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WO2017070012A1 - Lapachone derivatives containing two redox centers and methods of use thereof - Google Patents

Lapachone derivatives containing two redox centers and methods of use thereof Download PDF

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Publication number
WO2017070012A1
WO2017070012A1 PCT/US2016/056986 US2016056986W WO2017070012A1 WO 2017070012 A1 WO2017070012 A1 WO 2017070012A1 US 2016056986 W US2016056986 W US 2016056986W WO 2017070012 A1 WO2017070012 A1 WO 2017070012A1
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Prior art keywords
alkyl
hydrogen
substituted
compound
aryl
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PCT/US2016/056986
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French (fr)
Inventor
David A. Boothman
Eufranio N. DA SILVA JUNIOR
Eduardo H.g. DA CRUZ
Bruno C. CAVALCANTI
Antonio L. BRAGA
Claudia PESSOA
Molly SILVERS
Original Assignee
The Board Of Regents Of The University Of Texas System
Universidade Federal Do Ceara
Universidade Federal De Minas Gerais
Universidade Federal De Santa Catarina
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Priority to BR112018008076-4A priority Critical patent/BR112018008076B1/en
Publication of WO2017070012A1 publication Critical patent/WO2017070012A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/92—Naphthofurans; Hydrogenated naphthofurans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/04—1,2,3-Triazoles; Hydrogenated 1,2,3-triazoles
    • C07D249/06—1,2,3-Triazoles; Hydrogenated 1,2,3-triazoles with aryl radicals directly attached to ring atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/78—Ring systems having three or more relevant rings
    • C07D311/92—Naphthopyrans; Hydrogenated naphthopyrans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present disclosure relates generally to the field of chemotherapeutic compounds. More particularly, it concerns lapachone derivatives which contain two redox centers.
  • ⁇ -Lap had significant anti-tumor activity against Yoshida sarcoma and Walker 256 carcinoma cells in culture (Pink et al, 2000; Pink et al, 2000 and Planchon et al, 2001.)
  • the exact mechanism of action of ⁇ -lap was not known until recently, (Pink et al, 2000a; Pink et al, 2000b; Planchon et al, 2001) and the lack of a known intracellular target/mechanism previously limited development of ⁇ -lap as an antitumor agent.
  • ⁇ -Lap has a few characteristics that have limited its use as a chemotherapeutic agent, ⁇ -lap is highly hydrophobic and causes hemolysis in patients (Hartner et al, 2007).
  • the present disclosure provides lapachone derivatives which contain two redox centers.
  • these lapachone derivatives have a chalcogen redox center in addition to another redox center.
  • Xi is O or NRa, wherein:
  • Ra is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • X2 is ORb, wherein:
  • Rb is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or X2 is taken together with R2 as described below;
  • Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
  • Rc is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below;
  • R2 is hydrogen, halo, hydroxy, a selenium containing triazole group, or R2 is taken together with X2 as described below; or R2 is taken together with Ri as described below;
  • Yi is amino, cyano, halo, hydroxy, or nitro; or alkyl(c ⁇ 6), cycloalkyl(c ⁇ 6), acyl(c ⁇ 6), alkoxy(c ⁇ 6), acyloxy(c ⁇ 6), alkylamino(c ⁇ 6), dialkylamino(c ⁇ 6), amido(c ⁇ 6), or a substituted version of any of these groups; m is 0, 1, 2, 3, or 4;
  • X 3 is Cfo'I ' or O, wherein:
  • R3' and R4' are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c selenium containing triazole group;
  • R3, R4, and R5 are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; ni and n 2 are each independently 0, 1, 2, or 3; provided that when Ri and R2 are taken together, the compound is further defined by the formula:
  • X4 is CR(i'R7' or O, wherein: and R7' are each independently hydrogen, halo, alkyl(c ⁇ i2) substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or selenium containing triazole group; R.6, R.7, and Rs are each independently hydrogen, halo, alkyl(c ⁇ i2),
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 )rO(CH 2 ) s -, -(CH2)rNRd(CH 2 ) s -;
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • Xi is O or NRa, wherein:
  • Ra is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • X2 is ORb, wherein:
  • Rb is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
  • Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
  • Rc is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or
  • R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 ) r NRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4; Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • Xi is O or NRa, wherein:
  • Ra is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • X2 is ORb, wherein:
  • Rb is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
  • Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
  • Rc is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula: wherein:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 ) r NRd(CH 2 ) s -;
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i 2 ); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein: Rc is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); R.2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula: wherein:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 ) r NRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i 2 ); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • Xi is O or NRa, wherein:
  • Ra is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
  • Rc is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below; or
  • X3 is CR3'R4' or O, wherein:
  • R3' and R4' are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group;
  • R3, R4, and R5 are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; n is 0, 1 , 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 )rO(CH 2 ) s -, -(CH2)rNRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4; Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • X3 is CRj'I ' or O, wherein:
  • R3' and R4' are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group;
  • R3, R4, and R5 are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; ni and n 2 are each independently 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 ) r NRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i 2 ); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • R3, R.4, and R5 are each independently hydrogen, halo, alkyl(c ⁇ i 2 ), substituted alkyl(c ⁇ i 2 ), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; ni and n 2 are each independently 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula: wherein:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 ) r NRd(CH 2 ) s -;
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i 2 ); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined as:
  • Xi is O or NRa, wherein:
  • Ra is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • Rb is absent, hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
  • X4 is CR6'R7' or O, wherein:
  • R6' and R7' are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group;
  • R6, R7, and Rs are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 )rO(CH 2 ) s -, -(CH2)rNRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined:
  • X4 is CRe'Rj' or O, wherein:
  • R.6' and R7' are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group;
  • R.6, R7, and Rs are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
  • X5 is a covalent bond, -(CH 2 ) q - -(CH 2 )rO(CH 2 ) s -, -(CH 2 )rNRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2); or a pharmaceutically acceptable salt thereof.
  • the compounds are further defined:
  • R.6, R.7, and Rs are each independently hydrogen, halo, alkyl(c ⁇ i2), substituted alkyl(c ⁇ i2), aryl(c ⁇ i2), substituted aryl(c ⁇ i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 )rO(CH 2 ) s -, -(CH2)rNRd(CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
  • Y2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i2), alkenediyl(c ⁇ i2), arenediyl(c ⁇ i2), heteroarenediyl(c ⁇ i2), or a substituted version of any of these groups;
  • A is a group of the formula:
  • R.9 is aryl(c ⁇ i2) or substituted aryl(c ⁇ i2);
  • Xi is O.
  • X2 is ORb wherein Rb is absent.
  • Ri is hydrogen. In other embodiments, Ri is halo such as Ri is bromo or chloro. In other embodiments, Ri is a selenium containing triazole group.
  • R2 is hydrogen. In other embodiments, R2 is halo such as R2 is bromo or chloro. In some embodiments, R2 is a selenium containing triazole group. In some embodiments, m is 0.
  • X3 is O.
  • R3 is hydrogen.
  • R3 is halo such as R3 is chloro or bromo.
  • R3 is alkyl(c ⁇ 6) such as R3 is methyl.
  • R3 is a selenium containing triazole group.
  • R4 is hydrogen. In other embodiments, R4 is halo such as R4 is chloro or bromo. In other embodiments, R4 is alkyl(c ⁇ 6) such as R4 is methyl. In other embodiments, R4 is a selenium containing triazole group.
  • R5 is hydrogen. In other embodiments, R5 is a selenium containing triazole group. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2. In some embodiments, X4 is O. [0016] In some embodiments, R.6 is hydrogen. In other embodiments, R.6 is halo such as R.6 is chloro or bromo. In other embodiments, R.6 is alkyl(c ⁇ 6) such as R6 is methyl. In other embodiments, R6 is a selenium containing triazole group.
  • R7 is hydrogen. In other embodiments, R7 is halo such as R7 is chloro or bromo. In other embodiments, R7 is alkyl(c ⁇ 6) such as R7 is methyl. In other embodiments, R7 is a selenium containing triazole group.
  • Rs is hydrogen. In other embodiments, Rs is aryl(c ⁇ i2) such as R8 is phenyl or napthyl. In other embodiments, Rs is a selenium containing triazole group. In some embodiments, p is 1 or 2. In some embodiments, p is 1. In other embodiments, p is 2.
  • the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 )rNR d (CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • q 1, 2, 3, or 4;
  • r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups; and
  • R9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i2).
  • the selenium containing triazole group is further defined by the formula:
  • X 5 is a covalent bond, -(CH 2 ) q - -(CH 2 ) r O(CH 2 ) s - -(CH 2 )rNR d (CH 2 ) s -; wherein:
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6);
  • q 1, 2, 3, or 4;
  • r and s are each independently 0, 1, 2, 3, or 4;
  • Y 2 and Y3 are each independently a covalent bond, alkanediyl(c ⁇ i 2 ), alkenediyl(c ⁇ i 2 ), arenediyl(c ⁇ i 2 ), heteroarenediyl(c ⁇ i 2 ), or a substituted version of any of these groups; and
  • R9 is aryl(c ⁇ i 2 ) or substituted aryl(c ⁇ i2).
  • X5 is a covalent bond. In other embodiments, X5 is
  • Rd is hydrogen, alkyl(c ⁇ 6), or substituted alkyl(c ⁇ 6); r and s are each independently 0, 1, 2, 3, or 4.
  • X5 is -NH-. In other embodiments, X5 is -NHCH2-.
  • Y 2 is alkanediyl(c ⁇ 6) such as Y 2 is -CH 2 - In other embodiments, Y 2 is arenediyl(c ⁇ 6) such as Y 2 is benzenediyl.
  • Y3 is alkanediyl(c ⁇ 6) such as Y3 is -CH 2 -.
  • R9 is aryl(c ⁇ 8) such as R9 is phenyl.
  • the compound is further defined as:
  • compositions comprising:
  • the pharmaceutical composition is formulated for administration: orally, intraadiposally, intraarterially, intraarticularly, intracranially, intradermally, intralesionally, intramuscularly, intranasally, intraocularly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrarectally, intrathecally, intratracheally, intratumorally, intraumbilically, intravaginally, intravenously, intravesicularly, intravitreally, liposomally, locally, mucosally, parenterally, rectally, subconjunctival, subcutaneously, sublingually, topically, transbuccally, transdermally, vaginally, in cremes, in lipid compositions, via a catheter, via a lavage, via continuous infusion, via infusion, via inhalation, via injection, via local delivery, or via localized perfusion.
  • the pharmaceutical composition further comprises a cyclodextran.
  • the cyclodextran is a ⁇ -cyclodextran such as a hydropropyl- -cyclodextran.
  • the pharmaceutical composition is formulated as a micelle or a liposome such as a micelle or liposome is formed by poly(lactic acid) poly(ethylene glycol) (PLA PEG).
  • the present disclosure provides methods of treating a patient with a disease or disorder comprising administering a therapeutically effective amount of a compound or composition described herein to the patient in need thereof.
  • the disease or disorder is cancer.
  • the cancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma.
  • the cancer is of the bladder, blood, bone, brain, breast, central nervous system, cervix, colon, endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia, genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen, small intestine, large intestine, stomach, testicle, or thyroid.
  • the cancer overexpresses NAD(P)H:quinone oxidoreductase 1 (NQOl).
  • the cancer is leukemia, colon cancer, prostate cancer, brain cancer, ovarian cancer, or skin cancer.
  • the colon cancer is colon carcinoma.
  • the skin cancer is melanoma.
  • the brain cancer is a glioma.
  • the leukemia is acute myeloid leukemia.
  • the prostate cancer is an androgen independent prostate cancer.
  • the ovarian cancer is an ovarian adenocarcinoma.
  • the methods comprise administering a second anti- cancer therapy.
  • the second anti-cancer therapy is a second chemotherapeutic compound, radiation therapy, surgery, or immunotherapy.
  • the second chemotherapeutic compound is a PARP1 inhibitor, a glutamine/glutamate pathway inhibitor, or a DNA base excision repair (BER) inhibitor.
  • the patient is a mammal such as a human.
  • the methods comprise administering the compound once. In other embodiments, the methods comprise administering the compound two or more times.
  • the patient has been determined to have defective DNA repair, defective ability to maintain NAD(P)H levels by the glucose, glutamate/glutamine, or pyruvate pathways, or genetic defects causing synthetic lethality for precision therapy against specific NQ01+ human cancers.
  • the disease or disorder is a metabolic syndrome.
  • the metabolic syndrome exhibits elevated NAD(P)H/NAD(P) + levels.
  • the disease or disorder is trypanosomal disease.
  • FIGS. 1A-1G show the NQOl activity of compounds 21, 22, 50, 51, 52, 53, and ⁇ -lapachone.
  • Compound 53 showed the most dramatic loss of viability within a narrow therapeutic window, going from 93% viability at 0.8 ⁇ to 11% viability at 1.6 ⁇ .
  • 50-53 NQOl -specific compounds showed either similar or lower IC50 values than ⁇ -lapachone.
  • Compounds 21 and 22, selenium-containing quinones, with IC50 values 0.64 and 1.2, respectively, were the most active of this series and are also NQOl -dependent (see Table 2).
  • FIG. 2 shows the effect on phosphatidylserine externalization after 6 h-treated PC3 with 5 ⁇ of tested compounds.
  • the phosphatidylserine externalization was determined by flow cytometry using AnnV-FITC (YLW-HLog) and PI (RED-HLog). Viable cells are plotted at lower left quadrant, cells in early and late apoptosis with phosphatidylserine externalized are plotted at lower right and upper right quadrants, respectively, and necrotic cells are plotted at upper left quadrant.
  • *p ⁇ 0.05 compared to control by ANOVA followed by Newman-Keuls test. Data are presented as mean values ⁇ S.E.M. from three independent experiments in triplicate.
  • FIG. 3 shows the effect of compound 21 (5 ⁇ ) on ROS production on PC3 human prostate cancer cell line evaluated by H2DCFDA fluorescent probe after 1 h exposure in presence or absence of 5 mM NAC.
  • the present disclosure provides compounds which may be useful in the treatment of cancer which contain two different redox centers.
  • one of the redox centers in the lapachone derivatives contain a chalcogen atom as a redox center.
  • these compounds are useful in treating cancers including cancers which are NQOl dependent.
  • Chemical formulas used to represent compounds of the disclosure will typically only show one of possibly several different tautomers. For example, many types of ketone groups are known to exist in equilibrium with corresponding enol groups. Similarly, many types of imine groups exist in equilibrium with enamine groups. Regardless of which tautomer is depicted for a given compound, and regardless of which one is most prevalent, all tautomers of a given chemical formula are intended.
  • Compounds of the disclosure may also have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. , higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the indications stated herein or otherwise.
  • a better pharmacokinetic profile e.g. , higher oral bioavailability and/or lower clearance
  • atoms making up the compounds of the present disclosure are intended to include all isotopic forms of such atoms.
  • Isotopes include those atoms having the same atomic number but different mass numbers.
  • isotopes of hydrogen include tritium and deuterium
  • isotopes of carbon include 1 C and 14 C.
  • Compounds of the present disclosure may also exist in prodrug form. Since prodrugs are known to enhance numerous desirable qualities of pharmaceuticals (e.g. , solubility, bioavailability, manufacturing, etc.), the compounds employed in some methods of the disclosure may, if desired, be delivered in prodrug form. Thus, the disclosure contemplates prodrugs of compounds of the present disclosure as well as methods of delivering prodrugs. Prodrugs of the compounds employed in the disclosure may be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound.
  • prodrugs include, for example, compounds described herein in which a hydroxy, amino, or carboxy group is bonded to any group that, when the prodrug is administered to a subject, cleaves to form a hydroxy, amino, or carboxylic acid, respectively.
  • the particular anion or cation forming a part of any salt form of a compound provided herein is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (2002), which is incorporated herein by reference.
  • the present disclosure relates to compounds which contain a chalcogen atom which can act as a redox center.
  • a chalcogen is an element of Group 16 of the periodic table, including oxygen, sulfur, selenium and tellurium.
  • a "heavy chalcogen” as used herein refers to a chalcogen atom having an atomic weight heavier than oxygen, for example, sulfur or selenium.
  • selenium is the heavy chalcogen used herein.
  • Compounds containing a chalcogen atom are believed, without wishing to be bound by any theory, to function primarily as competitive substrates (e.g. as "sacrificial" antioxidants), which are preferentially attacked by oxidative species and thus useful in the treatment of cancer.
  • the chalcogen redox centers reduce the amount of reactive oxygen species present and/or reduce the oxidative stress experienced by cells.
  • the prototypical example is cancer.
  • cancer One of the key elements of cancer is that the cell's normal apoptotic cycle is interrupted and thus agents that interrupt the growth of the cells are important as therapeutic agents for treating these diseases.
  • the lapachone analogs described herein may be used to lead to decreased cell counts and as such can potentially be used to treat a variety of types of cancer lines. In some aspects, it is anticipated that the lapachone analogs described herein may be used to treat virtually any malignancy.
  • the compounds of the present disclosure may be used in the treatment of NQOl dependent cancers.
  • NQOl is known to regulate p53 activity and thus useful in the treatment of hyperproliferative diseases such as cancer. Additionally, NQOl acts as a detoxification protein which removes reactive oxygen species from the cell thus reducing oxidative damage and cancer. Modification of NQOl activity and expression is therefore useful in the treatment of cancer.
  • Some non-limiting examples of cancers which have been associated with mis-regulation of NQOl include but are not limited to bladder cancer, breast cancer, colorectal cancer, prostate cancer or acute lymphoblastic leukemia.
  • Cancer cells that may be treated with the compounds of the present disclosure include but are not limited to cells from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, pancreas, testis, tongue, cervix, or uterus.
  • the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acid
  • the tumor may comprise an osteosarcoma, angiosarcoma, rhabdosarcoma, leiomyosarcoma, Ewing sarcoma, glioblastoma, neuroblastoma, or leukemia.
  • compositions in a form appropriate for the intended application.
  • such formulation with the compounds of the present disclosure is contemplated.
  • this will entail preparing compositions that are essentially free of pyrogens, as well as other impurities that could be harmful to humans or animals.
  • compositions of the present disclosure comprise an effective amount of the vector to cells, dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium. Such compositions also are referred to as inocula.
  • pharmaceutically or pharmacologically acceptable refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • the use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the vectors or cells of the present disclosure, its use in therapeutic compositions is contemplated. Supplementary active ingredients also can be incorporated into the compositions.
  • the active lapachone analogs compositions of the present disclosure may include classic pharmaceutical preparations. Administration of these compositions according to the present disclosure will be via any common route so long as the target tissue is available via that route. Such routes include oral, nasal, buccal, rectal, vaginal or topical route. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intratumoral, intraperitoneal, or intravenous injection. Such compositions would normally be administered as pharmaceutically acceptable compositions, described supra. [0054] The active compounds may also be administered parenterally or intraperitoneally. Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
  • the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • a coating such as lecithin
  • surfactants for example, sodium sulfate, sodium sulfate, sodium sulfate, sodium sulfate, sodium sulfate, sodium sulfate, sodium sulfate, sodium sorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum-drying and freeze- drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • the use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • the lapachone analogs described herein may be incorporated with excipients and used in the form of non-ingestible mouthwashes and dentifrices.
  • a mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution).
  • the active ingredient may be incorporated into an antiseptic wash containing sodium borate, glycerin and potassium bicarbonate.
  • the active ingredient may also be dispersed in dentifrices, including: gels, pastes, powders and slurries.
  • the active ingredient may be added in a therapeutically effective amount to a paste dentifrice that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants.
  • the lapachone analog compositions of the present disclosure may be formulated in a neutral or salt form.
  • Pharmaceutically-acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.
  • solutions Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.
  • the formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like.
  • the solution For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose.
  • aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
  • sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure.
  • one dosage could be dissolved in 1 mL of isotonic NaCl solution and either added to 1000 mL of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences," 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.
  • compositions that may be used in treating microbial infections and cancer in a subject are disclosed herein.
  • the compositions described above are preferably administered to a mammal (e.g., rodent, human, non-human primates, canine, bovine, ovine, equine, feline, etc.) in an effective amount, that is, an amount capable of producing a desirable result in a treated subject (e.g., causing apoptosis of cancerous cells or killing bacterial cells).
  • Toxicity and therapeutic efficacy of the compositions utilized in methods of the disclosure can be determined by standard pharmaceutical procedures.
  • dosage for any one animal depends on many factors, including the subject's size, body surface area, body weight, age, the particular composition to be administered, time and route of administration, general health, the clinical symptoms of the infection or cancer and other drugs being administered concurrently.
  • a composition as described herein is typically administered at a dosage that inhibits the growth or proliferation of a bacterial cell, inhibits the growth of a biofilm, or induces death of cancerous cells (e.g., induces apoptosis of a cancer cell), as assayed by identifying a reduction in hematological parameters (complete blood count - CBC), or cancer cell growth or proliferation.
  • amounts of the lapachone analogs used to inhibit bacterial growth or induce apoptosis of the cancer cells is calculated to be from about 0.01 mg to about 10,000 mg/day. In some embodiments, the amount is from about 1 mg to about 1,000 mg/day. In some embodiments, these dosings may be reduced or increased based upon the biological factors of a particular patient such as increased or decreased metabolic breakdown of the drug or decreased uptake by the digestive tract if administered orally. Addtionally, the lapachone analogs may be more efficacious and thus a smaller dose is required to achieve a similar effect. Such a dose is typically administered once a day for a few weeks or until sufficient reducing in cancer cells has been achieved.
  • the therapeutic methods of the disclosure in general include administration of a therapeutically effective amount of the compositions described herein to a subject in need thereof, including a mammal, particularly a human.
  • Such treatment will be suitably administered to subjects, particularly humans, suffering from, having, susceptible to, or at risk for a disease, disorder, or symptom thereof. Determination of those subjects "at risk” can be made by any objective or subjective determination by a diagnostic test or opinion of a subject or health care provider (e.g., genetic test, enzyme or protein marker, marker (as defined herein), family history, and the like).
  • the disclosure provides a method of monitoring treatment progress.
  • the method includes the step of determining a level of changes in hematological parameters and/or cancer stem cell (CSC) analysis with cell surface proteins as diagnostic markers (which can include, for example, but are not limited to CD34, CD38, CD90, and CD 117) or diagnostic measurement (e.g. , screen, assay) in a subject suffering from or susceptible to a disorder or symptoms thereof associated with cancer (e.g., leukemia) in which the subject has been administered a therapeutic amount of a composition as described herein.
  • CSC cancer stem cell
  • diagnostic measurement e.g. , screen, assay
  • the level of marker determined in the method can be compared to known levels of marker in either healthy normal controls or in other afflicted patients to establish the subject's disease status.
  • a second level of marker in the subject is determined at a time point later than the determination of the first level, and the two levels are compared to monitor the course of disease or the efficacy of the therapy.
  • a pre-treatment level of marker in the subject is determined prior to beginning treatment according to the methods described herein; this pre-treatment level of marker can then be compared to the level of marker in the subject after the treatment commences, to determine the efficacy of the treatment.
  • the lapachone analogs described herein may be used in combination therapies with one or more cancer therapies or a compound which mitigates one or more of the side effects experienced by the patient. It is common in the field of cancer therapy to combine therapeutic modalities. The following is a general discussion of therapies that may be used in conjunction with the therapies of the present disclosure.
  • a tumor cell or subject with a compound and at least one other therapy.
  • These therapies would be provided in a combined amount effective to achieve a reduction in one or more disease parameter.
  • This process may involve contacting the cells/subjects with the both agents/therapies at the same time, e.g., using a single composition or pharmacological formulation that includes both agents, or by contacting the cell/subject with two distinct compositions or formulations, at the same time, wherein one composition includes the compound and the other includes the other agent.
  • the lapachone analogs described herein may precede or follow the other treatment by intervals ranging from minutes to weeks.
  • chemotherapeutic agent refers to the use of drugs to treat cancer.
  • a “chemotherapeutic agent” is used to connote a compound or composition that is administered in the treatment of cancer. These agents or drugs are categorized by their mode of activity within a cell, for example, whether and at what stage they affect the cell cycle. Alternatively, an agent may be characterized based on its ability to directly cross-link DNA, to intercalate into DNA, or to induce chromosomal and mitotic aberrations by affecting nucleic acid synthesis. Most chemotherapeutic agents fall into the following categories: alkylating agents, antimetabolites, antitumor antibiotics, mitotic inhibitors, and nitrosoureas.
  • chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duo
  • paclitaxel and docetaxel paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g.
  • topoisomerase inhibitor RFS 2000 difluorometlhylomithine (DMFO); retinoids such as retinoic acid; capecitabine; cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, paclitaxel, docetaxel, gemcitabien, navelbine, farnesyl-protein tansferase inhibitors, transplatinum, 5-fluorouracil, vincristin, vinblastin and methotrexate and pharmaceutically acceptable salts, acids or
  • Radiotherapy also called radiation therapy, is the treatment of cancer and other diseases with ionizing radiation. Ionizing radiation deposits energy that injures or destroys cells in the area being treated by damaging their genetic material, making it impossible for these cells to continue to grow. Although radiation damages both cancer cells and normal cells, the latter are able to repair themselves and function properly.
  • Radiation therapy used according to the present disclosure may include, but is not limited to, the use of ⁇ -rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors are also contemplated such as microwaves and UV-irradiation.
  • Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens.
  • Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.
  • Radiotherapy may comprise the use of radiolabeled antibodies to deliver doses of radiation directly to the cancer site (radioimmunotherapy).
  • Antibodies are highly specific proteins that are made by the body in response to the presence of antigens (substances recognized as foreign by the immune system). Some tumor cells contain specific antigens that trigger the production of tumor-specific antibodies. Large quantities of these antibodies can be made in the laboratory and attached to radioactive substances (a process known as radiolabeling). Once injected into the body, the antibodies actively seek out the cancer cells, which are destroyed by the cell-killing (cytotoxic) action of the radiation. This approach can minimize the risk of radiation damage to healthy cells.
  • Conformal radiotherapy uses the same radiotherapy machine, a linear accelerator, as the normal radiotherapy treatment but metal blocks are placed in the path of the x-ray beam to alter its shape to match that of the cancer. This ensures that a higher radiation dose is given to the tumor. Healthy surrounding cells and nearby structures receive a lower dose of radiation, so the possibility of side effects is reduced.
  • a device called a multi- leaf collimator has been developed and may be used as an alternative to the metal blocks.
  • the multi-leaf collimator consists of a number of metal sheets which are fixed to the linear accelerator. Each layer can be adjusted so that the radiotherapy beams can be shaped to the treatment area without the need for metal blocks. Precise positioning of the radiotherapy machine is very important for conformal radiotherapy treatment and a special scanning machine may be used to check the position of internal organs at the beginning of each treatment.
  • High-resolution intensity modulated radiotherapy also uses a multi-leaf collimator. During this treatment the layers of the multi-leaf collimator are moved while the treatment is being given. This method is likely to achieve even more precise shaping of the treatment beams and allows the dose of radiotherapy to be constant over the whole treatment area.
  • Radiosensitizers make the tumor cells more likely to be damaged, and radioprotectors protect normal tissues from the effects of radiation.
  • Hyperthermia the use of heat, is also being studied for its effectiveness in sensitizing tissue to radiation.
  • immunotherapeutics In the context of cancer treatment, immunotherapeutics, generally, rely on the use of immune effector cells and molecules to target and destroy cancer cells.
  • Trastuzumab (HerceptinTM) is such an example.
  • the immune effector may be, for example, an antibody specific for some marker on the surface of a tumor cell.
  • the antibody alone may serve as an effector of therapy or it may recruit other cells to actually affect cell killing.
  • the antibody also may be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and serve merely as a targeting agent.
  • the effector may be a lymphocyte carrying a surface molecule that interacts, either directly or indirectly, with a tumor cell target.
  • Various effector cells include cytotoxic T cells and NK cells. The combination of therapeutic modalities, i.e., direct cytotoxic activity and inhibition or reduction of ErbB2 would provide therapeutic benefit in the treatment of ErbB2 overexpressing cancers.
  • the tumor cell must bear some marker that is amenable to targeting, i.e. , is not present on the majority of other cells.
  • Common tumor markers include carcinoembryonic antigen, prostate specific antigen, urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor, laminin receptor, erb B and pi 55.
  • An alternative aspect of immunotherapy is to combine anticancer effects with immune stimulatory effects.
  • Immune stimulating molecules also exist including: cytokines such as IL-2, IL-4, IL-12, GM-CSF, ⁇ -IFN, chemokines such as MIP-1, MCP-1, IL-8 and growth factors such as FLT3 ligand.
  • cytokines such as IL-2, IL-4, IL-12, GM-CSF, ⁇ -IFN
  • chemokines such as MIP-1, MCP-1, IL-8
  • growth factors such as FLT3 ligand.
  • Combining immune stimulating molecules, either as proteins or using gene delivery in combination with a tumor suppressor has been shown to enhance antitumor effects (Ju et al , 2000).
  • antibodies against any of these compounds may be used to target the anti-cancer agents discussed herein.
  • immunotherapies currently under investigation or in use are immune adjuvants, e.g. , Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene and aromatic compounds (U.S. Patents 5,801,005 and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al, 1998), cytokine therapy, e.g.
  • interferons ⁇ , ⁇ , and ⁇ ; IL-1, GM-CSF and TNF (Bukowski et al, 1998; Davidson et al , 1998; Hellstrand et al , 1998) gene therapy, e.g., TNF, IL-1, IL-2, p53 (Qin et al, 1998; Austin-Ward and Villaseca, 1998; U.S. Patents 5,830,880 and 5,846,945) and monoclonal antibodies, e.g., anti-ganglioside GM2, anti-HER- 2, anti-pl85 (Pietras et al, 1998; Hanibuchi et al, 1998; U.S. Patent 5,824,311). It is contemplated that one or more anti-cancer therapies may be employed with the gene silencing therapies described herein.
  • an antigenic peptide, polypeptide or protein, or an autologous or allogenic tumor cell composition or "vaccine” is administered, generally with a distinct bacterial adjuvant (Ravindranath and Morton, 1991; Morton et al , 1992; Mitchell et al , 1990; Mitchell et al, 1993).
  • adoptive immunotherapy the patient's circulating lymphocytes, or tumor infiltrated lymphocytes, are isolated in vitro, activated by lymphokines such as IL-2 or transduced with genes for tumor necrosis, and readministered (Rosenberg et al, 1988; 1989).
  • Curative surgery is a cancer treatment that may be used in conjunction with other therapies, such as the treatment of the present disclosure, chemotherapy, radiotherapy, hormonal therapy, gene therapy, immunotherapy and/or alternative therapies.
  • Curative surgery includes resection in which all or part of cancerous tissue is physically removed, excised, and/or destroyed. Tumor resection refers to physical removal of at least part of a tumor.
  • treatment by surgery includes laser surgery, cryosurgery, electrosurgery, and microscopically controlled surgery (Mohs' surgery). It is further contemplated that the present disclosure may be used in conjunction with removal of superficial cancers, precancers, or incidental amounts of normal tissue.
  • a cavity may be formed in the body.
  • Treatment may be accomplished by perfusion, direct injection or local application of the area with an additional anti-cancer therapy.
  • Such treatment may be repeated, for example, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5 weeks or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.
  • These treatments may be of varying dosages as well.
  • an adjuvant treatment with a compound of the present disclosure is believe to be particularly efficacious in reducing the reoccurance of the tumor.
  • the compounds of the present disclosure can also be used in a neoadjuvant setting.
  • agents may be used with the present disclosure.
  • additional agents include immunomodulatory agents, agents that affect the upregulation of cell surface receptors and GAP junctions, cytostatic and differentiation agents, inhibitors of cell adhesion, agents that increase the sensitivity of the hyperproliferative cells to apoptotic inducers, or other biological agents.
  • Immunomodulatory agents include tumor necrosis factor; interferon alpha, beta, and gamma; IL-2 and other cytokines; F42K and other cytokine analogs; or MIP-1, ⁇ - ⁇ ⁇ , MCP-1, RANTES, and other chemokines.
  • cell surface receptors or their ligands such as Fas/Fas ligand, DR4 or DR5/TRAIL (Apo-2 ligand) would potentiate the apoptotic inducing abilities of the present disclosure by establishment of an autocrine or paracrine effect on hyperproliferative cells. Increases intercellular signaling by elevating the number of GAP junctions would increase the anti-hyperproliferative effects on the neighboring hyperproliferative cell population.
  • cytostatic or differentiation agents may be used in combination with the present disclosure to improve the anti-hyerproliferative efficacy of the treatments.
  • Inhibitors of cell adhesion are contemplated to improve the efficacy of the present disclosure.
  • cell adhesion inhibitors are focal adhesion kinase (FAKs) inhibitors and Lovastatin. It is further contemplated that other agents that increase the sensitivity of a hyperproliferative cell to apoptosis, such as the antibody c225, could be used in combination with the present disclosure to improve the treatment efficacy.
  • FAKs focal adhesion kinase
  • Lovastatin Lovastatin
  • hyperthermia is a procedure in which a patient's tissue is exposed to high temperatures (up to 106°F).
  • External or internal heating devices may be involved in the application of local, regional, or whole-body hyperthermia.
  • Local hyperthermia involves the application of heat to a small area, such as a tumor. Heat may be generated externally with high-frequency waves targeting a tumor from a device outside the body. Internal heat may involve a sterile probe, including thin, heated wires or hollow tubes filled with warm water, implanted microwave antennae, or radiofrequency electrodes.
  • a patient's organ or a limb is heated for regional therapy, which is accomplished using devices that produce high energy, such as magnets. Altematively, some of the patient's blood may be removed and heated before being perfused into an area that will be internally heated. Whole-body heating may also be implemented in cases where cancer has spread throughout the body. Warm- water blankets, hot wax, inductive coils, and thermal chambers may be used for this purpose.
  • the symbol “-” means a single bond
  • “ ⁇ ” means triple bond.
  • the symbol " " represents an optional bond, which if present is either single or double.
  • the covalent bond symbol when connecting one or two stereogenic atoms does not indicate any preferred stereochemistry. Instead, it covers all stereoisomers as well as mixtures thereof.
  • the symbol “ ⁇ ", when drawn perpendicularly across a bond indicates a point of attachment of the group.
  • the point of attachment is typically only identified in this manner for larger groups in order to assist the reader in unambiguously identifying a point of attachment.
  • the symbol “- ⁇ ” means a single bond where the group attached to the thick end of the wedge is “out of the page.”
  • the symbol “”HI” means a single bond where the group attached to the thick end of the wedge is “into the page”.
  • the symbol “ ⁇ vw” means a single bond where the geometry around a double bond (e.g., either E or Z) is undefined. Both options, as well as combinations thereof are therefore intended. Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to that atom.
  • a bold dot on a carbon atom indicates that the hydrogen attached to that carbon is oriented out of the plane of the paper.
  • R may replace any hydrogen atom attached to any of the ring atoms, including a depicted, implied, or expressly defined hydrogen, so long as a stable structure is formed.
  • a group "R” is depicted as a "floating group” on a fused ring system, as for example in the formula:
  • R may replace any hydrogen attached to any of the ring atoms of either of the fused rings unless specified otherwise.
  • Replaceable hydrogens include depicted hydrogens (e.g., the hydrogen attached to the nitrogen in the formula above), implied hydrogens (e.g. , a hydrogen of the formula above that is not shown but understood to be present), expressly defined hydrogens, and optional hydrogens whose presence depends on the identity of a ring atom (e.g., a hydrogen attached to group X, when X equals -CH-), so long as a stable structure is formed.
  • R may reside on either the 5-membered or the 6- membered ring of the fused ring system.
  • the number of carbon atoms in the group or class is as indicated as follows: "Cn” defines the exact number (n) of carbon atoms in the group/class. "C ⁇ n” defines the maximum number (n) of carbon atoms that can be in the group/class, with the minimum number as small as possible for the group/class in question, e.g. , it is understood that the minimum number of carbon atoms in the group “alkenyl(c ⁇ 8)” or the class “alkene(c ⁇ 8)” is two. Compare with “alkoxy(c ⁇ io)", which designates alkoxy groups having from 1 to 10 carbon atoms.
  • Cn-n defines both the minimum (n) and maximum number ( ⁇ ') of carbon atoms in the group.
  • alkyl(C2-io) designates those alkyl groups having from 2 to 10 carbon atoms. These carbon number indicators may precede or follow the chemical groups or class it modifies and it may or may not be enclosed in parenthesis, without signifying any change in meaning.
  • C5 olefin C5-olefin
  • olefin(C5) olefincs
  • saturated when used to modify a compound or chemical group means the compound or chemical group has no carbon-carbon double and no carbon-carbon triple bonds, except as noted below.
  • the term when used to modify an atom, it means that the atom is not part of any double or triple bond.
  • substituted versions of saturated groups one or more carbon oxygen double bond or a carbon nitrogen double bond may be present. And when such a bond is present, then carbon-carbon double bonds that may occur as part of keto-enol tautomerism or imine/enamine tautomerism are not precluded.
  • saturated when used to modify a solution of a substance, it means that no more of that substance can dissolve in that solution.
  • aliphatic when used without the "substituted” modifier signifies that the compound or chemical group so modified is an acyclic or cyclic, but non-aromatic hydrocarbon compound or group.
  • the carbon atoms can be joined together in straight chains, branched chains, or non-aromatic rings (alicyclic).
  • Aliphatic compounds/groups can be saturated, that is joined by single carbon-carbon bonds (alkanes/alkyl), or unsaturated, with one or more carbon-carbon double bonds (alkenes/alkenyl) or with one or more carbon-carbon triple bonds (alkynes/alkynyl).
  • aromatic when used to modify a compound or a chemical group atom means the compound or chemical group contains a planar unsaturated ring of atoms that is stabilized by an interaction of the bonds forming the ring.
  • alkyl when used without the “substituted” modifier refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, a linear or branched acyclic structure, and no atoms other than carbon and hydrogen.
  • alkanediyl when used without the "substituted” modifier refers to a divalent saturated aliphatic group, with one or two saturated carbon atom(s) as the point(s) of attachment, a linear or branched acyclic structure, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen.
  • the groups -CH2- (methylene), -CH2CH2-, -CH 2 C(CH 3 ) 2 CH 2 -, and -CH2CH2CH2- are non- limiting examples of alkanediyl groups.
  • An "alkane” refers to the class of compounds having the formula H-R, wherein R is alkyl as this term is defined above.
  • haloalkyl is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to halo (i.e.
  • -F, -CI, -Br, or -I such that no other atoms aside from carbon, hydrogen and halogen are present.
  • -CH2CI is a non-limiting example of a haloalkyl.
  • fluoroalkyl is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to fluoro such that no other atoms aside from carbon, hydrogen and fluorine are present.
  • the groups -CH2F, -CF3, and -CH2CF3 are non-limiting examples of fluoroalkyl groups.
  • cycloalkyl when used without the “substituted” modifier refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, said carbon atom forming part of one or more non-aromatic ring structures, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen.
  • Non-limiting examples include: -CH(CH2)2 (cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl (Cy).
  • cycloalkanediyl when used without the “substituted” modifier refers to a divalent saturated aliphatic group with two carbon atoms as points of attachment, no carbon- double or triple bonds, and no atoms other than carbon and hydrogen.
  • the group is a non-limiting example of cycloalkanediyl group.
  • a "cycloalkane” refers to the class of compounds having the formula H-R, wherein R is cycloalkyl as this term is defined above.
  • alkenyl when used without the "substituted” modifier refers to an monovalent unsaturated aliphatic group with a carbon atom as the point of attachment, a linear or branched, acyclic structure, at least one nonaromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen.
  • alkenediyl when used without the "substituted” modifier refers to a divalent unsaturated aliphatic group, with two carbon atoms as points of attachment, a linear or branched, a linear or branched acyclic structure, at least one nonaromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen.
  • alkenediyl group is aliphatic, once connected at both ends, this group is not precluded from forming part of an aromatic structure.
  • alkene and olefin are synonymous and refer to the class of compounds having the formula H-R, wherein R is alkenyl as this term is defined above.
  • terminal alkene and a-olefin are synonymous and refer to an alkene having just one carbon-carbon double bond, wherein that bond is part of a vinyl group at an end of the molecule.
  • one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH 2 , -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH 3 , -NHCH3, -NHCH2CH3, -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 )2, -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • alkynyl when used without the "substituted” modifier refers to a monovalent unsaturated aliphatic group with a carbon atom as the point of attachment, a linear or branched acyclic structure, at least one carbon-carbon triple bond, and no atoms other than carbon and hydrogen. As used herein, the term alkynyl does not preclude the presence of one or more non-aromatic carbon-carbon double bonds.
  • alkyne refers to the class of compounds having the formula H-R, wherein R is alkynyl.
  • one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, - H2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH 3 , -NHCH3, -NHCH2CH3, -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 ) 2 , -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • aryl when used without the "substituted” modifier refers to a monovalent unsaturated aromatic group with an aromatic carbon atom as the point of attachment, said carbon atom forming part of a one or more six-membered aromatic ring structure, wherein the ring atoms are all carbon, and wherein the group consists of no atoms other than carbon and hydrogen. If more than one ring is present, the rings may be fused or unfused. As used herein, the term does not preclude the presence of one or more alkyl or aralkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present.
  • Non-limiting examples of aryl groups include phenyl (Ph), methylphenyl, (dimethyl)phenyl, -C6H4CH2CH3 (ethylphenyl), naphthyl, and a monovalent group derived from biphenyl.
  • the term "arenediyl” when used without the “substituted” modifier refers to a divalent aromatic group with two aromatic carbon atoms as points of attachment, said carbon atoms forming part of one or more six-membered aromatic ring structure(s) wherein the ring atoms are all carbon, and wherein the monovalent group consists of no atoms other than carbon and hydrogen.
  • the term does not preclude the presence of one or more alkyl, aryl or aralkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present. If more than one ring is present, the rings may be fused or unfused. Unfused rings may be connected via one or more of the following: a covalent bond, alkanediyl, or alkenediyl groups (carbon number limitation permitting).
  • arenediyl groups include:
  • an "arene” refers to the class of compounds having the formula H-R, wherein R is aryl as that term is defined above. Benzene and toluene are non-limiting examples of arenes. When any of these terms are used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH 2 , -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH 3 , -NHCH3, -NHCH2CH3, -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 ) 2 , -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • acyl when used without the "substituted” modifier refers to the group -C(0)R, in which R is a hydrogen, alkyl, cycloalkyl, alkenyl, or aryl, as those terms are defined above.
  • the groups, -CHO, -C(0)CH 3 (acetyl, Ac), -C(0)CH 2 CH 3 , -C(0)CH 2 CH 2 CH 3 , -C(0)CH(CH 3 ) 2 , -C(0)CH(CH 2 ) 2 , -C(0)C 6 H 5 , -C(0)C 6 H 4 CH3, -C(0)CH2C6H5, -C(0)(imidazolyl) are non-limiting examples of acyl groups.
  • a “thioacyl” is defined in an analogous manner, except that the oxygen atom of the group -C(0)R has been replaced with a sulfur atom, -C(S)R.
  • aldehyde corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with a -CHO group.
  • one or more hydrogen atom (including a hydrogen atom directly attached to the carbon atom of the carbonyl or thiocarbonyl group, if any) has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH 3 , -NHCH3, -NHCH2CH3, -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 ) 2 , -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • the groups, -C(0)CH 2 CF 3 , -C0 2 H (carboxyl), -C0 2 CH 3 (methylcarboxyl), -C0 2 CH 2 CH 3 , -C(0)NH 2 (carbamoyl), and -CON(CH 3 ) 2 are non-limiting examples of substituted acyl groups.
  • alkoxy when used without the "substituted” modifier refers to the group -OR, in which R is an alkyl, as that term is defined above.
  • Non-limiting examples include: -OCH 3 (methoxy), -OCH 2 CH 3 (ethoxy), -OCH 2 CH 2 CH 3 , -OCH(CH 3 ) 2 (isopropoxy), -OC(CH 3 ) 3 (Yert-butoxy), -OCH(CH 2 ) 2 , -O-cyclopentyl, and -O-cyclohexyl.
  • cycloalkoxy refers to groups, defined as -OR, in which R is cycloalkyl, alkenyl, alkynyl, aryl, and acyl, respectively.
  • alkylthio and “acylthio” when used without the “substituted” modifier refers to the group -SR, in which R is an alkyl and acyl, respectively.
  • alcohol corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with a hydroxy group.
  • ether corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with an alkoxy group.
  • substituted one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH 2 , -NO2, -C0 2 H, -C0 2 CH 3 , -CN, -SH, -OCH 3 , -OCH 2 CH 3 , -C(0)CH 3 , -NHCH 3 , -NHCH 2 CH 3 , -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 ) 2 , -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • alkylamino when used without the "substituted” modifier refers to the group -NHR, in which R is an alkyl, as that term is defined above. Non-limiting examples include: -NHCH 3 and -NHCH 2 CH 3 .
  • dialkylamino when used without the "substituted” modifier refers to the group -NRR', in which R and R' can be the same or different alkyl groups, or R and R' can be taken together to represent an alkanediyl.
  • dialkylamino groups include: -N(CH 3 ) 2 and -N(CH 3 )(CH 2 CH 3 ).
  • cycloalkylamino refers to groups, defined as -NHR, in which R is cycloalkyl, alkenyl, alkynyl, aryl, and alkoxy, respectively.
  • a non-limiting example of an arylamino group is -NHC6H5.
  • ami do when used without the "substituted” modifier, refers to the group -NHR, in which R is acyl, as that term is defined above.
  • a non-limiting example of an amido group is -NHC(0)CH 3 .
  • R is an alkyl
  • one or more hydrogen atom attached to a carbon atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH 2 , -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH 3 , -NHCH3, -NHCH2CH3, -N(CH 3 ) 2 , -C(0)NH 2 , -C(0)NHCH 3 , -C(0)N(CH 3 ) 2 , -OC(0)CH 3 , -NHC(0)CH 3 , -S(0) 2 OH, or -S(0) 2 NH 2 .
  • IC50 refers to an inhibitory dose which is 50% of the maximum response obtained. This quantitative measure indicates how much of a particular drug or other substance (inhibitor) is needed to inhibit a given biological, biochemical or chemical process (or component of a process, i.e.
  • An "isomer" of a first compound is a separate compound in which each molecule contains the same constituent atoms as the first compound, but where the configuration of those atoms in three dimensions differs.
  • the term "patient” or “subj ect” refers to a living mammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat, mouse, rat, guinea pig, or transgenic species thereof.
  • the patient or subject is a primate.
  • Non-limiting examples of human subjects are adults, juveniles, infants and fetuses.
  • pharmaceutically acceptable refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and/or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio.
  • “Pharmaceutically acceptable salts” means salts of compounds of the present disclosure which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as 1 ,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-ene-l-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene- 1 -carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinn
  • Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present are capable of reacting with inorganic or organic bases.
  • Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide.
  • Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine and the like. It should be recognized that the particular anion or cation forming a part of any salt of this disclosure is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).
  • pharmaceutically acceptable carrier means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a chemical agent.
  • Prevention includes: (1) inhibiting the onset of a disease in a subj ect or patient which may be at risk and/or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease, and/or (2) slowing the onset of the pathology or symptomatology of a disease in a subject or patient which may be at risk and/or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease.
  • a “stereoisomer” or “optical isomer” is an isomer of a given compound in which the same atoms are bonded to the same other atoms, but where the configuration of those atoms in three dimensions differs.
  • “Enantiomers” are stereoisomers of a given compound that are mirror images of each other, like left and right hands.
  • “Diastereomers” are stereoisomers of a given compound that are not enantiomers.
  • Chiral molecules contain a chiral center, also referred to as a stereocenter or stereogenic center, which is any point, though not necessarily an atom, in a molecule bearing groups such that an interchanging of any two groups leads to a stereoisomer.
  • the chiral center is typically a carbon, phosphorus or sulfur atom, though it is also possible for other atoms to be stereocenters in organic and inorganic compounds.
  • a molecule can have multiple stereocenters, giving it many stereoisomers.
  • compounds whose stereoisomerism is due to tetrahedral stereogenic centers e.g. , tetrahedral carbon
  • the total number of hypothetically possible stereoisomers will not exceed 2 n , where n is the number of tetrahedral stereocenters.
  • Molecules with symmetry frequently have fewer than the maximum possible number of stereoisomers.
  • a 50:50 mixture of enantiomers is referred to as a racemic mixture.
  • a mixture of enantiomers can be enantiomerically enriched so that one enantiomer is present in an amount greater than 50%.
