for example, a foetus.
  • the subject/patient is a human.
  • the BPD compound While it is possible for the BPD compound to be administered alone, it is preferable to present it as a pharmaceutical formulation (e.g., composition, preparation, medicament) comprising at least one BPD compound, as described herein, together with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, including, but not limited to, pharmaceutically acceptable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, stabilisers, solubilisers, surfactants (e.g., wetting agents), masking agents, colouring agents, flavouring agents, and sweetening agents.
  • the formulation may further comprise other active agents, for example, other therapeutic or prophylactic agents.
  • the present invention further provides pharmaceutical compositions, as defined above, and methods of making a pharmaceutical composition comprising admixing at least one compound, as described herein, together with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, e.g., carriers, diluents, excipients, etc. If formulated as discrete units (e.g., tablets, etc.), each unit contains a predetermined amount (dosage) of the compound.
  • pharmaceutically acceptable refers to compounds, ingredients, materials, compositions, dosage forms, etc., which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of the subject in question (e.g., human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • Each carrier, diluent, excipient, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.
  • Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical texts, for example, Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients. 2nd edition, 1994.
  • the formulations may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing into association the compound with a carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compound with carriers (e.g., liquid carriers, finely divided solid carrier, etc.), and then shaping the product, if necessary.
  • carriers e.g., liquid carriers, finely divided solid carrier, etc.
  • the formulation may be prepared to provide for rapid or slow release; immediate, delayed, timed, or sustained release; or a combination thereof.
  • Formulations may suitably be in the form of liquids, solutions (e.g., aqueous, nonaqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, electuaries, mouthwashes, drops, tablets (including, e.g., coated tablets), granules, powders, lozenges, pastilles, capsules (including, e.g., hard and soft gelatin capsules), cachets, pills, ampoules, boluses, suppositories, pessaries, tinctures, gels, pastes, ointments, creams, lotions, oils, foams, sprays, mists, or aerosols.
  • Formulations may suitably be provided as a patch, adhesive plaster, bandage, dressing, or the like which is impregnated with one or more compounds and optionally one or more other pharmaceutically acceptable ingredients, including, for example, penetration, permeation, and absorption enhancers. Formulations may also suitably be provided in the form of a depot or reservoir.
  • the compound may be dissolved in, suspended in, or admixed with one or more other pharmaceutically acceptable ingredients.
  • the compound may be presented in a liposome or other microparticulate which is designed to target the compound, for example, to blood components or one or more organs.
  • Formulations suitable for oral administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, electuaries, tablets, granules, powders, capsules, cachets, pills, ampoules, boluses.
  • Formulations suitable for buccal administration include mouthwashes, lozenges, pastilles, as well as patches, adhesive plasters, depots, and reservoirs.
  • Lozenges typically comprise the compound in a flavored basis, usually sucrose and acacia or tragacanth.
  • Pastilles typically comprise the compound in an inert matrix, such as gelatin and glycerin, or sucrose and acacia.
  • Mouthwashes typically comprise the compound in a suitable liquid carrier.
  • Formulations suitable for sublingual administration include tablets, lozenges, pastilles, capsules, and pills.
  • Formulations suitable for oral transmucosal administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil- in-water, water-in-oil), mouthwashes, lozenges, pastilles, as well as patches, adhesive plasters, depots, and reservoirs.
  • solutions e.g., aqueous, non-aqueous
  • suspensions e.g., aqueous, non-aqueous
  • emulsions e.g., oil- in-water, water-in-oil
  • mouthwashes e.g., lozenges, pastilles, as well as patches, adhesive plasters, depots, and reservoirs.
  • Formulations suitable for non-oral transmucosal administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), suppositories, pessaries, gels, pastes, ointments, creams, lotions, oils, as well as patches, adhesive plasters, depots, and reservoirs.
  • solutions e.g., aqueous, non-aqueous
  • suspensions e.g., aqueous, non-aqueous
  • emulsions e.g., oil-in-water, water-in-oil
  • suppositories e.g., pessaries, gels, pastes, ointments, creams, lotions, oils, as well as patches, adhesive plasters, depots, and reservoirs.
  • Formulations suitable for transdermal administration include gels, pastes, ointments, 5 creams, lotions, and oils, as well as patches, adhesive plasters, bandages, dressings, depots, and reservoirs.
