WO2021211436A1 - Idebenone compounds - Google Patents

Idebenone compounds Download PDF

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WO2021211436A1
WO2021211436A1 PCT/US2021/026852 US2021026852W WO2021211436A1 WO 2021211436 A1 WO2021211436 A1 WO 2021211436A1 US 2021026852 W US2021026852 W US 2021026852W WO 2021211436 A1 WO2021211436 A1 WO 2021211436A1
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alkylene
compound
disorder
disease
pharmaceutically acceptable
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PCT/US2021/026852
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French (fr)
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Mahmood Ahmed
Sing Yeung Frankie MAK
David Lawrence Silver
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Travecta Therapeutics Pte. Ltd.
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C271/18—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by doubly-bound oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/22—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated the carbon skeleton being further substituted by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/30—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by doubly-bound oxygen atoms
    • C07C233/31—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by doubly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/47—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C235/12—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/70—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/72—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms
    • C07C235/76—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton
    • C07C235/78—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of an unsaturated carbon skeleton the carbon skeleton containing rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/04—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms
    • C07C275/20—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an unsaturated carbon skeleton
    • C07C275/22—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an unsaturated carbon skeleton containing rings other than six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
    • C07F9/02—Phosphorus compounds
    • C07F9/06—Phosphorus compounds without P—C bonds
    • C07F9/08—Esters of oxyacids of phosphorus
    • C07F9/09—Esters of phosphoric acids
    • C07F9/10—Phosphatides, e.g. lecithin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/14—Quaternary ammonium compounds, e.g. edrophonium, choline
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated

Definitions

  • the present application generally relates to idebenone compounds, in particular idebenone analogues that are useful for treating a disease or disorder in a subject in need thereof.
  • Idebenone is an antioxidant approved by European Medicines Agency (EMA) for adults and adolescents aged 12 years and over with Leber’s Hereditary Optic Neuropathy (LHON), which is a primary mitochondrial DNA (mtDNA) disorder characterised by bilateral sequential or simultaneous visual loss. Affected patients develop a dense central scotoma accompanied with rapid visual acuity deterioration. It has been postulated that idebenone prevents and/or reverses loss of vision in LHON patients by restoring mitochondrial function and preventing oxidative damage in retinal ganglion cells (e.g. see PLOS. 2012; 7(9): e45182). Idebenone has also been extensively studied in both pre-clinical and clinical setting as potential treatment for a range of neurological disorders (e.g. see J. Bioenerg Biomembr. 2015; 47(0): 111- 118).
  • L 1 is selected from the group consisting of C 8-20 alkylene and C 8-20 alkenylene, wherein the Cs-20 alkylene or Cs-20 alkenylene is optionally substituted by O ' , OH, Ci- 6 alkyl or halogen;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(Ci-e alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), O(Ci- 6 alkylene)0, OC(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(Ci- 6 alkylene), 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(Ci-e alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)
  • R 2 is a bond or a phosphate group
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1 -4 alkylene-CO 2 " , Ci- 4 alkylene-C0 2 H and Ci-6 alkyl, wherein the Ci- 6 alkyl is optionally substituted by O " , OH or halogen;
  • R 4 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C02 " , Ci-4 alkylene-C0 2 H and Ci-6 alkyl, wherein the Ci- 6 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5b is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C1-4 alkyl is optionally substituted by O " , OH or halogen;
  • R 5C is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O " , OH or halogen.
  • compositions comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carriers or excipients.
  • the present application also provides methods of treating a disease or disorder in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof or pharmaceutically compositions as defined herein.
  • the present application also discloses one or more dosage forms, comprising a compound of Formula I as defined herein, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent.
  • the present application provides a method of treating a disease or disorder in a subject comprising administering to the subject in need thereof one or more dosages forms as defined herein.
  • FIGs. 1A-6B show representative thin layer chromatography (TLC) images from the iodine (FIGs. 1 A, 2A, 3 A, 4A, 5 A and 6 A) and cupric acetate (FIGs. IB, 2B, 3B, 4B, 5B and 6B) stain analysis described in Examples 7 and 8.
  • TLC thin layer chromatography
  • FIGs. 7-12 show the concentration or peak area ratio of the compounds described in Examples 1-6 measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a, and empty vector (EV).
  • FIGs. 13A-13B show that Compound 1 (Example 1) exhibits cytoprotective properties in hiPSC-RPE cells.
  • the present application relates to idebenone derivatives that improve their Blood Brain Barrier (BBB) and/or Blood Retinal Barrier (BRB) penetrating profile.
  • BBB Blood Brain Barrier
  • BBB Blood Retinal Barrier
  • BBB Blood Brain Barrier
  • BBR Blood Retinal Barrier
  • the resulting compounds could advantageously present the key pharmacophore of idebenone with higher Central Nervous System (CNS) and/or ocular exposure, which may result in enhanced pharmacological activity and clinical benefit.
  • CNS Central Nervous System
  • L 1 is selected from the group consisting of Cs-2o alkylene and Cx-20 alkenylene, wherein the Cs-20 alkylene or Cs-20 alkenylene is optionally substituted by O " , OH, Ci- 6 alkyl, or halogen;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C I - 6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci.
  • Ci- 6 alkylene)C(0)NH 0(0)0, C(0)NH, C(0)NH(Ci_ 6 alkylene), 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(CI- 6 alkylene)C(0)0, NH(Ci-e alkylene)C(0)NH, NHC(0)(Ci_ 6 alkylene)C(0)0, and NHC(0)(Ci. 6 alkyl ene)C(0)NH, wherein each Ci- 6 alkylene is optionally substituted by O ' , OH, or halogen;
  • R 2 is a bond or a phosphate group
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C0 2 " , Ci-4 alkylene-C0 2 H and Ci-e alkyl, wherein the Ci- 6 alkyl is optionally substituted by O " , OH, or halogen;
  • R 4 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C0 2 " , Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci- 6 alkyl is optionally substituted by O " , OH, or halogen;
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the C M alkyl is optionally substituted by O ' , OH, or halogen;
  • R 5b is selected from the group consisting of H and C M alkyl, wherein the C M alkyl is optionally substituted by O ' , OH, or halogen; and R 5C is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C 1 - 4 alkyl is optionally substituted by O ' , OH, or halogen.
  • the present application provides a compound of Formula I, wherein:
  • L 1 is selected from the group consisting of C 8-20 alkylene and C 8-20 alkenylene, wherein the C 8-20 alkylene or C 8-2o alkenylene is optionally substituted by O " , OH, Ci- 6 alkyl, or halogen;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C I -6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, 0(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI- 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)0, and NHC(0)(CI-6
  • R 2 is a bond or a phosphate group
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C02 " , Ci- 4 alkylene-C0 2 H and Cue alkyl, wherein the Ci- 6 alkyl is optionally substituted by O ' , OH, or halogen;
  • R 4 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-COa " , Ci- 4 alkylene-C0 2 H and Ci- 6 alkyl, wherein the Ci-e alkyl is optionally substituted by O ' , OH, or halogen;
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O ' , OH, or halogen;
  • R 5b is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C 1 - 4 alkyl is optionally substituted by O ' , OH, or halogen;
  • R 5c is selected from the group consisting of H and C 1 - 4 alkyl, wherein the CM alkyl is optionally substituted by O ' , OH, or halogen.
  • L 1 is Cg- 20 alkylene. In some embodiments, L 1 is Cx- 12 alkylene. In some embodiments, L 1 is Cg alkylene, C 9 alkylene, C 10 alkylene, Cn alkylene, C 12 alkylene, C 13 alkylene, C M alkylene, C 15 alkylene, Cie alkylene, C 17 alkyl ene, Ci8 alkylene, C19 alkyl ene, or C20 alkyl ene. Unless specified otherwise, the Cs-zo alkylene or Cs-i 2 alkylene of the L 1 is a straight chain or a branched alkylene.
  • the Cs-zo alkylene or the Cg-iz alkylene of L 1 is a straight chain Cs-zo alkylene, a straight chain Cs-iz alkylene, a branched Cs-zo alkylene, or a branched Cs-i2 alkylene.
  • the straight chain or branched Cs- 20 alkylene of L 1 is optionally substituted by O ' , OH, Ci-e alkyl, halogen, or combinations thereof.
  • the straight chain or branched C 8 - 20 alkylene of L 1 may be substituted by one or more O " , OH, Cim alkyl, halogen, or combinations thereof.
  • the hydrogen atoms of Cs-zo alkylene may be substituted by O " and C1- 6 alkyl, O " , and halogen, OH and Ci-6 alkyl, OH and halogen, or O ' , OH, Ci-e alkyl, and halogen.
  • the straight chain or branched Cs-zo alkylene of L 1 is optionally substituted by 1, 2, 3, 4, 5, 6, 7, or 8 groups independently selected from the group consisting of O " , OH, Ci-e alkyl, and halogen.
  • the straight chain or branched Cs- 20 alkylene of L 1 is optionally substituted by 1, 2, 3, or 4 groups independently selected from the group consisting of O " , OH, Ci-e alkyl, and halogen.
  • L 1 is decanediyl.
  • L 1 is Cs-zo alkenylene. In some embodiments, L 1 is C 15 - 20 alkenylene. In some embodiments, L 1 is Cs alkenylene, C9 alkenylene, Cio alkenylene, Cn alkenylene, C 12 alkenylene, C 13 alkenylene, C 14 alkenylene, C 15 alkenylene, Cm alkenylene, C 17 alkenylene, Cis alkenylene, C 19 alkenylene, or C 20 alkylene. Unless specified otherwise, the Cs-zo alkenylene or C1 5 -2 0 alkenylene of L 1 may be a straight chain or a branched alkenylene.
  • the Cs-20 alkenylene or the C15-20 alkenylene of L 1 is a straight chain Cs-zo alkenylene, a straight chain C 15-20 alkenylene, a branched Cs-zo alkenylene, or a branched C 15-20 alkenylene.
  • the straight chain or branched Cs-zo alkenylene of L 1 is optionally substituted by O " , OH, Ci-e alkyl, halogen, or combinations thereof.
  • the straight chain or branched Cs- 20 alkenylene is substituted, the straight chain or branched Cs-zo alkenylene may be substituted by one or more O ' , OH, Ci-e alkyl, halogen, or combinations thereof.
  • the hydrogen atoms of C8-20 alkenylene may be substituted by O ' and C1-6 alkyl, O " and halogen, OH and Ci-e alkyl, OH and halogen, or O " , OH, Ci-e alkyl, and halogen.
  • the straight chain or branched C8-20 alkenylene of L 1 is optionally substituted by 1, 2, 3, 4, 5, 6, 7, or 8 groups independently selected from the group consisting of O " , OH, Ci- 6 alkyl, and halogen.
  • the straight chain or branched Cs-20 alkenylene of L 1 is optionally substituted by 1, 2, 3, or 4 groups independently selected from the group consisting of O ' , OH, Ci-e alkyl, and halogen.
  • the Cs-20 alkenylene of L 1 comprises 1, 2, 3 or 4 carbon-carbon double bonds. In some embodiments, the C 8-20 alkenylene of L 1 comprises 1 or 2 carbon-carbon double bonds. In some embodiments, the C 8-20 alkenylene of L 1 comprises 1 carbon-carbon double bond. In some embodiments, when the Cs-20 alkenylene of L 1 comprises 1 carbon-carbon double bond, L 1 is selected from the group consisting of tetradecenediyl, heptadecenediyl, and octadecenediyl. In some embodiments, when the C 8-20 alkenylene of the L 1 comprises 1 carbon-carbon double bond, L 1 is selected from the group consisting of heptadecenediyl and octadecenediyl.
  • L 1 when L 1 is tetradecenediyl, L 1 is selected from the group consisting of: o
  • the L 1 when L 1 is heptadecenediyl, the L 1 is selected from the group consisting of: w herein - indicates the bonds connecting L 1 to the moiety and R 1 .
  • the L 1 when the L 1 is octadecenediyl, the L 1 may be selected from the group consisting of the following structures: 9
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(CI-6 alkylene), 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(Ci- 6 alkylene)C(0)NH, 0(Ci-e alkylene)C(0)0, 0(Ci-e alkylene)C(0)NH, NH(CI- 6 alkylene)C(0)0, NH(CI- 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci_ 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(Ci- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(C I - 6 alkylene)C(0)0, NH(C I - 6 alkylene)C(0)NH, NHC(0)(C I - 6 alkylene)C(0)C(0), N
  • R 1 is selected from the group consisting of 0C(0)NH(CI-6 alkylene) and 0C(0)0(Ci-6 alkylene). In some embodiments, R 1 is selected from the group consisting of 0C(0)NH(C M alkylene) and 0C(0)0(Ci- 4 alkylene). In some embodiments, R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 and 0C(0)0CH 2 CH 2 .
  • R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0,
  • R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0, 0C(0)CH 2 C(0)NH, 0C(0)CH 2 CH 2 C(0)NH, 0C(0)CH 2 CH 2 CH 2 C(0)NH, C(0)0,
  • R 2 is a bond.
  • R 2 is a bond, it is to be understood that the R 1 is directly linked to -CH(R 3 )- moiety via a bond. Hence, the R 2 may be considered as being absent.
  • the R 2 is a phosphate group, wherein the phosphate group has the following structure: where - indicates the bonds connecting R 2 to R 1 and -CH(R 3 )- moiety.
  • R 3 is selected from the group consisting of H, C0 2 " , C0 2 H, CH 2 C0 2 " and CH 2 C0 2 H. In some embodiments, R 3 is H. In some embodiments, R 3 is C0 2 " . In some embodiments, R 3 is CH 2 C0 2 " .
  • R 4 is selected from the group consisting of H, CO2 " , C0 2 H, CH 2 C0 2 " and CH 2 C0 2 H. In some embodiments, R 4 is H. In some embodiments, R 4 is C0 2 ⁇ In some embodiments, R 4 is CH 2 C0 2 " .
  • R 4 when R 4 is H, R 3 is C0 2 " , C0 2 H, CH2CO2 " or CH 2 C0 2 H. In some embodiments, when R 3 is H, R 4 is C0 2 " , C0 2 H, CH 2 C0 2 " or
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O ' , OH or halogen. In some embodiments, R 5a is methyl, ethyl or n-propyl. In some embodiments, R 5a is methyl.
  • R 5b is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O ' , OH or halogen. In some embodiments, R 5b is methyl, ethyl or n-propyl. In some embodiments, R 5b is methyl.
  • R 5c is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O ' , OH or halogen. In some embodiments, R 5c is methyl, ethyl or n-propyl. In some embodiments, R 5c is methyl.
  • R 5a , R 5b , and R 5c are H. In some embodiments, R 5a , R 5b , and R 5c are each H. In some embodiments, two of R 5a , R 5b , and R 5c are methyl. In some embodiments, R 5a , R 5b , and R 5c are each methyl.
  • L 1 is C8-20 alkylene or Cg-2o alkenylene, wherein the Cs-2o alkylene or Cs-2o alkenylene is optionally substituted by O ' , OH, CM alkyl or halogen;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C M alkylene), NHC(0)NH, 0C(0)0(CM alkylene), 0(CM alkylene)0, 0C(0)(CM alkylene)C(0)0, 0C(0)(CM alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(CM alkylene)C(0)0, 0C(0)0(C M alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(CM alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH,
  • R 2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to -CH(R 3 )- moiety;
  • R 3 is H or CH2CO2 " ;
  • R 4 is H
  • R 5a is H or methyl
  • R 5b is H or methyl; and R 5C is H or methyl.
  • L 1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, tetradecenediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(Ci-e alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(Ci- 6 alkylene), 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(Ci-e alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)
  • R 2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to -CH(R 3 )- moiety;
  • R 3 is H or CH2CO2 " ;
  • R 4 is H
  • R 5a is H or methyl
  • R 5b is H or methyl; and R 5C is H or methyl.
  • L 1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C I - 6 alkylene), NHC(0)NH, 0C(0)0(Ci-e alkylene), 0(Ci-e alkylene)0, 0C(0)(Ci-e alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI- 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkyl ene)C(0)0, and NHC(0)
  • R 3 is H or CH2CO2 " ;
  • R 4 is H
  • R 5a is H or methyl
  • R 5b is H or methyl; and R 5C is H or methyl.
  • L 1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, tetradecenediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
  • R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0, 0C(0)CH 2 C(0)NH,
  • R 2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to -CH(R 3 )- moiety;
  • R 3 is H or CH2CO2 " ;
  • R 4 is H
  • R 5a is H or methyl
  • R 5b is H or methyl; and R 5C is H or methyl.
  • L 1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
  • R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0, 0C(0)CH 2 C(0)NH,
  • R 2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to -CH(R 3 )- moiety;
  • R 3 is H or CH2CO2 " ;
  • R 4 is H
  • R 5a is H or methyl
  • R 5b is H or methyl; and R 5C is H or methyl.
  • the compound of Formula I as defined above, or a pharmaceutically acceptable salt thereof is a compound of Formula II:
  • R 1 , R 2 , R 3 , R 5a , R 5b , and R 5c are as defined above.
  • the compound of Formula I as defined above, or a pharmaceutically acceptable salt thereof is a compound of Formula II:
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(C I - 6 alkylene)C(0)0, NH(C I - 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)0, and NHC(0)(
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C0 2 " , Ci- 4 alkylene-C0 2 H and Ci16 alkyl, wherein the C 1-6 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5a is selected from the group consisting of H and C4 alkyl, wherein the C 4 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5b is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5C is selected from the group consisting of H and C 4 alkyl, wherein the C 4 alkyl is optionally substituted by O ' , OH or halogen.
  • the R 1 is selected from the group consisting of
  • the compound of Formula I, or a pharmaceutically acceptable salt thereof is a compound of Formula III:
  • the compound of Formula I, or a pharmaceutically acceptable salt thereof is a compound of Formula III:
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C I - 6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(C I - 6 alkylene)C(0)0, NH(C I - 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)0, and
  • R 2 is a bond or a phosphate group; unless specified otherwise, when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to the -CH(R 3 )- moiety.
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C0 2 ' , Ci- 4 alkylene-C0 2 H and Ci-e alkyl, wherein the Ci- 6 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O ' , OH or halogen;
  • R 5b is selected from the group consisting of H and C 4 alkyl, wherein the C 4 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5C is selected from the group consisting of H and C 4 alkyl, wherein the C 4 alkyl is optionally substituted by O ' , OH or halogen.
  • the R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0, 0C(0)CH 2 C(0)NH, 0C(0)CH 2 CH 2 C(0)NH, 0C(0)CH 2 CH 2 CH 2 C(0)NH, C(0)0, C(0)NH, 0C(0)0CH 2 C(0)0, 0C(0)0CH 2 CH 2 C(0)0, 0C(0)0CH 2 CH 2 CH 2 C(0)0, 0C(0)0CH 2 CH 2 CH 2 C(0)0,
  • the compound of Formula I, or a pharmaceutically acceptable salt thereof is a compound of Formula IV:
  • R 1 , R 2 , R 3 , R 5a , R 5b , and R 5c are as defined above.
  • the compound of Formula I, or a pharmaceutically acceptable salt thereof is a compound of Formula IV:
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(C I - 6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(C I - 6 alkylene)C(0)0, NH(C I - 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)0, and N
  • R 2 is a bond or a phosphate group; unless specified otherwise, when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to the -CH(R 3 )- moiety.
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C1-4 alkylene-C02 " , Ci- 4 alkylene-C0 2 H and Ci-e alkyl, wherein the Ci-e alkyl is optionally substituted by O " , OH or halogen;
  • R 5a is selected from the group consisting of H and C1-4 alkyl, wherein the C M alkyl is optionally substituted by O ' , OH or halogen;
  • R 53 ⁇ 4 is selected from the group consisting of H and C M alkyl, wherein the C M alkyl is optionally substituted by O ' , OH or halogen; and R 5C is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C 1 - 4 alkyl is optionally substituted by O ' , OH or halogen.
  • the R 1 is selected from the group consisting of 0C(0)NHCH 2 CH 2 , 0C(0)0CH 2 CH 2 , NHC(0)0, NHC(0)NH, 0CH 2 CH(0H)CH 2 0, 0C(0)CH 2 C(0)0, 0C(0)CH 2 CH 2 C(0)0, 0C(0)CH 2 CH 2 CH 2 C(0)0, 0C(0)CH 2 C(0)NH, 0C(0)CH 2 CH 2 C(0)NH, 0C(0)CH 2 CH 2 CH 2 C(0)NH, C(0)0, C(0)NH, 0C(0)0CH 2 C(0)0, 0C(0)0CH 2 CH 2 C(0)0, 0C(0)0CH 2 CH 2 CH 2 C(0)0, 0C(0)0CH 2 C(0)NH, 0C(0)0CH 2 CH 2 C(0)NH, 0C(0)0CH 2 CH 2 CH 2 C(0)NH, 0C(0)NHCH 2 C(0)NH, 0C(0)0CH 2 CH 2 C(0)NH, 0C(0)0CH 2 CH 2 CH 2 C(0)NH, 0
  • the compound of Formula I or a pharmaceutically acceptable salt thereof is a compound of Formula V: v or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 5a , R 5b , and R 5c are as defined above.
  • the compound of Formula I, or a pharmaceutically acceptable salt thereof is a compound of Formula V:
  • R 1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI- 6 alkylene), NHC(0)NH, 0C(0)0(Ci- 6 alkylene), 0(Ci- 6 alkylene)0, 0C(0)(Ci- 6 alkylene)C(0)0, 0C(0)(Ci- 6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci- 6 alkylene)C(0)0, 0C(0)0(Ci- 6 alkylene)C(0)NH, 0C(0)NH(Ci- 6 alkylene)C(0)0, 0C(0)NH(CI- 6 alkylene)C(0)NH, 0(Ci- 6 alkylene)C(0)0, 0(Ci- 6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI- 6 alkylene)C(0)NH, NHC(0)(Ci- 6 alkylene)C(0)0, and NHC(0)(CI- 6
  • R 2 is a bond or a phosphate group; unless specified otherwise, when R 2 is a bond it is to be understood that R 2 may be considered as being absent such that R 1 is directly linked to the -CH(R 3 )- moiety.
  • R 3 is selected from the group consisting of H, CO2 " , CO2H, C 1 - 4 alkylene-C0 2 " , Ci -4 alkylene-C02H and Ci-e alkyl, wherein the Ci- 6 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5a is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C 1 - 4 alkyl is optionally substituted by O ' , OH or halogen;
  • R 5b is selected from the group consisting of H and C 1 - 4 alkyl, wherein the C 1 - 4 alkyl is optionally substituted by O ' , OH or halogen; and
  • R 5C is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O ' , OH or halogen.
  • the R 1 is selected from the group consisting of
  • the compound of Formula I as defined herein, or a pharmaceutically acceptable salt thereof is selected from the group consisting of compounds 1 to 43, where its chemical structure thereof is shown in Table A below.
  • the compounds of the present application may be prepared using known organic synthesis techniques and may be synthesized according to any of numerous possible synthetic routes.
  • the compounds provided herein can be prepared according to the general procedures shown in Schemes 1 to 9.