  • enantiomers and/or diastereomers can be resolved or separated using techniques known in the art. It is contemplated that that for any stereocenter or axis of chirality for which stereochemistry has not been defined, that stereocenter or axis of chirality can be present in its R form, S form, or as a mixture of the R and S forms, including racemic and non-racemic mixtures.
  • the phrase "substantially free from other stereoisomers" means that the composition contains ⁇ 15%, more preferably ⁇ 10%, even more preferably ⁇ 5%, or most preferably ⁇ 1% of another stereoisomer(s).
  • Treatment includes (1) inhibiting a disease in a subject or patient experiencing or displaying the pathology or symptomatology of the disease (e.g. , arresting further development of the pathology and/or symptomatology), (2) ameliorating a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease (e.g. , reversing the pathology and/or symptomatology), and/or (3) effecting any measurable decrease in a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease.
  • inhibiting a disease in a subject or patient experiencing or displaying the pathology or symptomatology of the disease e.g. , arresting further development of the pathology and/or symptomatology
  • ameliorating a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease e.g. , reversing the pathology and/or symptomatology
  • the first class of compounds prepared possessing two redox centers was selenium-containing dihydropyran naphthoquinones obtained from lapachol (1) (Scheme 1).
  • Two selenium-containing derivatives, 6 and 7, were synthesized from a-lapachone (2) which in turn was prepared by acid catalyzed cyclization from lapachol (1).
  • Compound 6 was prepared in moderate yield (75%) by copper(I) catalyzed click reaction (Rostovtsev et al, 2002.) between compound 4 and (azidomethyl)(phenyl)selane.
  • the intermediate compound 4 was obtained by the reaction of 3-ethynylaniline and the bromo derivative 3.
  • 1,4-naphthoquinone coupled to selenium-containing 1,2,3-triazole was also a subject of this study. From compounds 33-35 and 39, the respective triazolic derivatives, compounds 36-38 and 40, were prepared using methodology discussed previously (Scheme 5). Suitable crystals of compounds 35 and 38 were obtained, and the structures were solved by crystallographic methods.
  • a standard atmospheric pressure photoionization (APPI) source was used to generate the ions.
  • the sample was injected using a constant flow (3 ⁇ 7 ⁇ ).
  • the solvent was an acetonitrile/methanol mixture.
  • the APPI-Q-TOF MS instrument was calibrated in the mass range of 50-3000 m/z using an internal calibration standard (low concentration tuning mix solution) supplied by Agilent Technologies. Data were processed employing Bruker Data Analysis software version 4.0. Compounds were named following IUPAC rules as applied by ChemBioDraw Ultra (version 12.0). [00129] Lawsone were acquired from Sigma-Aldrich (St. Louis, MO, USA).
  • Lapachol (1) (2-hydroxy-3-(3'-methyl-2'-butenyl)-l,4-naphthoquinone) was extracted from the heartwood of Tabebuia sp. (Tecoma).
  • C-allyl lawsone (23) was prepared from lawsone as previously reported.
  • a saturated aqueous sodium carbonate solution was added to the sawdust of ipe tree.
  • hydrochloric acid was added, allowing the precipitation of lapachol.
  • the solution was filtered and a yellow solid was obtained. This solid was purified by recrystallizations with hexane. All chemicals were obtained from commercial sources and used without further purification. Solvents were distilled and when required were dried by distillation according to standard procedures.
  • PBMC Peripheral blood mononuclear cells
  • PBMC peripheral blood mononuclear cells
  • fetal bovine serum 2 mM L-glutamine
  • penicillin 100 IU/mL penicillin
  • streptomycin 100 ⁇ g/mL streptomycin at 37 °C with 5% CC .
  • PBMC cultures were also supplemented with 2% phytohaemagglutinin.
  • cytotoxicity experiments cells were plated in 96-well plates (0.1 ⁇ 10 6 cells/well for leukemia cells, 0.7 ⁇ 10 5 cells/well for solid tumor as well V79, L929 and MDCK cells, and 1 ⁇ 10 6 cells/well for PBMC). All tested compounds were dissolved with DMSO.
  • DMSO DMSO-dimethyl-2-thiazol
  • DMSO DMSO
  • the cell viability was determined by reduction of the yellow dye 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to a blue formazan product as described by Mosmann. (Mosmann, 1983.)
  • MTT 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide
  • NQOl- normal cells human peripheral blood mononucluear cells (PBMC), and murine fibroblast immortalized cell lines (V79 and L929) were used to evaluate the selectivity of the compounds.
  • Mechanistic aspects of selected compounds were also studied for NQOl -dependency using the fairly specific NQOl inhibitor, dicoumarol. As previously described, (Perez-Sacau et al , 2007) the compounds were classified according to their activity as highly active (IC50 ⁇ 2 ⁇ ), moderately active (2 ⁇ ⁇ IC50 ⁇ 10 ⁇ ), or inactive (IC50 >10 ⁇ ).
  • Naphthopyranquinones 5-7, 9 and 10 presented high to moderate activities (IC50 in the range of 0.92 to 5.46 ⁇ ) and the non-active compound 4 was the exception of this class.
  • the strategy of insertion of a second redox center was a success and these derivatives were more active than their naphthoquinoidal precursors.
  • the inventors reported the synthesis and antitumor activities of several a-lapachone-based 1,2,3-triazoles (da Cruz et al, 2014). It is important to highlight that the selenium-containing-based 1,2,3-triazole 7 displayed better activity than the compounds without the chalcogen.
  • Naphthofuranquinones were the second class of compounds evaluated.
  • Para-naphthoquinones 15 and 16 were active against all cancer cell lines studied. In the last few years, the inventors described nor-a-lapachone-based 1,2,3-triazoles obtained from lapachol (1) with IC50 values > 2 ⁇ . (da Cruz et al , 2014.) The strategy herein used to prepare compounds 15 and 16 with the presence of selenium improved the activities of nor-a- lapachone-based 1,2,3-triazoles and these derivatives presented IC50 values ranging from 0.68-1.71 ⁇ for 15 and 1.59-2.95 ⁇ for 16.
  • ⁇ - ⁇ -lapachone and derivatives are among the most potent compounds from the lapachol group, (da Silva Junior et al, 2007; da Silva Junior et al, 2009) Recently, the cytotoxicity and genetic toxicity of ⁇ - ⁇ -lapachone in human lymphocytes, HL-60 leukemia cells, and immortal normal murine V79 fibroblasts were demonstrated (Cavalcanti et al, 2011) at concentrations of 2.5 and 5 ⁇ . This compound failed to induce DNA damage in nontumor cells, but at the highest concentrations, it induced DNA single and double strand breaks and increased the frequency of chromosomal aberrations. The biological effects of ⁇ - ⁇ -lapachone are related to its ability to deplete reduced glutathione (GSH), which leads to a GSSG-dominant pro-oxidant cellular status that conribute to its antiproliferative properties.
  • GSH reduced glutathione
  • This compound contains the structural framework of the 3-arylamino-nor ⁇ -lapachone derivatives reported before, but with a second redox chalcogen center inserted by click chemistry reaction.
  • This substance was highly active against all cancer cell lines evaluated with IC50 values ranging from 0.07 to 0.38 ⁇ .
  • 21 exhibited a high selectivity index (SI) (SI represented by the ratio of the cytotoxicities against normal cells and cancer cell lines).
  • SI selectivity index
  • Table 2 shows the selectivity index of several compounds. For instance, PBMC vs. HL-60 is 19.8.
  • doxorubicin a standard clinically used drug against several types of cancers, the selectivity index value is 10.6.
  • compound 22 (IC50 in the range of 1.06 to 2.56 ⁇ ) was more active than ⁇ - ⁇ -lapachone-based 1,2,3-triazole without the chalcogen atom.
  • IC50 in the range of 1.06 to 2.56 ⁇
  • two important examples of successful preparation of potent antitumor quinones with two redox centers are reported.
  • A549 cells were plated into a 48-well plate with 10,000 cells/well in 500 of DMEM containing 10% FBS. The cells were allowed to attach and grow overnight. A stock of 5 mM of compounds or 10 mM ⁇ -lapachone, and 5 mM dicoumarol were made for the experiment. The 8 drug concentrations (0-3.2 ⁇ ) were prepared separately in 15 mL conical tubes with 7 mL of media each. The untreated control is DMSO. The media was removed from each well and 500 of each drug concentration was added to 6 wells (to produce sextuplet replicates for each concentration).
  • Compound 53 showed the most dramatic lethality within a narrow therapeutic window, going from 93% viability at 0.8 ⁇ to 11% viability at 1.6 ⁇ .
  • compounds 50-53 were NQOl -specific drugs exhibiting similar or lower IC50 values than ⁇ -lapachone.
  • Compounds 21 and 22, selenium-containing quinones, with IC50 values 0.64 and 1.2 ⁇ , respectively, were the most active of this series and were NQOl -dependent (Table 3).
  • the compounds showed tremendous therapeutic windows using DIC treatment as a surrogate for responses to NQOl- cells, such as that found for nearly all human normal tissue (Bey, et al , 2006).
  • the Annexin V cytometry assay was used to detect cell population in viable, early and late apoptosis stage as described by Cavalcanti and coworkers. After short exposure time (6 h) with compounds 21 and 22 at 5 ⁇ , PC3 cells were stained with fluorescein isothiocyanate (FITC) conjugated Annexin V (Guava Nexin kit, Guava Technologies, Inc., Hay ward, CA, USA) and PI (necrotic-cell indicator), and then they were subjected to flow cytometry (Guava EasyCyte Mini). Cells undergoing early and late apoptosis were detected by the emission of the fluorescence from only FITC and, both FITC and PI, respectively.
  • FITC fluorescein isothiocyanate
  • PI necrotic-cell indicator
  • H2DCFDA 2',7'-dichlorofluorescein diacetate
  • DCFH non-fluorescent dichlorofluorescein
  • DCF highly fluorescent 2',7'-dichlorofluorescein
  • the increased mean fluorescence intensity of DCF can be used as a probe for a broad range of oxidative events not limited to H2O2.
  • PC3 human prostate cancer cell line was exposed to the test compound (5 ⁇ ) for 1 h.
  • NAC 5 mM
  • the culture medium was then replaced by fresh serum-free medium containing 20 ⁇ FhDCFDA.
  • DCF fluorescence intensity was detected by flow cytometry using a Guava EasyCyte Mini (Guava Technologies, Inc., Hayward, CA, USA) and Guava Express Plus software.
  • phosphatidylserine is considered an important marker in the apoptotic process.
  • selected compounds 21 and 22 induced a significant increase on populations of PC3 cells with phosphatidylserine expressed on the cell surface (FIG. 2).
  • phosphatidylserine externalization was not observed in cultures pre-treated with NAC before 21 and 22 exposure or co-treated with dicoumarol (FIG. 2).
  • the data show that cytotoxic mechanisms of tested compounds may involve drug bioreduction by quinone reductase NQOl as well emphasizing the ROS contribution on the cytotoxicity suggesting that tested compounds-induced apoptosis is associated with ROS production.

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Abstract

Provided herein are compounds containing two redox centers including a chalcogen redox center of the formula: wherein: R1, R2, X1, X2, Y1, and m are as defined herein. Also provided herein are pharmaceutical composition of the present compounds and methods of treatment using the compounds including their use in the treatment of cancer.

Description

DESCRIPTION
LAPACHONE DERIVATIVES CONTAINING TWO REDOX CENTERS AND
METHODS OF USE THEREOF [0001] This application claims benefit of priority to U.S. Provisional Application
Serial No. 62/244,038, filed October 20, 2015, the entire contents of which are hereby incorporated by reference.
BACKGROUND
[0002] The invention was made with government support under Grant No. R01 CA102972 awarded by the National Institutes of Health. The government has certain rights in the invention.
1. Field
[0003] The present disclosure relates generally to the field of chemotherapeutic compounds. More particularly, it concerns lapachone derivatives which contain two redox centers.
2. Description of Related Art
[0004] Lapachol and β-lapachone (β-lap), compounds easily isolated from the heartwood of Tabebuia, are among the naturally occurring and most studied naphthoquinones (Arnaudon, 1858; Thomson, 1971 and DoCampo et. al, 1979) was the first to explore β-lap for its potential anti-tumor activity. Significant activity was found against S-180 cells in vitro, and in mice bearing S-180 tumors. β-Lap had significant anti-tumor activity against Yoshida sarcoma and Walker 256 carcinoma cells in culture (Pink et al, 2000; Pink et al, 2000 and Planchon et al, 2001.) However, the exact mechanism of action of β-lap was not known until recently, (Pink et al, 2000a; Pink et al, 2000b; Planchon et al, 2001) and the lack of a known intracellular target/mechanism previously limited development of β-lap as an antitumor agent. Furthermore, β-Lap has a few characteristics that have limited its use as a chemotherapeutic agent, β-lap is highly hydrophobic and causes hemolysis in patients (Hartner et al, 2007). Recently, Ohayon and coworkers (Ohayon et al, 2015) shed light onto the hypothesis of β-lap being able to act nonreversibly for inhibiting of deubiquitinases. Others have suggested that the therapeutic effect of β-lap could be also related with USP2 oxidation. [0005] A series of lapachones with C-ring modified with potent activity against several types of cancer lineages have been disclosure (de Castro et al, 2013). β-Lapachone- based 1,2,3-triazoles possess significant activity with IC50 values below 2 μΜ for MDA- MB435 cancer cell lines. These compounds can promote cell death by apoptosis and genotoxicity, by induction of apoptosis associated with ROS production (da Silva Junior et al, 2011.) The approach of insertion of the 1,2,3-triazole moiety in 1,4-naphthoquinones was also effective since this unit is known as a potent pharmacophoric group (Agalave et al, 2011 and Massarotti et al, 2014.)
[0006] Additionally, there is significant interest in research on the chemistry /biochemistry associated with organoselenium compounds (Alberto et al, 2010 and Santi et al, 2013; Iwaoka and Arai, 2013 and Alberto et al, 2012.). These compounds play important roles in the areas of biology and medicine. In particular, they show antioxidant, antitumor, antimicrobial, anti -neurodegenerative and antiviral properties. (Nogueira et al, 2004 and Braga and Rafique, 2014.) In this regard, results obtained with basic research and the use of animal models indicates that these compounds can provide protection against different types of cancer (de Souza et al, 2015). In the last few years, Jacob and co-workers (Shabaan et al, 2009; Doering et al, 2010 and Mecklenburg et al, 2009.) have demonstrated the potential of selenium-containing quinones capable to act as mimics of the human enzyme glutathione peroxidase (GPx), which target redox sensitive thiol proteins and enzymes, and at the same time, generate reactive oxygens species (ROS) in a critical threshold acting as ROS- users and ROS-enhancers multi-target drugs (Shaaban et al, 2012)
SUMMARY
[0007] In some aspects, the present disclosure provides lapachone derivatives which contain two redox centers. In some embodiments, these lapachone derivatives have a chalcogen redox center in addition to another redox center.
[0008] In some aspects, the present disclosure provides compounds of the formula:
Figure imgf000004_0001
wherein: Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or X2 is taken together with R2 as described below;
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below;
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group, or R2 is taken together with X2 as described below; or R2 is taken together with Ri as described below;
Yi is amino, cyano, halo, hydroxy, or nitro; or alkyl(c<6), cycloalkyl(c<6), acyl(c<6), alkoxy(c<6), acyloxy(c<6), alkylamino(c<6), dialkylamino(c<6), amido(c<6), or a substituted version of any of these groups; m is 0, 1, 2, 3, or 4;
provided that when X2 and R2 are taken together, the compound is further defined by the formula:
Figure imgf000006_0001
wherein:
X3 is Cfo'I ' or O, wherein:
R3' and R4' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and n2 are each independently 0, 1, 2, or 3; provided that when Ri and R2 are taken together, the compound is further defined by the formula:
Figure imgf000006_0002
wherein:
X4 is CR(i'R7' or O, wherein: and R7' are each independently hydrogen, halo, alkyl(c<i2) substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or selenium containing triazole group; R.6, R.7, and Rs are each independently hydrogen, halo, alkyl(c<i2),
substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1, 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000007_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-;
wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000007_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as:
Figure imgf000007_0003
(V) wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4; Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000009_0001
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as:
Figure imgf000009_0002
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000010_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-;
wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000010_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as:
Figure imgf000010_0003
wherein:
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein: Rc is hydrogen, alkyl(c<6), or substituted alkyl(c<6); R.2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000011_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000011_0002
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
[0009] In other embodiments, the compounds are further defined as:
Figure imgf000011_0003
wherein: Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below; or
X3 is CR3'R4' or O, wherein:
R3' and R4' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; n is 0, 1 , 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4; Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000013_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as:
Figure imgf000013_0002
wherein:
X3 is CRj'I ' or O, wherein:
R3' and R4' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and n2 are each independently 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein: X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000014_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as:
Figure imgf000014_0002
wherein:
R3, R.4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and n2 are each independently 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000015_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-;
wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000015_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In other embodiments, the compounds are further defined as:
Figure imgf000015_0003
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
ORb, wherein: Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
X4 is CR6'R7' or O, wherein:
R6' and R7' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R6, R7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000017_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined:
Figure imgf000017_0002
wherein:
X4 is CRe'Rj' or O, wherein:
R.6' and R7' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R.6, R7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000018_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined:
Figure imgf000018_0002
wherein:
R.6, R.7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000019_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2);
or a pharmaceutically acceptable salt thereof.
[0010] In some embodiments, Xi is O. In some embodiments, X2 is ORb wherein Rb is absent.
[0011] In some embodiments, Ri is hydrogen. In other embodiments, Ri is halo such as Ri is bromo or chloro. In other embodiments, Ri is a selenium containing triazole group.
[0012] In some embodiments, R2 is hydrogen. In other embodiments, R2 is halo such as R2 is bromo or chloro. In some embodiments, R2 is a selenium containing triazole group. In some embodiments, m is 0.
[0013] In some embodiments, X3 is O. In some embodiments, R3 is hydrogen. In other embodiments, R3 is halo such as R3 is chloro or bromo. In other embodiments, R3 is alkyl(c<6) such as R3 is methyl. In other embodiments, R3 is a selenium containing triazole group.
[0014] In some embodiments, R4 is hydrogen. In other embodiments, R4 is halo such as R4 is chloro or bromo. In other embodiments, R4 is alkyl(c<6) such as R4 is methyl. In other embodiments, R4 is a selenium containing triazole group.
[0015] In some embodiments, R5 is hydrogen. In other embodiments, R5 is a selenium containing triazole group. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2. In some embodiments, X4 is O. [0016] In some embodiments, R.6 is hydrogen. In other embodiments, R.6 is halo such as R.6 is chloro or bromo. In other embodiments, R.6 is alkyl(c<6) such as R6 is methyl. In other embodiments, R6 is a selenium containing triazole group.
[0017] In some embodiments, R7 is hydrogen. In other embodiments, R7 is halo such as R7 is chloro or bromo. In other embodiments, R7 is alkyl(c<6) such as R7 is methyl. In other embodiments, R7 is a selenium containing triazole group.
[0018] In some embodiments, Rs is hydrogen. In other embodiments, Rs is aryl(c<i2) such as R8 is phenyl or napthyl. In other embodiments, Rs is a selenium containing triazole group. In some embodiments, p is 1 or 2. In some embodiments, p is 1. In other embodiments, p is 2.
[0019] In some embodiments, the selenium containing triazole group is further defined by the formula:
Figure imgf000020_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
q is 1, 2, 3, or 4;
r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups; and
R9 is aryl(c<i2) or substituted aryl(c<i2).
In some embodiments, the selenium containing triazole group is further defined by the formula:
Figure imgf000020_0002
wherein: X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
q is 1, 2, 3, or 4;
r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups; and
R9 is aryl(c<i2) or substituted aryl(c<i2).
[0020] In some embodiments, X5 is a covalent bond. In other embodiments, X5 is
-(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); r and s are each independently 0, 1, 2, 3, or 4.
[0021] In some embodiments, X5 is -NH-. In other embodiments, X5 is -NHCH2-. In some embodiments, Y2 is alkanediyl(c<6) such as Y2 is -CH2- In other embodiments, Y2 is arenediyl(c<6) such as Y2 is benzenediyl. In some embodiments, Y3 is alkanediyl(c<6) such as Y3 is -CH2-. In some embodiments, R9 is aryl(c<8) such as R9 is phenyl.
[0022] In some embodiments, the compound is further defined as:
Figure imgf000021_0001
Figure imgf000022_0001
or a pharmaceutically acceptable salt thereof.
[0023] In still other aspects, the present disclosure provides compounds of the formula:
Figure imgf000023_0001
or a salt thereof.
[0024] In still yet another aspect, the present disclosure provides pharmaceutical compositions comprising:
(a) a compound described herein; and
(b) a pharmaceutically acceptable carrier.
[0025] In some embodiments, wherein the pharmaceutical composition is formulated for administration: orally, intraadiposally, intraarterially, intraarticularly, intracranially, intradermally, intralesionally, intramuscularly, intranasally, intraocularly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrarectally, intrathecally, intratracheally, intratumorally, intraumbilically, intravaginally, intravenously, intravesicularly, intravitreally, liposomally, locally, mucosally, parenterally, rectally, subconjunctival, subcutaneously, sublingually, topically, transbuccally, transdermally, vaginally, in cremes, in lipid compositions, via a catheter, via a lavage, via continuous infusion, via infusion, via inhalation, via injection, via local delivery, or via localized perfusion. In some embodiments, the pharmaceutical composition further comprises a cyclodextran. In some embodiments, the cyclodextran is a β-cyclodextran such as a hydropropyl- -cyclodextran. In some embodiments, the pharmaceutical composition is formulated as a micelle or a liposome such as a micelle or liposome is formed by poly(lactic acid) poly(ethylene glycol) (PLA PEG).
[0026] In yet another aspect, the present disclosure provides methods of treating a patient with a disease or disorder comprising administering a therapeutically effective amount of a compound or composition described herein to the patient in need thereof. In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma. In some embodiments, the cancer is of the bladder, blood, bone, brain, breast, central nervous system, cervix, colon, endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia, genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen, small intestine, large intestine, stomach, testicle, or thyroid. In some embodiments, the cancer overexpresses NAD(P)H:quinone oxidoreductase 1 (NQOl). In some embodiments, the cancer is leukemia, colon cancer, prostate cancer, brain cancer, ovarian cancer, or skin cancer.
[0027] In some embodiments, the colon cancer is colon carcinoma. In other embodiments, the skin cancer is melanoma. In other embodiments, the brain cancer is a glioma. In other embodiments, the leukemia is acute myeloid leukemia. In other embodiments, the prostate cancer is an androgen independent prostate cancer. In other embodiments, the ovarian cancer is an ovarian adenocarcinoma.
[0028] In some embodiments, the methods comprise administering a second anti- cancer therapy. In some embodiments, the second anti-cancer therapy is a second chemotherapeutic compound, radiation therapy, surgery, or immunotherapy. In some embodiments, the second chemotherapeutic compound is a PARP1 inhibitor, a glutamine/glutamate pathway inhibitor, or a DNA base excision repair (BER) inhibitor. In some embodiments, the patient is a mammal such as a human. In some embodiments, the methods comprise administering the compound once. In other embodiments, the methods comprise administering the compound two or more times. In some embodiments, the patient has been determined to have defective DNA repair, defective ability to maintain NAD(P)H levels by the glucose, glutamate/glutamine, or pyruvate pathways, or genetic defects causing synthetic lethality for precision therapy against specific NQ01+ human cancers. [0029] In other embodiments, the disease or disorder is a metabolic syndrome. In some embodiments, the metabolic syndrome exhibits elevated NAD(P)H/NAD(P)+ levels. In other embodiments, the disease or disorder is trypanosomal disease.
[0030] As used herein the specification, "a" or "an" may mean one or more. As used herein in the claim(s), when used in conjunction with the word "comprising", the words "a" or "an" may mean one or more than one. [0031] The use of the term "or" in the claims is used to mean "and/or" unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and "and/or." As used herein "another" may mean at least a second or more.
[0032] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.
[0033] Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating preferred embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.