  • Tablets may be made by conventional means, e.g., compression or moulding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared by
  • binders e.g., povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose
  • fillers or diluents e.g., lactose, microcrystalline cellulose, calcium hydrogen phosphate
  • lubricants e.g., magnesium stearate, talc, silica
  • disintegrants e.g., sodium starch glycolate, cross-linked povidone,
  • Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be
  • Tablets may optionally be provided with a coating, for example, to affect release, for example an enteric coating, to provide release in parts of the gut other than the stomach.
  • Ointments are typically prepared from the compound and a paraffinic or a water-miscible ointment base.
  • Creams are typically prepared from the compound and an oil-in-water cream base.
  • the aqueous phase of the cream base may include, for example, at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane-1 ,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof.
  • the topical formulations may desirably include a compound which enhances absorption or penetration of the compound through the skin or other affected 55 areas. Examples of such dermal penetration enhancers include dimethylsulfoxide and related analogues.
  • Emulsions are typically prepared from the compound and an oily phase, which may optionally comprise merely an emulsifier (otherwise known as an emulgent), or it may ⁇ 0 comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil.
  • an emulsifier otherwise known as an emulgent
  • a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabiliser. It is also preferred to include both an oil and a fat.
  • the emulsifier(s) with or without stabiliser(s) make up the so-called emulsifying wax
  • the wax together with the oil and/or fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.
  • Suitable emulgents and emulsion stabilisers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulphate.
  • Tween 60 Span 80
  • cetostearyl alcohol myristyl alcohol
  • glyceryl monostearate and sodium lauryl sulphate.
  • suitable oils or fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the compound in most oils likely to be used in
  • the cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers.
  • Straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,
  • esters 15 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.
  • UO Formulations suitable for intranasal administration, where the carrier is a liquid include, for example, nasal spray, nasal drops, and (for aerosol administration by nebuliser) aqueous or oily solutions of the compound.
  • Formulations suitable for intranasal administration include, .5 for example, those presented as a coarse powder having a particle size, for example, in the range of about 20 to about 500 microns which is administered in the manner in which snuff is taken, i.e., by rapid inhalation through the nasal passage from a container of the powder held close up to the nose.
  • Formulations suitable for pulmonary administration include those presented as an aerosol spray from a pressurised pack, with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide, or other suitable gases.
  • a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide, or other suitable gases.
  • Formulations suitable for ocular administration include eye drops wherein the compound is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the compound.
  • Formulations suitable for rectal administration may be presented as a suppository with a 0 suitable base comprising, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, for example, cocoa butter or a salicylate; or as a solution or suspension for treatment by enema.
  • a suitable base comprising, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, for example, cocoa butter or a salicylate; or as a solution or suspension for treatment by enema.
  • Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the compound, such carriers as are known in the art to be appropriate.
  • Formulations suitable for parenteral administration include aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (e.g., solutions, suspensions), in which the compound is dissolved, suspended, or otherwise provided (e.g., in a liposome or other microparticulate).
  • Such liquids may additionally contain other pharmaceutically acceptable ingredients, such as anti-oxidants, buffers, preservatives, stabilisers, bacteriostats, suspending agents, thickening agents, and solutes which render the formulation isotonic with the blood (or other relevant bodily fluid) of the intended recipient.
  • excipients include, for example, water, alcohols, polyols, glycerol, vegetable oils, and the like.
  • suitable isotonic carriers for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection.
  • concentration of the compound in the liquid is from about 1 ng/ml to about 10 ⁇ g/ml, for example from about 10 ng/ml to about 1 ⁇ g/ml.
  • the formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilised) disorder requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.
  • sterile liquid carrier for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.
  • appropriate dosages of the BPD compounds, and compositions comprising the BPD compounds can vary from patient to patient. Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleterious side effects.
  • the selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, and/or materials used in combination, the severity of the disorder, and the species, sex, age, weight, disorder, general health, and prior medical history of the patient.
  • the amount of compound and route of administration will ultimately be at the discretion of the physician, veterinarian, or clinician, although generally the dosage will be selected to achieve local concentrations at the site of action that achieve the desired effect without causing substantial harmful or deleterious side-effects.
  • Administration can be effected in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell(s) being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician, veterinarian, or clinician.