  • Suitable synthetic methods of starting materials, intermediates and products may be identified by reference to the literature, including reference sources such as: Advances in Heterocyclic Chemistry, Vols. 1-107 (Elsevier, 1963-2012); Journal of Heterocyclic Chemistry V o ⁇ s. 1-49 (Journal of Heterocyclic Chemistry, 1964-2012); Carreira, et al. (Ed.) Science of Synthesis, Vols. 1-48 (2001-2010) and Knowledge Updates KU2010/1-4; 2011/1-4; 2012/1-2 (Thieme, 2001-2012); Katritzky, et al.
  • Preparation of compounds described herein can involve the protection and deprotection of various chemical groups.
  • the need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by one skilled in the art.
  • the chemistry of protecting groups can be found, for example, in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., Wiley & Sons, Inc., New York (1999).
  • Reactions can be monitored according to any suitable method known in the art.
  • product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 'H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectroscopy (LCMS), or thin layer chromatography (TEC).
  • HPLC high performance liquid chromatography
  • LCMS liquid chromatography-mass spectroscopy
  • TEC thin layer chromatography
  • Compounds can be purified by those skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) and normal phase silica chromatography.
  • each divalent linking substituent include both the forward and backward forms of the linking substituent.
  • - NR(CR’R”) n - includes both -NR(CR’R”) n - and -(CR’R”) n NR-.
  • the Markush variables listed for that group are understood to be linking groups.
  • the phrase “optionally substituted” means unsubstituted or substituted.
  • substituted as used herein means that one or more hydrogen atoms are removed and replaced by one or more substituents. It is to be understood that substitution at a given atom is limited by valency.
  • C n - m indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include CM, CM, and the like.
  • C n - m alkyl refers to a saturated hydrocarbon group that may be straight-chain or branched, having n to m carbons.
  • alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, «-propyl, isopropyl, «-butyl, tert- butyl, isobutyl, sec-butyl; higher homologs such as 2 -methyl- 1 -butyl, «-pentyl, 3- pentyl, «-hexyl, 1 ,2,2-trimethylpropyl, and the like.
  • the alkyl group contains from 1 to 6 carbon atoms, from 1 to 4 carbon atoms, from 1 to 3 carbon atoms, or 1 to 2 carbon atoms.
  • Cn-m alkylene refers to a divalent alkyl linking group having n to m carbons.
  • alkylene groups include, but are not limited to, methylene, ethan-1,2- diyl, propan- 1, 3 -diyl, propan- 1,2-diyl, and the like.
  • the alkylene moiety contains 8 to 20, 10 to 20, 8 to 15, 1 to 6, 1 to 3, or 1 to 2 carbon atoms.
  • C n -m alkenyl refers to an alkyl group having one or more carbon-carbon double bonds and having n to m carbons.
  • Example alkenyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, heptenyl, octenyl, nonenyl, decenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, and the like.
  • the alkenyl moiety contains 5 to 25, 8 to 20, or 10 to 20 or carbon atoms.
  • C n - m alkenylene refers to a divalent alkenyl linking group having n to m carbons.
  • alkenylene groups include, ethen- 1,2-diyl, propen-1, 3-diyl, propen- 1 ,2- diyl , hexen-l,6-diyl, hepten-l,7-diyl, octen-l,8-diyl, nonen-l,9-diyl, decen-l,10-diyl, pentadecen- 1 , 15-diyl, hexadecene- 1 , 16-diyl, hexadecene-2, 16-diyl, heptadecen- 1,17- diyl, heptadecen-2, 17-diyl, and the like.
  • the alkenylene groups include, ethen- 1,2-diyl, propen-1
  • C n - m alkoxy refers to a group of formula -O- alkyl, wherein the alkyl group has n to m carbons.
  • Example alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), butoxy (e.g., n-butoxy and tert- butoxy), and the like.
  • the alkoxy group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.
  • halogen refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).
  • Tautomeric forms result from the swapping of a single bond with an adjacent double bond together with the concomitant migration of a proton.
  • Tautomeric forms include prototropic tautomers which are isomeric protonation states having the same empirical formula and total charge.
  • Example proto tropic tautomers include ketone — enol pairs, amide - imidic acid pairs, lactam - lactim pairs, enamine - imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, for example, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1 ,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole.
  • Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.
  • All compounds, and pharmaceutically acceptable salts thereof, can be found together with other substances such as water and solvents (e.g. hydrates and solvates) or can be isolated.
  • preparation of compounds can involve the addition of acids or bases to affect, for example, catalysis of a desired reaction or formation of salt forms such as acid addition salts.
  • Example acids can be inorganic or organic acids and include, but are not limited to, strong and weak acids.
  • Some example acids include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, 4- nitrobenzoic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, and nitric acid.
  • Some weak acids include, but are not limited to acetic acid, propionic acid, butanoic acid, benzoic acid, tartaric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, and decanoic acid.
  • Example bases include lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, and sodium bicarbonate.
  • Some example strong bases include, but are not limited to, hydroxide, alkoxides, metal amides, metal hydrides, metal dialkylamides and arylamines, wherein; alkoxides include lithium, sodium and potassium salts of methyl, ethyl and t-butyl oxides; metal amides include sodium amide, potassium amide and lithium amide; metal hydrides include sodium hydride, potassium hydride and lithium hydride; and metal dialkylamides include lithium, sodium, and potassium salts of methyl, ethyl, n-propyl, iso-propyl, n-butyl, tert- butyl, trimethylsilyl and cyclohexyl substituted amides.
  • the compounds and salts provided herein are substantially isolated.
  • substantially isolated is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected.
  • Partial separation can include, for example, a composition enriched in the compounds provided herein.
  • Substantial separation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compounds provided herein, or salt thereof. Methods for isolating compounds and their salts are routine in the art.
  • room temperature or “RT” as used herein, are understood in the art, and refer generally to a temperature (e.g., a reaction temperature) that is about the temperature of the room in which the reaction is carried out, for example, a temperature from about 20°C to about 30°C such as about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C or about 30°C.
  • the compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated.
  • Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms. Accordingly, the compounds of Formula I as defined herein or a pharmaceutically acceptable salt thereof, may be found as both cis and trans isomers. As can be seen from the exemplified compounds, the preferred isomers may be cis isomers.
  • An example method includes fractional recrystallizaion using a chiral resolving acid which is an optically active, salt-forming organic acid.
  • Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids such as ⁇ -camphorsulfonic acid.
  • resolving agents suitable for fractional crystallization methods include stereoisomerically pure forms of a-methylbenzylamine (e.g., S and R forms, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N- methylephedrine, cyclohexyl ethylamine, 1 ,2 -diaminocyclohexane, and the like.
  • Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine).
  • an optically active resolving agent e.g., dinitrobenzoylphenylglycine
  • Suitable elution solvent composition can be determined by one skilled in the art.
  • phrases “pharmaceutically acceptable” is employed herein to refer 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 of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • the present disclosure also provides pharmaceutically acceptable salts of the compounds as described above.
  • pharmaceutically acceptable salts refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form.
  • Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • the pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • the pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, non- aqueous media like ether, ethyl acetate, alcohols (e.g. , methanol, ethanol, iso-propanol, or butanol) or acetonitrile (MeCN) are preferred.
  • non- aqueous media like ether, ethyl acetate, alcohols (e.g. , methanol, ethanol, iso-propanol, or butanol) or acetonitrile (MeCN) are preferred.
  • the present disclosure further provides methods of treating a disease or disorder in a subject.
  • the term “subject,” refers to any animal, including mammals. Exemplary subjects include, but are not limited to, mice, rats, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, and humans.
  • the subject is a human.
  • the method comprises administering to the subject (e.g., a subject in need thereof) a therapeutically effective amount of a compound provided herein (e.g. , a compound of Formula I, II, III, IV or V as defined above), or a pharmaceutically acceptable salt thereof.
  • the disease or disorder is selected from the group consisting of mitochondrial disorder, pain, a pain-related disease or disorder, a mood disease or disorder, a disease or disorder of the central nervous system, an optical disease or disorder, cancer, a gastrointestinal disease or disorder, a renal disease or disorder, a renal-related disease or disorder, a cardiovascular disease or disorder, and a skin disease or disorder.
  • the disease or disorder is a mitochondrial disorder.
  • the mitochondrial disorder is selected from the group consisting of Leber's hereditary optic neuropathy (LHON), Duchenne muscular dystrophy, mitochondrial encephalopathy, lactic acidosis and stroke- like episodes (MELAS), mitochondrial myopathy, diabetes mellitus and deafness (DAD), Leigh syndrome, neuropathy, ataxia, and retinitis pigmentosa (NARP), ptosis, chronic progressive external ophthalmoplegia (CPEO), coenzyme Q10 deficiency, familial bilateral striatal necrosis, Friedreich ataxia, infantile-onset spinocerebellar ataxia (IOSCA), Kearns- Sayre syndrome, mitochondrial DNA depletion syndrome (MDS) and Rett syndrome (RTT).
  • LHON Leber's hereditary optic neuropathy
  • MELAS Duchenne muscular dystrophy
  • mitochondrial encephalopathy lactic acidosis and stroke- like episodes
  • the disease or disorder is pain or a pain-related disease or disorder.
  • the pain or a pain-related disease or disorder is selected from the group consisting of acute pain, chronic pain, neuropathic pain, nociceptive pain, inflammatory pain, cancer pain, fibromyalgia, rheumatoid arthritis, osteoarthritis, surgery-related pain, and osteoporosis.
  • the disease or disorder is selected from the group consisting of rheumatoid arthritis and osteoarthritis.
  • the disease or disorder is pain related to rheumatoid arthritis or pain related to osteoarthritis.
  • the disease or disorder is a mood disease or disorder.
  • the mood disease or disorder is selected from the group consisting of anxiety, depression, a sleeping disorder, an eating disorder, post- traumatic stress disorder, symptoms of drug or alcohol withdrawal or abuse, schizophrenia, obsessive-compulsive disorder, bipolar disorder, sexual dysfunction, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD).
  • the disease or disorder is a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, or an optical disease or disorder. In some embodiments, the disease or disorder is a disease or disorder of the central nervous system. In some embodiments, the disease or disorder is a disease or disorder of the peripheral nervous system. In some embodiments, the disease or disorder is an optical disease or disorder.
  • the disease or disorder of the central nervous system, peripheral nervous system, or optical disease or disorder is selected from the group consisting of a demyelinating disease, glaucoma, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), a cognitive disorder, Alzheimer’s disease, a movement disorder, Huntington’s disease, Huntington’s chorea, Tourette’s syndrome, Niemann-Pick disease, Parkinson's disease, epilepsy, a cerebrovascular disorder, ischemic stroke and brain injury.
  • AMD age-related macular degeneration
  • ALS amyotrophic lateral sclerosis
  • a cognitive disorder Alzheimer’s disease
  • a movement disorder Huntington’s disease
  • Huntington’s chorea Huntington’s chorea
  • Tourette’s syndrome Niemann-Pick disease
  • Parkinson's disease epilepsy
  • cerebrovascular disorder ischemic stroke and brain injury.
  • the demyelinating disease is selected from the group consisting of multiple sclerosis (MS), neuromyelitis optica (NMO), Devic’s disease, central nervous system neuropathy, central pontine myelinolysis, syphilitic myelopathy, leukoencephalopathies, leukodystrophies, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, anti-myelin-associated glycoprotein (MAG) peripheral neuropathy, Charcot-Marie-T ooth disease, peripheral neuropathy, myelopathy, optic neuropathy, progressive inflammatory neuropathy, optic neuritis, and transverse myelitis.
  • MS multiple sclerosis
  • NMO neuromyelitis optica
  • Devic Devic’s disease
  • central nervous system neuropathy central pontine myelinolysis
  • syphilitic myelopathy syphilitic myelopathy
  • leukoencephalopathies leukodystrophies
  • Guillain-Barre syndrome chronic inflammatory demy
  • the disease or disorder is cancer.
  • the cancer is selected from the group consisting of leukemia, mantle cell lymphoma, Hodgkin lymphoma, Non-Hodgkin lymphoma, hepatocellular carcinoma, ovarian cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, glioma, glioblastoma skin cancer, renal carcinoma, and lung cancer.
  • the disease or disorder is a gastrointestinal disease or disorder.
  • the gastrointestinal disease or disorder is selected from the group consisting of inflammatory bowel disease, gastroesophageal reflux disease, paralytic ileus, secretory diarrhoea, gastric ulcer, nausea, emesis, celiac disease, irritable bowel syndrome, and a liver disorder.
  • the liver disease is selected from the group consisting of acute liver failure, Alagille syndrome, hepatitis, enlarged liver, Gilbert’s syndrome, liver cyst, liver haemangioma, fatty liver disease, steatohepatitis, primary sclerosing cholangitis, fascioliasis, primary bilary cirrhosis, Budd-Chiari syndrome, hemochromatosis, Wilson’s disease, and transthyretin-related hereditary amyloidosis.
  • the disease or disorder is a renal disease or disorder or a renal-related disease or disorder.
  • the renal disease or disorder or a renal-related disease or disorder is selected from the group consisting of diabetes, diabetic nephropathy, acute inflammatory kidney injury, renal ischemia urinary incontinence, and overactive bladder.
  • the disease or disorder is a skin disease or disorder.
  • the skin disease or disorder is selected from the group consisting of atopic dermatitis, psoriasis, and lupus.
  • the disease or disorder is a cardiovascular disease or disorder.
  • the cardiovascular disease or disorder is selected from the group consisting of cardiovascular disease, vascular inflammation, idiopathic pulmonary fibrosis, cough, and hypertension.
  • the present disclosure also provides the compound described herein such as the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof, for use in treating the disease or disorder as previously described.
  • the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof for use in treating the mitochondrial disorder.
  • the present disclosure further provides use the compound as described herein (e.g. the compound of Formula I, II, III, IV or V) or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating the disease or disorder as previously described.
  • the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating the mitochondrial disorder.
  • the phrase “therapeutically effective amount” refers to the amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought in a tissue, system, animal, individual or human by a researcher, veterinarian, medical doctor or other clinician.
  • treating refers to one or more of (1) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology); and (2) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology) such as decreasing the severity of disease or reducing or alleviating one or more symptoms of the disease.
  • the present application further provides isotopically-labeled compounds provided herein.
  • An “isotopically” or “radio-labeled” compound is a compound wherein one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring).
  • Suitable radionuclides that may be incorporated in compounds of the present disclosure include but are not limited to 2 H (also written as D for deuterium), 3 H (also written as T for tritium), n C, 13 C, 14 C, 13 N, 15 N, 15 0, 17 0, and 18 0.
  • one or more hydrogen atoms in a compound of the present disclosure can be replaced by deuterium atoms (e.g., one or more hydrogen atoms of a Ci - 6 alkyl group as described herein can be optionally substituted with deuterium atoms, such as -CD3 being substituted for -CFL).
  • alkyl groups of the componuds provided herein can be perdeuterated.
  • the compound provided herein comprises at least one deuterium atom. In some embodiments, the compound provided herein comprises two or more deuterium atoms. In some embodiments, the compound provided herein comprises 1-2, 1-3, 1-4, 1-5, 1-6, 1-8, 1-10, 1-12, 1-14, 1-16, 1-18, or 1-20 deuterium atoms. In some embodiments, all of the hydrogen atoms in a compound provided herein are replaced or substituted by deuterium atoms.
  • the present application further provides synthetic methods for incorporating radio-isotopes into the compounds described herein. Synthetic methods for incorporating radio-isotopes into organic compounds are well known in the art, and an ordinary skill in the art will readily recognize the methods applicable for the compounds described herein.
  • substitution with heavier isotopes may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances, (see e.g., A. Kerekes et. al. J. Med. Chem. 2011, 54, 201-210; R. Xu et. al. J. Label Compd. Radiopharm. 2015, 58, 308-312).
  • substitution at one or more metabolism sites may afford one or more of the therapeutic advantages.
  • One or more additional therapeutic agents or other agents useful for treating the diseases or disorders as mentioned above can be used in combination with the compounds of Formula 1, 11, III, IV or V and salts thereof provided herein.
  • the additional therapeutic agent may be administered simultaneously with the compound or salt provided herein. In some embodiments, the additional therapeutic agent may be administered after administration of the compound or salt provided herein. In some embodiments, the additional therapeutic agent is administered prior to administration of the compound or salt provided herein. In some embodiments, the compound or salt provided herein is administered during a surgical procedure. In some embodiments, the compound or salt provided herein is administered in combination with an additional therapeutic agent during a surgical procedure.
  • compositions When employed as pharmaceuticals, the compounds and salts provided herein can be administered in the form of pharmaceutical compositions. These compositions can be prepared as described herein or elsewhere, and can be administered by a variety of routes, depending upon whether local or systemic treatment is desired and upon the area to be treated.
  • Administration may be topical (e.g. , transdermal, epidermal, ophthalmic and to mucous membranes including intranasal, vaginal and rectal delivery), pulmonary (e.g. , by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral, or parenteral.
  • Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular or injection or infusion; or intracranial (e.g., intrathecal or intraventricular, administration).
  • Parenteral administration can be in the form of a single bolus dose, or may be, for example, by a continuous perfusion pump.
  • the compounds provided herein are suitable for parenteral administration. In some embodiments, the compounds provided herein (e.g., the compounds of Formula I, II, III, IV and V) are suitable for intravenous administration.
  • compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • the pharmaceutical compositions provided herein are suitable for parenteral administration.
  • the compositions provided herein are suitable for intravenous or i.v. administration.
  • compositions comprising, as the active ingredient, a compound provided herein such as a compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers (or excipients).
  • the active ingredient is typically mixed with an excipient, diluted by an excipient or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or other container.
  • the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient.
  • compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.
  • excipients include, without limitation, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose.
  • the formulations can additionally include, without limitation, lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-b enzoates ; sweetening agents; flavoring agents, or combinations thereof.
  • reaction mixture was quenched with water at -78°C and extracted with EtO Ac (2 x 200 mL).
  • the total organic layer was dried over anhydrous NaaS0 4 and concentrated under reduced pressure to obtain the crude product, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 4%
  • Step 6 methyl (Z)-19-hydroxynonadec-l 1-enoate To a stirred solution of methyl (Z)-19-((tetrahydro-2H-pyran-2- yl)oxy)nonadec- 11 -enoate (6 g, 14.6 mmol, 1.0 eq.) in MeOH (150 mL) at 25°C was added /?-TSA (556 mg, 2.92 mmol, 0.2 eq.) The reaction was stirred for 16 hours and monitored by TLC.
  • Step 8 (Z)-2-(l 8-hydroxyoctadec-10-en-l-yl)-5, 6-dimethoxy-3-methylcyclohexa-2,5- diene-1, 4-dione
  • Triphenylphosphine 39 g, 148.28 mmol, 1.5 eq. was added to a stirred solution of methyl 9-bromononanoate (26 g, 98.85 mmol, 1.0 eq.) in acetonitrile (100 mL) at 25°C.
  • the reaction mixture was stirred at 110°C for 48 hours and monitored by TLC.
  • the reaction mixture was concentrated under vacuum to obtain the crude material.
  • the crude product was triturated with hexane (100 mL) to afford (9- methoxy-9-oxononyl)triphenylphosphonium bromide (50 g) as a colorless liquid. Yield: 98%.
  • reaction was stirred at 75°C for 2 hours and monitored by TLC.
  • the reaction mixture was poured into ice water and extracted with DCM (3 x 25 mL).
  • the combined organic layers were dried over anhydrous Na2SCL and concentrated under reduced pressure to obtain crude, which was purified by silica gel column chromatography (60-120 mesh) using gradient elution with 90-95% DCM/hexane to afford title compound (150 mg) as an orange liquid. Yield: 21%.
  • reaction mass was then cooled to 0°C, quenched with water (20 mL), and extracted with DCM (3 X 100 mL).
  • DCM 3 X 100 mL
  • the aqueous layer was purified by flash chromatography [Column: Biotage-C18, 60 g Duo-100 A 30 pm; Mobile phase: [ACN: Water + 0.1% TFA]; B%: B%: 0-8%, 0-20min / 8% 20-45 min.] to afford 2-
  • Step 4 (Z)-2-(( ((18-(4, 5 -dimethoxy-2 -methyl-3, 6-dioxocyclohexa- 1 , 4-dien-l- yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl (2-(trimethylammonio)ethyl) phosphate
  • the aqueous layer was purified from flash chromatography [Method : Column: Biotage-C18, 60 g Duo- 100 A 30 pm; Mobile phase: [ACN: Water + 0.1% TFA]; B%: B%: 0-8%, 0-20min / 8% 20-45 min.], pure fractions were lyophilized to afford 2-(benzyloxy)ethyl (2-
  • Step 3 2 -hydroxy ethyl (2-(trimethylammonio)ethyl) phosphate
  • reaction mixture was stirred for 6 hours and monitored by TLC.
  • the reaction mixture was concentrated under reduced pressure and crude was dissolved in DMF (5 mL).
  • DMAP (35.4 mg, 0.290 mmol, 1.3 eq.) was added, followed by 2-hydroxyethyl (2-(trimethylammonio)ethyl) phosphate (50.6 mg, 0.290 mmol, 1.0 eq.) at 25°C and left it to stir for 18 hours.
  • reaction mixture was diluted with isopropyl ether (10 mL) and the solvent was decanted to obtain crude, which was further purified according to PREP-HPLC Method-B: Gemini-C18, 250 X 21.2 mm, 5.0 pm; Buffer: [Acetonitrile, 0.1% of formic acid in water], time/B%: 0/30, 10/60, 20/80, 30/90 to afford title compound as a light brown semi solid.
  • reaction mixture was triturated with isopropyl ether (IPE) (20 mL) and was concentrated under reduced pressure to obtain crude, which was purified according to PREP-HPLC Method-B: Kinetex EVO, C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [0.1% of FA in Water: ACN]; time/ B%: 0/15, 20/40, 30/65 to afford title compound (10 mg) as a brown semi solid. Yield: 11.4%.
  • IPE isopropyl ether
  • Step 1 (((benzyloxy) carbonyl)amino) ethyl(2 ((tertbutoxycarbonyl) amino)ethyl) phosphate
  • Step 3 (18-(4, 5-dimethoxy-2-methyl-3, 6-dioxocyclohexa-l ,4-dien-l -yl)octadec-8- en-l-yl (2-(((2-((tert-butoxycarbonyl)amino)ethoxy)(hydroxy)phosphoryl) oxy)ethyl)carbamate
  • Step 4 (2-ammonioethyl (2-(((((18-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa- l,4-dien-l-yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl) phosphate
  • Step 1 (Z)-15-(4, 5-dimethoxy-2-methyl-3, 6-dioxocyclohexa- 1 ,4-dien-l -yl)pentadec-9- enoic acid
  • N -Hydroxysuccinimide (NHS) 45 mg, 0.392 mmol, 1.1 eq.