[0035] FIGS. 1A-1G show the NQOl activity of compounds 21, 22, 50, 51, 52, 53, and β-lapachone. The arylamine substituted ηοΓ-β-lapachones have predicted IC50 as follows: 50 = 2.6 μΜ, 51 = 1.8 μΜ, 52 = 2.4 μΜ and 53 = 1.3 μΜ. Compound 53 showed the most dramatic loss of viability within a narrow therapeutic window, going from 93% viability at 0.8 μΜ to 11% viability at 1.6 μΜ. Overall, 50-53 NQOl -specific compounds showed either similar or lower IC50 values than β-lapachone. Compounds 21 and 22, selenium-containing quinones, with IC50 values = 0.64 and 1.2, respectively, were the most active of this series and are also NQOl -dependent (see Table 2).
[0036] FIG. 2 shows the effect on phosphatidylserine externalization after 6 h-treated PC3 with 5 μΜ of tested compounds. The phosphatidylserine externalization was determined by flow cytometry using AnnV-FITC (YLW-HLog) and PI (RED-HLog). Viable cells are plotted at lower left quadrant, cells in early and late apoptosis with phosphatidylserine externalized are plotted at lower right and upper right quadrants, respectively, and necrotic cells are plotted at upper left quadrant. *p < 0.05 compared to control by ANOVA followed by Newman-Keuls test. Data are presented as mean values ± S.E.M. from three independent experiments in triplicate.
[0037] FIG. 3 shows the effect of compound 21 (5 μΜ) on ROS production on PC3 human prostate cancer cell line evaluated by H2DCFDA fluorescent probe after 1 h exposure in presence or absence of 5 mM NAC.
DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0038] In some aspects, the present disclosure provides compounds which may be useful in the treatment of cancer which contain two different redox centers. In some embodiments, one of the redox centers in the lapachone derivatives contain a chalcogen atom as a redox center. In some embodiments, these compounds are useful in treating cancers including cancers which are NQOl dependent.
I. Compounds of the Present Disclosure
[0039] The compounds provided by the present disclosure are shown, for example, above in the Summary and in the claims below. They may be made using the methods outlined in the Examples section. These methods can be further modified and optimized using the principles and techniques of organic chemistry as applied by a person skilled in the art. Such principles and techniques are taught, for example, in March 's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure (2007), which is incorporated by reference herein. [0040] Compounds of the disclosure may contain one or more asymmetrically- substituted carbon or nitrogen atoms, and may be isolated in optically active or racemic form. Thus, all chiral, diastereomeric, racemic form, epimeric form, and all geometric isomeric forms of a chemical formula are intended, unless the specific stereochemistry or isomeric form is specifically indicated. Compounds may occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. In some embodiments, a single diastereomer is obtained. The chiral centers of the compounds of the present disclosure can have the S or the R configuration.
[0041] Chemical formulas used to represent compounds of the disclosure will typically only show one of possibly several different tautomers. For example, many types of ketone groups are known to exist in equilibrium with corresponding enol groups. Similarly, many types of imine groups exist in equilibrium with enamine groups. Regardless of which tautomer is depicted for a given compound, and regardless of which one is most prevalent, all tautomers of a given chemical formula are intended.
[0042] Compounds of the disclosure may also have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. , higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the indications stated herein or otherwise.
[0043] In addition, atoms making up the compounds of the present disclosure are intended to include all isotopic forms of such atoms. Isotopes, as used herein, include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 1 C and 14C.
[0044] Compounds of the present disclosure may also exist in prodrug form. Since prodrugs are known to enhance numerous desirable qualities of pharmaceuticals (e.g. , solubility, bioavailability, manufacturing, etc.), the compounds employed in some methods of the disclosure may, if desired, be delivered in prodrug form. Thus, the disclosure contemplates prodrugs of compounds of the present disclosure as well as methods of delivering prodrugs. Prodrugs of the compounds employed in the disclosure may be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound. Accordingly, prodrugs include, for example, compounds described herein in which a hydroxy, amino, or carboxy group is bonded to any group that, when the prodrug is administered to a subject, cleaves to form a hydroxy, amino, or carboxylic acid, respectively. [0045] It should be recognized that the particular anion or cation forming a part of any salt form of a compound provided herein is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (2002), which is incorporated herein by reference. [0046] It will appreciated that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates." Where the solvent is water, the complex is known as a "hydrate." It will also be appreciated that many organic compounds can exist in more than one solid form, including crystalline and amorphous forms. All solid forms of the compounds provided herein, including any solvates thereof are within the scope of the present disclosure. [0047] In some aspects, the present disclosure relates to compounds which contain a chalcogen atom which can act as a redox center. A chalcogen is an element of Group 16 of the periodic table, including oxygen, sulfur, selenium and tellurium. A "heavy chalcogen" as used herein refers to a chalcogen atom having an atomic weight heavier than oxygen, for example, sulfur or selenium. In some embodiments, selenium is the heavy chalcogen used herein. Compounds containing a chalcogen atom are believed, without wishing to be bound by any theory, to function primarily as competitive substrates (e.g. as "sacrificial" antioxidants), which are preferentially attacked by oxidative species and thus useful in the treatment of cancer. In some embodiments, the chalcogen redox centers reduce the amount of reactive oxygen species present and/or reduce the oxidative stress experienced by cells.
II. Cancer and Other Hyperproliferative Diseases
[0048] While hyperproliferative diseases can be associated with any disease which causes a cell to begin to reproduce uncontrollably, the prototypical example is cancer. One of the key elements of cancer is that the cell's normal apoptotic cycle is interrupted and thus agents that interrupt the growth of the cells are important as therapeutic agents for treating these diseases. In this disclosure, the lapachone analogs described herein may be used to lead to decreased cell counts and as such can potentially be used to treat a variety of types of cancer lines. In some aspects, it is anticipated that the lapachone analogs described herein may be used to treat virtually any malignancy. [0049] In some aspects, the compounds of the present disclosure may be used in the treatment of NQOl dependent cancers. NQOl is known to regulate p53 activity and thus useful in the treatment of hyperproliferative diseases such as cancer. Additionally, NQOl acts as a detoxification protein which removes reactive oxygen species from the cell thus reducing oxidative damage and cancer. Modification of NQOl activity and expression is therefore useful in the treatment of cancer. Some non-limiting examples of cancers which have been associated with mis-regulation of NQOl include but are not limited to bladder cancer, breast cancer, colorectal cancer, prostate cancer or acute lymphoblastic leukemia.
[0050] Cancer cells that may be treated with the compounds of the present disclosure include but are not limited to cells from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, pancreas, testis, tongue, cervix, or uterus. In addition, the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; Paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w/squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; Sertoli cell carcinoma; Leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; malignant melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; Mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; Brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant; hemangiosarcoma; hemangioendothelioma, malignant; Kaposi's sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; Ewing's sarcoma; odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; and hairy cell leukemia. In certain aspects, the tumor may comprise an osteosarcoma, angiosarcoma, rhabdosarcoma, leiomyosarcoma, Ewing sarcoma, glioblastoma, neuroblastoma, or leukemia. III. Pharmaceutical Compositions, Formulations, and Routes of Administration
[0051] Where clinical applications are contemplated, it will be necessary to prepare pharmaceutical compositions in a form appropriate for the intended application. In some embodiments, such formulation with the compounds of the present disclosure is contemplated. Generally, this will entail preparing compositions that are essentially free of pyrogens, as well as other impurities that could be harmful to humans or animals.
[0052] One will generally desire to employ appropriate salts and buffers to render delivery vectors stable and allow for uptake by target cells. Buffers also will be employed when recombinant cells are introduced into a patient. Aqueous compositions of the present disclosure comprise an effective amount of the vector to cells, dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium. Such compositions also are referred to as inocula. The phrase "pharmaceutically or pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the vectors or cells of the present disclosure, its use in therapeutic compositions is contemplated. Supplementary active ingredients also can be incorporated into the compositions.
[0053] The active lapachone analogs compositions of the present disclosure may include classic pharmaceutical preparations. Administration of these compositions according to the present disclosure will be via any common route so long as the target tissue is available via that route. Such routes include oral, nasal, buccal, rectal, vaginal or topical route. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intratumoral, intraperitoneal, or intravenous injection. Such compositions would normally be administered as pharmaceutically acceptable compositions, described supra. [0054] The active compounds may also be administered parenterally or intraperitoneally. Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
[0055] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. [0056] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze- drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. [0057] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
[0058] For oral administration the lapachone analogs described herein may be incorporated with excipients and used in the form of non-ingestible mouthwashes and dentifrices. A mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution). Alternatively, the active ingredient may be incorporated into an antiseptic wash containing sodium borate, glycerin and potassium bicarbonate. The active ingredient may also be dispersed in dentifrices, including: gels, pastes, powders and slurries. The active ingredient may be added in a therapeutically effective amount to a paste dentifrice that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants.
[0059] The lapachone analog compositions of the present disclosure may be formulated in a neutral or salt form. Pharmaceutically-acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.
[0060] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this context, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 mL of isotonic NaCl solution and either added to 1000 mL of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences," 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.
IV. Methods of Treatment
[0061] In particular, the compositions that may be used in treating microbial infections and cancer in a subject (e.g., a human subject) are disclosed herein. The compositions described above are preferably administered to a mammal (e.g., rodent, human, non-human primates, canine, bovine, ovine, equine, feline, etc.) in an effective amount, that is, an amount capable of producing a desirable result in a treated subject (e.g., causing apoptosis of cancerous cells or killing bacterial cells). Toxicity and therapeutic efficacy of the compositions utilized in methods of the disclosure can be determined by standard pharmaceutical procedures. As is well known in the medical and veterinary arts, dosage for any one animal depends on many factors, including the subject's size, body surface area, body weight, age, the particular composition to be administered, time and route of administration, general health, the clinical symptoms of the infection or cancer and other drugs being administered concurrently. A composition as described herein is typically administered at a dosage that inhibits the growth or proliferation of a bacterial cell, inhibits the growth of a biofilm, or induces death of cancerous cells (e.g., induces apoptosis of a cancer cell), as assayed by identifying a reduction in hematological parameters (complete blood count - CBC), or cancer cell growth or proliferation. In some embodiments, amounts of the lapachone analogs used to inhibit bacterial growth or induce apoptosis of the cancer cells is calculated to be from about 0.01 mg to about 10,000 mg/day. In some embodiments, the amount is from about 1 mg to about 1,000 mg/day. In some embodiments, these dosings may be reduced or increased based upon the biological factors of a particular patient such as increased or decreased metabolic breakdown of the drug or decreased uptake by the digestive tract if administered orally. Addtionally, the lapachone analogs may be more efficacious and thus a smaller dose is required to achieve a similar effect. Such a dose is typically administered once a day for a few weeks or until sufficient reducing in cancer cells has been achieved.
[0062] The therapeutic methods of the disclosure (which include prophylactic treatment) in general include administration of a therapeutically effective amount of the compositions described herein to a subject in need thereof, including a mammal, particularly a human. Such treatment will be suitably administered to subjects, particularly humans, suffering from, having, susceptible to, or at risk for a disease, disorder, or symptom thereof. Determination of those subjects "at risk" can be made by any objective or subjective determination by a diagnostic test or opinion of a subject or health care provider (e.g., genetic test, enzyme or protein marker, marker (as defined herein), family history, and the like).
[0063] In one embodiment, the disclosure provides a method of monitoring treatment progress. The method includes the step of determining a level of changes in hematological parameters and/or cancer stem cell (CSC) analysis with cell surface proteins as diagnostic markers (which can include, for example, but are not limited to CD34, CD38, CD90, and CD 117) or diagnostic measurement (e.g. , screen, assay) in a subject suffering from or susceptible to a disorder or symptoms thereof associated with cancer (e.g., leukemia) in which the subject has been administered a therapeutic amount of a composition as described herein. The level of marker determined in the method can be compared to known levels of marker in either healthy normal controls or in other afflicted patients to establish the subject's disease status. In preferred embodiments, a second level of marker in the subject is determined at a time point later than the determination of the first level, and the two levels are compared to monitor the course of disease or the efficacy of the therapy. In certain preferred embodiments, a pre-treatment level of marker in the subject is determined prior to beginning treatment according to the methods described herein; this pre-treatment level of marker can then be compared to the level of marker in the subject after the treatment commences, to determine the efficacy of the treatment. V. Combination Therapies
[0064] It is envisioned that the lapachone analogs described herein may be used in combination therapies with one or more cancer therapies or a compound which mitigates one or more of the side effects experienced by the patient. It is common in the field of cancer therapy to combine therapeutic modalities. The following is a general discussion of therapies that may be used in conjunction with the therapies of the present disclosure.
[0065] To treat cancers using the methods and compositions of the present disclosure, one would generally contact a tumor cell or subject with a compound and at least one other therapy. These therapies would be provided in a combined amount effective to achieve a reduction in one or more disease parameter. This process may involve contacting the cells/subjects with the both agents/therapies at the same time, e.g., using a single composition or pharmacological formulation that includes both agents, or by contacting the cell/subject with two distinct compositions or formulations, at the same time, wherein one composition includes the compound and the other includes the other agent.
[0066] Alternatively, the lapachone analogs described herein may precede or follow the other treatment by intervals ranging from minutes to weeks. One would generally ensure that a significant period of time did not expire between the time of each delivery, such that the therapies would still be able to exert an advantageously combined effect on the cell/subject. In such instances, it is contemplated that one would contact the cell with both modalities within about 12-24 hours of each other, within about 6-12 hours of each other, or with a delay time of only about 1-2 hours. In some situations, it may be desirable to extend the time period for treatment significantly; however, where several days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) lapse between the respective administrations.
[0067] It also is conceivable that more than one administration of either the compound or the other therapy will be desired. Various combinations may be employed, where a compound of the present disclosure is "A," and the other therapy is "B," as exemplified below: A/B/A B/A/B B/B/A A/A/B B/A/A A/B/B B/B/B/A B/B/A/B
A/A/B/B A/B/A/B A/B/B/A B/B/A/A B/A/B/A B/A/A/B B/B/B/A A/A/A/B B/A/A/A A/B/A/A A/A/B/A A/B/B/B B/A/B/B B/B/A/B
Other combinations are also contemplated. The following is a general discussion of cancer therapies that may be used combination with the compounds of the present disclosure.
1. Chemotherapy
[0068] The term "chemotherapy" refers to the use of drugs to treat cancer. A "chemotherapeutic agent" is used to connote a compound or composition that is administered in the treatment of cancer. These agents or drugs are categorized by their mode of activity within a cell, for example, whether and at what stage they affect the cell cycle. Alternatively, an agent may be characterized based on its ability to directly cross-link DNA, to intercalate into DNA, or to induce chromosomal and mitotic aberrations by affecting nucleic acid synthesis. Most chemotherapeutic agents fall into the following categories: alkylating agents, antimetabolites, antitumor antibiotics, mitotic inhibitors, and nitrosoureas.
[0069] Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin γΐ and calicheamicin col ; dynemicin, including dynemicin A uncialamycin and derivatives thereof; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromophores, aclacinomysins, actinomycin, authrarnycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino- doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalarnycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5- fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharide complex); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2 ',2"- trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g. , paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g. , CPT-11); topoisomerase inhibitor RFS 2000; difluorometlhylomithine (DMFO); retinoids such as retinoic acid; capecitabine; cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, paclitaxel, docetaxel, gemcitabien, navelbine, farnesyl-protein tansferase inhibitors, transplatinum, 5-fluorouracil, vincristin, vinblastin and methotrexate and pharmaceutically acceptable salts, acids or derivatives of any of the above.
2. Radiotherapy
[0070] Radiotherapy, also called radiation therapy, is the treatment of cancer and other diseases with ionizing radiation. Ionizing radiation deposits energy that injures or destroys cells in the area being treated by damaging their genetic material, making it impossible for these cells to continue to grow. Although radiation damages both cancer cells and normal cells, the latter are able to repair themselves and function properly. [0071] Radiation therapy used according to the present disclosure may include, but is not limited to, the use of γ-rays, X-rays, and/or the directed delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors are also contemplated such as microwaves and UV-irradiation. It is most likely that all of these factors induce a broad range of damage on DNA, on the precursors of DNA, on the replication and repair of DNA, and on the assembly and maintenance of chromosomes. Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens. Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.
[0072] Radiotherapy may comprise the use of radiolabeled antibodies to deliver doses of radiation directly to the cancer site (radioimmunotherapy). Antibodies are highly specific proteins that are made by the body in response to the presence of antigens (substances recognized as foreign by the immune system). Some tumor cells contain specific antigens that trigger the production of tumor-specific antibodies. Large quantities of these antibodies can be made in the laboratory and attached to radioactive substances (a process known as radiolabeling). Once injected into the body, the antibodies actively seek out the cancer cells, which are destroyed by the cell-killing (cytotoxic) action of the radiation. This approach can minimize the risk of radiation damage to healthy cells.
[0073] Conformal radiotherapy uses the same radiotherapy machine, a linear accelerator, as the normal radiotherapy treatment but metal blocks are placed in the path of the x-ray beam to alter its shape to match that of the cancer. This ensures that a higher radiation dose is given to the tumor. Healthy surrounding cells and nearby structures receive a lower dose of radiation, so the possibility of side effects is reduced. A device called a multi- leaf collimator has been developed and may be used as an alternative to the metal blocks. The multi-leaf collimator consists of a number of metal sheets which are fixed to the linear accelerator. Each layer can be adjusted so that the radiotherapy beams can be shaped to the treatment area without the need for metal blocks. Precise positioning of the radiotherapy machine is very important for conformal radiotherapy treatment and a special scanning machine may be used to check the position of internal organs at the beginning of each treatment.
[0074] High-resolution intensity modulated radiotherapy also uses a multi-leaf collimator. During this treatment the layers of the multi-leaf collimator are moved while the treatment is being given. This method is likely to achieve even more precise shaping of the treatment beams and allows the dose of radiotherapy to be constant over the whole treatment area.
[0075] Although research studies have shown that conformal radiotherapy and intensity modulated radiotherapy may reduce the side effects of radiotherapy treatment, it is possible that shaping the treatment area so precisely could stop microscopic cancer cells just outside the treatment area being destroyed. This means that the risk of the cancer coming back in the future may be higher with these specialized radiotherapy techniques.
[0076] Scientists also are looking for ways to increase the effectiveness of radiation therapy. Two types of investigational drugs are being studied for their effect on cells undergoing radiation. Radiosensitizers make the tumor cells more likely to be damaged, and radioprotectors protect normal tissues from the effects of radiation. Hyperthermia, the use of heat, is also being studied for its effectiveness in sensitizing tissue to radiation.
3. Immunotherapy
[0077] In the context of cancer treatment, immunotherapeutics, generally, rely on the use of immune effector cells and molecules to target and destroy cancer cells. Trastuzumab (Herceptin™) is such an example. The immune effector may be, for example, an antibody specific for some marker on the surface of a tumor cell. The antibody alone may serve as an effector of therapy or it may recruit other cells to actually affect cell killing. The antibody also may be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and serve merely as a targeting agent. Alternatively, the effector may be a lymphocyte carrying a surface molecule that interacts, either directly or indirectly, with a tumor cell target. Various effector cells include cytotoxic T cells and NK cells. The combination of therapeutic modalities, i.e., direct cytotoxic activity and inhibition or reduction of ErbB2 would provide therapeutic benefit in the treatment of ErbB2 overexpressing cancers.
[0078] In one aspect of immunotherapy, the tumor cell must bear some marker that is amenable to targeting, i.e. , is not present on the majority of other cells. Many tumor markers exist and any of these may be suitable for targeting in the context of the present disclosure. Common tumor markers include carcinoembryonic antigen, prostate specific antigen, urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor, laminin receptor, erb B and pi 55. An alternative aspect of immunotherapy is to combine anticancer effects with immune stimulatory effects. Immune stimulating molecules also exist including: cytokines such as IL-2, IL-4, IL-12, GM-CSF, γ-IFN, chemokines such as MIP-1, MCP-1, IL-8 and growth factors such as FLT3 ligand. Combining immune stimulating molecules, either as proteins or using gene delivery in combination with a tumor suppressor has been shown to enhance antitumor effects (Ju et al , 2000). Moreover, antibodies against any of these compounds may be used to target the anti-cancer agents discussed herein.
[0079] Examples of immunotherapies currently under investigation or in use are immune adjuvants, e.g. , Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene and aromatic compounds (U.S. Patents 5,801,005 and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al, 1998), cytokine therapy, e.g. , interferons α, β, and γ; IL-1, GM-CSF and TNF (Bukowski et al, 1998; Davidson et al , 1998; Hellstrand et al , 1998) gene therapy, e.g., TNF, IL-1, IL-2, p53 (Qin et al, 1998; Austin-Ward and Villaseca, 1998; U.S. Patents 5,830,880 and 5,846,945) and monoclonal antibodies, e.g., anti-ganglioside GM2, anti-HER- 2, anti-pl85 (Pietras et al, 1998; Hanibuchi et al, 1998; U.S. Patent 5,824,311). It is contemplated that one or more anti-cancer therapies may be employed with the gene silencing therapies described herein.
[0080] In active immunotherapy, an antigenic peptide, polypeptide or protein, or an autologous or allogenic tumor cell composition or "vaccine" is administered, generally with a distinct bacterial adjuvant (Ravindranath and Morton, 1991; Morton et al , 1992; Mitchell et al , 1990; Mitchell et al, 1993). [0081] In adoptive immunotherapy, the patient's circulating lymphocytes, or tumor infiltrated lymphocytes, are isolated in vitro, activated by lymphokines such as IL-2 or transduced with genes for tumor necrosis, and readministered (Rosenberg et al, 1988; 1989).
4. Surgery
[0082] Approximately 60% of persons with cancer will undergo surgery of some type, which includes preventative, diagnostic or staging, curative, and palliative surgery. Curative surgery is a cancer treatment that may be used in conjunction with other therapies, such as the treatment of the present disclosure, chemotherapy, radiotherapy, hormonal therapy, gene therapy, immunotherapy and/or alternative therapies. [0083] Curative surgery includes resection in which all or part of cancerous tissue is physically removed, excised, and/or destroyed. Tumor resection refers to physical removal of at least part of a tumor. In addition to tumor resection, treatment by surgery includes laser surgery, cryosurgery, electrosurgery, and microscopically controlled surgery (Mohs' surgery). It is further contemplated that the present disclosure may be used in conjunction with removal of superficial cancers, precancers, or incidental amounts of normal tissue.
[0084] Upon excision of part or all of cancerous cells, tissue, or tumor, a cavity may be formed in the body. Treatment may be accomplished by perfusion, direct injection or local application of the area with an additional anti-cancer therapy. Such treatment may be repeated, for example, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5 weeks or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. These treatments may be of varying dosages as well.
[0085] In some particular embodiments, after removal of the tumor, an adjuvant treatment with a compound of the present disclosure is believe to be particularly efficacious in reducing the reoccurance of the tumor. Additionally, the compounds of the present disclosure can also be used in a neoadjuvant setting.
5. Other Agents
[0086] It is contemplated that other agents may be used with the present disclosure. These additional agents include immunomodulatory agents, agents that affect the upregulation of cell surface receptors and GAP junctions, cytostatic and differentiation agents, inhibitors of cell adhesion, agents that increase the sensitivity of the hyperproliferative cells to apoptotic inducers, or other biological agents. Immunomodulatory agents include tumor necrosis factor; interferon alpha, beta, and gamma; IL-2 and other cytokines; F42K and other cytokine analogs; or MIP-1, ΜΙΡ-Ι β, MCP-1, RANTES, and other chemokines. It is further contemplated that the upregulation of cell surface receptors or their ligands such as Fas/Fas ligand, DR4 or DR5/TRAIL (Apo-2 ligand) would potentiate the apoptotic inducing abilities of the present disclosure by establishment of an autocrine or paracrine effect on hyperproliferative cells. Increases intercellular signaling by elevating the number of GAP junctions would increase the anti-hyperproliferative effects on the neighboring hyperproliferative cell population. In other embodiments, cytostatic or differentiation agents may be used in combination with the present disclosure to improve the anti-hyerproliferative efficacy of the treatments. Inhibitors of cell adhesion are contemplated to improve the efficacy of the present disclosure. Examples of cell adhesion inhibitors are focal adhesion kinase (FAKs) inhibitors and Lovastatin. It is further contemplated that other agents that increase the sensitivity of a hyperproliferative cell to apoptosis, such as the antibody c225, could be used in combination with the present disclosure to improve the treatment efficacy.