  • a suitable dose of the compound is in the range of about 10 ⁇ g/m 2 to 1 g/m 2 per treatment (e.g., per day), more typically 1 mg/m 2 to 500 mg/m 2 per treatment (e.g., per day).
  • the compound is a salt, an ester, an amide, a prodrug, or the like
  • the amount administered is calculated on the basis of the parent compound and so the actual weight to be used is increased proportionately.
  • the BIAcore SPR (Surface Plasmon Resonance) method has been used to evaluate the binding affinity of the BPD compounds with G-quadruplex DNA. See, e.g.,
  • a native quadruplex forming sequence of htelo (d(biotin-[GTTA(GGGTTA) 4 GG]) and conserved c-kit (d(biotin-[C 3 G 3 CG 3 CGCGAG 3 AG 4 AG 2 ]) as well as a hairpin duplex DNA (d(biotin-[G 2 CATAGTGCGTG 3 CGT 2 AGC]) was used in this study.
  • BPD1 , BPD2, BPD3 and BPD4 moderately bind to G-quadruplex DNA of htelo and c-kit, but they also bind to duplex DNA with a weaker affinity.
  • BPD compounds BPD5, BPD6, BPD7 and BPD8 bind tightly in sub-micromolar binding affinity with G-quadruplex DNA of htelo and conserved c-kit, but they don't bind to duplex DNA, up to 200 ⁇ M.
  • some of the BPD compounds tested were found to be very selective to G-quadruplex DNA of conserved c-kit.
  • NB No binding observed at concentrations up to 200 ⁇ M of compound.
  • the data show that the compounds BPD5, BPD6, BPD10, and BPD11 are strong quadruplex binding agents.
  • the ability of the BPD compounds to stabilize G-quadruplex DNA was assessed by a fluorescence resonance energy transfer (FRET) melting assay, in order to measure the melting transitions.
  • FRET fluorescence resonance energy transfer
  • Figure 1 illustrates the circular dichroism (CD) spectral data (ellipticity (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 (40 ⁇ M) with G-quadruplex DNA of htelo (4 ⁇ M), in K + and Na + containing buffer (50 mM Tris.HCI, pH 7.4, 100 mM KCI, and 100 mM NaCI):
  • Figure 2 illustrates the circular dichroism (CD) spectral data (ellipticity (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 with G-quadruplex DNA of htelo in K + containing buffer (50 mM Tris.HCI, pH 7.4, 100 mM KCI): (a) folded in the presence of 100 mM potassium without ligand (O 1 open hexagon) ("HteloKIOO");
  • Figure 3 illustrates the circular dichroism (CD) spectral data (ellipticity 20 (mdeg) versus wavelength (nm)) obtained in the study of the interaction of BPD5 (40 ⁇ M) with G-quadruplex DNA of c-kit (4 ⁇ M) in K + containing buffer (50 mM Tris.HCI, pH 7.4, 10O mM KCI):
  • MCF-7 a breast cancer cell line
  • BPD5 and BPD8 BPD8-activated c-kit gene expression
  • Quantification of c-kit gene expression by real-time PCR Human MCF-7 cells were grown in DMEM + 10% Fetal Calf Serum, seeded at a cell density of 3.4x10 5 cells per 6- well plate (9.5 cm 2 growth area). At time point 0, the medium was supplemented with test compound. Cells were harvested after several hours incubation and total RNA was extracted (Rneasy Mini Kit, Qiagen). RNA (200 ng) was reverse transcribed with SuperScriptlll (Invitrogen) according to the manufacturer's instructions. The cDNA was quantified using primers specific to human c-kit and beta-actin genes in a Roche LC480 LightCylcler, using the SYBR Green Master Mix. Crossing point values (Cp) were
  • BPD5 and BPD8 (0.5 ⁇ M and 5 ⁇ M concentration) were incubated with MCF-7 cells for 3 hours and 6 hours.
  • the results showed significant c-kit gene expression changes with
  • Figure 4 is bar graph showing the percentage gene expression of c-kit in MCF-7 cells
  • the figure shows the levels of expression for control cells treated with 10% DMSO in water (100% expression, blank bar), and for cells treated with the isoalloxazines BPD5 or BPD8 at 6 hours at 5 ⁇ M concentration (shaded bars).
  • the BIAcore SPR (Surface Plasmon Resonance) method has also been used to evaluate the differences in dissociation constant of the BPD compounds binding to c-kit and c-kit1, as well as htelo.