  • DCC ⁇ , ⁇ '- Dicyclohexylcarbodiimide
  • Step 3 -3-((l 5-(4,5-dimethoxy-2-methyl-3 ,6-dioxocyclohexa-l ,4-dien- 1 - yl)pentadec-9-enoyl)oxy)-4-(trimethylammonio)butanoate
  • the reaction mixture was triturated with isopropyl ether (IPE) (2 x 5 mL) and dried under vacuum to obtain crude, which was purified by PREP -HP LC Method-D: Gemini-C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [ACN: Water]; B%: 0/10, 10/50, 20/60, 25/95 to afford title compound (1.8 mg) as a brown semi solid. Yield: 8.5%.
  • IPE isopropyl ether
  • the reaction mixture was triturated with isopropyl ether (IPE) (2 x 5 mL) and was dried under vacuum to obtain crude, which was purified by PREP-HPLC Method-D: Gemini-C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [ACN: Water]; B%: 0/10, 10/50, 20/60, 25/95 to afford title compound (14 mg) as a yellow solid. Yield: 14.4%.
  • IPE isopropyl ether
  • the assay was conducted in a low throughput 6-well format with HEK293 cells prepared and then transfected in duplicate wells with plasmids containing either the wildtype (WT) version of hMfsd2a, the D97A mutant version, or an empty vector (EV) as control. Uptake into the cells was assessed by both thin-layer chromatography (TLC) and ultrahigh performance liquid chromatography hyphenated mass spectrometry.
  • TLC thin-layer chromatography
  • EV empty vector
  • HEK293 cells were seeded at 6.25 x 10 s per 6-well in 2 mL of DMEM with 10% FBS and 1% penicillin-streptomycin (P/S) media (Sigma) and incubated overnight at 37°C in 5% C0 2 . Cells were checked for confluency the next morning. On a per well basis the following lipid mix was generated; 6 pL of Lipofectamine
  • OptiMEM 2000 was added dropwise to 200 pL of OptiMEM, this was left to stand for 5 minutes at room temperature (RT).
  • RT room temperature
  • One pg of hMfsd2a WT, D97A or empty plasmid was prepared in 200 pL of OptiMEM as appropriate for each well; the Lipofectamine 2000 in OptiMEM solution was then added dropwise to a total volume of 400 pL (this can be scaled to support the number of wells/plates to be assayed). This transfection preparation was then incubated at RT for 20 minutes.
  • DMEM with 10% FBS and no P/S media was warmed to 37°C
  • the HEK293 plate media was changed and the cells washed carefully with 1 mL of the warmed DMEM with 10% FBS no P/S media, 1.6 mL of the warmed DMEM with 10% FBS no P/S media was then added to each well.
  • Four hundred pL of the transfection preparation was then added dropwise to each well as appropriate and the plate was gently swirled in a circular motion. The plate was then incubated overnight at 37°C in 5% C0 2 .
  • Compound stock solutions were prepared in a 12% BSA in PBS solution such that a 40 pL spike into 2 mL of plain DMEM would yield a concentration of 50 pM of test compound (the compound treated media). Remaining compound stock solution in 12% BSA in PBS was frozen at -20°C to allow for media stability testing. The HEK293 6-well plate was removed from the incubator and the wells gently rinsed with 1 mL of plain DMEM that had been prewarmed to 37°C. 2 mL of the compound treated media was then added to each well.
  • the remaining media was then removed from the HEK293 6-well plate, the wells were gently rinsed twice with 1 mL of 0.5% BSA in DMEM, the media was then removed and the 6-well plate allowed to dry completely at RT. 1 mL of 3:2 Hexane: Isopropanol (HIP) was added to each well in a fume cupboard and the plate allowed to stand for 30 minutes at RT without shaking. The HIP solution was then transferred to 2 mL Eppendorf tubes, and the process was repeated with a second 1 mL aliquot of HIP and the two aliquots combined. The HIP samples were then dried down under a nitrogen stream.
  • HIP Isopropanol
  • Silica gel plates were prepared in a fume cupboard by initially drawing a line 1.5 cm from the bottom edge of the plate and then drawing sample lanes with a width of 1 cm and a separation of 0.5 cm between lanes.
  • TLC buffer for phospholipids was prepared as a 31:62:7 solvent mix of MethanokChloroform: Ammonium Hydroxide. Plates were pre-run in a humid chamber containing 200 mL of the TLC buffer until the solvent front was 1.5 cm from the plate edge, the plate was then allowed to dry. HIP samples prepared as described above were reconstituted in 50 pL of chloroform, briefly vortex ed 3 times and then kept on ice.
  • Iodine crystals were added to a new chamber which was sealed to allow the iodine vapor to saturate the container, the plate was then exposed to the iodine vapor in the chamber to allow visualization of bands of unsaturated fatty acids, once the plate was developed a second image was taken using Bio-Rad Image lab 6.0. The plate was then air-dried to remove the iodine. The plate was then saturated using a spray bottle with cupric acetate solution consisting of 3% cupric acetate by weight, 8% phosphoric acid by volume made up in an aqueous solution.
  • the plate was allowed to dry for 5 minutes at RT and then heated in a fume cupboard using a hot air gun to make the bands more visible.
  • a final image was acquired using the Bio-Rad Image lab 6.0.
  • the difference in intensity between the bands generated from hMfsd2a (WT) or D97A transfected HEK293 cells were compared to the empty vector (EV) transfected cells, allowed for uptake into the cells driven by hMfsd2a to be identified against the reference (REF). Results of the TEC analysis are shown in FIGs. 1A-3B and 5A-6B.
  • FIGs. 1A-1B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 1 (Example 1) - lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 1 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector, thus confirming the compound was advantageously transported via Mfsd2a.
  • FIGs. 2A-2B show the TEC images using the iodine and cupric acetate staining, respectively as described above with compound of Compound 2 (Example 2) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 2 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector, thus confirming the compound was advantageously transported via Mfsd2a.
  • FIGs. 3A-3B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 40 (Example 3) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It was unclear from the TEC images if Compound 40 was transported via Mfsd2a.
  • FIGs. 5A-5B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 42 (Example 5) - lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It is unclear from the TEC images if the Compound 42 was transported via Mfsd2a.
  • FIGs. 6A-6B show the TEC images using the iodine and cupric acetate staining respectively as described above with Compound 43 (Example 6) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It was unclear from the TEC images if Compound 43 was transported via Mfsd2a.
  • HEP samples prepared as described above were reconstituted in 100 pL of MeCN, vortex mixed and inverted multiple times to ensure all surfaces of the Eppendorf tube were rinsed with the MeCN and finally pulse centrifuged. A 50 pL aliquot of the MeCN reconstitution solution was then taken as a non-diluted HIP extract sample and added to the 96-well plate, alongside this a 1:10 dilution sample was prepared by taking a 5 pL aliquot and diluting with 45 pL of MeCN; 50 pL of Millipore water was added to each sample.
  • a bioanalytical calibration line was prepared to cover a range of concentration from 0.0001 to 10 pM by spiking 2 pL of a 0.5 mM DMSO stock of the test compound into 98 pL of MeCN to generate a 10 pM top standard that was then serial diluted with MeCN to produce 6 calibration standard stocks.
  • Fifty pL of each calibration standard stock was added to the 96-well plate and diluted with 50 pL of Millipore water.
  • Fifty pL of an appropriate internal standard in MeCN was then added to each of the wells in the 96-well plate that contained either a sample or calibration standard, the plate was sealed and transferred to the UPLC-MS-MS system for analysis.
  • FIG. 7 shows the concentration of Compound 1 (Example 1) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 1 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
  • FIG. 8 shows the peak area ratio of Compound 2 (Example 2) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 2 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageous! y transported via Mfsd2a.
  • Example 9 shows the concentration of Compound 40 (Example 3) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 40 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
  • FIG. 11 shows the concentration of Compound 42 (Example 5) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 42 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
  • FIG. 12 shows the concentration of Compound 43 (Example 6) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 43 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
  • Bioanalytical samples were prepared according to the procedures described above for LC-MS-MS analysis. The samples were analyzed by LC-MS/MS utilizing the AB Sciex QTRAP 5500. The instrument was set to operate in positive ion mode for all analyses and the parameters are shown below in Table below.
  • the assay was conducted in a low throughput 6-well format with HEK293 cells prepared and then transfected in duplicate wells with plasmids containing either the wildtype (WT) version of hMfsd2a, the D97A mutant version, or an empty vector (EV) as control. Uptake into the cells was assessed by both thin-layer chromatography (TEC) and ultrahigh performance liquid chromatography hyphenated mass spectrometry.
  • HEK293 cells were seeded at 6.8 x 10 5 per 6-well in 2 ml. of DMEM with 10% FBS and 1% penicillin-streptomycin (P/S) media (Sigma) and incubated overnight at 37°C in 5% C0 2 . Cells were checked for confluency the next morning. On a per well basis the following lipid mix was generated; 6 pL of Lipofectamine 2000 was added dropwise to 200 pL of OptiMEM, this was left to stand for 5 minutes at room temperature (RT).
  • RT room temperature
  • hMfsd2a WT, D97A or empty plasmid was prepared in 200 pL of OptiMEM as appropriate for each well; the Lipofectamine 2000 in OptiMEM solution was then added dropwise to a total volume of 400 pL (this can be scaled to support the number of wells/plates to be assayed). This transfection preparation was then incubated at RT for 20 minutes.
  • DMEM with 10% FBS and no P/S media was warmed to 37°C
  • the HEK293 plate media was changed and the cells washed carefully with 1 mL of the warmed DMEM with 10% FBS no P/S media, 1.6 mL of the warmed DMEM with 10% FBS no P/S media was then added to each well. 400 pL of the transfection preparation was then added dropwise to each well as appropriate and the plate was gently swirled in a circular motion. The plate was then incubated overnight at 37°C in 5% CO,.
  • Compound stock solutions were prepared in a 12% BSA in PBS solution such that a 40 pL spike into 2 mL of plain DMEM would yield a concentration of 25 ⁇ of test compound (the compound treated media). Remaining compound stock solution in 12% BSA in PBS was frozen at -20°C to allow for future transport assays. The HEK293 6-well plate was removed from the incubator and the wells gently rinsed with 1 mL of plain DMEM that had been prewarmed to 37°C. 2 mL of the compound treated media was then added to each well.
  • the remaining media was then removed from the HEK293 6-well plate, the wells were gently rinsed twice with 1 mL of 0.5% BSA in DMEM, the media was then removed and the 6-well plate allowed to dry completely at RT. 1 mL of 3:2 Hexane:Isopropanol (HIP) was added to each well in a fume cupboard and the plate allowed to stand for 30 minutes at RT without shaking. The HIP solution was then transferred to 2 mL Eppendorf tubes, and the process was repeated with a second 1 mL aliquot of HIP and the two aliquots combined. The HIP samples were then dried down in a CentriVap. For media stability testing, 2 mL of diluted compound solution was added to new 6 well plate and 100 pL was sampled at the time point of 0 minutes, 15 minutes, 30 minutes and 60 minutes of incubation at 37°C in 5% CO2
  • Silica gel plates were prepared in a fume cupboard by initially drawing a line 1.5 cm from the bottom edge of the plate and then drawing sample lanes with a width of 1 cm and a separation of 0.5 cm between lanes.
  • TLC buffer for phospholipids was prepared as a 31:62:7 solvent mix of Methanol: Chloroform: Ammonium Hydroxide . Plates were pre-run in a humid chamber containing 150 mL of the TLC buffer until the solvent front was 1.5 cm from the plate edge, the plate was then allowed to dry. HIP samples prepared as described above were reconstituted in 50 pL of chloroform, briefly vortexed 3 times and then kept on ice.
  • Samples were loaded onto the plate (along with reference compound) by streaking gently with a pipette tip, samples were allowed to dry between streaks.
  • the plate was run in a sealed humid chamber containing the TLC buffer as described above for approximately 1.5 hours or until the solvent front had nearly reached the top of the plate. The plate was removed from the chamber and dried. An initial image was taken using camera.
  • Iodine crystals were added to a new chamber which was sealed to allow the iodine vapor to saturate the container, the plate was then exposed to the iodine vapor in the chamber to allow visualization of bands of unsaturated fatty acids, once the plate was developed a second image was taken using camera. The plate was then air- dried to remove the iodine. The plate was then saturated using a spray bottle with cupric acetate solution consisting of 3% cupric acetate by weight, 8% phosphoric acid by volume made up in an aqueous solution.
  • the plate was allowed to dry for 5 minutes at RT and then heated in an oven at 130°C for 10 minutes to make the bands more visible. A final image was acquired using the camera. Quantitative analysis of the band signals was performed by ImageJ. The difference in intensity between the bands generated from hMfsd2a (WT) or D97A transfected HEK293 cells were compared to the empty vector (EV) transfected cells, allowed for uptake into the cells driven by hMfsd2a to be identified against the reference (REF). Results of the TLC analysis are shown inFIGs. 4A-4B.
  • FIGs. 4A-4B show the TLC images using the iodine and cupric acetate staining, respectively as described above with Compound 41 (Example 4) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 41 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector (iodine stain image), thus confirming the compound was advantageously transported via Mfsd2a.
  • UPLC-MS-MS Analysis HIP samples prepared as described above were reconstituted in 100 pL of MeCN, vortex mixed and inverted multiple times to ensure all surfaces of the Eppendorf tube were rinsed with the MeCN and finally pulse centrifuged. A 50 pL aliquot of the MeCN reconstitution solution was then taken as a non-diluted HIP extract sample and added to the 96-well plate, alongside this a 1:10 dilution sample was prepared by taking a 5 pL aliquot and diluting with 45 pL of MeCN; 50 pL of Millipore water was added to each sample.
  • a bioanalytical calibration line was prepared to cover a range of concentration from 0.001 to 1 pM by spiking 1 pL of a 2 mM MeOH stock of the test compound into 199 pL of MeCN to generate a 10 pM top standard that was then serial diluted with MeCN to produce 7 calibration standard stocks. 50pL of each calibration standard stock was added to the 96- well plate and diluted with 50 pL of Millipore water. 50 pL of an appropriate internal standard in H20 was then added to each of the wells in the 96-well plate that contained either a sample or calibration standard, the plate was sealed and transferred to the UPLC-MS-MS system for analysis.
  • FIG. 10 shows the concentration of Compound 41 (Example 4) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 41 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
  • Bioanalytical samples were prepared according to the procedures described above for LC -MS/MS analysis. The samples were analyzed by LC-MS/MS utilizing the Waters Xevo TQ-S micro. The instrument was set to operate in positive ion mode for all analyses and the parameters are shown below in Table below.
  • Example 9 In vivo AD ME Protocol 1 : IV terminal rat PK study at 3 mg/kg
  • Compound 1 (Example 1) was orally dosed at 3 mg/kg to a group of 18 individually housed male Sprague Dawley rats that were fasted overnight and fed 4 hours post-dose. Dosing was performed with 2 mL/kg dosing volumes with 5% DMSO 95% water used as a dosing vehicle. Terminal plasma samples were taken from groups of 3 animals at each of 5 time-points post dose (0.167 hours, 0.5 hours, 1 hour, 2 hours, 4 hours and 8 hours) by cardiac puncture under CO2 into lithium heparin coated tubes and centrifuged to obtain plasma.
  • PBS phosphate buffered saline
  • Protocol 2 Bioanalvtical samples preparation for plasma samples
  • Plasma samples were defrosted and sample preparation was carried out using liquid-liquid extraction. All the samples were extracted and the supernatants analysed by LC-MS/MS according to the following procedures:
  • the tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
  • Protocol 3 Bioanalvtical samples preparation for brain samples
  • Brain homogenates were prepared from whole frozen brains harvested from Protocol 1 using ACN and water. Harvested whole brains were weighed and 4x weight equivalent volume ACN : water (1:1 v/v) was added before homogenization. Analysis of the brain samples was conducted according to the following procedures and the data are shown in Table 2.
  • the tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
  • Protocol 4 Bioanalytical samples preparation for eve samples
  • Eye homogenates were prepared from whole frozen eyes harvested from Protocol 1 using ACN and water. Harvested whole eyes were weighed and 9x weight equivalent volume ACN: water (1:1 v/v) was added before homogenization. Analysis of the eye samples was conducted according to the following procedures and the data are shown in Table 3.
  • the tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
  • Example 1 demonstrates the cytoprotective effects of Compound 1 10 (Example 1) in human induced pluripotent stem cell (iPSC) derived retinal pigment epithelium (RPE) cells.
  • Human iPSC-RPE cells were differentiated on 96 -well plates for 16 days. Cells were exposed to the compound at 5 ⁇ in quadruplicates for 48 hours before induction of oxidative stress (day 19) with teri-butyl hydroperoxide solution (tBHP) (0-10 mM) at 37°C 5% CO2. Lactate dehydrogenase (LDH) was 15 measured on samples collected after 24 hours tBHP -incubation as a readout of the cytoprotective effect of the compound.
  • iPSC human induced pluripotent stem cell
  • RPE retinal pigment epithelium
  • iPSC-RPE cells acquired their characteristic polygonal morphology and were pigmented at passage 3 in Matrigel ® coated cell culture dishes. Cells were passaged (passage 4) onto Matrigel ® coated 96-well plates at a density of 100,000 cells/cm 2 , and cultured until fully differentiated into RPE cells for 16 days.
  • Compound 1 was dissolved in 100% DMSO (Sigma- Aldrich, USA) at concentration 10 mM. Thereafter, dilutions of study compounds were prepared in RPE medium with B-27 AO neg. (Gibco Thermo Fischer Scientific, USA, 100 mM) and sterile filtered through 0.22 pm. Sterile aliquots were stored at -20°C. Final working dilutions of study compounds (5 pM, 0.1% DMSO) were prepared fresh in RPE medium with B-27 AO neg. Vehicle was 0.1% DMSO in RPE medium with B- 27 AO neg. RPE cells were pre-treated with each of the study compounds or vehicle 48 hours before induction of oxidative stress. LDH Assay Protocol
  • Human iPSC-RPE cells were pre-incubated with the compound at 5 pM in quadmplicates for 48 hours before exposure to whole concentration range of tBHP at 37°C for 24 hours (co-incubation period during which iPSC-RPE cells will be continually exposed to the compound).
  • LDH quantification was performed on samples collected after 6 hours tBHP incubation, and again after the 24 hours tBHP- incubation, according to the previously published protocol (see e.g., Kaja et al. J. Pharmacol. Toxicol. Methods, 2015, 73:1-6).
  • Compound 1 (Example 1) displayed cytoprotective effects.
  • the compound showed lower LDH release at 0.6 mM tBHP compared to the vehicle, indicating that this compound has cytoprotective properties in hiPSC-RPE cells.
  • Compound 1 also demonstrated a trend towards better cell viability (lower LDH release at 6 hours) than vehicle at high tBHP concentrations (1- 10 mM) indicating acute (0-6 hours) protective effects against oxidative stress (FIGs. 13A-13B).

Abstract

The present application provides idebenone derivatives or analogues useful for treating a disease or disorder in a subject in need thereof. Pharmaceutical compositions comprising the compounds and methods of treating the disease or disorder are also provided.

Description

IDEBENONE COMPOUNDS
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims benefit of Singapore Provisional Application No. 1020200337 IQ, filed on April 13, 2020, the disclosure of which is incorporated herein by reference in its entirety.
TECHNICAL FIELD
The present application generally relates to idebenone compounds, in particular idebenone analogues that are useful for treating a disease or disorder in a subject in need thereof.
BACKGROUND
Idebenone is an antioxidant approved by European Medicines Agency (EMA) for adults and adolescents aged 12 years and over with Leber’s Hereditary Optic Neuropathy (LHON), which is a primary mitochondrial DNA (mtDNA) disorder characterised by bilateral sequential or simultaneous visual loss. Affected patients develop a dense central scotoma accompanied with rapid visual acuity deterioration. It has been postulated that idebenone prevents and/or reverses loss of vision in LHON patients by restoring mitochondrial function and preventing oxidative damage in retinal ganglion cells (e.g. see PLOS. 2012; 7(9): e45182). Idebenone has also been extensively studied in both pre-clinical and clinical setting as potential treatment for a range of neurological disorders (e.g. see J. Bioenerg Biomembr. 2015; 47(0): 111- 118).
SUMMARY
The present application provides, inter alia, compounds of Formula I:
Figure imgf000002_0001
I or a pharmaceutically acceptable salt thereof, wherein: L1 is selected from the group consisting of C8-20 alkylene and C8-20 alkenylene, wherein the Cs-20 alkylene or Cs-20 alkenylene is optionally substituted by O', OH, Ci-6 alkyl or halogen;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(Ci-e alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), O(Ci-6 alkylene)0, OC(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(Ci-6 alkylene), 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(Ci-e alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(Ci-6 alkyl ene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O", OH or halogen;
R2 is a bond or a phosphate group;
R3 is selected from the group consisting of H, CO2", CO2H, C1 -4 alkylene-CO2 ", Ci-4 alkylene-C02H and Ci-6 alkyl, wherein the Ci-6 alkyl is optionally substituted by O", OH or halogen;
R4 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02", Ci-4 alkylene-C02H and Ci-6 alkyl, wherein the Ci-6 alkyl is optionally substituted by O', OH or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH or halogen;
R5b is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O", OH or halogen; and
R5C is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O", OH or halogen.
Further, the present application provides pharmaceutical compositions comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carriers or excipients.
The present application also provides methods of treating a disease or disorder in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof or pharmaceutically compositions as defined herein. The present application also discloses one or more dosage forms, comprising a compound of Formula I as defined herein, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent.
Further, the present application provides a method of treating a disease or disorder in a subject comprising administering to the subject in need thereof one or more dosages forms as defined herein.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Methods and materials are described herein for use in the present disclosure; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.
BRIEF DESCRIPTION OF DRAWINGS
FIGs. 1A-6B show representative thin layer chromatography (TLC) images from the iodine (FIGs. 1 A, 2A, 3 A, 4A, 5 A and 6 A) and cupric acetate (FIGs. IB, 2B, 3B, 4B, 5B and 6B) stain analysis described in Examples 7 and 8.
FIGs. 7-12 show the concentration or peak area ratio of the compounds described in Examples 1-6 measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a, and empty vector (EV).
FIGs. 13A-13B show that Compound 1 (Example 1) exhibits cytoprotective properties in hiPSC-RPE cells.
DETAILED DESCRIPTION
The present application relates to idebenone derivatives that improve their Blood Brain Barrier (BBB) and/or Blood Retinal Barrier (BRB) penetrating profile. The resulting compounds could advantageously present the key pharmacophore of idebenone with higher Central Nervous System (CNS) and/or ocular exposure, which may result in enhanced pharmacological activity and clinical benefit. Compounds
Accordingly, the present application provides a compound of Formula I:
Figure imgf000005_0001
I or a pharmaceutically acceptable salt thereof, wherein:
L1 is selected from the group consisting of Cs-2o alkylene and Cx-20 alkenylene, wherein the Cs-20 alkylene or Cs-20 alkenylene is optionally substituted by O", OH, Ci-6 alkyl, or halogen;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci.6 alkylene)C(0)NH, 0(0)0, C(0)NH, C(0)NH(Ci_6 alkylene), 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(Ci-e alkylene)C(0)NH, NHC(0)(Ci_6 alkylene)C(0)0, and NHC(0)(Ci.6 alkyl ene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O', OH, or halogen;
R2 is a bond or a phosphate group;
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02 ", Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci-6 alkyl is optionally substituted by O", OH, or halogen;
R4 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02 ", Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci-6 alkyl is optionally substituted by O", OH, or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH, or halogen;
R5b is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O', OH, or halogen; and R5C is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH, or halogen.