[0087] There have been many advances in the therapy of cancer following the introduction of cytotoxic chemotherapeutic drugs. However, one of the consequences of chemotherapy is the development/acquisition of drug-resistant phenotypes and the development of multiple drug resistance. The development of drug resistance remains a major obstacle in the treatment of such tumors and therefore, there is an obvious need for alternative approaches such as gene therapy.
[0088] Another form of therapy for use in conjunction with chemotherapy, radiation therapy or biological therapy includes hyperthermia, which is a procedure in which a patient's tissue is exposed to high temperatures (up to 106°F). External or internal heating devices may be involved in the application of local, regional, or whole-body hyperthermia. Local hyperthermia involves the application of heat to a small area, such as a tumor. Heat may be generated externally with high-frequency waves targeting a tumor from a device outside the body. Internal heat may involve a sterile probe, including thin, heated wires or hollow tubes filled with warm water, implanted microwave antennae, or radiofrequency electrodes. [0089] A patient's organ or a limb is heated for regional therapy, which is accomplished using devices that produce high energy, such as magnets. Altematively, some of the patient's blood may be removed and heated before being perfused into an area that will be internally heated. Whole-body heating may also be implemented in cases where cancer has spread throughout the body. Warm- water blankets, hot wax, inductive coils, and thermal chambers may be used for this purpose.
[0090] The skilled artisan is directed to "Remington's Pharmaceutical Sciences" 15th Edition, chapter 33, in particular pages 624-652. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.
[0091] It also should be pointed out that any of the foregoing therapies may prove useful by themselves in treating cancer. VI. Definitions
[0092] When used in the context of a chemical group: "hydrogen" means -H; "hydroxy" means -OH; "oxo" means =0; "carbonyl" means -C(=0)-; "carboxy" means -C(=0)OH (also written as -COOH or -CO2H); "halo" means independently -F, -CI, -Br or -I; "amino" means -NH2; "hydroxyamino" means -NHOH; "nitro" means -NO2; imino means =NH; "cyano" means -CN; "isocyanate" means -N=C=0; "azido" means -N3; in a monovalent context "phosphate" means -OP(0)(OH)2 or a deprotonated form thereof; in a divalent context "phosphate" means -OP(0)(OH)0- or a deprotonated form thereof; "mercapto" means -SH; and "thio" means =S; "sulfonyl" means -S(0)2-; "hydroxysulfonyl" means -S(0)20H; "sulfonamide" means -S(0)2NH2; and "sulfinyl" means -S(O)-. [0093] In the context of chemical formulas, the symbol "-" means a single bond, "=" means a double bond, and "≡" means triple bond. The symbol " " represents an optional bond, which if present is either single or double. The symbol "==" represents single bond or a and
Figure imgf000044_0001
double bond. Furthermore, it is noted that the covalent bond symbol when connecting one or two stereogenic atoms, does not indicate any preferred stereochemistry. Instead, it covers all stereoisomers as well as mixtures thereof. The symbol " ^ ", when drawn perpendicularly across a bond (e.g. , j— CH3 for methyl) indicates a point of attachment of the group. It is noted that the point of attachment is typically only identified in this manner for larger groups in order to assist the reader in unambiguously identifying a point of attachment. The symbol "-^ " means a single bond where the group attached to the thick end of the wedge is "out of the page." The symbol " ""HI " means a single bond where the group attached to the thick end of the wedge is "into the page". The symbol " ^vw " means a single bond where the geometry around a double bond (e.g., either E or Z) is undefined. Both options, as well as combinations thereof are therefore intended. Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to that atom. A bold dot on a carbon atom indicates that the hydrogen attached to that carbon is oriented out of the plane of the paper. [0094] When a group "R" is depicted as a "floating group" on a ring system, for example, in the formula:
Figure imgf000045_0001
then R may replace any hydrogen atom attached to any of the ring atoms, including a depicted, implied, or expressly defined hydrogen, so long as a stable structure is formed. When a group "R" is depicted as a "floating group" on a fused ring system, as for example in the formula:
Figure imgf000045_0002
then R may replace any hydrogen attached to any of the ring atoms of either of the fused rings unless specified otherwise. Replaceable hydrogens include depicted hydrogens (e.g., the hydrogen attached to the nitrogen in the formula above), implied hydrogens (e.g. , a hydrogen of the formula above that is not shown but understood to be present), expressly defined hydrogens, and optional hydrogens whose presence depends on the identity of a ring atom (e.g., a hydrogen attached to group X, when X equals -CH-), so long as a stable structure is formed. In the example depicted, R may reside on either the 5-membered or the 6- membered ring of the fused ring system. In the formula above, the subscript letter "y" immediately following the group "R" enclosed in parentheses, represents a numeric variable. Unless specified otherwise, this variable can be 0, 1, 2, or any integer greater than 2, only limited by the maximum number of replaceable hydrogen atoms of the ring or ring system.
[0095] For the chemical groups and compound classes, the number of carbon atoms in the group or class is as indicated as follows: "Cn" defines the exact number (n) of carbon atoms in the group/class. "C≤n" defines the maximum number (n) of carbon atoms that can be in the group/class, with the minimum number as small as possible for the group/class in question, e.g. , it is understood that the minimum number of carbon atoms in the group "alkenyl(c≤8)" or the class "alkene(c≤8)" is two. Compare with "alkoxy(c≤io)", which designates alkoxy groups having from 1 to 10 carbon atoms. "Cn-n"' defines both the minimum (n) and maximum number (η') of carbon atoms in the group. Thus, "alkyl(C2-io)" designates those alkyl groups having from 2 to 10 carbon atoms. These carbon number indicators may precede or follow the chemical groups or class it modifies and it may or may not be enclosed in parenthesis, without signifying any change in meaning. Thus, the terms "C5 olefin", "C5-olefin", "olefin(C5)", and "olefincs" are all synonymous.
[0096] The term "saturated" when used to modify a compound or chemical group means the compound or chemical group has no carbon-carbon double and no carbon-carbon triple bonds, except as noted below. When the term is used to modify an atom, it means that the atom is not part of any double or triple bond. In the case of substituted versions of saturated groups, one or more carbon oxygen double bond or a carbon nitrogen double bond may be present. And when such a bond is present, then carbon-carbon double bonds that may occur as part of keto-enol tautomerism or imine/enamine tautomerism are not precluded. When the term "saturated" is used to modify a solution of a substance, it means that no more of that substance can dissolve in that solution.
[0097] The term "aliphatic" when used without the "substituted" modifier signifies that the compound or chemical group so modified is an acyclic or cyclic, but non-aromatic hydrocarbon compound or group. In aliphatic compounds/groups, the carbon atoms can be joined together in straight chains, branched chains, or non-aromatic rings (alicyclic). Aliphatic compounds/groups can be saturated, that is joined by single carbon-carbon bonds (alkanes/alkyl), or unsaturated, with one or more carbon-carbon double bonds (alkenes/alkenyl) or with one or more carbon-carbon triple bonds (alkynes/alkynyl).
[0098] The term "aromatic" when used to modify a compound or a chemical group atom means the compound or chemical group contains a planar unsaturated ring of atoms that is stabilized by an interaction of the bonds forming the ring.
[0099] The term "alkyl" when used without the "substituted" modifier refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, a linear or branched acyclic structure, and no atoms other than carbon and hydrogen. The groups -CH3 (Me), -CH2CH3 (Et), -CH2CH2CH3 (w-Pr or propyl), -CH(CH3)2 (z-Pr, ;Pr or isopropyl), -CH2CH2CH2CH3 (w-Bu), -CH(CH3)CH2CH3 (sec-butyl), -CH2CH(CH3)2 (isobutyl), -C(CH3)3 (fert-butyl, i-butyl, t-Bu or ¾u), and -CH2C(CH3)3 («eo-pentyl) are non-limiting examples of alkyl groups. The term "alkanediyl" when used without the "substituted" modifier refers to a divalent saturated aliphatic group, with one or two saturated carbon atom(s) as the point(s) of attachment, a linear or branched acyclic structure, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. The groups -CH2- (methylene), -CH2CH2-, -CH2C(CH3)2CH2-, and -CH2CH2CH2- are non- limiting examples of alkanediyl groups. The term "alkylidene" when used without the "substituted" modifier refers to the divalent group =CRR' in which R and R' are independently hydrogen or alkyl. Non-limiting examples of alkylidene groups include: =CH2, =CH(CH2CH3), and
Figure imgf000047_0001
An "alkane" refers to the class of compounds having the formula H-R, wherein R is alkyl as this term is defined above. When any of these terms is used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, - H2, -NO2, -CO2H, -C02CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2. The following groups are non-limiting examples of substituted alkyl groups: -CH2OH, -CH2CI, -CF3, -CH2CN, -CH2C(0)OH, -CH2C(0)OCH3, -CH2C(0)NH2, -CH2C(0)CH3, -CH2OCH3, -CH2OC(0)CH3, -CH2NH2, -CH2N(CH3)2, and -CH2CH2CI. The term "haloalkyl" is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to halo (i.e. -F, -CI, -Br, or -I) such that no other atoms aside from carbon, hydrogen and halogen are present. The group, -CH2CI is a non-limiting example of a haloalkyl. The term "fluoroalkyl" is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to fluoro such that no other atoms aside from carbon, hydrogen and fluorine are present. The groups -CH2F, -CF3, and -CH2CF3 are non-limiting examples of fluoroalkyl groups.
[00100] The term "cycloalkyl" when used without the "substituted" modifier refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, said carbon atom forming part of one or more non-aromatic ring structures, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. Non-limiting examples include: -CH(CH2)2 (cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl (Cy). The term "cycloalkanediyl" when used without the "substituted" modifier refers to a divalent saturated aliphatic group with two carbon atoms as points of attachment, no carbon- double or triple bonds, and no atoms other than carbon and hydrogen. The group
Figure imgf000048_0001
is a non-limiting example of cycloalkanediyl group. A "cycloalkane" refers to the class of compounds having the formula H-R, wherein R is cycloalkyl as this term is defined above. When any of these terms is used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2.
[00101] The term "alkenyl" when used without the "substituted" modifier refers to an monovalent unsaturated aliphatic group with a carbon atom as the point of attachment, a linear or branched, acyclic structure, at least one nonaromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen. Non-limiting examples include: -CH=CH2 (vinyl), -CH=CHCH3, -CH=CHCH2CH3, -CH2CH=CH2 (allyl), -CH2CH=CHCH3, and -CH=CHCH=CH2. The term "alkenediyl" when used without the "substituted" modifier refers to a divalent unsaturated aliphatic group, with two carbon atoms as points of attachment, a linear or branched, a linear or branched acyclic structure, at least one nonaromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen. The groups -CH=CH-, -CH=C(CH3)CH2- -CH=CHCH2- and -CH2CH=CHCH2- are non-limiting examples of alkenediyl groups. It is noted that while the alkenediyl group is aliphatic, once connected at both ends, this group is not precluded from forming part of an aromatic structure. The terms "alkene" and "olefin" are synonymous and refer to the class of compounds having the formula H-R, wherein R is alkenyl as this term is defined above. Similarly the terms "terminal alkene" and "a-olefin" are synonymous and refer to an alkene having just one carbon-carbon double bond, wherein that bond is part of a vinyl group at an end of the molecule. When any of these terms are used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2. The groups -CH=CHF, -CH=CHC1 and -CH=CHBr are non-limiting examples of substituted alkenyl groups. [00102] The term "alkynyl" when used without the "substituted" modifier refers to a monovalent unsaturated aliphatic group with a carbon atom as the point of attachment, a linear or branched acyclic structure, at least one carbon-carbon triple bond, and no atoms other than carbon and hydrogen. As used herein, the term alkynyl does not preclude the presence of one or more non-aromatic carbon-carbon double bonds. The groups -C≡CH, -C≡CCH3, and -CH2C≡CCH3 are non-limiting examples of alkynyl groups. An "alkyne" refers to the class of compounds having the formula H-R, wherein R is alkynyl. When any of these terms are used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, - H2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2.
[00103] The term "aryl" when used without the "substituted" modifier refers to a monovalent unsaturated aromatic group with an aromatic carbon atom as the point of attachment, said carbon atom forming part of a one or more six-membered aromatic ring structure, wherein the ring atoms are all carbon, and wherein the group consists of no atoms other than carbon and hydrogen. If more than one ring is present, the rings may be fused or unfused. As used herein, the term does not preclude the presence of one or more alkyl or aralkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present. Non-limiting examples of aryl groups include phenyl (Ph), methylphenyl, (dimethyl)phenyl, -C6H4CH2CH3 (ethylphenyl), naphthyl, and a monovalent group derived from biphenyl. The term "arenediyl" when used without the "substituted" modifier refers to a divalent aromatic group with two aromatic carbon atoms as points of attachment, said carbon atoms forming part of one or more six-membered aromatic ring structure(s) wherein the ring atoms are all carbon, and wherein the monovalent group consists of no atoms other than carbon and hydrogen. As used herein, the term does not preclude the presence of one or more alkyl, aryl or aralkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present. If more than one ring is present, the rings may be fused or unfused. Unfused rings may be connected via one or more of the following: a covalent bond, alkanediyl, or alkenediyl groups (carbon number limitation permitting). Non-limiting examples of arenediyl groups include:
Figure imgf000050_0001
An "arene" refers to the class of compounds having the formula H-R, wherein R is aryl as that term is defined above. Benzene and toluene are non-limiting examples of arenes. When any of these terms are used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2.
[00104] The term "acyl" when used without the "substituted" modifier refers to the group -C(0)R, in which R is a hydrogen, alkyl, cycloalkyl, alkenyl, or aryl, as those terms are defined above. The groups, -CHO, -C(0)CH3 (acetyl, Ac), -C(0)CH2CH3, -C(0)CH2CH2CH3, -C(0)CH(CH3)2, -C(0)CH(CH2)2, -C(0)C6H5, -C(0)C6H4CH3, -C(0)CH2C6H5, -C(0)(imidazolyl) are non-limiting examples of acyl groups. A "thioacyl" is defined in an analogous manner, except that the oxygen atom of the group -C(0)R has been replaced with a sulfur atom, -C(S)R. The term "aldehyde" corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with a -CHO group. When any of these terms are used with the "substituted" modifier one or more hydrogen atom (including a hydrogen atom directly attached to the carbon atom of the carbonyl or thiocarbonyl group, if any) has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2. The groups, -C(0)CH2CF3, -C02H (carboxyl), -C02CH3 (methylcarboxyl), -C02CH2CH3, -C(0)NH2 (carbamoyl), and -CON(CH3)2, are non-limiting examples of substituted acyl groups.
[00105] The term "alkoxy" when used without the "substituted" modifier refers to the group -OR, in which R is an alkyl, as that term is defined above. Non-limiting examples include: -OCH3 (methoxy), -OCH2CH3 (ethoxy), -OCH2CH2CH3, -OCH(CH3)2 (isopropoxy), -OC(CH3)3 (Yert-butoxy), -OCH(CH2)2, -O-cyclopentyl, and -O-cyclohexyl. The terms "cycloalkoxy", "alkenyloxy", "alkynyloxy", "aryloxy", and "acyloxy", when used without the "substituted" modifier, refers to groups, defined as -OR, in which R is cycloalkyl, alkenyl, alkynyl, aryl, and acyl, respectively. The term "alkylthio" and "acylthio" when used without the "substituted" modifier refers to the group -SR, in which R is an alkyl and acyl, respectively. The term "alcohol" corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with a hydroxy group. The term "ether" corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with an alkoxy group. When any of these terms is used with the "substituted" modifier one or more hydrogen atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -C02H, -C02CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2. [00106] The term "alkylamino" when used without the "substituted" modifier refers to the group -NHR, in which R is an alkyl, as that term is defined above. Non-limiting examples include: -NHCH3 and -NHCH2CH3. The term "dialkylamino" when used without the "substituted" modifier refers to the group -NRR', in which R and R' can be the same or different alkyl groups, or R and R' can be taken together to represent an alkanediyl. Non- limiting examples of dialkylamino groups include: -N(CH3)2 and -N(CH3)(CH2CH3). The terms "cycloalkylamino", "alkenylamino", "alkynylamino", "arylamino", and "alkoxyamino" when used without the "substituted" modifier, refers to groups, defined as -NHR, in which R is cycloalkyl, alkenyl, alkynyl, aryl, and alkoxy, respectively. A non-limiting example of an arylamino group is -NHC6H5. The term "ami do" (acylamino), when used without the "substituted" modifier, refers to the group -NHR, in which R is acyl, as that term is defined above. A non-limiting example of an amido group is -NHC(0)CH3. The term "alkylimino" when used without the "substituted" modifier refers to the divalent group =NR, in which R is an alkyl, as that term is defined above. When any of these terms is used with the "substituted" modifier one or more hydrogen atom attached to a carbon atom has been independently replaced by -OH, -F, -CI, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(0)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(0)NH2, -C(0)NHCH3, -C(0)N(CH3)2, -OC(0)CH3, -NHC(0)CH3, -S(0)2OH, or -S(0)2NH2. The groups -NHC(0)OCH3 and -NHC(0)NHCH3 are non-limiting examples of substituted amido groups.
[00107] The use of the word "a" or "an," when used in conjunction with the term "comprising" in the claims and/or the specification may mean "one," but it is also consistent with the meaning of "one or more," "at least one," and "one or more than one."
[00108] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.
[00109] The terms "comprise," "have" and "include" are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as "comprises," "comprising," "has," "having," "includes" and "including," are also open-ended. For example, any method that "comprises," "has" or "includes" one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps.
[00110] The term "effective," as that term is used in the specification and/or claims, means adequate to accomplish a desired, expected, or intended result. "Effective amount," "Therapeutically effective amount" or "pharmaceutically effective amount" when used in the context of treating a patient or subject with a compound means that amount of the compound which, when administered to a subject or patient for treating a disease, is sufficient to effect such treatment for the disease. [00111] As used herein, the term "IC50" refers to an inhibitory dose which is 50% of the maximum response obtained. This quantitative measure indicates how much of a particular drug or other substance (inhibitor) is needed to inhibit a given biological, biochemical or chemical process (or component of a process, i.e. an enzyme, cell, cell receptor or microorganism) by half. [00112] An "isomer" of a first compound is a separate compound in which each molecule contains the same constituent atoms as the first compound, but where the configuration of those atoms in three dimensions differs.
[00113] As used herein, the term "patient" or "subj ect" refers to a living mammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat, mouse, rat, guinea pig, or transgenic species thereof. In certain embodiments, the patient or subject is a primate. Non-limiting examples of human subjects are adults, juveniles, infants and fetuses.
[00114] As generally used herein "pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and/or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio.
[00115] "Pharmaceutically acceptable salts" means salts of compounds of the present disclosure which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as 1 ,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-ene-l-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene- 1 -carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, gly colic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, / chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, />-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiarybutylacetic acid, trimethylacetic acid, and the like. Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present are capable of reacting with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine and the like. It should be recognized that the particular anion or cation forming a part of any salt of this disclosure is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).
[00116] The term "pharmaceutically acceptable carrier," as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a chemical agent.
[00117] "Prevention" or "preventing" includes: (1) inhibiting the onset of a disease in a subj ect or patient which may be at risk and/or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease, and/or (2) slowing the onset of the pathology or symptomatology of a disease in a subject or patient which may be at risk and/or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease.
[00118] A "stereoisomer" or "optical isomer" is an isomer of a given compound in which the same atoms are bonded to the same other atoms, but where the configuration of those atoms in three dimensions differs. "Enantiomers" are stereoisomers of a given compound that are mirror images of each other, like left and right hands. "Diastereomers" are stereoisomers of a given compound that are not enantiomers. Chiral molecules contain a chiral center, also referred to as a stereocenter or stereogenic center, which is any point, though not necessarily an atom, in a molecule bearing groups such that an interchanging of any two groups leads to a stereoisomer. In organic compounds, the chiral center is typically a carbon, phosphorus or sulfur atom, though it is also possible for other atoms to be stereocenters in organic and inorganic compounds. A molecule can have multiple stereocenters, giving it many stereoisomers. In compounds whose stereoisomerism is due to tetrahedral stereogenic centers (e.g. , tetrahedral carbon), the total number of hypothetically possible stereoisomers will not exceed 2n, where n is the number of tetrahedral stereocenters. Molecules with symmetry frequently have fewer than the maximum possible number of stereoisomers. A 50:50 mixture of enantiomers is referred to as a racemic mixture. Alternatively, a mixture of enantiomers can be enantiomerically enriched so that one enantiomer is present in an amount greater than 50%. Typically, enantiomers and/or diastereomers can be resolved or separated using techniques known in the art. It is contemplated that that for any stereocenter or axis of chirality for which stereochemistry has not been defined, that stereocenter or axis of chirality can be present in its R form, S form, or as a mixture of the R and S forms, including racemic and non-racemic mixtures. As used herein, the phrase "substantially free from other stereoisomers" means that the composition contains < 15%, more preferably < 10%, even more preferably < 5%, or most preferably < 1% of another stereoisomer(s).
[00119] "Treatment" or "treating" includes (1) inhibiting a disease in a subject or patient experiencing or displaying the pathology or symptomatology of the disease (e.g. , arresting further development of the pathology and/or symptomatology), (2) ameliorating a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease (e.g. , reversing the pathology and/or symptomatology), and/or (3) effecting any measurable decrease in a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease.
[00120] The above definitions supersede any conflicting definition in any reference that is incorporated by reference herein. The fact that certain terms are defined, however, should not be considered as indicative that any term that is undefined is indefinite. Rather, all terms used are believed to describe the disclosure in terms such that one of ordinary skill can appreciate the scope and practice the present disclosure.
VII. Examples
[00121] The following examples are included to demonstrate preferred embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the disclosure, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure. Example 1 - Chemical Synthesis
A. Synthesis
[00122] The first class of compounds prepared possessing two redox centers was selenium-containing dihydropyran naphthoquinones obtained from lapachol (1) (Scheme 1). Two selenium-containing derivatives, 6 and 7, were synthesized from a-lapachone (2) which in turn was prepared by acid catalyzed cyclization from lapachol (1). Compound 6 was prepared in moderate yield (75%) by copper(I) catalyzed click reaction (Rostovtsev et al, 2002.) between compound 4 and (azidomethyl)(phenyl)selane. The intermediate compound 4 was obtained by the reaction of 3-ethynylaniline and the bromo derivative 3. As previously reported, (Guimaraes et al, 2013) 4-azide-a-lapachone (5) was easily synthesized from the reaction of 3 with sodium azide in dichloromethane. The reaction of 5 and phenyl propargyl selenide affords selenium-containing α-lapachone 1,2,3-triazole 7. Finally, from the azide derivative 9, prepared as reported by us, (da Silva Junior et al, 2010) β-lapachone-based 1,2,3-triazole 10 containing the chalcogen was obtained as a red solid. Compounds 3-10 are racemic. However, compounds 9 and 10 are single diastereomers, the relative stereochemistry is trans. The frara-stereochemistry was confirmed by comparison with previously reported data (da Silva Junior, et al , 2010; Jardim, et al , 2015).
Figure imgf000056_0001
Scheme 1. Synthesis of selenium-containing β-lapachone and a-lapachone-based 1,2,3- triazoles.
[00123] The synthesis of selenium-containing dihydrofuran naphthoquinones, the second class of compounds, was begun initially by synthesizing nor-a-lapachone derivatives 15 and 16. Since the synthesis of arylamino substituted lapachones and azidoquinones have been previously developed (da Silva Junior et al., 2008), compounds 13 and 14 were prepared as shown in Scheme 2. Following click methodology, compounds 13 and 14 were reacted with selenium-containing azide and alkyne, respectively, to furnish the naphthoquinones 15 and 16 in 70% and 80% yield, respectively.
Figure imgf000057_0001
Scheme 2. Synthesis of selenium-containing nor-a-lapachone-based 1,2,3-triazoles.
[00124] From nor-lapachol (17), the bromo intermediate 18 was synthesized following the methodology described by Pinto and co-workers (see Scheme 3). (Pinto et al, 1982.) Synthesis of various antitumor compounds from 18 was reported, as for instance, arylamino and alkoxy substituted ηοΓ-β-lapachone (da Silva Junior et al, 2010) , lapachones in the presence of 1,2,3-triazole moiety (da Silva Junior, 2011) and hybrids with chalcones. (Jardim et al., 2015.) The unpublished arylamino substituted lapachone 19 bearing a terminal alkyne group was prepared based on the previously described compounds possessing activity against cancer cell lines. The formation of the selenium-containing 1,2,3-triazole 21 from 19 herein described, allowed access to a product designed with two redox centers. Using the same strategy discussed above, compound 22 was obtained from the azide derivative 20 (Scheme 3) (da Silva Junior et al, 2008.).