  • the sequences and methods described in Study 1 were used in this study along with the c-kit1 promoter G-quadruplex sequence ⁇ (DiOtJn-[AG 3 AG 3 CGCTG 3 AG 2 AG 3 ])).
  • the c-kit sequence used in this study (and Study 1 ) is also known in the art as the c-kit2 promoter G-quadruplex sequence.
  • the ability of the BPD compounds to stabilize G-quadruplex DNA of c-kit1 was assessed by a fluorescence resonance energy transfer (FRET) melting assay, as described in Study 2, in order to measure the melting transitions.
  • FRET fluorescence resonance energy transfer
  • HGC-27 is a gastrointestinal stromal tumour cell line which shows much higher basal levels of c-kit oncogene expression compared with MCF-7 (-16-fold higher expression).
  • Human HGC-27 cells were quantified in a manner identical to the quantification of MCF-7 cells, as described above in Study 4.
  • BPD8 (5 ⁇ M concentration) was incubated with HGC-27 cells for 2, 6 and 24 hours. The results showed significant c-kit gene expression changes with BPD8.
  • Figure 5 is a bar graph showing the percentage gene expression of c-kit in HGC-27 cells treated with BPD8. The figure shows the levels of expression for control cells treated with 10% DMSO in water (100% expression, blank bar), and for cells treated with BPD8 at 2, 4 and 8 hours at 5 ⁇ M concentration (shaded bars).
  • telomere-binding protein RP1 binds to and 15 promotes the formation of DNA quadruplexes in telomeric DNA.

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Abstract

L'invention concerne de manière générale le domaine des ligands G-quadruplex, et plus particulièrement certains composés 10H-benzo[g]ptéridine-2,4-dione (composés BPD), tels que décrits dans le présent document, qui se lient (sélectivement) à (et stabilisent) les G-quadruplex, entre autres. La présente invention concerne également des compositions pharmaceutiques qui comprennent de tels composés et l'utilisation de tels composés et de telles compositions, à la fois in vivo et in vitro, pour se lier (sélectivement) à (et stabiliser) les G-quadruplex, pour inhibiter la télomérase, pour réguler la prolifération cellulaire et pour le traitement de troubles prolifératifs, tels que le cancer.
PCT/GB2008/000962 2007-03-22 2008-03-18 Composés de 10h-benzo[g]ptéridine-2,4-dione pour le traitement de troubles prolifératifs Ceased WO2008114009A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012175729A1 (fr) * 2011-06-22 2012-12-27 TriOpto Tec GmbH Dérivés de 10h-benzo[g]ptéridine-2,4-dione, procédé de production et d'utilisation desdits dérivés

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999040093A2 (fr) * 1998-02-04 1999-08-12 Board Of Regents, The University Of Texas System Synthese d'analogues de la quinobenzoxazine par interaction avec la topoisomerase ii et avec des quadruplex, pouvant etre utilises comme agents antineoplasiques

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999040093A2 (fr) * 1998-02-04 1999-08-12 Board Of Regents, The University Of Texas System Synthese d'analogues de la quinobenzoxazine par interaction avec la topoisomerase ii et avec des quadruplex, pouvant etre utilises comme agents antineoplasiques

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Title
BEJUGAM ET AL.: "Trisubstituted Isoalloxazines as a New Class of G-Quadruplex Binding Ligands: Small Molecule Regulation of c-kit Oncogene Expression", J. AM. CHEM. SOC., vol. 129, no. 43, 10 May 2007 (2007-05-10), pages 12926 - 12927, XP002479922 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012175729A1 (fr) * 2011-06-22 2012-12-27 TriOpto Tec GmbH Dérivés de 10h-benzo[g]ptéridine-2,4-dione, procédé de production et d'utilisation desdits dérivés
US20140212459A1 (en) * 2011-06-22 2014-07-31 Tim Maisch 10h-benzo[g]pteridine-2,4-dione derivatives, method for the production thereof, and use thereof
US9241995B2 (en) 2011-06-22 2016-01-26 Trioptotec Gmbh 10H-benzo[G]pteridine-2,4-dione derivatives, method for the production thereof, and use thereof
US9745302B2 (en) 2011-06-22 2017-08-29 Trioptotec Gmbh 10H-benzo[g]pteridine-2,4-dione derivatives, method for the production thereof, and use thereof

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