In some embodiments, the present application provides a compound of Formula I, wherein:
L1 is selected from the group consisting of C8-20 alkylene and C8-20 alkenylene, wherein the C8-20 alkylene or C8-2o alkenylene is optionally substituted by O", OH, Ci-6 alkyl, or halogen;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, 0(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O", OH, or halogen;
R2 is a bond or a phosphate group;
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02", Ci-4 alkylene-C02H and Cue alkyl, wherein the Ci-6 alkyl is optionally substituted by O', OH, or halogen;
R4 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-COa", Ci-4 alkylene-C02H and Ci-6 alkyl, wherein the Ci-e alkyl is optionally substituted by O', OH, or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH, or halogen;
R5b is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH, or halogen; and
R5c is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH, or halogen.
In some embodiments, L1 is Cg-20 alkylene. In some embodiments, L1 is Cx-12 alkylene. In some embodiments, L1 is Cg alkylene, C9 alkylene, C10 alkylene, Cn alkylene, C12 alkylene, C13 alkylene, CM alkylene, C15 alkylene, Cie alkylene, C17 alkyl ene, Ci8 alkylene, C19 alkyl ene, or C20 alkyl ene. Unless specified otherwise, the Cs-zo alkylene or Cs-i2 alkylene of the L1 is a straight chain or a branched alkylene.
In some embodiments, the Cs-zo alkylene or the Cg-iz alkylene of L1 is a straight chain Cs-zo alkylene, a straight chain Cs-iz alkylene, a branched Cs-zo alkylene, or a branched Cs-i2 alkylene.
In some embodiments, the straight chain or branched Cs-20 alkylene of L1 is optionally substituted by O', OH, Ci-e alkyl, halogen, or combinations thereof. When the straight chain or branched C 8-20 alkylene of L1 is substituted, the straight chain or branched Cs-20 alkylene may be substituted by one or more O", OH, Cim alkyl, halogen, or combinations thereof. For example, at different positions, the hydrogen atoms of Cs-zo alkylene may be substituted by O" and C1-6 alkyl, O", and halogen, OH and Ci-6 alkyl, OH and halogen, or O', OH, Ci-e alkyl, and halogen. In some embodiments, the straight chain or branched Cs-zo alkylene of L1 is optionally substituted by 1, 2, 3, 4, 5, 6, 7, or 8 groups independently selected from the group consisting of O", OH, Ci-e alkyl, and halogen. In some embodiments, the straight chain or branched Cs-20 alkylene of L1 is optionally substituted by 1, 2, 3, or 4 groups independently selected from the group consisting of O", OH, Ci-e alkyl, and halogen.
In some embodiments, L1 is decanediyl.
In some embodiments, L1 is Cs-zo alkenylene. In some embodiments, L1 is C15- 20 alkenylene. In some embodiments, L1 is Cs alkenylene, C9 alkenylene, Cio alkenylene, Cn alkenylene, C12 alkenylene, C13 alkenylene, C 14 alkenylene, C15 alkenylene, Cm alkenylene, C 17 alkenylene, Cis alkenylene, C 19 alkenylene, or C20 alkylene. Unless specified otherwise, the Cs-zo alkenylene or C15-20 alkenylene of L 1 may be a straight chain or a branched alkenylene.
In some embodiments, the Cs-20 alkenylene or the C15-20 alkenylene of L1 is a straight chain Cs-zo alkenylene, a straight chain C 15-20 alkenylene, a branched Cs-zo alkenylene, or a branched C 15-20 alkenylene.
In some embodiments, the straight chain or branched Cs-zo alkenylene of L1 is optionally substituted by O", OH, Ci-e alkyl, halogen, or combinations thereof. When the straight chain or branched Cs-20 alkenylene is substituted, the straight chain or branched Cs-zo alkenylene may be substituted by one or more O', OH, Ci-e alkyl, halogen, or combinations thereof. For example, at different positions, the hydrogen atoms of C8-20 alkenylene may be substituted by O' and C1-6 alkyl, O" and halogen, OH and Ci-e alkyl, OH and halogen, or O", OH, Ci-e alkyl, and halogen. In some embodiments, the straight chain or branched C8-20 alkenylene of L1 is optionally substituted by 1, 2, 3, 4, 5, 6, 7, or 8 groups independently selected from the group consisting of O", OH, Ci-6 alkyl, and halogen. In some embodiments, the straight chain or branched Cs-20 alkenylene of L1 is optionally substituted by 1, 2, 3, or 4 groups independently selected from the group consisting of O', OH, Ci-e alkyl, and halogen.
In some embodiments, the Cs-20 alkenylene of L1 comprises 1, 2, 3 or 4 carbon-carbon double bonds. In some embodiments, the C8-20 alkenylene of L 1 comprises 1 or 2 carbon-carbon double bonds. In some embodiments, the C8-20 alkenylene of L1 comprises 1 carbon-carbon double bond. In some embodiments, when the Cs-20 alkenylene of L1 comprises 1 carbon-carbon double bond, L1 is selected from the group consisting of tetradecenediyl, heptadecenediyl, and octadecenediyl. In some embodiments, when the C8-20 alkenylene of the L1 comprises 1 carbon-carbon double bond, L1 is selected from the group consisting of heptadecenediyl and octadecenediyl.
In some embodiments, when L1 is tetradecenediyl, L1 is selected from the group consisting of:
Figure imgf000008_0001
o
-O-
Ό' L wherein - indicates the bonds connecting L1 to the o moiety and R1.
In some embodiments, when L1 is heptadecenediyl, the L1 is selected from the group consisting of: w
Figure imgf000009_0002
herein - indicates the bonds connecting L1 to the moiety and R1.
In some embodiments, when the L1 is octadecenediyl, the L1 may be selected from the group consisting of the following structures:
Figure imgf000009_0001
9
Figure imgf000010_0001
wherein - indicates the bonds connecting the L1 to the
Figure imgf000010_0002
moiety and R1.
In some embodiments, R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(CI-6 alkylene), 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-e alkylene)C(0)0, 0(Ci-e alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH.
In some embodiments, R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci_6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(CI-6 alkylene)C(0)0, and NHC(0)(Ci-6 alkylene)C(0)NH, wherein each Ci- 6 alkyl ene is substituted by O", OH, or halogen (fluorine, chlorine, bromine).
In some embodiments, R1 is selected from the group consisting of 0C(0)NH(CI-6 alkylene) and 0C(0)0(Ci-6 alkylene). In some embodiments, R1 is selected from the group consisting of 0C(0)NH(CM alkylene) and 0C(0)0(Ci-4 alkylene). In some embodiments, R1 is selected from the group consisting of 0C(0)NHCH2CH2 and 0C(0)0CH2CH2.
In some embodiments, R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0,
0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, C(0)NHCH2, C(0)NHCH2CH2, C(0)NHCH2CH2CH2, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH, 0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0, 0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, MHC(0)CH2CH2CH2C(0)NH,
NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0,
C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH,
0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, R2 is a bond. When the R2 is a bond, it is to be understood that the R1 is directly linked to -CH(R3)- moiety via a bond. Hence, the R2 may be considered as being absent. In some embodiments, the R2 is a phosphate group, wherein the phosphate group has the following structure:
Figure imgf000012_0001
where - indicates the bonds connecting R2 to R1 and -CH(R3)- moiety.
In some embodiments, R3 is selected from the group consisting of H, C02 ", C02H, CH2C02 " and CH2C02H. In some embodiments, R3 is H. In some embodiments, R3 is C02 ". In some embodiments, R3 is CH2C02 ".
In some embodiments, R4 is selected from the group consisting of H, CO2", C02H, CH2C02 " and CH2C02H. In some embodiments, R4 is H. In some embodiments, R4 is C02\ In some embodiments, R4 is CH2C02 ".
In some embodiments, when R4 is H, R3 is C02 ", C02H, CH2CO2" or CH2C02H. In some embodiments, when R3 is H, R4 is C02 ", C02H, CH2C02 " or
CH2CO2H. In some embodiments, R5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen. In some embodiments, R5a is methyl, ethyl or n-propyl. In some embodiments, R5a is methyl.
In some embodiments, R5b is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen. In some embodiments, R5b is methyl, ethyl or n-propyl. In some embodiments, R5b is methyl.
In some embodiments, R5c is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen. In some embodiments, R5c is methyl, ethyl or n-propyl. In some embodiments, R5c is methyl.
In some embodiments, two of R5a, R5b, and R5c are H. In some embodiments, R5a, R5b, and R5c are each H. In some embodiments, two of R5a, R5b, and R5c are methyl. In some embodiments, R5a, R5b, and R5c are each methyl.
In some embodiments:
L1 is C8-20 alkylene or Cg-2o alkenylene, wherein the Cs-2o alkylene or Cs-2o alkenylene is optionally substituted by O', OH, CM alkyl or halogen;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CM alkylene), NHC(0)NH, 0C(0)0(CM alkylene), 0(CM alkylene)0, 0C(0)(CM alkylene)C(0)0, 0C(0)(CM alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(CM alkylene)C(0)0, 0C(0)0(CM alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(CM alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH,
NH(CI-6 alkylene)C(0)0, NH(CM alkylene)C(0)NH, NHC(0)(CM alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by OH;
R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to -CH(R3)- moiety;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
In some embodiments: L1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, tetradecenediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(Ci-e alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(Ci-6 alkylene), 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(Ci-e alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(Ci-6 alkyl ene)C(0)NH, wherein each C1-6 alkylene is optionally substituted by OH;
R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to -CH(R3)- moiety;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
In some embodiments:
L1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-e alkylene), 0(Ci-e alkylene)0, 0C(0)(Ci-e alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkyl ene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by OH; R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to -CH(R3)- moiety;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
In some embodiments:
L1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, tetradecenediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH,
0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, C(0)NHCH2, C(0)NHCH2CH2, C(0)NHCH2CH2CH2, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH, 0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0, 0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH,
NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH; R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to -CH(R3)- moiety;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
In some embodiments:
L1 is selected from the group consisting of octanediyl, nonanediyl, decanediyl, undecanediyl, dodecanediyl, hexadecenediyl, heptadecenediyl, octadecenediyl and nonadecenediyl;
R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH,
0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH, 0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0,
NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH; R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to -CH(R3)- moiety;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
In some embodiments, the compound of Formula I as defined above, or a pharmaceutically acceptable salt thereof, is a compound of Formula II:
Figure imgf000017_0001
II or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, R5a, R5b, and R5c are as defined above.
In some embodiments, the compound of Formula I as defined above, or a pharmaceutically acceptable salt thereof, is a compound of Formula II:
Figure imgf000017_0002
II or a pharmaceutically acceptable salt thereof; wherein:
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O', OH, or halogen; R2 is a bond or a phosphate group, wherein said phosphate group is as defined above; unless specified otherwise, when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to the -CH(R3)- moiety.
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02 ", Ci-4 alkylene-C02H and Ci16 alkyl, wherein the C1-6 alkyl is optionally substituted by O', OH or halogen;
R5a is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O', OH or halogen;
R5b is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O', OH or halogen; and
R5C is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O', OH or halogen.
In some embodiments of the compound of Formula II, or a pharmaceutically acceptable salt thereof, the R1 is selected from the group consisting of
0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH,
0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH,
0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is a compound of Formula III:
Figure imgf000019_0001
Ill or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, R5a, R5b, and R5c are as defined above.
In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is a compound of Formula III:
Figure imgf000019_0002
III or a pharmaceutically acceptable salt thereof; wherein:
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-e alkylene is optionally substituted by O', OH, or halogen;
R2 is a bond or a phosphate group; unless specified otherwise, when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to the -CH(R3)- moiety. R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02 ', Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci-6 alkyl is optionally substituted by O', OH or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen;
R5b is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O', OH or halogen; and
R5C is selected from the group consisting of H and C4 alkyl, wherein the C4 alkyl is optionally substituted by O', OH or halogen.
In some embodiments of the compound of Formula III, or a pharmaceutically acceptable salt thereof, the R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0,
0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0, 0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0,
0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is a compound of Formula IV:
Figure imgf000021_0001
IV or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, R5a, R5b, and R5c are as defined above.
In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is a compound of Formula IV:
Figure imgf000021_0002
IV or a pharmaceutically acceptable salt thereof; wherein:
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O', OH, or halogen;
R2 is a bond or a phosphate group; unless specified otherwise, when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to the -CH(R3)- moiety.
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02", Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci-e alkyl is optionally substituted by O", OH or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen;
R5¾ is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen; and R5C is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH or halogen.
In some embodiments of the compound of Formula IV, or a pharmaceutically acceptable salt thereof, the R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof, is a compound of Formula V:
Figure imgf000022_0001
v or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, R5a, R5b, and R5c are as defined above.
In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is a compound of Formula V:
Figure imgf000023_0001
V or a pharmaceutically acceptable salt thereof; wherein:
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O', OH, or halogen;
R2 is a bond or a phosphate group; unless specified otherwise, when R2 is a bond it is to be understood that R2 may be considered as being absent such that R1 is directly linked to the -CH(R3)- moiety.
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02 ", Ci -4 alkylene-C02H and Ci-e alkyl, wherein the Ci-6 alkyl is optionally substituted by O', OH or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH or halogen;
R5b is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH or halogen; and
R5C is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O', OH or halogen.
In some embodiments of the compound of Formula V, or a pharmaceutically acceptable salt thereof, the R1 is selected from the group consisting of
0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH,
0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2 CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2CH2C(0)NH.
In some embodiments, the compound of Formula I as defined herein, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of compounds 1 to 43, where its chemical structure thereof is shown in Table A below.
Table A.
Compound 1 or a pharmaceutically acceptable salt thereof
Figure imgf000024_0001
Compound 2 or a pharmaceutically acceptable salt thereof
Figure imgf000024_0002
Compound 3 or a pharmaceutically acceptable salt thereof
Figure imgf000024_0003
Compound 4 or a pharmaceutically acceptable salt thereof o
Figure imgf000025_0001
Compound 5 or a pharmaceutically acceptable salt thereof o o OH o. O COO"
Ό o
Compound 6 or a pharmaceutically acceptable salt thereof
Figure imgf000025_0002
Compound 7 or a pharmaceutically acceptable salt thereof
O
O o
.0,
’"’O
Figure imgf000025_0003
O
Compound 8 or a pharmaceutically acceptable salt thereof o o
Ό o o o k. COO"
Compound 9 or a pharmaceutically acceptable salt thereof
Figure imgf000025_0004
Compound 10 or a pharmaceutically acceptable salt thereof
Figure imgf000025_0005
Figure imgf000026_0001
Figure imgf000027_0001
Figure imgf000028_0001
Figure imgf000029_0001
Figure imgf000030_0001
Synthesis
As will be appreciated, the compounds of the present application (for e.g. compounds of Formula I, II, III, IV and V provided herein), including salts thereof, may be prepared using known organic synthesis techniques and may be synthesized according to any of numerous possible synthetic routes. For example, the compounds provided herein can be prepared according to the general procedures shown in Schemes 1 to 9.
Scheme 1.
Figure imgf000031_0001
Figure imgf000031_0002
Scheme 2.
Figure imgf000032_0001
Scheme 3.
Figure imgf000032_0002
Scheme 4.
Figure imgf000032_0003
Scheme 5.
Figure imgf000033_0001
Scheme 6.
Figure imgf000033_0002
Scheme 7.
Figure imgf000033_0003
Scheme 8.
Figure imgf000034_0001
It will be appreciated by one skilled in the art that the processes described above are not the exclusive means by which compounds provided herein may be synthesized and that a broad repertoire of synthetic organic reactions is available to be potentially employed in synthesizing compounds provided herein. The person skilled in the art knows how to select and implement appropriate synthetic routes.
Suitable synthetic methods of starting materials, intermediates and products may be identified by reference to the literature, including reference sources such as: Advances in Heterocyclic Chemistry, Vols. 1-107 (Elsevier, 1963-2012); Journal of Heterocyclic Chemistry V o\s. 1-49 (Journal of Heterocyclic Chemistry, 1964-2012); Carreira, et al. (Ed.) Science of Synthesis, Vols. 1-48 (2001-2010) and Knowledge Updates KU2010/1-4; 2011/1-4; 2012/1-2 (Thieme, 2001-2012); Katritzky, et al.
(Ed.) Comprehensive Organic Functional Group Transformations, (Pergamon Press, 1996); Katritzky et al. (Ed.); Comprehensive Organic Functional Group Transformations II (Elsevier, 2nd Edition, 2004); Katritzky et al. (Ed.),
Comprehensive Heterocyclic Chemistry (Pergamon Press, 1984); Katritzky et al., Comprehensive Heterocyclic Chemistry II, (Pergamon Press, 1996); Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6th Ed. (Wiley, 2007); Trost et al. (Ed.), Comprehensive Organic Synthesis (Pergamon Press, 1991).
Preparation of compounds described herein can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., Wiley & Sons, Inc., New York (1999).
Reactions can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 'H or 13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectroscopy (LCMS), or thin layer chromatography (TEC). Compounds can be purified by those skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) and normal phase silica chromatography.
Definitions
At various places in the present specification, divalent linking substituents are described. It is specifically intended that each divalent linking substituent include both the forward and backward forms of the linking substituent. For example, - NR(CR’R”)n- includes both -NR(CR’R”)n- and -(CR’R”)nNR-. Where the structure clearly requires a linking group, the Markush variables listed for that group are understood to be linking groups.
As used herein, the phrase “optionally substituted” means unsubstituted or substituted. The term “substituted” as used herein means that one or more hydrogen atoms are removed and replaced by one or more substituents. It is to be understood that substitution at a given atom is limited by valency.
Throughout the definitions, the term “Cn-m” indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include CM, CM, and the like.
As used herein, the term “Cn-m alkyl”, employed alone or in combination with other terms, refers to a saturated hydrocarbon group that may be straight-chain or branched, having n to m carbons. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, «-propyl, isopropyl, «-butyl, tert- butyl, isobutyl, sec-butyl; higher homologs such as 2 -methyl- 1 -butyl, «-pentyl, 3- pentyl, «-hexyl, 1 ,2,2-trimethylpropyl, and the like. In some embodiments, the alkyl group contains from 1 to 6 carbon atoms, from 1 to 4 carbon atoms, from 1 to 3 carbon atoms, or 1 to 2 carbon atoms.
As used herein, the term “Cn-m alkylene”, employed alone or in combination with other terms, refers to a divalent alkyl linking group having n to m carbons. Examples of alkylene groups include, but are not limited to, methylene, ethan-1,2- diyl, propan- 1, 3 -diyl, propan- 1,2-diyl, and the like. In some embodiments, the alkylene moiety contains 8 to 20, 10 to 20, 8 to 15, 1 to 6, 1 to 3, or 1 to 2 carbon atoms.
As used herein, the term “Cn-m alkenyl”, employed alone or in combination with other terms, refers to an alkyl group having one or more carbon-carbon double bonds and having n to m carbons. Example alkenyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, heptenyl, octenyl, nonenyl, decenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, and the like. In some embodiments, the alkenyl moiety contains 5 to 25, 8 to 20, or 10 to 20 or carbon atoms.
As used herein, the term “Cn-m alkenylene”, employed alone or in combination with other terms, refers to a divalent alkenyl linking group having n to m carbons. Examples of alkenylene groups include, ethen- 1,2-diyl, propen-1, 3-diyl, propen- 1 ,2- diyl , hexen-l,6-diyl, hepten-l,7-diyl, octen-l,8-diyl, nonen-l,9-diyl, decen-l,10-diyl, pentadecen- 1 , 15-diyl, hexadecene- 1 , 16-diyl, hexadecene-2, 16-diyl, heptadecen- 1,17- diyl, heptadecen-2, 17-diyl, and the like. In some embodiments, the alkenylene moiety contains 5 to 25, 8 to 20, or 10 to 20 or carbon atoms.
As used herein, the term “Cn-m alkoxy”, employed alone or in combination with other terms, refers to a group of formula -O- alkyl, wherein the alkyl group has n to m carbons. Example alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), butoxy (e.g., n-butoxy and tert- butoxy), and the like. In some embodiments, the alkoxy group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.
As used herein, the term “halogen” refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).
The term “compound” as used herein is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.
Compounds provided herein also include tautomeric forms. Tautomeric forms result from the swapping of a single bond with an adjacent double bond together with the concomitant migration of a proton. Tautomeric forms include prototropic tautomers which are isomeric protonation states having the same empirical formula and total charge. Example proto tropic tautomers include ketone — enol pairs, amide - imidic acid pairs, lactam - lactim pairs, enamine - imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, for example, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1 ,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.
All compounds, and pharmaceutically acceptable salts thereof, can be found together with other substances such as water and solvents (e.g. hydrates and solvates) or can be isolated.
In some embodiments, preparation of compounds can involve the addition of acids or bases to affect, for example, catalysis of a desired reaction or formation of salt forms such as acid addition salts.
Example acids can be inorganic or organic acids and include, but are not limited to, strong and weak acids. Some example acids include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, 4- nitrobenzoic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, and nitric acid. Some weak acids include, but are not limited to acetic acid, propionic acid, butanoic acid, benzoic acid, tartaric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, and decanoic acid.
Example bases include lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, and sodium bicarbonate. Some example strong bases include, but are not limited to, hydroxide, alkoxides, metal amides, metal hydrides, metal dialkylamides and arylamines, wherein; alkoxides include lithium, sodium and potassium salts of methyl, ethyl and t-butyl oxides; metal amides include sodium amide, potassium amide and lithium amide; metal hydrides include sodium hydride, potassium hydride and lithium hydride; and metal dialkylamides include lithium, sodium, and potassium salts of methyl, ethyl, n-propyl, iso-propyl, n-butyl, tert- butyl, trimethylsilyl and cyclohexyl substituted amides.
In some embodiments, the compounds and salts provided herein are substantially isolated. By “substantially isolated” is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation can include, for example, a composition enriched in the compounds provided herein. Substantial separation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compounds provided herein, or salt thereof. Methods for isolating compounds and their salts are routine in the art.
The term, “room temperature” or “RT” as used herein, are understood in the art, and refer generally to a temperature (e.g., a reaction temperature) that is about the temperature of the room in which the reaction is carried out, for example, a temperature from about 20°C to about 30°C such as about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C or about 30°C.
The compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of Formula I described in the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically inactive starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms. Accordingly, the compounds of Formula I as defined herein or a pharmaceutically acceptable salt thereof, may be found as both cis and trans isomers. As can be seen from the exemplified compounds, the preferred isomers may be cis isomers.