Figure imgf000058_0001
Scheme 3. Synthesis of selenium-containing ηοΓ-β-lapachone-based 1,2,3-triazoles.
[00125] At this juncture, the synthesis of lapachones obtained from lapachol (1) and nor-lapachol (17), their inferior homologue, was described. Recently, the synthesis of a new class of naphthoquinone compounds has been reported, containing a pendant 1,2,3- triazole motif from C-allyl lawsone (23) (Jardim et al, 2015). The iodination of 23 affords compounds 24 and 27 in 68% yield and 1 : 1 ratio (Scheme 4) which were easily separated by column chromatography. From these compounds, the respective azide derivatives, compounds 25 and 28, were synthesized by the reaction of sodium azide in dimethylformamide. The respective selenium derivatives, compounds 26 and 29, were prepared by Cu-catalyzed azide-alkyne cycloaddition (Scheme 4).
Figure imgf000059_0001
Scheme 4. Synthesis of 26 and 29 from C-allyl-lawsone (23).
[00126] 1,4-naphthoquinone coupled to selenium-containing 1,2,3-triazole was also a subject of this study. From compounds 33-35 and 39, the respective triazolic derivatives, compounds 36-38 and 40, were prepared using methodology discussed previously (Scheme 5). Suitable crystals of compounds 35 and 38 were obtained, and the structures were solved by crystallographic methods.
Figure imgf000059_0002
Scheme 5. Synthesis of selenium-containing /¾zra-quinone-based 1,2,3-triazoles. [00127] The structures of the novel compounds 4, 6, 7, 10, 13, 15, 16, 19, 21, 22, 26, 29, 36, 37, 38, and 40 were determined by ¾ NMR, 13C NMR, and 2D NMR spectra (COSY, HMBC and HSQC). Electrospray ionization mass spectra were also obtained to confirm compound identities. B. Characterization
[00128] Melting points were obtained on a Thomas Hoover melting point apparatus and are uncorrected. Column chromatography was performed on silica gel (SiliaFlash G60 UltraPure 60-200 μιτι, 60 A). Infrared spectra were recorded on an FTIR Spectrometer IR Prestige-21-Shimadzu. ¾ and 1 C NMR were recorded at room temperature using a Bruker AVANCE DRX200 and DRX400 MHz, in the solvents indicated, with tetramethylsilane (TMS) as internal reference. Chemical shifts (δ) are given in parts per million (ppm) and coupling constants (J) in Hertz (Hz). The mass spectrometer was operated in the positive ion mode. A standard atmospheric pressure photoionization (APPI) source was used to generate the ions. The sample was injected using a constant flow (3 μΙ7ιηίη). The solvent was an acetonitrile/methanol mixture. The APPI-Q-TOF MS instrument was calibrated in the mass range of 50-3000 m/z using an internal calibration standard (low concentration tuning mix solution) supplied by Agilent Technologies. Data were processed employing Bruker Data Analysis software version 4.0. Compounds were named following IUPAC rules as applied by ChemBioDraw Ultra (version 12.0). [00129] Lawsone were acquired from Sigma-Aldrich (St. Louis, MO, USA).
Lapachol (1) (2-hydroxy-3-(3'-methyl-2'-butenyl)-l,4-naphthoquinone) was extracted from the heartwood of Tabebuia sp. (Tecoma). C-allyl lawsone (23) was prepared from lawsone as previously reported. A saturated aqueous sodium carbonate solution was added to the sawdust of ipe tree. Upon observing rapid formation of lapachol sodium salt, hydrochloric acid was added, allowing the precipitation of lapachol. Then, the solution was filtered and a yellow solid was obtained. This solid was purified by recrystallizations with hexane. All chemicals were obtained from commercial sources and used without further purification. Solvents were distilled and when required were dried by distillation according to standard procedures. [00130] For the synthesis of (azidomethyl)(phenyl)selane (PhSeCH2N3): Initially
(chloromethyl)(phenyl)selane was prepared by the reaction of a solution of diphenyl diselenide (3.0 mmol) in THF (6.0 mL) with NaBH4 (2 eq.) in EtOH (6.0 mL) and CH2CI2 (30 mL). The mixture was kept under reflux in inert atmosphere and stirred for 12 h. After this period and subsequently extraction with H2O, the organic phase were combined, dried over MgS04, and concentrated under vacuum. The residue was purified by flash chromatography on silica gel using hexane as the eluent. To a solution of (chloromethyl)(phenyl)selane (PhSeCH2Cl) (3.0 mmol) in CH3CN (5.0 mL), sodium azide (4.5 mmol) and 18-crown-6 (0.60 mmol) were added at room temperature. Then the reaction mixture was stirred for 48 h under nitrogen atmosphere. After this time, 30 mL of H2O was added and the organic phase was extracted with CH2CI2. The organic layers were combined, dried over MgS04, and concentrated under vacuum. The residue was purified by flash chromatography on silica gel using hexane as the eluent. The product was obtained in 90% yield.
[00131] For the synthesis of phenyl propargyl selenide: To a solution of diphenyl diselenide (1.0 mmol) in THF (8.0 mL) with NaBH4 (2 eq.) in EtOH (4 mL). The mixture was kept under agitation at temperatue of 0 °C under inert atmosphere and, then, propargyl bromide (2.0 mmol) in THF (4 mL) was added. After 10 minutes, 30 mL of H2O was added and the organic phase was extracted with ethyl acetate. The organic phases were combined, dried over MgS04, and concentrated under vacuum. The residue was purified by flash chromatography on silica gel using hexane as the eluent. The product was obtained in 85% yield. 1. Procedures to prepare arylamino lapachones 4, 13 and 19
[00132] To prepare 4 and 13: Compounds 3 and 12 (1.0 mmol) were dissolved in 25 mL of CH2CI2 and an excess of 3-ethynylaniline (117 mg, 1.2 mmol) was added. The mixture was left under stirring overnight, followed by the addition of 50 mL of water. The organic phase was extracted with CH2CI2, washed with 10% HC1 (3 x 50 mL), dried over sodium sulfate, and filtered. The solvent from the crude was evaporated under reduced pressure and it was purified by column chromatography on silica-gel, using eluents with an increasing polarity gradient mixture of hexane and ethyl acetate (9/1 to 7/3).
[00133] To prepare 19: To a solution of nor-lapachol (17) (228 mg, 1.0 mmol) in 25 mL of chloroform, 2 mL of bromine was added. The bromo intermediate 18 precipitated immediately as an orange solid. After removal of bromine, by adding dichloromethane and then removing the organic solvent with dissolved bromine by rotary evaporator, an excess of 3-ethynylaniline (117 mg, 1.2 mmol) was added in CH2CI2 and the mixture was stirred overnight. The crude reaction mixture was poured into 50 mL of water. The organic phase was separated and washed with 10% HC1 (3 x 50 mL), dried over sodium sulfate, filtered, and evaporated under reduced pressure. The product 19 was obtained after purification by column chromatography in silica-gel, eluted with an increasing polarity gradient mixture of hexane and ethyl acetate (9/1 to 7/3).
2. General procedures to prepare the azide derivatives
[00134] The previously published azido derivatives 5, 9, 14, 20, 25, 39, 46 were prepared as described in the literature (Guimaraes et al, 2013; da Silva Junior et al, 2010; da Silva Junior et al, 2008; da Silva Junior et al, 2008 and Jardim et al, 2015). Compound 28 (1.0 mmol) was prepared from 27 in the presence of sodium azide (120 mg, 1.85 mmol) in 2 mL of dimethylformamide (DMF). The mixture was stirred at room temperature until product formation was complete as determined by thin layer chromatography. After extraction with CH2CI2, the residue was dried over anhydrous Na2S04 and concentrated under reduced pressure. Compound 28 was obtained after purification by column chromatography on silica gel eluting with a gradient mixture of hexane: ethyl acetate with increasing polarity. The azide derivative 47 was prepared following the same procedure previously described. (Nair et al, 2014.)
3. General procedures for the preparation of 1,2,3-triazole derivatives
[00135] The azidoquinones (1.0 mmol) or quinone with terminal alkynes (1.0 mmol) were reacted with CUSO4 5H2O (0.04 mmol) and sodium ascorbate (0.11 mmol) and the phenyl propargyl selenide (195 mg, 1.0 mmol) or (azidomethyl)(phenyl)selane (212 mg, 1.0 mmol) in a mixture of CH2C12:H20 (12 mL, 1 : 1, v/v). The mixture was stirred at room temperature until product formation was complete as determined by thin layer chromatography. The aqueous phase was extracted with CH2CI2, dried over anhydrous Na2S04 and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with a gradient mixture of hexane: ethyl acetate with increasing polarity. 4-((3-ethynylphenyl)amino)-2,2-dimethyl-3,4-dihydro-2H-benzo[^]chromene-5,10-dione
(4)
Figure imgf000063_0001
[00136] Yield: 70%; mp 196-197 °C; Brown solid. ¾ NMR (200 MHz, CDCh) δ 8.11-8.03 (m, 2H), 7.75-7.68 (m, 2H), 7.16 (t, J = 7.5 Hz, 1H), 6.95-6.85 (m, 2H), 6.71 (d, J = 8.0 Hz, 1H), 4.68 (t, J = 3.7 Hz, 1H), 3.03 (s, 1H), 2.27 (dd, J = 3.5 and 14.4 Hz, 1H), 2.02 (dd, J = 5.5 and 14.4 Hz, 1H), 1.54 (s, 6H); 13C NMR (CDCh, 50 MHz) δ 183.7, 180.0, 155.5, 146.3, 134.6, 133.3, 132.1, 131.0, 129.4, 126.5, 126.3, 123.0, 122.8, 118.4, 117.1, 115.2, 84.1, 79.2, 43.7, 37.4, 28.7, 26.3; HRMS (ES+) calculated for C23H20NO3 [M+H]+: 358.1443; found: 358.1488.
2,2-dimethyl-4-((3-(l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)phenyl)amino)-3,4- dihydro-2H-benzo[^]chromene-5,10-dione (6)
Figure imgf000063_0002
[00137] Yield: 75%; mp 101-102 °C; Red solid. ¾ NMR (400 MHz, CDCh) δ 8.11 (dd, J = 7.1 and 1.7 Hz, H9), 8.07 (dd, J = 7.1 and 1.6 Hz, He), 7.72 (td, J = 7.1, 7.1 and 1.6 Hz, He), 7.68 (td, J = 7.1, 7.1 and 1.7 Hz, Hv), 7.62 (s, Hs-triazole), 7.50 (d, J = 7.4 Hz, H2 /6"), 7.33-7.39 (m, H ), 7.26-7.28 (m, H2 ), 7.28-7.33 (m, H3 /5"), 7.20-7.26 (m, H5 ), 7.05 (d, J = 7.7 Hz, H ), 6.69 (dd, J = 8.0 and 1.8 Hz, He ), 5.71 (s, H12), 4.78 (dd, J = 5.6 and 3.4 Hz, H4), 2.32 (dd, J = 14.3 and 3.4 Hz, H3a or Ha), 2.06 (dd, J = 14.3 and 5.6 Hz, H3a or H3b), 1.55 (s, Hir or H11 ), 1.54 (s, Hir or H11 ); 1 C NMR (CDCh, 100 MHz) δ 183.5 (Cs), 180.0 (Cio), 155.3 (Cioa), 148.5 (C4-triazole), 147.4 (Cr), 134.8 (Crve-), 134.3 (Cs), 133.1 (Cv), 132.1 (C9a), 131.3 (Cr), 130.9 (Csa), 129.8 (Cs-), 129.6 (C3 /5 ), 129.0 (C4 ), 127.4 (Cr ), 126.4 (C9), 126.2 (Ce), 119.4 (Cs-triazole), 118.9 (C4a), 115.7 (C4 ), 113.6 (Ce ), 110.7 (C2 , 79.1 (C2), 44.8 (C12), 43.2 (C4), 37.6 (Cs), 29.0 (Cir or Cii"), 26.1 (Cir or Cii"); HRMS (ES+) calculated for C3oH27N 03Se [M+H]+: 571.1248; found: 571.1234. 2,2-dimethyl-4-(4-((phenylselanyl)methyl)-lH-l,2,3-triazol-l-yl)-3,4-dihyd
benzo[g]chromene-5,10-dione (7)
Figure imgf000064_0001
[00138] Yield: 70%; mp 168-169 °C; Yellow solid. ¾ NMR (200 MHz, CDCh) δ 8.18-8.09 (m, 1H), 8.04-7.96 (m, 1H), 7.80-7.68 (m, 2H), 7.50-7.39 (m, 2H), 7.24-7.10 (m, 4H), 5.71 (t, J = 6.3 Hz, 1H), 4.13 (s, 2H), 2.73 (dd, J = 5.5 and 14.4 Hz, 1H), 2.29 (dd, J = 6.3 and 14.4 Hz, 1H), 1.50 (s, 3H), 1.21 (s, 3H); 13C NMR (CDCh, 50 MHz) δ 182.7, 179.5, 156.3, 145.3, 134.7, 133.7, 131.8, 131.1, 129.7, 129.1, 127.5, 126.8, 126.6, 122.0, 115.0, 79.4, 49.6, 39.0, 27.0, 26.5, 20.8; HRMS (ES+) calculated for C24H2iN303SeNa [M+Na]+: 502.0646; found: 502.0643.
3-bromo-2,2-dimethyl-4-(4-((phenylselanyl)methyl)-lH-l,2,3-triazol-l-yl)-3,4-dihydro-
2H-benzo[A]chromene-5,6-dione (10)
Figure imgf000064_0002
[00139] Yield: 90%; mp 105-106 °C; Orange solid. ¾ NMR (400 MHz, CDCh) δ 8.11 (dd, J = 7.6 and 1.4 Hz, Hv), 7.90 (dd, J = 7.6 and 1.2 Hz, Hio), 7.73 (td, J = 7.6, 7.6 and 1.2 Hz, He), 7.63 (td, J = 7.6, 7.6 and 1.4 Hz, H9), 7.48-7.52 (m, Η2 /6·), 7.48 (s, Hs-triazole), 7.22-7.25 (m, H375 ), 7.22-7.25 (m, H ), 5.55 (d, J = 8.9 Hz, H4), 4.93 (d, J = 8.9 Hz, H3), 4.17 (d, J = 13.3 Hz, Hi2a), 4.13 (d, J = 13.3 Hz, Hub), 1.71 (s, Hir or H11 ), 1.64 (s, Hir or H11 ); 1 C NMR (CDCh, 100 MHz) δ 177.8 (Ce), 176.4 (Cs), 162.7 (Ciob), 144.5 (C4- triazole), 135.2 (Cs), 134.1 (Cr/e'), 132.3 (C9), 130.6 (Cioa), 130.5 (C6a), 129.4 (Cr), 129.2 (Cv), 129.1 (Cr/y), 127.6 (C4-), 125.3 (Cio), 125.0 (Cs-triazole), 110.3 (C4a), 83.4 (C2), 54.4 (Cs), 58.9 (C4), 20.7 (Cir or C11 ), 27.4 (Cir or C11 ), 20.7 (C12); HRMS (ES+) calculated for C24H2iBrN303Se [M+H]+: 557.9931; found: 557.9923. 3-((3-ethynylphenyl)amino)-2,2-dimethyl-2,3-dihydronaphtho[2,3-Z>]furan-4,9-dione (13)
Figure imgf000065_0001
[00140] Yield: 65%; mp 165-167 °C; Brown solid. ¾ NMR (400 MHz, CDCh) δ 8.10 (dd, J = 7.4 and 1.4 Hz, He), 8.07 (dd, J = 7.4 and 1.6 Hz, Hs), 7.73 (td, J = 7.4, 7.4 and 1.6 Hz, Hv), 7.68 (td, J = 7.4, 7.4 and 1.4 Hz, He), 7.60 (s, Hs-triazole), 7.50 (d, J = 7.5 Hz, H2 /6"), 7.27-7.39 (m, Hy/s"), 7.27-7.39 (m, H ), 7.22 (t, J = 7.8 Hz, H5 ), 7.20 (s, H2 ), 7.04 (d, J = 7.8 Hz, H4 ), 6.61 (dd, J = 7.8 and 1.8 Hz, He ), 5.71 (s, H11), 4.96 (s, H3), 4.05 (si, NH), 1.67 (s, Η10· or Hio ), 1 -57 (s, Η10· or H10 ··); 1 C NMR (CDCh, 100 MHz) δ 181.8 (C4), 178.6 (C9), 159.8 (C9a), 148.3 (C4-triazole), 147.5 (Cr), 134.8 (C2 /6 ), 134.5 (Cv), 133.2 (C8a), 133.1 (Ce), 131.9 (Cr), 131.6 (C4a), 129.9 (C5 ), 129.6 (C3 /5 ), 129.0 (C4 ), 127.4 (Ci ), 126.5 (Cs), 126.2 (C5), 122.3 (C3a), 119.4 (Cs-triazole), 115.9 (C4 ), 113.1 (Ce ), 110.5 (C2 , 95.5 (C2), 62.3 (C3), 44.7 (C11), 27.2 (Cio- or C10"), 21.6 (Cio- or C10"); HRMS (ES+) calculated for C29H25N 03Se [M+H]+: 557.1092; found: 557.1101.
2,2-dimethyl-3-((3-(l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)phenyl)amino)-2,3- dihydronaphtho[2,3-Z>]furan-4,9-dione (15)
Figure imgf000065_0002
[00141] Yield: 70%; mp 120-122 °C; Red solid. ¾ NMR (400 MHz, CDCh) δ 8.10-8.03 (m, 2H), 7.76-7.65 (m, 2H), 7.59 (s, 1H), 7.52-7.47 (m, 2H), 7.37-7.27 (m, 3H), 7.23-7.17 (m, 2H), 7.01 (d, J = 7.6 Hz, 1H), 6.60 (dd, J = 2.1 and 7.6 Hz, 1H), 5.69 (s, 2H), 4.96 (s, 1H), 4.11 (s, 1H), 1.66 (s, 3H), 1.56 (s, 3H); 13C NMR (CDCh, 100 MHz) δ 181.9, 178.6, 159.7, 148.3, 147.5, 134.8, 134.6, 129.9, 129.6, 129.0, 126.5, 126.2, 119.4, 115.8, 113.0, 110.4, 95.1, 62.1, 44.8, 27.2, 21.6; HRMS (ES+) calculated for C29H24N 03SeH [M+H]+: 557.1092; found: 557.1101. 2,2-dimethyl-3-(4-((phenylselanyl)methyl)- 1H- 1,2,3-triazol- l-yl)-2,3- dihydronaphtho[2,3-Z>]furan-4,9-dione (16)
Figure imgf000066_0001
[00142] Yield: 80%; mp 172-174 °C; Yellow solid. ¾ NMR (400 MHz, CDCh) δ 8.18 (dd, J = 7.4 and 1.6 Hz, He), 8.08 (dd, J = 7.4 and 1.6 Hz, Hs), 7.80 (td, J = 7.4, 7.4 and 1.6 Hz, Hv), 7.76 (td, J = 7.4, 7.4 and 1.6 Hz, He), 7.40 (dd, J = 7.9 and 1.4 Hz, Η2 /6·), 7.08- 7.19 (m, H3 /5 ), 7.08-7.19 (m, H ), 6.98 (s, H5-triazole), 5.92 (s, H3), 4.14 (d, J = 13.5 Hz, H11), 4.09 (d, J = 13.5 Hz, H11), 1.67 (s, Η10· or H10 ), 1 02 (s, Η10· or H10 ··); 1 C NMR (CDCh, 100 MHz) δ 180.5 (C4), 177.8 (C9), 151.1 (C9a), 145.8 (C4-triazole), 134.9 (Cv), 133.9 (C2V6'), 133.6 (Ce), 132.7 (Cua), 131.6 (C4a), 129.13 (Cr), 129.06 (Csvs-), 127.5 (C4-), 126.8 (Cs), 126.5 (Cs), 121.2 (Cs-triazole), 118.3 (Csa), 94.5 (C2), 67.3 (Cs), 27.4 (Cio- or Cio"), 20.7 (Cio- or Cio"), 20.5 (C11); HRMS (ES+) calculated for C23Hi9N303SeNa [M+Na]+: 488.0489; found: 488.0486.
3-((3-ethynylphenyl)amino)-2, tho[l,2-Z>]furan-4,5-dione (19)
Figure imgf000066_0002
[00143] Yield: 70%; mp 205-206 °C; Red solid. ¾ NMR (200 MHz, CDCh) δ 8.10 (dd, J = 2.0 and 8.0 Hz, 1H), 7.76-7.58 (m, 3H), 7.12 (t, J= 7.8 Hz, 1H), 6.88 (d, J= 7.8 Hz, 1H), 6.69 (s, 1H), 6.58 (dd, J = 2.0 and 8.0 Hz, 1H), 4.79 (s, 1H), 3.02 (s, 1H), 1.68 (s, 3H), 1.57 (s, 3H); 13C NMR (CDCh, 50 MHz) δ 175.4, 169.7, 147.1, 134.7, 132.7, 131.2, 129.6, 129.4, 128.6, 125.2, 122.9, 122.2, 116.1, 115.0, 114.1, 96.8, 84.1, 61.5, 27.4, 21.8; HRMS (ES+) calculated for C22Hi7N03Na [M+Na]+: 366.1106; found: 366.1108. 2,2-dimethyl-3-((3-(l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)phenyl)amino)-2,3- dihydronaphtho[l,2-Z>]furan-4,5-dione (21)
Figure imgf000067_0001
[00144] Yield: 50%; mp 135-137 °C; Red solid. ¾ NMR (400 MHz, CDCh) δ 8.12 (d, J = 7.4 Hz, He), 7.70 (td, J = 7.4, 7.4 and 1.1 Hz, Hv), 7.74 (dd, J = 7.4 and 1.1 Hz, H9), 7.64 (td, J = 7.4, 7.4 and 1.6 Hz, He), 7.59 (s, H5-triazole), 7.50 (d, J = 6.9 Hz, H2 /6 ), 7.28-7.40 (m, H3 /5 ), 7.28-7.40 (m, H ), 7.14 (s, H2 ), 7.20 (dd, J = 8.1 and 7.7 Hz, H5 ), 7.02 (d, J = 7.7 Hz, H4 ), 6.56 (dd, J = 8.1 and 2.2 Hz, He ), 5.51 (s, H11), 4.89 (s, H3), 3.70- 4.37 (si, NH), 1.60 (s, Η10· or H10 ··), 1.17 (s, Η10· or H10 ··); 1 C NMR (CDCh, 100 MHz) δ 181.0 (Cs), 175.4 (C4), 169.6 (C»), 148.3 (C4-triazole), 147.8 (Cr), 134.8 (C2 /6 ), 134.6 (Cv), 132.5 (Cs), 131.3 (Cr), 131.2 (C9a), 129.8 (Cs-), 129.6 (Cs-vs-), 129.5 (Ce), 128.9 (C4-), 127.5 (Csa), 127.4 (Cr), 125.1 (C9), 119.5 (Cs-triazole), 115.5 (C4 ), 115.1 (Csa), 113.0 (Ce ), 110.2 (C2 , 96.9 (C2), 61.6 (Cs), 44.8 (C11), 27.5 (Cnr or C10 ··), 21.8 (Cio- or C10 ); HRMS (ES+) calculated for C29H24N 03SeNa [M+Na]+: 579.0911 ; found: 579.0890.