Resolution of racemic mixtures of compounds can be carried out by any of numerous methods known in the art. An example method includes fractional recrystallizaion using a chiral resolving acid which is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids such as β -camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include stereoisomerically pure forms of a-methylbenzylamine (e.g., S and R forms, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N- methylephedrine, cyclohexyl ethylamine, 1 ,2 -diaminocyclohexane, and the like.
Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent composition can be determined by one skilled in the art.
The phrase “pharmaceutically acceptable” is employed herein to refer 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 of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. The present disclosure also provides pharmaceutically acceptable salts of the compounds as described above. The term “pharmaceutically acceptable salts”, as used herein, refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form.
Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
The pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, non- aqueous media like ether, ethyl acetate, alcohols (e.g. , methanol, ethanol, iso-propanol, or butanol) or acetonitrile (MeCN) are preferred.
Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977). Conventional methods for preparing salt forms are described, for example, in Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH, 2002.
Methods of Use
The present disclosure further provides methods of treating a disease or disorder in a subject. As used herein, the term “subject,” refers to any animal, including mammals. Exemplary subjects include, but are not limited to, mice, rats, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, and humans.
In some embodiments, the subject is a human. In some embodiments, the method comprises administering to the subject (e.g., a subject in need thereof) a therapeutically effective amount of a compound provided herein (e.g. , a compound of Formula I, II, III, IV or V as defined above), or a pharmaceutically acceptable salt thereof. In some embodiments, the disease or disorder is selected from the group consisting of mitochondrial disorder, pain, a pain-related disease or disorder, a mood disease or disorder, a disease or disorder of the central nervous system, an optical disease or disorder, cancer, a gastrointestinal disease or disorder, a renal disease or disorder, a renal-related disease or disorder, a cardiovascular disease or disorder, and a skin disease or disorder.
In some embodiments, the disease or disorder is a mitochondrial disorder. In some embodiments, the mitochondrial disorder is selected from the group consisting of Leber's hereditary optic neuropathy (LHON), Duchenne muscular dystrophy, mitochondrial encephalopathy, lactic acidosis and stroke- like episodes (MELAS), mitochondrial myopathy, diabetes mellitus and deafness (DAD), Leigh syndrome, neuropathy, ataxia, and retinitis pigmentosa (NARP), ptosis, chronic progressive external ophthalmoplegia (CPEO), coenzyme Q10 deficiency, familial bilateral striatal necrosis, Friedreich ataxia, infantile-onset spinocerebellar ataxia (IOSCA), Kearns- Sayre syndrome, mitochondrial DNA depletion syndrome (MDS) and Rett syndrome (RTT).
In some embodiments, the disease or disorder is pain or a pain-related disease or disorder. In some embodiments, the pain or a pain-related disease or disorder is selected from the group consisting of acute pain, chronic pain, neuropathic pain, nociceptive pain, inflammatory pain, cancer pain, fibromyalgia, rheumatoid arthritis, osteoarthritis, surgery-related pain, and osteoporosis. In some embodiments, the disease or disorder is selected from the group consisting of rheumatoid arthritis and osteoarthritis. In some embodiments, the disease or disorder is pain related to rheumatoid arthritis or pain related to osteoarthritis.
In some embodiments, the disease or disorder is a mood disease or disorder. In some embodiments, the mood disease or disorder is selected from the group consisting of anxiety, depression, a sleeping disorder, an eating disorder, post- traumatic stress disorder, symptoms of drug or alcohol withdrawal or abuse, schizophrenia, obsessive-compulsive disorder, bipolar disorder, sexual dysfunction, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD).
In some embodiments, the disease or disorder is a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, or an optical disease or disorder. In some embodiments, the disease or disorder is a disease or disorder of the central nervous system. In some embodiments, the disease or disorder is a disease or disorder of the peripheral nervous system. In some embodiments, the disease or disorder is an optical disease or disorder.
In some embodiments, the disease or disorder of the central nervous system, peripheral nervous system, or optical disease or disorder is selected from the group consisting of a demyelinating disease, glaucoma, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), a cognitive disorder, Alzheimer’s disease, a movement disorder, Huntington’s disease, Huntington’s chorea, Tourette’s syndrome, Niemann-Pick disease, Parkinson's disease, epilepsy, a cerebrovascular disorder, ischemic stroke and brain injury.
In some embodiments, the demyelinating disease is selected from the group consisting of multiple sclerosis (MS), neuromyelitis optica (NMO), Devic’s disease, central nervous system neuropathy, central pontine myelinolysis, syphilitic myelopathy, leukoencephalopathies, leukodystrophies, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, anti-myelin-associated glycoprotein (MAG) peripheral neuropathy, Charcot-Marie-T ooth disease, peripheral neuropathy, myelopathy, optic neuropathy, progressive inflammatory neuropathy, optic neuritis, and transverse myelitis.
In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is selected from the group consisting of leukemia, mantle cell lymphoma, Hodgkin lymphoma, Non-Hodgkin lymphoma, hepatocellular carcinoma, ovarian cancer, colorectal cancer, pancreatic cancer, prostate cancer, breast cancer, glioma, glioblastoma skin cancer, renal carcinoma, and lung cancer.
In some embodiments, the disease or disorder is a gastrointestinal disease or disorder. In some embodiments, the gastrointestinal disease or disorder is selected from the group consisting of inflammatory bowel disease, gastroesophageal reflux disease, paralytic ileus, secretory diarrhoea, gastric ulcer, nausea, emesis, celiac disease, irritable bowel syndrome, and a liver disorder.
In some embodiments, the liver disease is selected from the group consisting of acute liver failure, Alagille syndrome, hepatitis, enlarged liver, Gilbert’s syndrome, liver cyst, liver haemangioma, fatty liver disease, steatohepatitis, primary sclerosing cholangitis, fascioliasis, primary bilary cirrhosis, Budd-Chiari syndrome, hemochromatosis, Wilson’s disease, and transthyretin-related hereditary amyloidosis.
In some embodiments, the disease or disorder is a renal disease or disorder or a renal-related disease or disorder. In some embodiments, the renal disease or disorder or a renal-related disease or disorder is selected from the group consisting of diabetes, diabetic nephropathy, acute inflammatory kidney injury, renal ischemia urinary incontinence, and overactive bladder.
In some embodiments, the disease or disorder is a skin disease or disorder. In some embodiments, the skin disease or disorder is selected from the group consisting of atopic dermatitis, psoriasis, and lupus.
In some embodiments, the disease or disorder is a cardiovascular disease or disorder. In some embodiments, the cardiovascular disease or disorder is selected from the group consisting of cardiovascular disease, vascular inflammation, idiopathic pulmonary fibrosis, cough, and hypertension.
The present disclosure also provides the compound described herein such as the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof, for use in treating the disease or disorder as previously described. In some embodiments, there is provided the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof, for use in treating the mitochondrial disorder.
The present disclosure further provides use the compound as described herein (e.g. the compound of Formula I, II, III, IV or V) or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating the disease or disorder as previously described. In some embodiments, there is provided use the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating the mitochondrial disorder.
As used herein, the phrase “therapeutically effective amount” refers to the amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought in a tissue, system, animal, individual or human by a researcher, veterinarian, medical doctor or other clinician.
As used herein, the term “treating” or “treatment” refers to one or more of (1) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology); and (2) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology) such as decreasing the severity of disease or reducing or alleviating one or more symptoms of the disease.
Labeled Compounds
The present application further provides isotopically-labeled compounds provided herein. An “isotopically” or “radio-labeled” compound is a compound wherein one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated in compounds of the present disclosure include but are not limited to 2H (also written as D for deuterium), 3H (also written as T for tritium), nC, 13C, 14C, 13N, 15N, 150, 170, and 180. For example, one or more hydrogen atoms in a compound of the present disclosure can be replaced by deuterium atoms (e.g., one or more hydrogen atoms of a Ci -6 alkyl group as described herein can be optionally substituted with deuterium atoms, such as -CD3 being substituted for -CFL). In some embodiments, alkyl groups of the componuds provided herein can be perdeuterated.
One or more constituent atoms of the compounds presented herein can be replaced or substituted with isotopes of the atoms in natural or non-natural abundance. In some embodiments, the compound provided herein comprises at least one deuterium atom. In some embodiments, the compound provided herein comprises two or more deuterium atoms. In some embodiments, the compound provided herein comprises 1-2, 1-3, 1-4, 1-5, 1-6, 1-8, 1-10, 1-12, 1-14, 1-16, 1-18, or 1-20 deuterium atoms. In some embodiments, all of the hydrogen atoms in a compound provided herein are replaced or substituted by deuterium atoms.
Synthetic methods for including isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, N.Y., Appleton-Century-Crofts, 1971; The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in various studies such as NMR spectroscopy, metabolism experiments, and/or assays. In some embodiments, the present application further provides synthetic methods for incorporating radio-isotopes into the compounds described herein. Synthetic methods for incorporating radio-isotopes into organic compounds are well known in the art, and an ordinary skill in the art will readily recognize the methods applicable for the compounds described herein.
Substitution with heavier isotopes, such as deuterium, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances, (see e.g., A. Kerekes et. al. J. Med. Chem. 2011, 54, 201-210; R. Xu et. al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolism sites may afford one or more of the therapeutic advantages.
Combination Therapies
One or more additional therapeutic agents or other agents useful for treating the diseases or disorders as mentioned above can be used in combination with the compounds of Formula 1, 11, III, IV or V and salts thereof provided herein.
In some embodiments, the additional therapeutic agent may be administered simultaneously with the compound or salt provided herein. In some embodiments, the additional therapeutic agent may be administered after administration of the compound or salt provided herein. In some embodiments, the additional therapeutic agent is administered prior to administration of the compound or salt provided herein. In some embodiments, the compound or salt provided herein is administered during a surgical procedure. In some embodiments, the compound or salt provided herein is administered in combination with an additional therapeutic agent during a surgical procedure. Pharmaceutical Formulations
When employed as pharmaceuticals, the compounds and salts provided herein can be administered in the form of pharmaceutical compositions. These compositions can be prepared as described herein or elsewhere, and can be administered by a variety of routes, depending upon whether local or systemic treatment is desired and upon the area to be treated.
Administration may be topical (e.g. , transdermal, epidermal, ophthalmic and to mucous membranes including intranasal, vaginal and rectal delivery), pulmonary (e.g. , by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular or injection or infusion; or intracranial (e.g., intrathecal or intraventricular, administration). Parenteral administration can be in the form of a single bolus dose, or may be, for example, by a continuous perfusion pump.
In some embodiments, the compounds provided herein (e.g., compounds of Formula I, II, III, IV and V) are suitable for parenteral administration. In some embodiments, the compounds provided herein (e.g., the compounds of Formula I, II, III, IV and V) are suitable for intravenous administration.
Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. In some embodiments, the pharmaceutical compositions provided herein are suitable for parenteral administration. In some embodiments, the compositions provided herein are suitable for intravenous or i.v. administration.
Also provided are pharmaceutical compositions comprising, as the active ingredient, a compound provided herein such as a compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers (or excipients). In making the compositions provided herein, the active ingredient is typically mixed with an excipient, diluted by an excipient or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.
Some examples of suitable excipients include, without limitation, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include, without limitation, lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-b enzoates ; sweetening agents; flavoring agents, or combinations thereof.
EXAMPLES
The present disclosure will be described in greater detail by way of specific examples as shown below. However, it is to be understood that the following examples are offered for illustrative purposes and are not intended to limit the present disclosure in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same results. Analytical methods described throughout the Examples were performed according to the following procedures:
LC-MS - Method A
25
Figure imgf000047_0001
LC-MS - Method B
Figure imgf000048_0001
LC-MS - Method C
Figure imgf000048_0002
LC-MS - Method D
Figure imgf000048_0003
LC-MS - Method E
Figure imgf000048_0004
Figure imgf000049_0001
LC-MS - Method F
Figure imgf000049_0002
LC-MS - Method G
Figure imgf000049_0003
LC-MS - Method H
Figure imgf000049_0004
Figure imgf000050_0001
PREP-HPLC Method A
Figure imgf000050_0002
PREP-HPLC Method B
Figure imgf000050_0003
PREP-HPLC Method C
Figure imgf000050_0004
PREP-HPLC Method D
Figure imgf000051_0003
Intermediate 1. (Z)-2-(18-hydroxyoctadec-10-en-l-yl)-5,6-dimethoxy-3- methylcyclohexa-2,5-diene-l,4-dione
Figure imgf000051_0001
Step 1. methyl 11-bromoundecanoate
Figure imgf000051_0002
To a stirred solution of 11 -bromoundecanoic acid (51 g, 192.4mmol, 1.0 eq.) in MeOH (500 mL) at 0°C was added H2SO4 (20 mL, 384.9 mmol, 2 eq.). The reaction was refluxed for 2 hours and was monitored by thin layer chromatography (TLC). The reaction mixture was concentrated, quenched with ice water (50 mL) and saturated NaHCOg solution, and extracted with EtOAc (3 x 200 mL). The total organic layer was dried over anhydrous Na2SC>4, filtered, and concentrated under reduced pressure to obtain crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 10% EtO Ac/hexane to afford methyl 11- bromoundecanoate (51 g) as a colorless liquid. 'H-NMR (400 MHz, CDCI3): δ 3.66 (s, 3H), 3.40 (t, .7=6.8 Hz, 2H), 2.30 (t, .7=7.2 Hz, 2H), 1.88-1.81 (m, 2H), 1.66-1.59 (m, 2H), 1.46-1.37 (m, 2H), 1.34-1.25 (m, 10H).
Step 2. (11-methoxy-l l-oxoundecyl)triphenylphosphonium bromide
Figure imgf000052_0001
To a stirred solution of methyl 11-bromoundecanoate (51 g, 182.7 mmol, 1.0 eq.) in acetonitrile (150 mL) at 25°C was added triphenylphosphine (71.8 g, 274.1 mmol, 1.5 eq.). The reaction was stirred at 110°C for 48 hours and was monitored by TLC. The reaction mixture was concentrated under reduced pressure and the crude was triturated with 20% EtO Ac/Hexane (100 mL) to give (11-methoxy-l 1- oxoundecyl)triphenylphosphonium bromide (90 g) as a white solid. 1H-NMR (400 MHz, CDCI3): δ 7.93-7.87 (m, 3H), 7.84-7.73 (m, 12H), 3.62-3.51 (m, 5H), 2.27 (t, .7=7.2 Hz, 2H), 1.57-1.39 (m, 6H), 1.29-1.16 (m, 10H).
Step 3. 8-((tetrahydro-2H-pyran-2-yl)oxy)octan-l-ol
Figure imgf000052_0002
To a stirred solution of octane- 1 ,8-diol (20 g, 137.9 mol, 1.0 eq.) in Toluene/DCM (4:1) (500 mL) at 25°C was added DHP (13.4 mL, 157.2 mmol, 1.14 eq.) and Dowex 50WX8 Hydrogen form (5 g). The reaction was stirred at 25°C for 16 hours and monitored by TLC. The reaction mixture was quenched with ice water (50 mL) and extracted with EtOAc (3 x 200 mL). The total organic layer was dried over anhydrous NazSCL and concentrated under reduced pressure to obtain the crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 10% EtO Ac/hexane to afford 8 -((tetrahydro-2H-pyran-2-yl)oxy)octan- 1 - ol (20 g) as a colorless liquid. Ή-NMR (400 MHz, CDC13): δ 4.59-4.55 (m, 1H), 3.90-3.83 (m, 1H), 3.76-3.69 (m, 1H), 3.66-3.60 (m, 2H), 3.53-3.46 (m, 1H), 3.41- 3.34 (m, 1H), 1.88-1.78 (m, 1H), 1.75-1.69 (m, 1H), 1.63-1.48 (m, 8H), 1.39-1.29 (m,
8H).
Step 4. 8-((tetrahydro-2H-pyran-2-yl)oxy)octanal
Figure imgf000052_0003
To a stirred solution of 8-((tetrahydro-2H-pyran-2-yl)oxy)octan- 1 -ol (15 g, 65.2 mmol, 1.0 eq.) in DCM (500 mL) at 0°C was added pyridinium chlorochromate (PCC, 21 g, 97.8 mmol, 1.5 eq.) and Celite (15 g) mixture. The reaction was stirred at 25°C for 2 hours and monitored by TLC. The reaction mixture was filtered through Celite pad and washed with DCM (100 mL). The filtrate was dried over anhydrous Na2$04 and concentrated under reduced pressure to obtain crude solid, which was purified by silica gel flash chromatography (60-120 mesh) with gradient elution with 10% EtO Ac/Hexane to afford 8-((tetrahydro-2H-pyran-2-yl)oxy)octanal (11 g) as a colorless liquid. ¾-NMR (400 MHz, CDC13): δ 9.76 (t, J-2.0 Hz, 1H), 4.59-4.55 (m, 1H), 3.90-3.83 (m, 1H), 3.76-3.69 (m, 1H), 3.53-3.46 (m, 1H), 3.41-3.34 (m, 1H), 2.45-2.38 (m, 2H), 1.88-1.78 (m, IH), 1.75-1.47 (m, 10H), 1.41-1.29 (m, 6H).
Step 5. methyl (Z)-19-((tetrahydro-2H-pyran-2-yl)oxy)nonadec-l 1-enoate
Figure imgf000053_0001
To a stirred solution of (11 -methoxy- 11 -oxoundecyl)triphenylphosphonium bromide (52 g, 96.4 mmol, 2.0 eq.) in dry THE (180 mL) at -40°C was added NaHMDS (41 mL, 82.0 mmol, 1.7 eq.) stirred for 1 hour while warming to 0°C. The reaction was then cooled to -78°C. 8-((tetrahydro-2H-pyran-2-yl)oxy)octanal (11 g, 48.2 mmol, 1.0 eq.) in 10 ml of dry THE was then added drop wise. The reaction was stirred at -78°C for 30 minutes and monitored by TLC.
The reaction mixture was quenched with water at -78°C and extracted with EtO Ac (2 x 200 mL). The total organic layer was dried over anhydrous NaaS04 and concentrated under reduced pressure to obtain the crude product, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 4%
EtO Ac/hexane to afford methyl (Z)- 19-((tetrahydro-2H-pyran-2-yl)oxy)nonadec- 11- enoate (6 g) as a colorless liquid. 'H-NMR (400 MHz, CDCI3): S 5.39-5.31 (m, 2H), 4.59-4.55 (m, 1H), 3.90-3.83 (m, 1H), 3.76-3.69 (m, 1H), 3.66 (s, 3H), 3.53-3.46 (m,
1H), 3.41-3.34 (m, 1H), 2.30 (t, J=7.2 Hz, 2H), 2.05-1.93 (m, 4H), 1.88-1.78 (m, 1H), 1.76-1.67 (m, 1H), 1.66-1.47 (m, 10H), 1.39-1.21 (m, 20H).
Step 6. methyl (Z)-19-hydroxynonadec-l 1-enoate
Figure imgf000053_0002
To a stirred solution of methyl (Z)-19-((tetrahydro-2H-pyran-2- yl)oxy)nonadec- 11 -enoate (6 g, 14.6 mmol, 1.0 eq.) in MeOH (150 mL) at 25°C was added /?-TSA (556 mg, 2.92 mmol, 0.2 eq.) The reaction was stirred for 16 hours and monitored by TLC. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (3 x 50 mL), The total organic layer was dried over anhydrous Na2SC>4 and concentrated under reduced pressure to obtain the crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 20% EtO Ac/hexane to afford methyl (Z)- 19-hydroxynonadec- 11 -enoate (3.8 g) as a colorless liquid. 'H-NMR (400 MHz, CDCb): δ 5.38-5.33 (m, 2H), 3.66 (s, 3H), 3.65- 3.61 (m, 2H), 2.30 (t, J=7.2 Hz, 2H), 2.06-1.93 (m, 4H), 1.65-1.53 (m, 5H), 1.39-1.23
(m, 20H).
Step 7. (Z)-l 9-hydroxynonadec- 11 -enoic acid
Figure imgf000054_0001
To a stirred solution of methyl (Z)-l 9-hydroxynonadec- 11 -enoate (3.8 g, 11.65 mmol, 1.0 eq.) in THF/H2O (3:1) (50 mL) at 0°C was added Li0H.5H20 (1.95 mg, 46.6 mmol, 4.0 eq.) and MeOH (15 mL). The reaction was stirred for 16 hours and monitored by TLC. The reaction mixture was acidified to pH 4 with saturated KHSO4 solution and extracted with EtOAc (3 x 50 mL). The total organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to afford crude (Z)- 19-hydroxynonadec- 11 -enoic acid (2.8 g) as a white color solid that was used directly for the next step without any further purification. 1 H-NMR (400 MHz, DMSO-De): δ 5.38-5.28 (m, 2H), 3.36 (t, 7=6.4 Hz, 2H), 3.33-3.27 (m, 2H), 2.17 (t, «7=7.6 Hz, 2H), 2.02-1.93 (m, 4H), 1.51-1.44 (m, 2H), 1.43-1.34 (m, 2H), 1.33-1.21
(m, 20H).
Step 8. (Z)-2-(l 8-hydroxyoctadec-10-en-l-yl)-5, 6-dimethoxy-3-methylcyclohexa-2,5- diene-1, 4-dione
Figure imgf000055_0001
To a stirred solution of (Z)- 19-hydroxynonadec- 11 -enoic acid (500 mg, 1.60 mmol, 1.0 eq.) and 2,3 -dimethoxy-5-methylcyclohexa-2, 5-diene- 1 ,4-dione (437 mg, 2.40 mmol, 1.5 eq.) in CH3CN/H2O (1 : 1) (20 mL) at 25°C was added AgNCb (342 mg, 2.10 mmol, 1.26 eq.). The reaction was heated to 70°C and K2S2O8 was added portion-wise. The reaction was stirred for 2 hours and monitored by TLC. The reaction mixture was poured into ice water and extracted with DCM (3 x 50 mL), The total organic layer was dried over anhydrous Na2SC>4 and concentrated under reduced pressure to afford crude, which was purified by silica gel chromatography (100-200 mesh) using gradient elution with 15% EtO Ac/hexane to afford (Z)-2-(18- hydroxyoctadec - 10-en- 1 -yl)- 5 ,6-dimethoxy-3 -methylcyclohexa-2 , 5 -diene- 1 ,4-dione (160 mg) as a light orange solid. Mass [m/z]: 449 [M+H]+.(LCMS Method A) 1 II- NMR (400 MHz, CDCb): δ 5.39-5.31 (m, 2H), 3.98 (s, 6H), 3.64 (t, .7=6.4 Hz, 2H), 2.44 (t, 7=6.8 Hz, 2H), 2.04-1.96 (m, 7H), 1.64-1.51 (m, 4H), 1.41-1.23 (m, 20H).
Intermediate 2. Methyl (Z)-15-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4- dien-l-yl)pentadec-9-enoate
Figure imgf000055_0002
Step 1. (9-methoxy-9-oxononyl)triphenylphosphonium bromide
Figure imgf000055_0003
Triphenylphosphine (39 g, 148.28 mmol, 1.5 eq.) was added to a stirred solution of methyl 9-bromononanoate (26 g, 98.85 mmol, 1.0 eq.) in acetonitrile (100 mL) at 25°C. The reaction mixture was stirred at 110°C for 48 hours and monitored by TLC. The reaction mixture was concentrated under vacuum to obtain the crude material. The crude product was triturated with hexane (100 mL) to afford (9- methoxy-9-oxononyl)triphenylphosphonium bromide (50 g) as a colorless liquid. Yield: 98%. JH-NMR (400 MHz, CDCh): δ 7.86-7.81 (m, 7H), 7.76-7.70 (m, 5H), 7.36-7.30 (m, 3H), 3.75-3.66 (m, 2H), 3.64 (s, 3H), 2.25 (t, 7=7.6 Hz, 2H), 2.01 (s, 2H), 1.63-1.62 (m, 2H), 1.56-1.53 (m, 2H), 1.31-1.22 (m, 6H).
Step 2. 7-((tert-butyldiphenylsilyl)oxy)heptan-l-ol
TBDPSO OH
To a stirred solution of NaH (4.5 g, 0.114 mol, 1.0 eq.) in THF (200 mL) at 25°C was added heptane- 1 ,7-diol (15 g, 0.113 mol, 1.0 eq.) dissolved in THF (100 mL). The reaction was stirred for one hour and a dissolved solution of tert- Butyl(chloro)diphenylsilane (TBDPSC1, 29.5 mL, 0.113 mol, 1.0 eq.) in THF (100 mL) was added. The reaction mixture was stirred at 25 °C for 48 hours and monitored by TLC. The reaction mixture was quenched with ice water (50 mL) and extracted with EtO Ac (3 x 200 mL). The combined organic layers were concentrated under vacuum to obtain the crude material. The crude product was purified by silica gel chromatography (60-120 mesh) using 15% EtO Ac/hexane to furnish 7-((tert- butyldiphenylsilyl)oxy)heptan- 1 -ol (117 g) as a colorless liquid. Yield: 40%. 'H-
NMR (400 MHz, CDC13): δ 7.67-7.65 (m, 4H), 7.43-7.34 (m, 6H), 3.66-3.60 (m, 4H), 1.59-1.50 (m, 4H), 1.39-1.25 (m, 7H), 1.06 (s, 9H).
Step 3. 7-((tert-butyldiphenylsilyl)oxy)heptanal
TBDPSO Ό
Pyridinium chlorochromate (PCC) (8.7 g, 40.43 mmol, 1.5 eq.) and Celite (10 g) mixture was added to a stirred solution of 7-((tert-butyldiphenylsilyl)oxy)heptan- 1 - ol (10 g, 26.95 mmol, 1.0 eq.) in DCM (200 mL) at 0°C. The reaction mixture was stirred at 25°C for 2 hours and monitored by TLC. The reaction mixture was filtered through Celite pad and washed with DCM (50 mL). The filtrate was dried over anhydrous NazSCL and concentrated under vacuum to obtain the crude material. The crude product was purified by silica gel chromatography (60-120 mesh) using 10% EtO Ac/hexane to furnish 7-((tert-butyldiphenylsilyl)oxy)heptanal (8.1 g) as a colorless liquid. Yield: 81%. 'H-NMR (400 MHz, CDCb): S 9.75 (s,lH), 7.67-7.64 (m, 4H), 7.44-7.35 (m, 6H), 3.65 (t, 7=6.4 Hz, 2H), 2.41-2.37 (m, 2H), 1.64-1.52 (m,
4H), 1.39-1.28 (m, 5H), 1.07 (s, 9H). Step 4. Methyl (Z)-16-((tert-butyldiphenylsilyl)oxy)hexadec-9-enoate
Figure imgf000057_0001
NaHMDS (46.7 mL, 93.54 mmol, 1.7 eq.) was added to a stirred solution of (9-methoxy-9-oxononyl)triphenylphosphonium bromide (58 g, 110.05 mmol, 2.0 eq.) in anhydrous THF (200 mL) at -40°C and the resulting mixture was slowly warmed for 0°C for 1 hour. The reaction mixture was cooled to -78°C and 7-((tert- butyldiphenylsilyl)oxy)heptanal (20 g, 55.02 mmol, 1.0 eq.) dissolved in 30 mL of dry THF was added dropwise and stirred for 30 minutes at -78°C. The reaction was monitored by TLC. The reaction mixture was quenched with water at -78°C and extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous Na2S04 and concentrated under vacuum to obtain the crude material. The crude product was purified by silica gel chromatography (60-120 mesh) using 2% EtOAc/hexane to furnish methyl (Z)-16-((tert-butyldiphenylsilyl)oxy)hexadec-9- enoate (16 g) as a colorless liquid. Yield: 55%. 'H-NMR (400 MHz, CDCI3): ό 7.67- 7.65 (m, 4H), 7.43-7.38 (m, 10H), 5.33 (t, 7=4.4 Hz, 2H), 5.29 (s, 1H), 3.66-3.63 (m,
5H), 2.30 (t, 7=3.6 Hz, 2H), 2.00-1.97 (m, 3H), 1.63-1.52 (m, 3H), 1.42-1.26 (m, 14H), 1.05 (s, 9H).
Step 5. Methyl (Z)-16-hydroxyhexadec-9-enoate
Figure imgf000057_0002
To a stirred solution of methyl (Z)- 16-((tert-butyldiphenylsilyl)oxy)hexadec-9- enoate (16 g, 29.96 mmol, 1.0 eq.) in anhydrous THF (130 mL) at 20°C was added TBAF [1M in THF] (38.9 mL, 38.95 mmol, 1.3 eq.) dropwise. The reaction was stirred at 25°C for 2 hours and monitored by TLC. The reaction mixture was quenched with water (40 mL) and extracted with EtOAc (3 x 70 mL). The combined organic layers were dried over anhydrous NaaStAi and concentrated under vacuum to obtain the crude material. The crude product was purified by silica gel chromatography (60-120 mesh) using 20% EtOAc/hexane to furnish methyl (Z)-16- hydroxyhexadec-9-enoate (4.5 g) as a colorless liquid. Yield: 51%. *H-NMR (400 MHz, CDCb): δ 5.35-5.33 (m, 2H), 3.66 (s, 3H), 3.64 (t, <7=6.4 Hz, 2H,), 2.30 (t, .7=7.6 Hz, 2H), 2.04-2.00 (m, 4H), 1.61-1.55 (m, 4H), 1.34-1.25 (m, 14H).
Step 6. (Z)-l 6-methoxy- 16-oxohexadec-7 -enoic acid
Figure imgf000058_0001
CrOg (417 mg, 4.22 mmol, 2.4 eq.) was added to a stirred solution of methyl (Z)-16-hydroxyhexadec-9-enoate (500 mg, 1.76 mmol, 1.0 eq.) in acetone (10 mL) and acetic acid (5 mL) at 0°C. The reaction was stirred for 2 hours at 25°C and monitored by TLC. The reaction mixture was quenched with sat. Na2S20s solution (50 mL) and stirred for 18 hours at 25 °C. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (2 x 10 mL). The combined organic layers were dried over anhydrous Na2SC>4 and concentrated under vacuum to obtain crude, which was purified by silica gel column chromatography (60-120 mesh) using gradient elution with 15-20% EtO Ac/Hexane to afford (Z)- 16-methoxy- 16- oxohexadec-7 -enoic acid (220 mg) as a colorless liquid. Yield: 42%. Ή-ΝΜΚ (400 MHz, CDClg): δ 5.38-5.29 (m, 2H), 3.67 (t, J=10.4, Hz, 3H), 2.37 (m, 4H), 2.03-1.98 (m, 4H), 1.68-1.60 (m, 4H), 1.40-1.32 (m, 12H).
Step 7. (Z)-16-methoxy-16-oxohexadec-7-enoic acid
Figure imgf000058_0002
AgNOg (359 mg, 2.11 mmol, 1.26 eq.) was added to a stirred solution of (Z)- 16-methoxy- 16-oxohexadec-7-enoic acid (500 mg, 1.68 mmol, 1.0 eq.) and 2,3- dimethoxy-5-methylcyclohexa-2, 5-diene- 1 ,4-dione (458 mg, 2.51 mmol, 1.5 eq.) in CH3CN/H2O (1:1) (40 mL) at 25°C. The reaction temperature was slowly raised to 75°C and was added a solution of K2S2O8 (1.36 g, 5.03 mmol, 3.0 eq.) dissolved in water (20 mL). The reaction was stirred at 75°C for 2 hours and monitored by TLC. The reaction mixture was poured into ice water and extracted with DCM (3 x 25 mL). The combined organic layers were dried over anhydrous Na2SCL and concentrated under reduced pressure to obtain crude, which was purified by silica gel column chromatography (60-120 mesh) using gradient elution with 90-95% DCM/hexane to afford title compound (150 mg) as an orange liquid. Yield: 21%. 'H-NMR (400 MHz, CDCb): δ 5.37-5.32 (m, 2H), 3.99 (s, 6H), 3.66 (s, 3H), 2.47-2.43 (m, 2H), 2.30 (t, J =7.2 Hz, 2H), 2.04-1.96 (m, 7H), 1.62 (t, J= 6.4 Hz, 2H), 1.42-1.24 (m, 14H).
Example 1. (Z)-2-((((18-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4-dien-l- yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl (2-(trimethylammonio)ethyl) phosphate (Compound 1)
Figure imgf000059_0001
Step 1. benzyl (2-hydroxyethyl)carbamate
Figure imgf000059_0003
To a stirred solution of 2-aminoethan- 1 -ol (50 g, 0.819 mol, 1.0 eq.) in DCM (1.5 L) at 0°C was added EtsN (137 mL, 0.983 mol, 1.2 eq.) dropwise. The reaction was stirred for 10 minutes at 0°C and benzyl carbonochloridate (Cbz-Cl; 50 %; 302 mL, 1.064 mol, 1.2 eq) was added drop wise. The reaction was stirred for 3 hours at 0°C and monitored by TLC. The reaction mixture was quenched with water (500 mL) and extracted with DCM (3 X 500 mL). The total organic layer was dried over anhydrous NaiSCL and concentrated under reduced pressure to obtain crude compound, which was purified by silica gel column chromatography (60-120 mesh) using gradient elution with 3% MeOH/DCM to afford benzyl (2- hydroxyethyl)carbamate (95 g) as a white solid. Mass [m/z]: 196.09 [M+H]+.
Step 2. 2-(( (benzyloxy)carbonyl)amino)ethyl(2- ( trimethylammonio)ethyl)phosphate
Figure imgf000059_0002
To a stirred solution of benzyl (2-hydroxyethyl)carbamate (5 g, 25.641 mmol,
1 eq.) in chloroform (100 mL) was added EtgN (5.5 mL, 38.461 mmol, 1.5 eq.) followed by POCI3 (2.65 mL, 28.205 mmol, 1.1 eq.) at -10°C. Then the reaction mixture was stirred for 1 hour at 25 °C and monitored by TLC. After 1 hour, pyridine (17.5 mL, 220.512 mmol, 8.6 eq.) and choline tosylate (10.55 g, 38.461 mmol, 1.5 eq) were added at -10 °C and the resulting mixture was stirred at 25°C for 18 hours. The reaction mass was then cooled to 0°C, quenched with water (20 mL), and extracted with DCM (3 X 100 mL). The aqueous layer was purified by flash chromatography [Column: Biotage-C18, 60 g Duo-100 A 30 pm; Mobile phase: [ACN: Water + 0.1% TFA]; B%: B%: 0-8%, 0-20min / 8% 20-45 min.] to afford 2-
(((benzyloxy)carbonyl)amino)ethyl(2-(trimethylammonio)ethyl)phosphate (2.4 g) as a colorless liquid. Mass [m/z]: 361.01 [M+H]+.
Step 3. 2-aminoethyl(2-(trimethylammonio)ethyl)phosphate
Figure imgf000060_0001
To a stirred solution of 2-(((benzyloxy)carbonyl)amino)ethyl(2- (trimethylammonio)ethyl)phosphate (2.3 g, 0.638 mmol, 1.0 eq.) in isopropyl alcohol (20 mL) at 25 °C was added 10% Pd/C (50% wet; 500 mg). The reaction was stirred under hydrogen pressure at 25°C for 18 hours. The resulting mixture was filtered through a Celite pad and washed with IPA. The filtrate was concentrated under reduced pressure to afford 2-aminoethyl(2-(trimethylammonio)ethyl)phosphate (1.6 g) as a colorless liquid that was used without further purification. Mass [m/z]: 227.5 [M+H]+.
Step 4. (Z)-2-(( ((18-(4, 5 -dimethoxy-2 -methyl-3, 6-dioxocyclohexa- 1 , 4-dien-l- yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl (2-(trimethylammonio)ethyl) phosphate
To a stirred solution of (Z)-2-(l 8-hydroxyoctadec- 10-en- 1 -yl)-5,6-dimethoxy- 3-methylcyclohexa-2, 5-diene- 1 ,4-dione (20 mg, 0.044 mmol, 1.0 eq.) in DCM (5 mL) at 25°C was added pyridine (7 pL, 0.089 mmol, 2.0 eq.) and 4-nitro phenyl chloro formate (10.7 mg, 0.053 mmol, 1.2 eq.). The reaction was stirred at 25°C for 6 hours and was concentrated under reduced pressure to dryness. The crude residue was dissolved in DMF (2 mL) and was added DMAP (7 mg, 0.057 mmol, 1.3 eq.) and followed by 2-aminoethyl(2-(trimethylammonio)ethyl)phosphate (10 mg, 0.044 mmol, 1.0 eq.) at 25°C and stirred for 18 hours. The reaction mixture was diluted with isopropyl ether (10 mL) and the solvent was decanted to obtain crude, which was purified according to PREP-HPLC Method- A: Gemini-C18, 250 X 21.2 mm, 5.0 pm; Buffer: [Acetonitrile, 0.1% of formic acid in water]; time/B%: 0/30, 10/60, 30/90 to afford title compound as a light brown semi solid. (2.4 mg) Mass [m/z]: 702.5 [M+H]+. (LCMS Method B) 'H-NMR (400 MHz, DMSO-D6): δ 5.38-5.27 (m, 2H), 4.14 (t, 5.2 Hz, 2H), 4.08-3.98 (m, 4H), 3.87 (s, 6H), 3.82-3.75 (m, 2H), 3.52-3.47
(m, 2H), 3.12 (s, 9H), 3.41-3.34 (m, 2H), 2.01-1.94 (m, 4H), 1.92 (s, 3H), 1.61-1.54 (m, 2H), 1.35-1.19 (m, 22H). 31P-NMR (400 MHz, DMSO-De): δ -1.09.
Example 2. (Z)-2-((((18-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4-dien-l- yl)octadec-8-en-l-yl)oxy)carbonyl)oxy)ethyl (2-(trimethylammonio)ethyl) phosphate (Compound 2)
Figure imgf000061_0001
Step 1. 2-(benzyloxy)ethan-l-ol
Figure imgf000061_0002
To a stirred solution of NaH (1.93 g, 80.645 mmol, 1.0 eq.) in THF (50 mL) at 0°C was added ethylene glycol (5 g, 80.645 mmol, 1.0 eq.) and catalytic amount of TBAI (100 mg). The reaction was stirred for 10 minutes at 0°C and then warmed to RT over 1 hour. After 1 hour, benzyl bromide (13.7mL, 80.645 mmol, 1.0 eq.) was added drop wise to the reaction mixture at 0°C. The reaction was stirred at 25 °C for 18 hours and monitored by TLC. The reaction mixture was quenched with cold water (250 mL) and extracted with EtOAc (3 x 250 mL). The total organic layer was dried over anhydrous Na2S04 and concentrated under reduced pressure to obtain crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 30% EtOAc/Hexane to afford 2-(benzyloxy)ethan- 1 -ol (2.5 g) as a pale- yellow oil. 1H NMR (CDCb): δ 7.38-7.30 (m, 5H), 4.56 (s, 2H), 3.78-3.74 (m, 2H), 3.61-3.59 (m, 2H), 2.06 (t, J= 6.4 Hz, 1H).
Step 2. 2-(benzyloxy) ethyl (2-(trimethylammonio)ethyl) phosphate
Figure imgf000062_0002
To a stirred solution of 2-(benzyloxy)ethan- 1 -ol (1 g, 6.535 mmol, 1 eq.) in chloroform (10 mL) at -10°C was added EtgN (1.18 mL, 8.169 mmol, 1.25 eq.) and POCI3 (0.78 mL, 8.169 mmol, 1.25 eq.). The reaction mixture was stirred for 1 hour and monitored by TLC. After 1 hour, pyridine (2.5 mL, 56.208 mmol, 8.6 eq.) was added, followed by choline tosylate (2.6 g, 9.803 mmol, 1.5 eq) at -10°C and the reaction was stirred at 25 °C for 18 hours. The reaction mixture was cooled to 0°C and quenched with water (5 mL) and extracted with DCM (3 x 50 mL). The aqueous layer was purified from flash chromatography [Method : Column: Biotage-C18, 60 g Duo- 100 A 30 pm; Mobile phase: [ACN: Water + 0.1% TFA]; B%: B%: 0-8%, 0-20min / 8% 20-45 min.], pure fractions were lyophilized to afford 2-(benzyloxy)ethyl (2-
(trimethylammonio)ethyl) phosphate (1.1 g) as a colorless liquid. 'H NMR (CDCI3): δ 7.34-7.28 (m, 5H), 4.46 (s, 2H), 4.25 (bs, 2H), 4.04 (bs, 2H), 3.64 (bs, 2H), 3.52 (bs, 2H), 3.03 (s, 9H). 31P NMR: δ -2.23.
Step 3. 2 -hydroxy ethyl (2-(trimethylammonio)ethyl) phosphate
Figure imgf000062_0001
To a stirred solution of 2-(benzyloxy)ethyl (2-(trimethylammonio)ethyl) phosphate (300 mg, 0.946 mmol, 1.0 eq.) in EP A (3 mL) was added 10% Pd/C (50% wet, 50 mg) at 25°C. The reaction mixture was stirred under hydrogen pressure at 25°C for 18 hours and monitored by LCMS. The reaction mixture was filtered through Celite pad and washed with IPA. The filtrate was concentrated under reduced pressure to afford 2 -hydroxyethyl (2 -(trimethylammonio)ethyl) phosphate (200 mg) as a colorless liquid. Mass [m/z]: 227.5 [M+H]+ (LCMS Method C) 1 H NMR (CD3OD): 4.32-4.28 (m, 2H), 3.96-3.92 (m, 2H), 3.70 (t, J= 4.8 Hz, 2H), 3.65-3.63 (m, 2H), 3.22 (s, 9H). 31P NMR: δ -0.17. Step 4. (Z)-2-(( ((18-(4, 5 -dimethoxy-2 -methyl-3, 6-dioxocyclohexa-l , 4-dien-l- yl)octadec-8-en-l-yl)oxy)carbonyl)oxy)ethyl (2-(trimethylammonio)ethyl) phosphate To a stirred solution of (Z)-2-(l 8-hydroxyoctadec- 10-en- 1 -yl)-5,6-dimethoxy- 3-methylcyclohexa-2, 5-diene- 1 ,4-dione (100 mg, 0.223 mmol, 1.0 eq.) in DCM (5 ml) at 25°C was added pyridine (35 μΤ, 0.446 mmol, 2.0 eq.) and 4-nitrophenyl chloro formate (53.8 mg, 0.267 mmol, 1.2 eq.). The reaction mixture was stirred for 6 hours and monitored by TLC. The reaction mixture was concentrated under reduced pressure and crude was dissolved in DMF (5 mL). DMAP (35.4 mg, 0.290 mmol, 1.3 eq.) was added, followed by 2-hydroxyethyl (2-(trimethylammonio)ethyl) phosphate (50.6 mg, 0.290 mmol, 1.0 eq.) at 25°C and left it to stir for 18 hours. The reaction mixture was diluted with isopropyl ether (10 mL) and the solvent was decanted to obtain crude, which was further purified according to PREP-HPLC Method-B: Gemini-C18, 250 X 21.2 mm, 5.0 pm; Buffer: [Acetonitrile, 0.1% of formic acid in water], time/B%: 0/30, 10/60, 20/80, 30/90 to afford title compound as a light brown semi solid. Mass [m/z]: 702.5 [M+H]+ (LCMS Method D) 'H-NMR (400 MHz, DMSO-De): S 5.38-5.27 (m, 2H), 4.14 (t, J= 5.2 Hz, 2H), 4.08-3.98 (m, 4H), 3.87 (s, 6H), 3.82-3.75 (m, 2H), 3.52-3.47 (m, 2H), 3.12 (s, 9H), 3.41-3.34 (m, 2H), 2.01-1.94 (m, 4H), 1.92 (s, 3H), 1.61-1.54 (m, 2H), 1.35-1.19 (m, 22H). 31P-NMR (400 MHz, DMSO-De): -1.09.
Example 3. 2-((((10-(4,5-dimethoxy-2-methyl-3, 6-dioxocyclohexa-l, 4-dien-l- yl)decyl)oxy)carbonyl)amino)ethyl (2-(trimethylammonio)ethyl) phosphate (Compound 40)
Figure imgf000063_0001
Pyridine (0.23 mL, 0.290 mmol, 2.0 eq.) and 4-nitrophenyl chloroformate (36 mg, 0.174 mmol, 1.2 eq.) were added to a stirred solution of idebenone (50 mg, 0.145 mmol, 1.0 eq.) in DCM (2.0 mL) at 25°C. The reaction was stirred at 25°C for 6 hours and monitored by TLC. Following the formation of the nitrophenyl intermediate, the reaction was concentrated under reduced pressure to obtain crude residue, which was subsequently dissolved in DMF (4.0 mL).
DMAP (53 mg, 0.435 mmol, 3.0 eq.) and 2-aminoethyl(2 - (trimethylammonio)ethyl)phosphate (dissolved in 1 mL of DMF) (65 mg, 0.290 mmol, 2.0 eq.) were added to the reaction mixture at 25°C. The reaction was further stirred at 25°C for 18 hours and monitored by LC-MS. The reaction mixture was triturated with isopropyl ether (IPE) (20 mL) and was concentrated under reduced pressure to obtain crude, which was purified according to PREP-HPLC Method-B: Kinetex EVO, C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [0.1% of FA in Water: ACN]; time/ B%: 0/15, 20/40, 30/65 to afford title compound (10 mg) as a brown semi solid. Yield: 11.4%. Mass [m/z]: 591.2 [M+H]+ (LCMS Method E) 'H-NMR (400 MHz, DMSO-de): δ 7.48 (t, J = 5.2 Hz, 1H), 4.02 (bs, 2H), 3.91-3.87 (m, 8H), 3.66-3.62 (m, 2H), 3.51-3.48 (m, 2H), 3.13-3.08 (m, 11H), 2.36 (t, J= 7.6 Hz, 2H), 1.93 (s, 3H), 1.55-1.48 (m, 2H), 1.31-1.22 (m, 14H). 31P-NMR (162 MHz, DMSO- de): δ -0.31.