2,2-dimethyl-3-(4-((phenylselanyl)methyl)- 1H- 1,2,3-triazol- l-yl)-2,3- dihydronaphtho[l,2-Z>]furan-4,5-dione (22)
Figure imgf000067_0002
[00145] Yield: 80%; mp 186-188 °C; Yellow solid. ¾ NMR (200 MHz, CDCh) δ 8.19 (d, J = 7.3 Hz, He), 7.68-7.82 (m, Hv), 7.68-7.82 (m, He), 7.68-7.82 (m, H9), 7.41 (d, J = 7.1 Hz, Η2 /6·), 7.08-7.21 (m, H3 /5 ), 7.08-7.21 (m, H ), 7.00 (s, Hs-triazole), 5.86 (s, H3), 4.05-4.16 (m, H11), 1.70 (s, Η10· or H10 ), 1 05 (s, Η10· or H10 ··); 1 C NMR (CDCh, 50 MHz) δ 180.0 (Cs), 174.5 (C4), 171.1 (C»), 145.5 (C4-triazole), 134.9 (Cv), 134.0 (C2ve ), 133.4 (Cs), 131.4 (C9a), 129.9 (Ce), 129.2 (Cr), 129.1 (Csvs-), 127.5 (C4-), 126.6 (Csa), 125.6 (C9), 121.2 (Cs-triazole), 111.2 (Csa), 95.9 (C2), 66.7 (Cs), 27.6 (Cio- or C10"), 20.9 (Cio- or C10"), 20.6 (C11); HRMS (ES+) calculated for C23Hi9N303SeNa [M+Na]+: 488.0489; found: 488.0482. 2-((4-((phenylselanyl)methyl)-lH-l,2,3-triazol-l-yl)methyl)-2,3-dihydronaphtho[l,2-
Z>]furan-4,5-dione (26)
Figure imgf000068_0001
8.00 (dd, J = 7.4 and 1.5 Hz, He), 7.59 (td, J = 7.4, 7.4 and 1.4 Hz, Hv), 7.53 (td, J = 7.4, 7.4 and 1.5 Hz, He), 7.44 (dd, J = 7.4 and 1.1 Hz, H9), 7.33-7.41 (m, Η2 /6·), 7.31 (s, Hs-triazole), 7.13-7.18 (m, H375 ), 7.13-7.18 (m, H4 ), 5.33-5.42 (m, H2), 4.55 (dd, J = 14.6 and 7.3 Hz, Hioa), 4.64 (dd, J = 14.6 and 3.9 Hz, Hiob), 4.11 (s, H11), 3.23 (dd, J = 15.8 and 10.2 Hz, H3a), 2.83 (dd, J = 15.8 and 7.0 Hz, ¾b); 1 C NMR (CDCh, 100 MHz) δ 180.4 (C5), 175.2 (C4), 186.6 (C9b), 146.4 (C4-triazole), 134.7 (C7), 132.9 (C276'), 132.3 (C8), 130.6 (C9a), 129.8 (C6), 129.8 (CI '), 129.2 (C375'), 127.5 (C4'), 126.9 (C5a), 124.4 (C9), 122.7 (C5-triazole), 114.7 (C3a), 84.3 (C2), 53.3 (CIO), 29.6 (C3), 20.2 (Cl l); HRMS (ES+) calculated for C22Hi8N303Se [M+H]+: 452.0513; found: 452.0896.
2-(azidomethyl)-2,3-dihydronaphtho [2,3-Z>]furan-4,9-dione (28)
Figure imgf000068_0002
[00147] Yield: 90%; mp 116-117 °C; Yellow solid. ¾ NMR (200 MHz, CDCh) δ 8.06-7.94 (m, 2H), 7.71-7.57 (m, 2H), 5.23-5.09 (m, 1H), 3.75 (dd, J = 3.7 and 13.3 Hz, 1H), 3.56 (dd, J = 5.0 and 13.3 Hz, 1H), 3.26 (dd, J = 10.8 and 17.4 Hz, 1H), 3.02 (dd, J = 7.5 and 17.4 Hz, 1H); 13C NMR (CDCh, 50 MHz) δ 181.8, 177.1, 159.4, 134.1, 133.0, 132.6, 131.2, 126.1, 125.8, 124.0, 83.6, 53.5, 30.0; HRMS (ES+) calculated for C13H10N3O3 [M+H]+: 256.0722; found: 256.0716. 2-((4-((phenylselanyl)methyl)-lH-l,2,3-triazol-l-yl)methyl)-2,3-dihydronaphtho[2,3-
Z>]furan-4,9-dione (29)
Figure imgf000069_0001
[00148] Yield: 90%; mp 169-171 °C; Yellow solid. ¾ NMR (400 MHz, CDCh) δ 8.00 (dd, J = 7.4 and 1.5 Hz, He), 7.93 (dd, J = 7.4 and 1.6 Hz, H5), 7.66 (td, J = 7.4, 7.4 and 1.6 Hz, Hv), 7.62 (td, J = 7.4, 7.4 and 1.5 Hz, He), 7.32-7.37 (m, Hs-triazole), 7.32-7.37 (m, Η2 /6·), 7.11-7.17 (m, H3 /5 ), 7.11-7.17 (m, H4 ), 5.23-5.32 (m, H2), 4.64 (dd, J = 14.7 and 3.9 Hz, Hiob), 4.57 (dd, J = 14.7 and 5.6 Hz, Hioa), 4.05 (s, H11), 3.26 (dd, J = 17.4 and 10.6 Hz, H3b), 2.95 (dd, J = 17.4 and 8.1 Hz, H3b); 1 C NMR (CDCh, 50 MHz) δ 181.6 (C4), 177.2 (C9), 159.0 (C9a), 146.4 (C4-triazole), 134.4 (Cv), 133.1 (C2 /6 ), 132.7 (Cua), 133.2 (Ce), 131.3 (C4a), 129.7 (Cr), 129.1 (Csvs-), 127.4 (C4-), 126.4 (Cs), 126.2 (C5), 124.1 (C3a), 123.1 (C5- triazole), 83.0 (C2), 52.9 (Cio), 30.1 (C3), 20.3 (C11); HRMS (ES+) calculated for C22Hi8N303Se [M+H]+: 452.0513; found: 452.0512.
2-chloro-3-(((l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)methyl)amino)-
Figure imgf000069_0002
[00149] Yield: 65%; mp 127-128 °C; Orange solid. ¾ NMR (200 MHz, CDCh) δ 8.14 (dd, J = 1.2 and 7.5 Hz, 1H), 8.03 (dd, J = 1.2 and 7.5 Hz, 1H), 7.97-7.57 (m, 2H), 7.51- 7.40 (m, 3H), 7.33-7.23 (m, 3H), 6.49 (si, 1H), 5.68 (s, 2H), 5.10 (d, J = 6.3 Hz, 2H); 13C NMR (CDCh, 50 MHz) δ 180.0, 176.7, 144.8, 143.6, 134.8, 134.7, 132.5, 132.3, 129.7, 129.5, 129.0, 126.9, 126.8, 126.7, 121.8, 111.6, 43.6, 40.0; HRMS (ES+) calculated for C2oHi6ClN402Se [M+H]+: 459.0127; found: 459.0128. 2-bromo-3-(((l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)methyl)amino)-l,4- naphthoquinone (37)
Figure imgf000070_0001
[00150] Yield: 60%; mp 124-126 °C; Orange solid. ¾ NMR (200 MHz, CDCh) δ 8.10 (dd, J = 7.6 and 1.3 Hz, He), 8.06 (dd, J = 7.6 and 1.3 Hz, Hs), 7.74 (td, J = 7.6, 7.6 and 1.3 Hz, He), 7.64 (td, J = 7.6, 7.6 and 1.3 Hz, Hv), 7.46 (dd, J = 7.8 and 1.5 Hz, Η2 /6·), 7.39 (s, Hs-triazole), 7.27-7.35 (m, H375 ), 7.27-7.35 (m, H ), 6.22-6.35 (m, NH), 5.77 (s, H3), 5.68 (s, H10), 4.45 (d, J = 5.8 Hz, H9); 1 C NMR (CDCh, 50 MHz) δ 183.0 (Ci), 181.6 (C4), 147.4 (C4-triazole), 143.2 (C2), 134.82 (Cr/v), 134.78 (Ce), 133.4 (C4a), 132.2 (Cv), 130.4 (Csa), 129.7 (C3 /5 ), 129.1 (C4 ), 127.0 (Cr), 126.3 (Cs), 126.2 (Cs), 121.7 (Cs-triazole), 101.7 (Cs), 44.7 (Cio), 38.1 (C9); HRMS (ES+) calculated for C2oHivN402Se [M+H]+: 425.0517; found: 425.0512.
2-(((l-((phenylselanyl)methyl)-lH-l,2,3-triazol-4-yl)methyl)amino)-l,4-naphthoquinone
(38)
Figure imgf000070_0002
[00151] Yield: 60%; mp 162-164 °C; Red solid. ¾ NMR (200 MHz, CDCh) δ 8.10 (dd, J = 7.6 and 1.3 Hz, Hs), 8.06 (dd, J = 7.6 and 1.3 Hz, H5), 7.74 (td, J = 7.6, 7.6 and 1.3 Hz, He), 7.64 (td, J = 7.6, 7.6 and 1.3 Hz, Hv), 7.46 (dd, J = 7.8 and 1.5 Hz, Η2 /6·), 7.39 (s, Hs-triazole), 7.27-7.35 (m, H3 /5 ), 7.27-7.35 (m, H4 ), 6.22-6.35 (m, NH), 5.77 (s, H3), 5.68 (s, H10), 4.45 (d, J = 5.8 Hz, H9); 1 C NMR (CDCh, 50 MHz) δ 183.0 (Ci), 181.6 (C4), 147.4 (C4-triazole), 143.2 (C2), 134.82 (C2V6-), 134.78 (Ce), 133.4 (C4a), 132.2 (Cv), 130.4 (Csa), 129.7 (C3 /5 ), 129.1 (C4 ), 127.0 (Cr), 126.3 (Cs), 126.2 (Cs), 121.7 (Cs-triazole), 101.7 (Cs), 44.7 (Cio), 38.1 (C9); HRMS (ES+) calculated for C2oHivN 02Se [M+H]+: 425.0517; found: 425.0512. ((phenylselanyl)methyl)- 1H- 1 ,2,3-triazol- 1-yl)- 1 ,4-naph th oq u iiione (40)
Figure imgf000071_0001
[00152] Yield: 60%; mp 117-120 °C; Brown solid. ¾ NMR (200 MHz, CDCh) δ 8.36 (s, 1H), 8.24-8.12 (m, 2H), 7.88-7.79 (m, 2H), 7.72 (s, 1H), 7.59-7.48 (m, 2H), 7.29- 7.26 (m, 2H), 4.25 (s, 2H), 1.6 (s, 1H); 13C NMR (CDCh, 50 MHz) δ 183.6, 179.1, 146.8, 139.1, 134.9, 134.2, 133.4, 131.3, 130.9, 129.3, 129.1, 127.6, 127.1, 126.4, 126.2, 123.8, 20.2; HRMS (ES+) calculated for CuHwNsChSe [M+H]+: 396.0251; found: 396.0242.
Example 2 - Biological Activity
A. Antitumor Activity
1. Methods
[00153] Compounds were tested for antitumor activity in cell culture in vitro using several human cancer cell lines obtained from the National Cancer Institute, NCI (Bethesda, MD). The L929 cells (mouse fibroblast L cells NCTC clone 929) were obtained from the American Type Culture Collection (Manassas, VA), MDCK cells were purchased from the Rio de Janeiro Cell Bank (Rio de Janeiro, Brazil), and the Chinese hamster lung fibroblasts (V79 cells) were kindly provided by Dr. JAP Henriques (UFRGS, Brazil). Peripheral blood mononuclear cells (PBMC) were isolated from heparinized blood from healthy, non-smoker donors who had not taken any medication at least 15 days prior to sampling by a standard method of density-gradient centrifugation on Histopaque-1077 (Sigma Aldrich Co., St. Louis, MO/USA). All cancer cell lines and PBMC were maintained in RPMI 1640 medium. The L929, MDCK and V79 cells were cultivated under standard conditions in DMEM with Earle's salts. All culture media were supplemented with 20% (PBMC) or 10% (cancer, L929, MDCK and V79 cells) fetal bovine serum, 2 mM L-glutamine, 100 IU/mL penicillin, 100 μg/mL streptomycin at 37 °C with 5% CC . PBMC cultures were also supplemented with 2% phytohaemagglutinin. In cytotoxicity experiments, cells were plated in 96-well plates (0.1 χ 106 cells/well for leukemia cells, 0.7 χ 105 cells/well for solid tumor as well V79, L929 and MDCK cells, and 1 χ 106 cells/well for PBMC). All tested compounds were dissolved with DMSO. The final concentration of DMSO in the culture medium was kept constant (0.1%, v/v). Doxorubicin (0.001-1.10 μΜ) was used as the positive control, and negative control groups received the same amount of vehicle (DMSO). The cell viability was determined by reduction of the yellow dye 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to a blue formazan product as described by Mosmann. (Mosmann, 1983.) At the end of the incubation time (72 h), the plates were centrifuged and the medium was replaced by fresh medium (200 μί) containing 0.5 mg/mL MTT. Three hours later, the MTT formazan product was dissolved in DMSO (150 μί) and the absorbance was measured using a multiplate reader (Spectra Count, Packard, Ontario, Canada). Drug effect was quantified as the percentage of control absorbance of the reduced dye at 550 nm. All cell treatments were performed with three replicates. All cells were mycoplasma-free. 2. Results
[00154] All of the selenium-containing quinone-based 1,2,3-triazoles described (Schemes 1-5) and their synthetic precursors were evaluated in vitro using the MTT assay against six cancer cell lines: HL-60 (human promyelocytic leukemia cells, NQ01-), HCT- 116 (human colon carcinoma cells, NQOl), PC3 (human prostate cells, NQ01+), SF295 (human glioblastoma cells, NQ01+), MDA-MB-435 (melanoma cells, NQ01+) and OVCAR-8 (human ovarian carcinoma cells, NQ01+). β-Lapachone and doxorubicin were used as positive controls (Table 1). NQOl- normal cells, human peripheral blood mononucluear cells (PBMC), and murine fibroblast immortalized cell lines (V79 and L929) were used to evaluate the selectivity of the compounds. Mechanistic aspects of selected compounds were also studied for NQOl -dependency using the fairly specific NQOl inhibitor, dicoumarol. As previously described, (Perez-Sacau et al , 2007) the compounds were classified according to their activity as highly active (IC50 < 2 μΜ), moderately active (2 μΜ < IC50 < 10 μΜ), or inactive (IC50 >10 μΜ).
[00155] The results showed most of compounds were highly active against all cancer cell lines evaluated with IC50 values < 2 μΜ. In general terms, ort/20-quinoidal compounds were more active than /¾zra-quinones. However, a-lapachone derivatives with potent antitumor activities were also identified.
[00156] Naphthopyranquinones 5-7, 9 and 10 presented high to moderate activities (IC50 in the range of 0.92 to 5.46 μΜ) and the non-active compound 4 was the exception of this class. For the selenium-containing quinones 6 and 10, the strategy of insertion of a second redox center was a success and these derivatives were more active than their naphthoquinoidal precursors. Recently, the inventors reported the synthesis and antitumor activities of several a-lapachone-based 1,2,3-triazoles (da Cruz et al, 2014). It is important to highlight that the selenium-containing-based 1,2,3-triazole 7 displayed better activity than the compounds without the chalcogen. [00157] Naphthofuranquinones were the second class of compounds evaluated.
Para-naphthoquinones 15 and 16 were active against all cancer cell lines studied. In the last few years, the inventors described nor-a-lapachone-based 1,2,3-triazoles obtained from lapachol (1) with IC50 values > 2 μΜ. (da Cruz et al , 2014.) The strategy herein used to prepare compounds 15 and 16 with the presence of selenium improved the activities of nor-a- lapachone-based 1,2,3-triazoles and these derivatives presented IC50 values ranging from 0.68-1.71 μΜ for 15 and 1.59-2.95 μΜ for 16.
[00158] ΝθΓ-β-lapachone and derivatives are among the most potent compounds from the lapachol group, (da Silva Junior et al, 2007; da Silva Junior et al, 2009) Recently, the cytotoxicity and genetic toxicity of ηοΓ-β-lapachone in human lymphocytes, HL-60 leukemia cells, and immortal normal murine V79 fibroblasts were demonstrated (Cavalcanti et al, 2011) at concentrations of 2.5 and 5 μΜ. This compound failed to induce DNA damage in nontumor cells, but at the highest concentrations, it induced DNA single and double strand breaks and increased the frequency of chromosomal aberrations. The biological effects of ηοΓ-β-lapachone are related to its ability to deplete reduced glutathione (GSH), which leads to a GSSG-dominant pro-oxidant cellular status that conribute to its antiproliferative properties.
[00159] In this context, ηοΓ-β-lapachones with potent antitumor activity have been described and (da Silva Junior et al, 2007; da Silva Junior et al, 2009; da Silva Junior et al, 2010 and da Cruz et al, 2014.) ηοΓ-β-lapachone with arylamino groups were the most active lapachones described, (da Silva Junior et al, 2007 and da Silva Junior et al, 2009.) These compounds present significant antiproliferative effects in human myeloid leukemia cell lines and induce oxidative DNA damage by ROS generation. The compounds also impair DNA repair activity triggering apoptosis. (Cavalcanti et al, 2013.) This compound contains the structural framework of the 3-arylamino-nor^-lapachone derivatives reported before, but with a second redox chalcogen center inserted by click chemistry reaction. This substance was highly active against all cancer cell lines evaluated with IC50 values ranging from 0.07 to 0.38 μΜ. Moreover, 21 exhibited a high selectivity index (SI) (SI represented by the ratio of the cytotoxicities against normal cells and cancer cell lines). Table 2 shows the selectivity index of several compounds. For instance, PBMC vs. HL-60 is 19.8. By comparison, doxorubicin, a standard clinically used drug against several types of cancers, the selectivity index value is 10.6. In the same way, compound 22 (IC50 in the range of 1.06 to 2.56 μΜ) was more active than ηοΓ-β-lapachone-based 1,2,3-triazole without the chalcogen atom. Herein, two important examples of successful preparation of potent antitumor quinones with two redox centers are reported.
[00160] Compound 26, another nor-lapachone derivative, obtained from C-allyl lawsone (23), also exhibited potent antitumor activity. This drug was considered highly active with IC50 values ranging from 0.07 to 0.29 μΜ, suggesting a highly active structure. Compounds 21 and 26 presented similar antitumor activities, showing the importance of the ort/20-naphthofuranquinone moiety and the triazole selenium-containing, group that potentially works together in the same two redox center structures. 1,4-Naphthoquinones 36- 40 were also evaluated and the compounds were considered moderately active with exception of compound 40, inactive against all cancer cells examined. The selectivity index for the most active compounds is summarized in Table 2.
Table 1. Cytotoxic activity expressed by IC50 μΜ (95% CI) of compounds 4-7, 9-10, 13-16, 19-22, 25, 26, 28, 29, 36-38, 39, and 40 in cancer and normal cell lines after 72 h exposure, obtained by nonlinear regression for all cell lines from three independent experiments.
Figure imgf000074_0001
MDA-
Comp HCT- OVCAR PBM
HL-60 PC3 SF295 MB- V79 L929 d 116 -8 C
435
1.43 2.48 1.68 1.93 1.54 2.24 3.58 2.40 2.04
9
(1.32- (2.32- (1.52- (1.76- (1.46- (1.99- (3.34- (2.29- (1.74- 1.54)a 2.68) 2.02) 2.12)a 1.62)a 2.70) 3.86)a 2.51) 2.43)
1.22 1.11 1.90 1.52 1.31 0.92 1.87 2.17 1.74
10
(1.13- (0.84- (1.60- (1.44- (1.22- (0.88- (1.45- (2.03- (1.67- 1.35) 1.26) 2.01) 1.67) 1.52) 1.22) 2.26) 2.31) 1.94)
2.59 4.60 5.15 3.93 7.13 6.46 6.06 5.33
>14.5
13
(2.30- (3.78- (4.63- 6 (3.44- (6.12- (5.50- (5.36- (5.15- 2.94) 5.59) 5.80) 4.40) 8.30) 7.10) 6.55) 5.53)
2.23 3.30 4.09 3.04 2.52 3.53 3.16 4.20 3.68
14
(1.78- (2.90- (3.49- (2.64- (2.23- (3.34- (2.86- (3.82- (3.45- 2.67) 3.68) 4.57) 3.82) 2.75) 3.75) 3.49) 4.90) 4.34)
1.08 1.28 1.71 1.08 0.88 0.68 3.33 3.78 3.38
15
(0.86- (1.04- (1.44- (0.94- (0.76- (0.43- (2.94- (3.73- (3.06- 1.49) 1.57) 2.03) 1.46) 1.06) 1.06) 3.56) 3.94) 3.65)
1.59 2.54 2.95 2.65 2.15 2.80 3.23 4.03 3.77
16
(1.34- (1.55- (2.22- (2.39- (2.00- (2.45- (2.43- (3.29- (3.06- 1.87) 2.88) 3.47) 3.34) 2.32) 3.34) 3.96) 4.63) 4.48)
0.26 1.48 1.86 2.59 2.04 2.53 2.80 2.16 2.50
19
(0.20- (1.34- (1.69- (2.36- (1.78- (2.33- (2.30- (1.83- (2.33- 0.35) 1.69) 2.39) 2.85) 2.18) 2.74) 3.35) 2.68) 2.77)
3.08 0.89 1.74 3.23 1.19 1.34 3.27 2.67
20 nd
1.48- (0.82- (1.22- 2.38- 1.08- (1.15- (2.97- (2.26- 6.46b 1.00) 2.15) 4.38b 1.34b 1.45) 3.53) 3.49)
0.07 0.14 0.38 0.34 0.23 0.20 1.39 1.13 0.94
21
(0.02- (0.11- (0.29- (0.20- (0.16- (0.14- (1.12- (1.03- (0.86- 0.16) 0.25) 0.67) 0.45) 0.38) 0.43) 1.53) 1.26) 1.15) MDA-
Comp HCT- OVCAR PBM
HL-60 PC3 SF295 MB- V79 L929 d 116 -8
435
1.29 2.52 2.43 1.31 1.06 2.26 1.76 2.65 2.30
22
(1.03- (2.00- (1.87- (1.10- (0.99- (1.66- (1.44- (1.76- (1.89-
1.59) 2.88) 3.21) 1.59) 1.12) 3.06) 2.11) 3.16) 2.71)
0.82 0.63 0.51 0.39 0.39 0.35 1.80 0.82 1.06
25
(0.55- (0.43- (0.24- (0.31- (0.27- (0.12- (1.57- (0.67- (0.94-
1.45) 0.86) 0.59) 0.43) 0.59) 0.47) 2.15) 1.14) 1.18)
0.13 0.24 0.24 0.22 0.29 0.07 1.31 0.44 0.62
26
(0.04- (0.18- (0.20- (0.20- (0.22- (0.02- (1.11- (0.22- (0.49-
0.29) 0.31) 0.27) 0.24) 0.33) 0.13) 1.40) 0.60) 0.78)
0.31 0.24 0.39 0.35 0.20 0.35 3.21 2.47 2.15
28
(0.20- (0.16- (0.16- (0.12- (0.04- (0.27- (2.58- (2.27- (1.88-
0.43) 0.39) 0.63) 0.47) 0.27) 0.43) 4.27) 3.02) 2.70)
0.82 2.42 1.40 0.98 0.62 0.91 2.80 2.02 2.15
29
(0.58- (1.80- (1.13- (0.69- (0.42- (0.75- (2.53- (1.64- (2.04-
1.09) 3.26) 1.71) 1.44) 1.02) 1.11) 3.04) 2.44) 2.53)
2.14 3.25 2.25 2.93 2.01 2.58 6.34 2.51 3.43
36
(1.99- (3.01- (1.92- (2.77- (1.64- (2.21- (5.70- (2.03- (3.01-
2.49) 3.76) 2.40) 3.08) 2.47) 2.97) 6.66) 2.86) 3.78)
1.11 1.67 2.29 1.61 1.43 2.37 3.86 3.03 3.30
37
(0.96- (1.43- (2.07- (1.43- (1.17- (1.87- (3.78- (2.87- (2.97-
1.23) 2.05) 2.53) 1.89) 1.81) 2.49) 4.04) 3.40) 3.62)
1.13 1.72 2.08 0.97 3.09 1.89 4.75 3.99 3.83
38
(0.78- (1.42- (1.84- (0.85- (2.72- (1.46- (4.44- (3.40- (3.76-
1.20) 1.98) 2.41) 1.28) 3.76) 2.67) 5.06) 4.49) 3.97)
2.21 4.27 2.16 3.41 2.36 3.51 8.64 5.77 5.17
39
(1.91- (4.22- (1.76- (3.11- (1.91- (3.16- (7.53- (4.62- (4.27-
2.56) 4.52) 2.71) 3.62) 2.91) 4.27) 9.69) 6.73) 5.97)
>12.6 >12.6 >12.6 >12.6 >12.6 >12.6 >12.6
40 >12.68 >12.68
8 8 8 8 8 8 8 MDA-
Comp HCT- OVCAR PBM
HL-60 PC3 SF295 MB- V79 L929 d 116 -8 C
435
1.57 0.87 1.65 0.95 0.25
1.16
β-lap.
1.11- 0.74- 1.40- 0.70- >20.63
0.16-
0.97-1.25
1.69 0.95 1.94 1.03 0.33
0.06 0.15 0.02 0.51 0.96 0.55 0.28 0.23
0.41
DOXO 0.02- 0.09- 0.02- 0.43- 0.87- 0.41- 0.21- 0.15-
0.36-0.49
0.09 0.21 0.04 0.58 1.10 0.58 0.36 0.30
(a) da Silva Junior, et al, 2011
(b) da Silva Junior, et al, 2007 and da Silva Junior, et al, 2009
nd, not determined.
Table 2: Selectivity index for several active compounds
Figure imgf000077_0001
B. NQOl Activity
1. Methods
[00161] A549 cells were plated into a 48-well plate with 10,000 cells/well in 500 of DMEM containing 10% FBS. The cells were allowed to attach and grow overnight. A stock of 5 mM of compounds or 10 mM β-lapachone, and 5 mM dicoumarol were made for the experiment. The 8 drug concentrations (0-3.2 μΜ) were prepared separately in 15 mL conical tubes with 7 mL of media each. The untreated control is DMSO. The media was removed from each well and 500 of each drug concentration was added to 6 wells (to produce sextuplet replicates for each concentration). After aliquoting the drug-containing media, 40 of dicoumarol was added to each remaining 15 mL conical tube (there remains 4 mL left of each drug concentration) to give a final concentration of 50 μΜ dicoumarol. The media was removed from a 2nd 48-well plate and 500 μΐ. of the remaining drug + DIC media was aliquoted/well. The plates were gently shaken to mix and placed in the incubator for 2 h. After 2 h, all media was aspirated from the wells and 1 mL of fresh media was aliquoted into each well. The plates were then left in the incubator for 7 days, or until there was 100% confluency for the untreated control. Once the control was confluent, the media was discarded and 500 μίΛ εΙΙ of l x PBS was added to wash the wells. The PBS was discarded and 250 μΐ. of dLhO/well was added. The plates were then put in the -80 °C freezer overnight. The next day, the plates were thawed completely and 500 μΐ. of Hoechst staining buffer (50 μΐ, of Hoechst 33258 in 50 mL of l TNE buffer) was added to each well. The plates were incubated in the dark at RT for 2 h. After two hours, the plates were read on a PerkinElmer Victor X3 plate reader and the readings were plotted as the treated/control (T/C) ± SEM.
2. Results
[00162] Previous studies have revealed that compounds 50-53 can be considered as prototypes possessing potent antitumor activities against diverse cancer cells, (da Silva Junior et al, 2007; da Silva Junior et al, 2009 and da Silva Junior et al, 2010.) To better understand the mechanism of action of 50-53 and compare the previously reported structures with selenium-containing lapachone-based 1,2,3-triazoles, their potential NQOl -dependent cytotoxicity was examined. The cytotoxicity was measured using a set two-hour exposure, with or without the NQOl inhibitor, dicoumarol. Such exposures take advantage of elevated NQOl levels specifically in most solid cancers compared to associated human tissue (Bey, et al , 2007). Within the class of selenium-containing quinones, compounds 21 and 22 were selected to evaluate their characteristics by an NQOl -dependent mechanism.
Figure imgf000079_0001
Selected compounds for NQOl-speciflc studies.
[00163] Within the drug concentration range tested, the compounds showed activity against human A549 non-small cell lung adenocarcinoma, an alveolar basal epithelial cell line. These cells express high levels of NQOl (3000 + 300 enzymatic units). Cell death for many of the compounds tested were NQOl -specific, since addition of dicoumarol (DIC, an NQOl inhibitor) spared their lethality. Based on the survival curves (FIG. 1A-1G), previously reported arylamine substituted ηοΓ-β-lapachones have predicted IC50 as follows: Compound 50 = 2.6 μΜ, Compound 51 = 1.8 μΜ, Compound 52 = 2.4 μΜ and Compound 53 = 1.3 μΜ. Compound 53 showed the most dramatic lethality within a narrow therapeutic window, going from 93% viability at 0.8 μΜ to 11% viability at 1.6 μΜ. Overall, compounds 50-53 were NQOl -specific drugs exhibiting similar or lower IC50 values than β-lapachone. Compounds 21 and 22, selenium-containing quinones, with IC50 values = 0.64 and 1.2 μΜ, respectively, were the most active of this series and were NQOl -dependent (Table 3). The compounds showed tremendous therapeutic windows using DIC treatment as a surrogate for responses to NQOl- cells, such as that found for nearly all human normal tissue (Bey, et al , 2006). These responses are indicative of NQOl -dependent futile redox cycling of these drugs that create massive ROS, specifically H2O2, that ultimately cause PARPl hyperactivation and programmed necrosis (Huang, et al , 2012; Moore, et al, 2015; Bey, et al, 2013).
Table 3. Quinones NQOl specific and IC50 values in the absence and presence of DIC
Compound Activity NQOl Specific IC50 (μΜ) ICso (μΜ) + DIC
DIC Protection
21 Yes Yes 0.64 >3.2 Yes
22 Yes Yes 1.2 >3.2 Yes
50 Yes Yes 2.6 >3.2 Yes Compound Activity NQOl Specific ICso (μΜ) ICso (μΜ) + DIC
DIC Protection
51 Yes Yes 1.8 >3.2 Yes
52 Yes Yes 2.4 >3.2 Yes
53 Yes Yes 1.3 >3.2 Yes β-lapachone Yes Yes 3.4 >10 Yes
C. Cytotoxicity
1. Methods
i. Cellular Viability
[00164] The Annexin V cytometry assay was used to detect cell population in viable, early and late apoptosis stage as described by Cavalcanti and coworkers. After short exposure time (6 h) with compounds 21 and 22 at 5 μΜ, PC3 cells were stained with fluorescein isothiocyanate (FITC) conjugated Annexin V (Guava Nexin kit, Guava Technologies, Inc., Hay ward, CA, USA) and PI (necrotic-cell indicator), and then they were subjected to flow cytometry (Guava EasyCyte Mini). Cells undergoing early and late apoptosis were detected by the emission of the fluorescence from only FITC and, both FITC and PI, respectively. To determine whether ROS are involved with tested compounds- induced cytotoxicity, cultures were pre-exposed (24 h) to 5 mM N-acetylcysteine (NAC), a widely used thiol-containing antioxidant that is a precursor of GSH which protects against oxidative stress-induced cell death. Also, cultures were co-treated with dicoumarol (50 μΜ) in order to evaluate the role of NQOl on compounds bioactivation. A total of 10,000 events was evaluated per experiment (n = 3) and cellular debris was omitted from analysis. ii. ROS
[00165] Levels of intracellular ROS were estimated following treatment with compound 21 using 2',7'-dichlorofluorescein diacetate (H2DCFDA) as a fluorescent probe. H2DCFDA readily diffuses through the cell membrane and is hydrolyzed by intracellular esterases to non-fluorescent dichlorofluorescein (DCFH), which is then rapidly oxidized to highly fluorescent 2',7'-dichlorofluorescein (DCF) by a broad range of intracellular oxidative stresses in addition to H2O2 (Hempel, et al, 1999; LeBel, et al , 1992). Therefore, the increased mean fluorescence intensity of DCF can be used as a probe for a broad range of oxidative events not limited to H2O2. Briefly, PC3 human prostate cancer cell line was exposed to the test compound (5 μΜ) for 1 h. In parallel, to emphasize the pro-oxidative effect of the test compound, cells were pre-treated with NAC (5 mM) and then exposed to the test compound (5 μΜ) for 1 h. After treatments, the culture medium was then replaced by fresh serum-free medium containing 20 μΜ FhDCFDA. In DCF fluorescence intensity was detected by flow cytometry using a Guava EasyCyte Mini (Guava Technologies, Inc., Hayward, CA, USA) and Guava Express Plus software. In general, the DCF fluorescence intensity is proportional to the amount of intracellular ROS (LeBel, et al, 1992). A total of 10,000 events was evaluated per experiment (n = 3) and cellular debris was omitted from analysis.
2. Results
[00166] The externalization of phosphatidylserine is considered an important marker in the apoptotic process. After treatments, selected compounds 21 and 22 induced a significant increase on populations of PC3 cells with phosphatidylserine expressed on the cell surface (FIG. 2). On the other hand, phosphatidylserine externalization was not observed in cultures pre-treated with NAC before 21 and 22 exposure or co-treated with dicoumarol (FIG. 2). The data show that cytotoxic mechanisms of tested compounds may involve drug bioreduction by quinone reductase NQOl as well emphasizing the ROS contribution on the cytotoxicity suggesting that tested compounds-induced apoptosis is associated with ROS production. Finally, corroborating these studies, short exposure (1 h) to compound 21 was observed to led to the generation of intracellular ROS. In other hand, in cultures pre-exposed with NAC, compound 21 was not able to generate ROS, which may be explained due the antioxidant protection exercised by NAC (FIG. 3).
* * *
[00167] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this disclosure have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the disclosure. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the disclosure as defined by the appended claims. REFERENCES
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U.S. Patent 5,739,169
U.S. Patent 5,801,005
U.S. Patent 5,824,311
U.S. Patent 5,830,880
U.S. Patent 5,846,945

Claims

WHAT IS CLAIMED IS:
1. A compound of the formula:
Figure imgf000085_0001
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6); X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or X2 is taken together with R2 as described below;
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below;
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group, or R2 is taken together with X2 as described below; or R2 is taken together with Ri as described below;
Yi is amino, cyano, halo, hydroxy, or nitro; or alkyl(c<6), cycloalkyl(C<6), acyl(c<6), alkoxy(C<6), acyloxycc<6), alkylamino(c<6), dialkylamino(c<6), amido(c<6), or a substituted version of any of these groups; m is 0, 1, 2, 3, or 4; provided that when X2 and R2 are taken together, the compound is further defined by the formula:
Figure imgf000086_0001
wherein:
X3 is CRJ'R or O, wherein:
R3' and R4' are each independently hydrogen, halo, alkyl(c≤i2), substituted alkyl(c<i2), aryl(c≤i2). substituted aryl(c a selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and are each independently 0, 1, 2, or 3; provided that when Ri and R2 are taken together, the compound is further defined by the formula:
Figure imgf000086_0002
wherein: X4 is CRe'Rj' or O, wherein:
R.6' and R7' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R.6, R7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000087_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000087_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
2. The compound of claim 1 further defined as:
Figure imgf000088_0001
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000088_0002
wherein: X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000089_0001
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. The compound of either claim 1 or claim 2 further defined as:
Figure imgf000089_0002
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6); X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000090_0001
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
4. The compound according to any one of claims 1-3 further defined as:
Figure imgf000091_0001
wherein:
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
R2 is hydrogen, halo, hydroxy, a selenium containing triazole group; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000091_0002
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000091_0003
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. The compound of claim 1 further defined as:
Figure imgf000092_0001
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
Ri is hydrogen, halo, a selenium containing triazole group, or ORc, wherein:
Rc is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rc is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rc is absent; or Ri is taken together with R2 as described below; or
X3 is CR3'R4' or O, wherein:
R3' and R4' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; n is 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000093_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000093_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
6. The compound of either claim 1 or claim 5 further defined as:
Figure imgf000093_0003
wherein:
CR3'R4' or O, wherein: R3' and R4' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R3, R4, and R5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and n2 are each independently 0, 1 , 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000094_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000094_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof. The compound according to any one of claims 1, 5, and 6 further defined
Figure imgf000095_0001
wherein:
R.3, R.4, and R.5 are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; ni and n2 are each independently 0, 1, 2, or 3; wherein the selenium containing triazole group is further defined by the formula:
-X5-Y2-A-Y3-Se-R9 (IV) wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000096_0001
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
The compound of claim 1 further defined as:
Figure imgf000096_0002
wherein:
Xi is O or NRa, wherein:
Ra is hydrogen, alkyl(c<6), or substituted alkyl(c<6);
X2 is ORb, wherein:
Rb is absent, hydrogen, alkyl(c<6), or substituted alkyl(c<6); provided that when Rb is absent, the atom to which it is bound is part of a double bond and when the atom to which it is bound is part of a double bond, then Rb is absent; or
X4 is CR6'R7' or O, wherein:
R6' and R7' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c selenium containing triazole group;
R6, R7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1, 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000097_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1, 2, 3, or 4; r and s are each independently 0, 1, 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000097_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
9. The compound of either claim 1 or claim 8 further defined:
Figure imgf000097_0003
wherein:
X4 is CR6'R7' or O, wherein: R.6' and Rj' are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group;
R6, R7, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000098_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000098_0002
R9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
10. The compound according to any one of claims 1 , 8, and 9 further defined:
Figure imgf000099_0001
wherein:
R.6, Rj, and Rs are each independently hydrogen, halo, alkyl(c<i2), substituted alkyl(c<i2), aryl(c<i2), substituted aryl(c<i2), or a selenium containing triazole group; p is 0, 1 , 2, or 3; and wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000099_0002
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups;
A is a group of the formula:
Figure imgf000099_0003
R.9 is aryl(c<i2) or substituted aryl(c<i2); or a pharmaceutically acceptable salt thereof.
11. The compound according to any one of claims 1-10, wherein Xi is O.
12. The compound according to any one of claims 1-11, wherein X2 is ORb wherein Rb is absent.
13. The compound according to any one of claims 1-12, wherein Ri is hydrogen.
14. The compound according to any one of claims 1-12, wherein Ri is halo.
15. The compound of claim 14, wherein Ri is bromo or chloro.
16. The compound according to any one of claims 1-12, wherein Ri is a selenium containing triazole group.
17. The compound according to any one of claims 1-12, wherein R2 is hydrogen.
18. The compound according to any one of claims 1-12, wherein R2 is halo.
19. The compound of claim 14, wherein R2 is bromo or chloro.
20. The compound according to any one of claims 1-16, wherein R2 is a selenium containing triazole group.
21. The compound according to any one of claims 1-20, wherein m is 0.
22. The compound according to any one of claims 1-21, wherein X3 is O.
23. The compound according to any one of claims 1-21, wherein R3 is hydrogen.
24. The compound according to any one of claims 1-21, wherein R3 is halo.
25. The compound of claim 24, wherein R3 is chloro or bromo.
26. The compound according to any one of claims 1-21, wherein R3 is alkyl(c<6).
27. The compound of claim 26, wherein R3 is methyl.
28. The compound according to any one of claims 1-21, wherein R3 is a selenium containing triazole group.
29. The compound according to any one of claims 1-28, wherein R is hydrogen.
30. The compound according to any one of claims 1-28, wherein R is halo.
31. The compound of claim 30, wherein R is chloro or bromo.
32. The compound according to any one of claims 1-28, wherein R is alkyl(c<6).
33. The compound of claim 32, wherein R is methyl.
34. The compound according to any one of claims 1-28, wherein R is a selenium containing triazole group.
35. The compound according to any one of claims 1-34, wherein R5 is hydrogen.
36. The compound according to any one of claims 1-34, wherein R5 is a selenium containing triazole group.
37. The compound according to any one of claims 1-36, wherein n is 1 or 2.
38. The compound of claim 37, wherein n is 1.
39. The compound of claim 38, wherein n is 2.
40. The compound according to any one of claims 1-39, wherein X4 is O.
41. The compound according to any one of claims 1-40, wherein R6 is hydrogen.
42. The compound according to any one of claims 1-40, wherein R6 is halo.
43. The compound of claim 42, wherein R6 is chloro or bromo.
44. The compound according to any one of claims 1-40, wherein R6 is alkyl(c<6).
45. The compound of claim 26, wherein R6 is methyl.
46. The compound according to any one of claims 1-40, wherein R6 is a selenium containing triazole group.
47. The compound according to any one of claims 1-46, wherein Rj is hydrogen. 48. The compound according to any one of claims 1-46, wherein R7 is halo. 49. The compound of claim 48, wherein R7 is chloro or bromo. 50. The compound according to any one of claims 1-46, wherein R7 is alkyl(c<6). 51. The compound of claim 50, wherein R7 is methyl. 52. The compound according to any one of claims 1-46, wherein R7 is a selenium containing triazole group.
53. The compound according to any one of claims 1-52, wherein Rs is hydrogen. 54. The compound according to any one of claims 1-52, wherein Rs is aryl(c<i2). 55. The compound of claim 54, wherein Rs is phenyl or napthyl. 56. The compound according to any one of claims 1-52, wherein Rs is a selenium containing triazole group.
57. The compound according to any one of claims 1-56, wherein p is 1 or 2. 58. The compound of claim 57, wherein p is 1. 59. The compound of claim 57, wherein p is 2. 60. The compound according to any one of claims 1-59, wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000102_0001
wherein: is a covalent bond, (CH2)q- (CH2)rO(CH2)s- -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups; and
R.9 is aryl(c<i2) or substituted aryl(c<i2).
The compound according to any one of claims 1 -59, wherein the selenium containing triazole group is further defined by the formula:
Figure imgf000103_0001
wherein:
X5 is a covalent bond, -(CH2)q- -(CH2)rO(CH2)s-, -(CH2)rNRd(CH2)s-; wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); q is 1 , 2, 3, or 4; r and s are each independently 0, 1 , 2, 3, or 4;
Y2 and Y3 are each independently a covalent bond, alkanediyl(c<i2), alkenediyl(c<i2), arenediyl(c<i2), heteroarenediyl(c<i2), or a substituted version of any of these groups; and
R.9 is aryl(c<i2) or substituted aryl(c<i2).
62. The compound of either claim 60 or claim 61 , wherein X5 is a covalent bond.
63. The compound of either claim 60 or claim 61, wherein X5 is -(CH2)rNRd(CH2)s-;
wherein:
Rd is hydrogen, alkyl(c<6), or substituted alkyl(c<6); r and s are each independently 0, 1, 2, 3, or 4.
64. The compound of claim 64, wherein X5 is -NH-. 65. The compound of claim 64, wherein X5 is -NHCH2-. 66. The compound according to any one of claims 60-65, wherein Y2 is alkanediyl(c<6). 67. The compound of claim 66, wherein Y2 is -CH2- 68. The compound according to any one of claims 60-65. wherein Y2 is arenediyl(c<6). 69. The compound of claim 68, wherein Y2 is benzenediyl. 70. The compound according to any one of claims 60-69, wherein Y3 is alkanediyl(c<6). 71. The compound of claim 70, wherein Y3 is -CH2-. 72. The compound according to any one of claims 60-71, wherein R9 is aryl(c<8). 73. The compound of claim 72, wherein R9 is phenyl. 74. The compound according to any one of claims 1 -73. wherein the compound is further defined as:
Figure imgf000104_0001
Figure imgf000105_0001
or a pharmaceutically acceptable salt thereof.
A compound of the formula:
Figure imgf000106_0001
or a salt thereof.
A pharmaceutical composition comprising:
(a) a compound according to any one of claims 1 -75; and
(b) a pharmaceutically acceptable carrier.
The pharmaceutical composition of claim 76, wherein the pharmaceutical composition is formulated for administration: orally, intraadiposally, intraarterially, intraarticularly, intracranially, intradermally, intralesionally, intramuscularly, intranasally, intraocularly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrarectally, intrathecally, intratracheally, intratumorally, intraumbilically, intravaginally, intravenously, intravesicularly, intravitreally, liposomally, locally, mucosally, parenterally, rectally, subconjunctival, subcutaneously, sublingually, topically, transbuccally, transdermally, vaginally, in cremes, in lipid compositions, via a catheter, via a lavage, via continuous infusion, via infusion, via inhalation, via injection, via local delivery, or via localized perfusion.
The pharmaceutical composition of claim 77, wherein the pharmaceutical composition further comprises a cyclodextran.
The pharmaceutical composition of claim 78, wherein the cyclodextran is a β- cyclodextran.
80. The pharmaceutical composition of claim 78, wherein the cyclodextran is hydropropyl- -cyclodextran.
81. The pharmaceutical composition of claim 76, wherein the pharmaceutical composition is formulated as a micelle or a liposome.
82. The pharmaceutical composition of claim 81 , wherein the micelle or liposome is formed by poly(lactic acid) poly(ethylene glycol) (PLA PEG).
83. A method of treating a patient with a disease or disorder comprising administering a therapeutically effective amount of a compound or composition according to any one of claims 1-82 to the patient in need thereof.
84. The method of claim 83, wherein the disease or disorder is cancer.
85. The method of claim 84, wherein the cancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma.
86. The method of claim 84, wherein the cancer is of the bladder, blood, bone, brain, breast, central nervous system, cervix, colon, endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia, genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen, small intestine, large intestine, stomach, testicle, or thyroid.
87. The method of claim 84, wherein the cancer overexpresses NAD(P)H: quinone oxidoreductase 1 (NQOl).
88. The method according to any one of claims 84-87, wherein the cancer is leukemia, colon cancer, prostate cancer, brain cancer, ovarian cancer, or skin cancer.
89. The method of claim 88, wherein the colon cancer is colon carcinoma.
90. The method of claim 88, wherein the skin cancer is melanoma.
91. The method of claim 88, wherein the brain cancer is a glioma.
92. The method of claim 88, wherein the leukemia is acute myeloid leukemia.
93. The method of claim 88, wherein the prostate cancer is an androgen independent prostate cancer.
94. The method of claim 88, wherein the ovarian cancer is an ovarian adenocarcinoma.
95. The method according to any one of claims 83-94, wherein the method comprises administering a second anti-cancer therapy.
96. The method of claim 95, wherein the second anti-cancer therapy is a second chemotherapeutic compound, radiation therapy, surgery, or immunotherapy.
97. The method of claim 96, wherein the second chemotherapeutic compound is a PARP1 inhibitor, a glutamine/glutamate pathway inhibitor, or a DNA base excision repair (BER) inhibitor.
98. The method according to any one of claims 83-96, wherein the patient is a mammal.
99. The method of claim 98, wherein the patient is a human.
100. The method according to any one of claims 83-99, wherein the method comprises administering the compound once.
101. The method according to any one of claims 83-99, wherein the method comprises administering the compound two or more times.
102. The method according to any one of claims 83-99, wherein the patient has been determined to have defective DNA repair, defective ability to maintain NAD(P)H levels by the glucose, glutamate/glutamine, or pyruvate pathways, or genetic defects causing synthetic lethality for precision therapy against specific NQ01+ human cancers.
103. The method of claim 83, wherein the disease or disorder is a metabolic syndrome.
104. The method of claim 103, wherein the metabolic syndrome exhibits elevated NAD(P)H/NAD(P)+ levels.
105. The method of claim 83, wherein the disease or disorder is trypanosomal disease.
PCT/US2016/056986 2015-10-20 2016-10-14 Lapachone derivatives containing two redox centers and methods of use thereof WO2017070012A1 (en)

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CN109705095A (en) * 2019-02-25 2019-05-03 山东大学 A kind of fused ring p-benzoquinone CDC25 protein phosphatase inhibitor containing triazole ring and preparation method and application thereof
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Publication number Priority date Publication date Assignee Title
CN109020986A (en) * 2018-08-28 2018-12-18 中南大学 Quinoline quinone and oxa- ring derivatives and preparation method thereof and the application on anti-tumor drug
CN109705095A (en) * 2019-02-25 2019-05-03 山东大学 A kind of fused ring p-benzoquinone CDC25 protein phosphatase inhibitor containing triazole ring and preparation method and application thereof
CN109705095B (en) * 2019-02-25 2021-06-08 山东大学 Fused ring p-benzoquinone CDC25 protein phosphatase inhibitor containing triazole ring, and preparation method and application thereof
CN113368087A (en) * 2020-03-10 2021-09-10 复旦大学附属华山医院 Juglone derivatives and application thereof in preparation of drugs for treating brain tumors through chemotherapy
CN115215837A (en) * 2022-06-29 2022-10-21 山东大学 Fused ring p-benzoquinone dimer derivative containing triazole ring and preparation method and application thereof

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