Example 4. (Z)-2-ammonioethyl (2-((((18-(4,5-dimethoxy-2-methyl-3,6- dioxocyclohexa-l,4-dien-l-yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl) phosphate (Compound 41)
Figure imgf000064_0002
Step 1. 2(((benzyloxy) carbonyl)amino) ethyl(2 ((tertbutoxycarbonyl) amino)ethyl) phosphate
Figure imgf000064_0001
To a stirred solution of benzyl (2-hydroxyethyl)carbamate (1 g, 5.128 mmol, 1 eq.) in chloroform (20 mL) was added EtgN (1.1 mL, 7.692 mmol, 1.5 eq.) followed by POCh (0.52 mL, 5.641 mmol, 1.1 eq.) at -10°C. The reaction mixture was then stirred for 1 hour at 25 °C, followed by the addition of pyridine (3.6 mL, 44.102 mmol, 8.6 eq.) and N-Boc ethanol amine (1.23 g, 7.692 mmol, 1.5 eq.) at -10°C. The reaction mixture was stirred for 18 hours at 25°C. The mixture was then cooled to 0°C, water was added (20 mL), and the mixture was extracted with dichloromethane (DCM; 3 x 30 mL). The organic layer was concentrated and purified by flash chromatography Column: Biotage-C18, 30g Duo-100 A 30 pm; Mobile phase: ACN: Water + 0.015% NH4HCO3; min/B%: 0/0, 5/5, 30/5, 35/14, 55/14, 58/100. The pure fractions were lyophilized to afford the 2(((benzyloxy) carbonyl)amino) ethyl(2((tertbutoxycarbonyl) amino)ethyl) phosphate (200 mg) as off-white solid. [M- H]+ (m/z): 417.2; 'H-NMR (400 MHz, DMSO-d6): S 7.69 (hr s, 1H), 7.38-7.27 (m, 5H), 7.09 (hr s, 1H), 4.99 (s, 2H), 3.72-3.57 (m, 4H), 3.14-3.10 (m, 2H), 3.05-3.01 (m, 2H), 1.36 (9H). 31P NMR (400 MHz, DMSO-d6): 0.14.
Step 2. 2-aminoethyl (2-((tert-butoxycarbonyl)amino)ethyl) phosphate
Figure imgf000065_0001
10% Pd/C (50% wet) (15 mg) was added to a stirred solution of 2(((benzyloxy) carbonyl)amino) ethyl(2((tertbutoxycarbonyl) amino) ethyl) phosphate (50 mg, 0.119 mmol, 1.0 eq.) in isopropanol (IP A; 2 mL) at 25°C. The reaction mixture was stirred under hydrogen (about 1 atm) at 25 °C for 2 hours and was filtered through C elite pad, and washed with IP A (5 mL). The filtrate was concentrated and dried under vacuum to afford 2-aminoethyl (2 -((tert-butoxycarbonyl) amino) ethyl) phosphate (40 mg, crude) as off-white solid, which was used for the next step without purification. Mass [m/z]: 285.2 [M+H]+. 'H-NMR (400 MHz, DMSO-de): S 8.36 (br s, 2H), 6.96 (br s, 1H), 3.87-3.79 (m, 2H), 3.68-3.60 (m, 2H), 3.10-3.02 (m, 2H), 2.97-2.92 (m, 2H), 1.37 (s, 9H). 31P NMR (400 MHz, DMSO-de): 0.82.
Step 3. (Z)-18-(4, 5-dimethoxy-2-methyl-3, 6-dioxocyclohexa-l ,4-dien-l -yl)octadec-8- en-l-yl (2-(((2-((tert-butoxycarbonyl)amino)ethoxy)(hydroxy)phosphoryl) oxy)ethyl)carbamate
Figure imgf000065_0002
Pyridine (36 pL, 0.446 mmol, 2.0 eq.) and 4-nitro phenyl chloroformate (54 mg, 0.256 mmol, 1.2 eq.) were added to a stirred solution of (Z)-2-(18- hydroxyoctadec - 10-en- 1 -yl)- 5 ,6-dimethoxy-3 -methylcyclohexa-2 , 5 -diene- 1 ,4-dione (100 mg, 0.223 mmol, 1.0 eq.) in 10 mL of DCM at 25°C. The reaction was stirred at 25°C for 6 hours and monitored by TLC. The reaction was concentrated under reduced pressure to dryness. 2-aminoethyl (2-((tert-butoxycarbonyl)amino)ethyl) phosphate (95 mg, 0.334 mmol, 1.5 eq.) and DMAP (76 mg, 0.624 mmol, 2.8 eq.) were added to a stirred solution of the residue dissolved in DMF (5 mL) at 25°C. The reaction was stirred at 25 °C for 18 hours. The reaction mixture was concentrated under reduced pressure to obtain crude (Z)-18-(4,5-dimethoxy-2 -methyl-3, 6- dioxocyclohexa- 1 ,4-dien- 1 -yl)octadec-8-en- 1 -yl (2-(((2-((tert- butoxycarbonyl)amino)ethoxy)(hydroxy)phosphoryl) oxy)ethyl)carbamate (255 mg) as a light brown liquid. The crude was used for the next step without further purification.
Step 4. (Z)-2-ammonioethyl (2-((((18-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa- l,4-dien-l-yl)octadec-8-en-l-yl)oxy)carbonyl)amino)ethyl) phosphate
TFA (0.2 ml) was added to a stirred solution of (Z)-18-(4,5-dimethoxy-2- methyl-3,6-dioxocyclohexa- 1 ,4-dien- 1 -yl)octadec-8-en- 1 -yl (2-(((2-((tert- butoxycarbonyl)amino)ethoxy)(hydroxy)phosphoryl) oxy)ethyl)carbamate (255 mg, 0.336 mmol, 1.0 eq.) in DCM (10 mL) at 0°C. The reaction mixture was stirred at 25°C for 18 hours and monitored by LC-MS. The reaction mixture was concentrated under reduced pressure to obtain crude, which was purified by PREP-HPLC Method- C: Kinetix Evo C18, 250 x 21.2 mm, 5.0 pm; Buffer: [Acetonitrile, 0.1% of formic acid in water]; time/B%: 0/30, 10/60, 30/90 to afford title compound (54 mg) as a yellow solid. Yield: 24%. Mass [m/z]: 659.4 [M+H]+ (LCMS Method F) 'H-NMR (400 MHz, DMSO-De): δ 8.37 (s, 2H), 7.25 (t, J=4.8 Hz, 1H), 5.37-5.29 (m, 2H),
3.93-3.82 (m, 10H), 3.66 (q, J=7.6 Hz, 2H), 3.12 (q, J=5.6 Hz, 2H), 2.95 (s, 2H), 2.39-2.35 (m, 2H), 2.02-1.94 (m, 4H), 1.93 (s, 3H), 1.56-1.47 (m, 2H), 1.34-1.19 (m, 22H). 31P-NMR (162 MHz, DMSO-D6): δ 0.66. Example 5. (Z)-3-((15-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4-dien-l- yl)pentadec-9-enoyl)oxy)-4-(trimethylammonio)butanoate (Compound 42)
Figure imgf000067_0001
Step 1. (Z)-15-(4, 5-dimethoxy-2-methyl-3, 6-dioxocyclohexa- 1 ,4-dien-l -yl)pentadec-9- enoic acid
Figure imgf000067_0002
2N HC1 (4.0 ml, 3 vol.) was added to a stirred solution of methyl (Z)-15-(4,5- dimethoxy-2-methyl-3 , 6-dioxocyclohexa- 1 ,4-dien- 1 -yl)pentadec-9-enoate (150 mg, 1.13 mmol, 1.0 eq.) in 1,4-dioxane (8 mL) at 0°C. The reaction was stirred at 40°C for 18 hours and monitored by TLC. The reaction mixture was concentrated under reduced pressure, diluted with water (5 mL) and extracted with EtOAc (2 x 25 mL). The combined organic layers were dried over anhydrous NaaSCL and concentrated under reduced pressure to obtain crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 20-25% EtOAc: Hexane to afford (Z)- 15-(4,5-dimethoxy-2-methyl-3 , 6-dioxocyclohexa- 1 ,4-dien- 1 -yl)pentadec- 9-enoic acid (120 mg) as an orange semi solid. Yield: 83%. *H-NMR (400 MHz, CDCh): δ 5.37-5.32 (m, 2H), 3.99 (s, 6H), 2.45-2.42 (m, 2H), 2.35 (t, J =7.6 Hz, 2H), 2.04-1.96 (m, 7H), 1.64 (t, J =6.4 Hz, 2H), 1.44-1.28 (m, 14H).
Step 2. 2,5-dioxopyrrolidin-l-yl (Z)-15-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa- l,4-dien-l-yl)pentadec-9-enoate
Figure imgf000067_0003
N -Hydroxysuccinimide (NHS) (45 mg, 0.392 mmol, 1.1 eq.) and Ν,Ν'- Dicyclohexylcarbodiimide (DCC) (80 mg, 0.392 mmol, 1.1 eq.) were added to a stirred solution of (Z)- 15-(4,5-dimethoxy-2-methyl-3 , 6-dioxocyclohexa- 1 ,4-dien- 1 - yl)pentadec -9 -enoic acid (150 mg, 0.357 mmol, 1.0 eq.) in ethyl acetate (3.0 mL) at 25°C. The reaction mixture was stirred at the 25°C for 18 hours and monitored by
TLC. The reaction mixture was filtered through celite pad and the filtrate was concentrated under reduced pressure to obtain crude, which was purified by silica gel chromatography (60-120 mesh) using gradient elution with 20-25% EtOAc:Hexane to afford 2,5-dioxopyrrolidin-l-yl (Z)-15-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa- 1 ,4-dien- 1 -yl)pentadec-9-enoate (170 mg) as an orange semi solid. Yield: 92%. *H- NMR (400 MHz, CDCb): δ 5.37-5.34 (m, 2H), 3.98 (s, 6H), 2.84 (bs, 4H), 2.60 (t, J= 6.4 Hz, 2H), 2.48-2.42 (m, 2H), 2.17-2.00 (m, 7H), 1.78-1.70 (m, 2H), 1.42-1.32 (m, 14H).
Step 3. (Z)-3-((l 5-(4,5-dimethoxy-2-methyl-3 ,6-dioxocyclohexa-l ,4-dien- 1 - yl)pentadec-9-enoyl)oxy)-4-(trimethylammonio)butanoate
EtsN (0.011 mL, 0.077 mmol, 2.0 eq.) and L-Camitine HC1 (12.3 mg, 0.077 mmol, 2.0 eq.) were added to a stirred solution of 2,5-dioxopyrrolidin-l-yl (Z)-15- (4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4-dien-l-yl)pentadec-9-enoate (20 mg, 0.038 mmol, 1.0 eq.) in DMF (2.0 mL) at 25°C. The reaction was stirred at 25°C for 18 hours and monitored by TLC and LC-MS. The reaction mixture was triturated with isopropyl ether (IPE) (2 x 5 mL) and dried under vacuum to obtain crude, which was purified by PREP -HP LC Method-D: Gemini-C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [ACN: Water]; B%: 0/10, 10/50, 20/60, 25/95 to afford title compound (1.8 mg) as a brown semi solid. Yield: 8.5%. Mass [m/z]: 564.4 [M+H]+ (LCMS Method G) ¾-NMR (400 MHz, DMSO-de): δ 5.38-5.30 (m, 3H), 3.91-3.59 (m, 8H), 3.07 (s, 9H), 2.37-2.34 (m, 2H), 2.33-2.23 (m, 4H), 2.05-1.92 (m, 7H), 1.54-1.40 (m, 2H), 1.32-1.22 (m, 14H).
Example 6. (Z)-2-(15-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-l,4-dien-l- yl)pentadec-9-enamido)ethyl(2-(trimethylammonio)ethyl)phosphate (Compound
43)
Figure imgf000068_0001
TEA (0.04 mL, 0.301 mmol, 2.0 eq.) and 2-aminoethyl(2- (trimethylammonio)ethyl)phosphate (50.8 mg, 0.225 mmol, 1.5 eq.) were added to a stirred solution of (Z)- 15-(4,5-dimethoxy-2-methyl-3 ,6-dioxocyclohexa- 1 ,4-dien- 1 - yl)pentadec-9-enoate (80 mg, 0.150 mmol, 1.0 eq.) in DMF (3.0 mL) at 25°C. The reaction mixture was stirred at 25 °C for 18 hours and monitored by TLC and LC-MS. The reaction mixture was triturated with isopropyl ether (IPE) (2 x 5 mL) and was dried under vacuum to obtain crude, which was purified by PREP-HPLC Method-D: Gemini-C18 X 250 X 21.2 mm, 5 pm; Mobile phase: [ACN: Water]; B%: 0/10, 10/50, 20/60, 25/95 to afford title compound (14 mg) as a yellow solid. Yield: 14.4%. Mass [m/z]: 629.2 [M+H]+ (LCMS Method H) 'H-NMR (400 MHz, DMSO-d6): δ 8.29 (t, J =5.2 Hz, 1H), 5.37-5.29 (m, 2H), 4.05-4.01 (m, 2H), 3.87 (s, 3H), 3.86 (s, 3H), 3.66- 3.61 (m, 2H), 3.52-3.48 (m, 2H), 3.16-3.10 (m, 11H), 2.36 (t, J= 7.2 Hz, 2H), 2.03- 1.92 (m, 9H), 1.46 (t, J= 7.2 Hz, 2H), 1.35-1.18 (m, 14H). 31P-NMR (162 MHz,
DMSO-de): δ -0.19.
It will be appreciated that all other compounds disclosed in the present application can be prepared according to the synthetic routes or procedures described in Examples 1-6 and/or Schemes 1-9, using appropriately substituted materials.
Example 7. In vitro Mfsdla Transport (Compounds 1, 2, 40, 42, and 43)
The following assay was used for analysis of Compounds 1. 2. 40. 42. and 43.
Methodology
The assay was conducted in a low throughput 6-well format with HEK293 cells prepared and then transfected in duplicate wells with plasmids containing either the wildtype (WT) version of hMfsd2a, the D97A mutant version, or an empty vector (EV) as control. Uptake into the cells was assessed by both thin-layer chromatography (TLC) and ultrahigh performance liquid chromatography hyphenated mass spectrometry.
Cell Transfection
HEK293 cells were seeded at 6.25 x 10s per 6-well in 2 mL of DMEM with 10% FBS and 1% penicillin-streptomycin (P/S) media (Sigma) and incubated overnight at 37°C in 5% C02. Cells were checked for confluency the next morning. On a per well basis the following lipid mix was generated; 6 pL of Lipofectamine
2000 was added dropwise to 200 pL of OptiMEM, this was left to stand for 5 minutes at room temperature (RT). One pg of hMfsd2a WT, D97A or empty plasmid was prepared in 200 pL of OptiMEM as appropriate for each well; the Lipofectamine 2000 in OptiMEM solution was then added dropwise to a total volume of 400 pL (this can be scaled to support the number of wells/plates to be assayed). This transfection preparation was then incubated at RT for 20 minutes. DMEM with 10% FBS and no P/S media was warmed to 37°C, the HEK293 plate media was changed and the cells washed carefully with 1 mL of the warmed DMEM with 10% FBS no P/S media, 1.6 mL of the warmed DMEM with 10% FBS no P/S media was then added to each well. Four hundred pL of the transfection preparation was then added dropwise to each well as appropriate and the plate was gently swirled in a circular motion. The plate was then incubated overnight at 37°C in 5% C02.
Compound Incubation and Preparation of Analysis Samples
Compound stock solutions were prepared in a 12% BSA in PBS solution such that a 40 pL spike into 2 mL of plain DMEM would yield a concentration of 50 pM of test compound (the compound treated media). Remaining compound stock solution in 12% BSA in PBS was frozen at -20°C to allow for media stability testing. The HEK293 6-well plate was removed from the incubator and the wells gently rinsed with 1 mL of plain DMEM that had been prewarmed to 37°C. 2 mL of the compound treated media was then added to each well.
A 100 pL sample of the compound treated media was sampled to represent a T = 0 hour sample; 5 pL of this sample (the remainder was reserved and frozen in case of re-analysis being required) was diluted with 45 pL of DMEM and crashed with 50 pL MeCN in a 96-well plate which was sealed and kept on ice. The HEK293 6-well plate was then incubated at 37°C in 5% C02 for 1 hour, the plate was then removed from the incubator and a 100 pL sample of media taken from each well to represent a T = 1 hour sample; 5 pL of this sample (the remainder was reserved and frozen in case of re-analysis being required) was diluted with 45 pL of DMEM and crashed with 50 pL MeCN into the 96-well plate which was re-sealed and stored at -20°C.
The remaining media was then removed from the HEK293 6-well plate, the wells were gently rinsed twice with 1 mL of 0.5% BSA in DMEM, the media was then removed and the 6-well plate allowed to dry completely at RT. 1 mL of 3:2 Hexane: Isopropanol (HIP) was added to each well in a fume cupboard and the plate allowed to stand for 30 minutes at RT without shaking. The HIP solution was then transferred to 2 mL Eppendorf tubes, and the process was repeated with a second 1 mL aliquot of HIP and the two aliquots combined. The HIP samples were then dried down under a nitrogen stream.
Thin Laver Chromatography (TLC) Analysis
Silica gel plates were prepared in a fume cupboard by initially drawing a line 1.5 cm from the bottom edge of the plate and then drawing sample lanes with a width of 1 cm and a separation of 0.5 cm between lanes. TLC buffer for phospholipids was prepared as a 31:62:7 solvent mix of MethanokChloroform: Ammonium Hydroxide. Plates were pre-run in a humid chamber containing 200 mL of the TLC buffer until the solvent front was 1.5 cm from the plate edge, the plate was then allowed to dry. HIP samples prepared as described above were reconstituted in 50 pL of chloroform, briefly vortex ed 3 times and then kept on ice.
Samples were loaded onto the plate (along with reference compound) by streaking gently with a pipette tip, samples were allowed to dry between streaks. On completion of the sample loading the plate was run in a sealed humid chamber containing the TLC buffer as described above for approximately 1.5 hours or until the solvent front had nearly reached the top of the plate. The plate was removed from the chamber and dried. An initial image was taken using Bio-Rad Image lab 6.0.
Iodine crystals were added to a new chamber which was sealed to allow the iodine vapor to saturate the container, the plate was then exposed to the iodine vapor in the chamber to allow visualization of bands of unsaturated fatty acids, once the plate was developed a second image was taken using Bio-Rad Image lab 6.0. The plate was then air-dried to remove the iodine. The plate was then saturated using a spray bottle with cupric acetate solution consisting of 3% cupric acetate by weight, 8% phosphoric acid by volume made up in an aqueous solution.
The plate was allowed to dry for 5 minutes at RT and then heated in a fume cupboard using a hot air gun to make the bands more visible. A final image was acquired using the Bio-Rad Image lab 6.0. The difference in intensity between the bands generated from hMfsd2a (WT) or D97A transfected HEK293 cells were compared to the empty vector (EV) transfected cells, allowed for uptake into the cells driven by hMfsd2a to be identified against the reference (REF). Results of the TEC analysis are shown in FIGs. 1A-3B and 5A-6B.
FIGs. 1A-1B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 1 (Example 1) - lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 1 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector, thus confirming the compound was advantageously transported via Mfsd2a.
FIGs. 2A-2B show the TEC images using the iodine and cupric acetate staining, respectively as described above with compound of Compound 2 (Example 2) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 2 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector, thus confirming the compound was advantageously transported via Mfsd2a.
FIGs. 3A-3B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 40 (Example 3) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It was unclear from the TEC images if Compound 40 was transported via Mfsd2a.
FIGs. 5A-5B show the TEC images using the iodine and cupric acetate staining, respectively as described above with Compound 42 (Example 5) - lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It is unclear from the TEC images if the Compound 42 was transported via Mfsd2a.
FIGs. 6A-6B show the TEC images using the iodine and cupric acetate staining respectively as described above with Compound 43 (Example 6) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector. It was unclear from the TEC images if Compound 43 was transported via Mfsd2a. UPLC-MS-MS Analysis
HEP samples prepared as described above were reconstituted in 100 pL of MeCN, vortex mixed and inverted multiple times to ensure all surfaces of the Eppendorf tube were rinsed with the MeCN and finally pulse centrifuged. A 50 pL aliquot of the MeCN reconstitution solution was then taken as a non-diluted HIP extract sample and added to the 96-well plate, alongside this a 1:10 dilution sample was prepared by taking a 5 pL aliquot and diluting with 45 pL of MeCN; 50 pL of Millipore water was added to each sample.
A bioanalytical calibration line was prepared to cover a range of concentration from 0.0001 to 10 pM by spiking 2 pL of a 0.5 mM DMSO stock of the test compound into 98 pL of MeCN to generate a 10 pM top standard that was then serial diluted with MeCN to produce 6 calibration standard stocks. Fifty pL of each calibration standard stock was added to the 96-well plate and diluted with 50 pL of Millipore water. Fifty pL of an appropriate internal standard in MeCN was then added to each of the wells in the 96-well plate that contained either a sample or calibration standard, the plate was sealed and transferred to the UPLC-MS-MS system for analysis. Uptake of the test compound into the HEK293 cells determined from the HIP sample analysis with the impact of hMfsd2a assessed by comparing the ratio of the concentration or peak area ratio of test compound in the hMfsd2a and D97A transfected cells to the empty vector transfected cells, as shown in FIGs. 7-9, 11 and
12.
FIG. 7 shows the concentration of Compound 1 (Example 1) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 1 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
FIG. 8 shows the peak area ratio of Compound 2 (Example 2) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 2 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageous! y transported via Mfsd2a. FIG. 9 shows the concentration of Compound 40 (Example 3) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 40 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
FIG. 11 shows the concentration of Compound 42 (Example 5) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 42 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
FIG. 12 shows the concentration of Compound 43 (Example 6) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 43 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
Bioanalytical Analysis
Bioanalytical samples were prepared according to the procedures described above for LC-MS-MS analysis. The samples were analyzed by LC-MS/MS utilizing the AB Sciex QTRAP 5500. The instrument was set to operate in positive ion mode for all analyses and the parameters are shown below in Table below.
MS/MS TUNE PARAMETERS
Figure imgf000074_0001
ADDITIONAL MS/MS PARAMETERS
Figure imgf000074_0002
Figure imgf000075_0001
Example 8. In vitro Mfsd2a Transport (Compound 41)
The following assay was used for analysis of Compound 41.
Methodology
The assay was conducted in a low throughput 6-well format with HEK293 cells prepared and then transfected in duplicate wells with plasmids containing either the wildtype (WT) version of hMfsd2a, the D97A mutant version, or an empty vector (EV) as control. Uptake into the cells was assessed by both thin-layer chromatography (TEC) and ultrahigh performance liquid chromatography hyphenated mass spectrometry.
Cell Transfection
Before cell seeding, 6-well plates were coated with Poly-D lysine (Sigma). HEK293 cells were seeded at 6.8 x 105 per 6-well in 2 ml. of DMEM with 10% FBS and 1% penicillin-streptomycin (P/S) media (Sigma) and incubated overnight at 37°C in 5% C02. Cells were checked for confluency the next morning. On a per well basis the following lipid mix was generated; 6 pL of Lipofectamine 2000 was added dropwise to 200 pL of OptiMEM, this was left to stand for 5 minutes at room temperature (RT).
1 pg of hMfsd2a WT, D97A or empty plasmid was prepared in 200 pL of OptiMEM as appropriate for each well; the Lipofectamine 2000 in OptiMEM solution was then added dropwise to a total volume of 400 pL (this can be scaled to support the number of wells/plates to be assayed). This transfection preparation was then incubated at RT for 20 minutes. DMEM with 10% FBS and no P/S media was warmed to 37°C, the HEK293 plate media was changed and the cells washed carefully with 1 mL of the warmed DMEM with 10% FBS no P/S media, 1.6 mL of the warmed DMEM with 10% FBS no P/S media was then added to each well. 400 pL of the transfection preparation was then added dropwise to each well as appropriate and the plate was gently swirled in a circular motion. The plate was then incubated overnight at 37°C in 5% CO,.
Compound Incubation and Preparation of Analysis Samples
Compound stock solutions were prepared in a 12% BSA in PBS solution such that a 40 pL spike into 2 mL of plain DMEM would yield a concentration of 25 μΜ of test compound (the compound treated media). Remaining compound stock solution in 12% BSA in PBS was frozen at -20°C to allow for future transport assays. The HEK293 6-well plate was removed from the incubator and the wells gently rinsed with 1 mL of plain DMEM that had been prewarmed to 37°C. 2 mL of the compound treated media was then added to each well.
A 100 pL sample of the compound treated media was sampled to represent a T = 0 hour sample; The HEK293 6-well plate was then incubated at 37°C in 5% C02 for 1 hour, the plate was then removed from the incubator and a 100 pL sample of media taken from each well to represent a T = 1 hour sample
The remaining media was then removed from the HEK293 6-well plate, the wells were gently rinsed twice with 1 mL of 0.5% BSA in DMEM, the media was then removed and the 6-well plate allowed to dry completely at RT. 1 mL of 3:2 Hexane:Isopropanol (HIP) was added to each well in a fume cupboard and the plate allowed to stand for 30 minutes at RT without shaking. The HIP solution was then transferred to 2 mL Eppendorf tubes, and the process was repeated with a second 1 mL aliquot of HIP and the two aliquots combined. The HIP samples were then dried down in a CentriVap. For media stability testing, 2 mL of diluted compound solution was added to new 6 well plate and 100 pL was sampled at the time point of 0 minutes, 15 minutes, 30 minutes and 60 minutes of incubation at 37°C in 5% CO2
Thin Laver Chromatography (TLC) Analysis
Silica gel plates were prepared in a fume cupboard by initially drawing a line 1.5 cm from the bottom edge of the plate and then drawing sample lanes with a width of 1 cm and a separation of 0.5 cm between lanes. TLC buffer for phospholipids was prepared as a 31:62:7 solvent mix of Methanol: Chloroform: Ammonium Hydroxide . Plates were pre-run in a humid chamber containing 150 mL of the TLC buffer until the solvent front was 1.5 cm from the plate edge, the plate was then allowed to dry. HIP samples prepared as described above were reconstituted in 50 pL of chloroform, briefly vortexed 3 times and then kept on ice.
Samples were loaded onto the plate (along with reference compound) by streaking gently with a pipette tip, samples were allowed to dry between streaks. On completion of the sample loading the plate was run in a sealed humid chamber containing the TLC buffer as described above for approximately 1.5 hours or until the solvent front had nearly reached the top of the plate. The plate was removed from the chamber and dried. An initial image was taken using camera.
Iodine crystals were added to a new chamber which was sealed to allow the iodine vapor to saturate the container, the plate was then exposed to the iodine vapor in the chamber to allow visualization of bands of unsaturated fatty acids, once the plate was developed a second image was taken using camera. The plate was then air- dried to remove the iodine. The plate was then saturated using a spray bottle with cupric acetate solution consisting of 3% cupric acetate by weight, 8% phosphoric acid by volume made up in an aqueous solution.
The plate was allowed to dry for 5 minutes at RT and then heated in an oven at 130°C for 10 minutes to make the bands more visible. A final image was acquired using the camera. Quantitative analysis of the band signals was performed by ImageJ. The difference in intensity between the bands generated from hMfsd2a (WT) or D97A transfected HEK293 cells were compared to the empty vector (EV) transfected cells, allowed for uptake into the cells driven by hMfsd2a to be identified against the reference (REF). Results of the TLC analysis are shown inFIGs. 4A-4B.
FIGs. 4A-4B show the TLC images using the iodine and cupric acetate staining, respectively as described above with Compound 41 (Example 4) — lane from left: reference compound, HIP sample from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with bands corresponding to Compound 41 showing higher intensity in WT cells compared with cells transfected with D97A mutant Mfsd2a and/or empty vector (iodine stain image), thus confirming the compound was advantageously transported via Mfsd2a.
UPLC-MS-MS Analysis HIP samples prepared as described above were reconstituted in 100 pL of MeCN, vortex mixed and inverted multiple times to ensure all surfaces of the Eppendorf tube were rinsed with the MeCN and finally pulse centrifuged. A 50 pL aliquot of the MeCN reconstitution solution was then taken as a non-diluted HIP extract sample and added to the 96-well plate, alongside this a 1:10 dilution sample was prepared by taking a 5 pL aliquot and diluting with 45 pL of MeCN; 50 pL of Millipore water was added to each sample.
A bioanalytical calibration line was prepared to cover a range of concentration from 0.001 to 1 pM by spiking 1 pL of a 2 mM MeOH stock of the test compound into 199 pL of MeCN to generate a 10 pM top standard that was then serial diluted with MeCN to produce 7 calibration standard stocks. 50pL of each calibration standard stock was added to the 96- well plate and diluted with 50 pL of Millipore water. 50 pL of an appropriate internal standard in H20 was then added to each of the wells in the 96-well plate that contained either a sample or calibration standard, the plate was sealed and transferred to the UPLC-MS-MS system for analysis. Uptake of the test compound into the HEK293 cells determined from the HIP sample analysis with the impact of hMfsd2a assessed by comparing the ratio of the concentration or peak area ratio of test compound in the hMfsd2a and D97A transfected cells to the empty vector transfected cells, as shown in FIG. 10.
FIG. 10 shows the concentration of Compound 41 (Example 4) measured in HIP samples from WT cells, cells transfected with D97A mutant Mfsd2a and empty vector, with more Compound 41 detected in WT cells compared to cells transfected with D97A mutant Mfsd2a and/or empty vector, confirming the compound was advantageously transported via Mfsd2a.
Bioanalytical Analysis
Bioanalytical samples were prepared according to the procedures described above for LC -MS/MS analysis. The samples were analyzed by LC-MS/MS utilizing the Waters Xevo TQ-S micro. The instrument was set to operate in positive ion mode for all analyses and the parameters are shown below in Table below.
MS/MS TUNE PARAMETERS
Figure imgf000078_0001
Figure imgf000079_0001
ADDITIONAL MS/MS PARAMETERS
Figure imgf000079_0002
Example 9. In vivo AD ME Protocol 1 : IV terminal rat PK study at 3 mg/kg
Compound 1 (Example 1) was orally dosed at 3 mg/kg to a group of 18 individually housed male Sprague Dawley rats that were fasted overnight and fed 4 hours post-dose. Dosing was performed with 2 mL/kg dosing volumes with 5% DMSO 95% water used as a dosing vehicle. Terminal plasma samples were taken from groups of 3 animals at each of 5 time-points post dose (0.167 hours, 0.5 hours, 1 hour, 2 hours, 4 hours and 8 hours) by cardiac puncture under CO2 into lithium heparin coated tubes and centrifuged to obtain plasma. Immediately following collection of plasma samples, whole body perfusion was performed using phosphate buffered saline (PBS) and the brain and eyes excised, rinsed in ice-cold PBS, blotted dry, weighed, and snap frozen in liquid nitrogen. Collected tissue samples were stored in the freezer at -70°C until bioanalysis.
Protocol 2: Bioanalvtical samples preparation for plasma samples
Plasma samples were defrosted and sample preparation was carried out using liquid-liquid extraction. All the samples were extracted and the supernatants analysed by LC-MS/MS according to the following procedures:
1. 50 pL of plasma was transferred to a clean 1.5 mL tube. 2. 5 pL of MeOH was added to each tube.
3. 20 pL of 100 ng/mL internal standard (IS) in MeOH:ACN (1:1 v/v) was added to each tube.
4. 800 pL of ethyl acetate was added to each tube.
5. The tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
6. 700 pL of supernatant was aliquoted out and evaporated to dryness under nitrogen gas.
7. Dried residues were reconstituted in 200 pL MeOH: ACN (1:1 v/v).
8. Reconstituted samples were analysed for the compound using LC-MS/MS. The resulting PK parameters of the compound in plasma are shown below in Table 1.
Table 1.
Figure imgf000080_0001
Protocol 3: Bioanalvtical samples preparation for brain samples
Brain homogenates were prepared from whole frozen brains harvested from Protocol 1 using ACN and water. Harvested whole brains were weighed and 4x weight equivalent volume ACN : water (1:1 v/v) was added before homogenization. Analysis of the brain samples was conducted according to the following procedures and the data are shown in Table 2.
1. 50 pL of brain homogenate was transferred to a clean 1.5 mL tube.
2. 5 pL of MeOH was added to each tube.
3. 20 pL of 100 ng/mL IS in MeOH: ACN (1 : 1 v/v) was added to each tube.
4. 800 pL of ethyl acetate was added to each tube.
5. The tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
6. 700 pL of supernatant was aliquoted out and evaporated to dryness under nitrogen gas. 7. Dried residues were reconstituted in 200 pL MeOHrACN (1:1 v/v).
8. Reconstituted samples were analysed for the compound using LC -MS/MS. The resulting PK parameters of the compound in brain are shown below in Table 2.
Table 2.
Figure imgf000081_0001
Protocol 4: Bioanalytical samples preparation for eve samples
Eye homogenates were prepared from whole frozen eyes harvested from Protocol 1 using ACN and water. Harvested whole eyes were weighed and 9x weight equivalent volume ACN: water (1:1 v/v) was added before homogenization. Analysis of the eye samples was conducted according to the following procedures and the data are shown in Table 3.
1. 50 pL of eye homogenate was transferred to a clean 1.5 mL tube.
2. 5 pL of MeOH was added to each tube.
3. 20 pL of 100 ng/mL IS in MeOH: ACN (1:1 v/v) was added to each tube.
4. 800 pL of ethyl acetate was added to each tube.
5. The tubes were mixed using a vortex for 1 minute and were centrifuged for 15 minutes at 4000 rpm.
6. 700 pL of supernatant was aliquoted out and evaporated to dryness under nitrogen gas.
7. Dried residues were reconstituted in 200 pL MeOH: ACN (1 : 1 v/v).
8. Reconstituted samples were analysed for the compound using LC-MS/MS. The resulting PK parameters of the compound in eye are shown below in Table 3.
Table 3.
Figure imgf000081_0002
Protocol 5: Bioanalytical Analysis Bioanalytical samples were prepared according to the procedures described above for LC -MS/MS analysis. The samples were analyzed by LC-MS/MS utilizing the AB Sciex QTRAP 6500+ coupled to a UPLC system. The LC-MS/MS conditions used are shown below in Table 4.
5
Table 4.
Figure imgf000082_0001
Example 10. Cytoprotective Effects
This Example demonstrates the cytoprotective effects of Compound 1 10 (Example 1) in human induced pluripotent stem cell (iPSC) derived retinal pigment epithelium (RPE) cells. Human iPSC-RPE cells were differentiated on 96 -well plates for 16 days. Cells were exposed to the compound at 5 μΜ in quadruplicates for 48 hours before induction of oxidative stress (day 19) with teri-butyl hydroperoxide solution (tBHP) (0-10 mM) at 37°C 5% CO2. Lactate dehydrogenase (LDH) was 15 measured on samples collected after 24 hours tBHP -incubation as a readout of the cytoprotective effect of the compound.
Materials and Methods hiPSC derived Retinal Pigment Epithelium
20 Human iPSC-RPE cells (PCi-RPE, p2, Phenocell SAS, France) were cultured according to manufacturer’s instructions. Cells (passage 2) were expanded for 14 days in culture medium containing 70% DMEM, high glucose (Gibco Thermo Fischer Scientific, USA), 30% Ham’s F12 Nutrient Mix (Gibco Thermo Fischer Scientific, USA), 2% B-27® Supplement (Gibco Thermo Fischer Scientific, USA), 1% Antibiotic-Antimycotic (Gibco Thermo Fischer Scientific, USA) in Matrigel® coated cell culture dishes (Coming, USA, final density of Matrigel 8-10 pg/cm2) at 37°C 5% CO2. iPSC-RPE cells acquired their characteristic polygonal morphology and were pigmented at passage 3 in Matrigel® coated cell culture dishes. Cells were passaged (passage 4) onto Matrigel® coated 96-well plates at a density of 100,000 cells/cm2, and cultured until fully differentiated into RPE cells for 16 days.
Compounds. Delivery, Storage , and Administration
Compound 1 was dissolved in 100% DMSO (Sigma- Aldrich, USA) at concentration 10 mM. Thereafter, dilutions of study compounds were prepared in RPE medium with B-27 AO neg. (Gibco Thermo Fischer Scientific, USA, 100 mM) and sterile filtered through 0.22 pm. Sterile aliquots were stored at -20°C. Final working dilutions of study compounds (5 pM, 0.1% DMSO) were prepared fresh in RPE medium with B-27 AO neg. Vehicle was 0.1% DMSO in RPE medium with B- 27 AO neg. RPE cells were pre-treated with each of the study compounds or vehicle 48 hours before induction of oxidative stress. LDH Assay Protocol
Human iPSC-RPE cells were pre-incubated with the compound at 5 pM in quadmplicates for 48 hours before exposure to whole concentration range of tBHP at 37°C for 24 hours (co-incubation period during which iPSC-RPE cells will be continually exposed to the compound). LDH quantification was performed on samples collected after 6 hours tBHP incubation, and again after the 24 hours tBHP- incubation, according to the previously published protocol (see e.g., Kaja et al. J. Pharmacol. Toxicol. Methods, 2015, 73:1-6). Samples (50 pL) of cell culture supernatants were collected (quadmplicates) and incubated in 1 mM INT (Iodonitrotetrazolium Chloride, Sigma- Aldrich, USA), 1.6 mM NAD (Beta- Nicotinotinamide Adenine Dinucleotide Sodium Salt, Sigma-Aldrich, USA), 80 mM lithium L-lactate (Sigma-Aldrich, USA), 7.5 pM MPMS ( 1 -Methoxyphenazine methosulfate, Sigma-Aldrich, USA) in 0.2 M Tris-HCl (Sigma-Aldrich, USA), pH 8.2 for 30 minutes. The reaction was stopped using 1 M acetic acid (Fisher, USA). Absorbance at 490 nm was measured using Cytation 3 multi-mode reader (BioTek, Winooski, VT, USA).
Data Analysis
All the assay data was normalized to cell viability of vehicle treated control cells (not induced with tBHP). All values are presented as mean ± standard deviation (SD). Parametric data were analyzed using One-way- ANOV A followed by Dunnett’s multiple comparisons posthoc test to compare each group to the vehicle group. Differences were considered to be statistically significant at P<0.05.
Cvtoyrotection of human iPSC-RPE cells readout
As can be seen from FIGs. 13A-13B, Compound 1 (Example 1) displayed cytoprotective effects. The compound showed lower LDH release at 0.6 mM tBHP compared to the vehicle, indicating that this compound has cytoprotective properties in hiPSC-RPE cells. Compound 1 also demonstrated a trend towards better cell viability (lower LDH release at 6 hours) than vehicle at high tBHP concentrations (1- 10 mM) indicating acute (0-6 hours) protective effects against oxidative stress (FIGs. 13A-13B).
OTHER EMBODIMENTS
It is to be understood that while the present disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. It should be appreciated by those persons having ordinary skill in the art(s) to which the present disclosure relates that any of the features described herein in respect of any particular aspect and/or embodiment of the present disclosure can be combined with one or more of any of the other features of any other aspects and/or embodiments of the present disclosure described herein, with modifications as appropriate to ensure compatibility of the combinations. Such combinations are considered to be part of the present invention contemplated by this disclosure.

Claims

WHAT IS CLAIMED IS:
1. A compound of Formula I:
Figure imgf000086_0001
I or a pharmaceutically acceptable salt thereof, wherein:
L1 is selected from the group consisting of Cs-20 alkylene and Cs-zo alkenylene, wherein the Cg-2o alkylene or Cs-20 alkenylene is optionally substituted by O', OH, Ci-e alkyl or halogen;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, C(0)NH(Ci-6 alkylene), 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci_6 alkylene)C(0)NH, 0C(0)NH(CI-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci_6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(Ci-6 alkyl ene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by O", OH, or halogen;
R2 is a bond or a phosphate group;
R3 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-COi", Ci-4 alkylene-COaH and Ci-e alkyl, wherein the Ci-6 alkyl is optionally substituted by O", OH, or halogen;
R4 is selected from the group consisting of H, CO2", CO2H, C1-4 alkylene-C02", Ci-4 alkylene-C02H and Ci-e alkyl, wherein the Ci-e alkyl is optionally substituted by O", OH, or halogen;
R5a is selected from the group consisting of H and C1-4 alkyl, wherein the CM alkyl is optionally substituted by O", OH, or halogen;
R5b is selected from the group consisting of H and CM alkyl, wherein the CM alkyl is optionally substituted by O", OH, or halogen; and R5C is selected from the group consisting of H and C1-4 alkyl, wherein the C1-4 alkyl is optionally substituted by O', OH, or halogen.
2. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein L1 is Cs-20 alkylene.
3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein L1 is Cs-i2 alkylene.
4. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L1 is decanediyl.
5. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein L1 is Cs-2o alkenylene.
6. The compound of claim 1 or 5, or a pharmaceutically acceptable salt thereof, wherein L1 is C 15-20 alkenylene.
7. The compound of any one of claims 1, 5, and 6, or a pharmaceutically acceptable salt thereof, wherein the alkenylene of L1 comprises 1 or 2 carbon-carbon double bonds.
8. The compound of any one of claims 1, 5, and 6, or a pharmaceutically acceptable salt thereof, wherein the alkenylene of L1 comprises 1 carbon-carbon double bond.
9. The compound of any one of claims 1 and 5 to 8, or a pharmaceutically acceptable salt thereof, wherein L1 is selected from the group consisting of tetradecenediyl , heptadecenediyl and octadecenediyl.
10. The compound of any one of claims 1 to 9, wherein R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(Ci-e alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(Ci-6 alkylene)C(0)NH, 0(Ci-e alkylene)C(0)0, 0(Ci-e alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(CI-6 alkylene)C(0)NH, NHC(0)(CI-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-e alkylene is optionally substituted by O", OH, or halogen.
11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, OCH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2C(0)NH.
12. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of 0C(0)1NH(CI-6 alkylene) and 0C(0)0(Ci-6 alkylene).
13. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of 0C(0)NH(CI-4 alkylene) and 0C(0)0(Ci-4 alkylene).
14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of 0C(0)NHCH2CH2 and 0C(0)0CH2CH2.
15. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein R2 is a bond.
16. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein R2 is a phosphate group.
17. The compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein R3 is selected from the group consisting of H, CO2", CO2H, CH2CO2" and CH2CO2H.
18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein R3 is H.
19. The compound of any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, wherein R4 is selected from the group consisting of H, CO2", CO2H, CH2CO2" and CH2CO2H.
20. The compound of any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, wherein R4 is H.
21. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R5a is H.
22. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R5a is methyl.
23. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R5b is H.
24. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R5b is methyl.
25. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R5c is H.
26. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R5c is methyl.
27. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein two of R5a, R5b, and R5c are H.
28. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R5a, R5b, and R5c are each H.
29. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein two of R5a, R5b, and R5c are methyl.
30. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R5a, R5b, and R5c are each methyl.
31. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein :
L1 is C8-20 alkylene or Cg-20 alkenylene, wherein the Cg-20 alkylene or Cs-2o alkenylene is optionally substituted by O", OH, Ci-e alkyl or halogen; R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, C(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(Ci-6 alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkyiene)C(0)NH, wherein each Ci-6 alkylene is optionally substituted by OH;
R2 is a bond or a phosphate group;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and
R5C is H or methyl.
32. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein:
L1 is selected from the group consisting of decanediyl, heptadecenediyl and octadecenediyl;
R1 is selected from the group consisting of NHC(0)0, 0C(0)NH(CI-6 alkylene), NHC(0)NH, 0C(0)0(Ci-6 alkylene), 0(Ci-6 alkylene)0, 0C(0)(Ci-6 alkylene)C(0)0, 0C(0)(Ci-6 alkylene)C(0)NH, 0(0)0, C(0)NH, 0C(0)0(Ci-6 alkylene)C(0)0, 0C(0)0(Ci-6 alkylene)C(0)NH, 0C(0)NH(Ci-6 alkylene)C(0)0, 0C(0)NH(CI-6 alkylene)C(0)NH, 0(Ci-6 alkylene)C(0)0, 0(Ci-6 alkylene)C(0)NH, NH(CI-6 alkylene)C(0)0, NH(Ci-e alkylene)C(0)NH, NHC(0)(Ci-6 alkylene)C(0)0, and NHC(0)(CI-6 alkylene)C(0)NH, wherein each Ci-e alkylene is optionally substituted by OH;
R2 is a bond or a phosphate group;
R3 is H or CH2CO2";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and R5C is H or methyl.
33. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:
L1 is selected from the group consisting of decanediyl, tetradecenediyl, heptadecenediyl and octadecenediyl;
R1 is selected from the group consisting of 0C(0)NHCH2CH2, 0C(0)0CH2CH2, NHC(0)0, NHC(0)NH, 0CH2CH(0H)CH20, 0C(0)CH2C(0)0, 0C(0)CH2CH2C(0)0, 0C(0)CH2CH2CH2C(0)0, 0C(0)CH2C(0)NH, 0C(0)CH2CH2C(0)NH, 0C(0)CH2CH2CH2C(0)NH, C(0)0, C(0)NH, 0C(0)0CH2C(0)0, 0C(0)0CH2CH2C(0)0, 0C(0)0CH2CH2CH2C(0)0, 0C(0)0CH2C(0)NH, 0C(0)0CH2CH2C(0)NH, 0C(0)0CH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)NH, 0C(0)NHCH2CH2C(0)NH,
0C(0)NHCH2CH2CH2C(0)NH, 0C(0)NHCH2C(0)0, 0C(0)NHCH2CH2C(0)0, 0C(0)NHCH2CH2CH2C(0)0, 0CH2C(0)0, 0CH2CH2C(0)0,
0CH2CH2CH2C(0)0, 0CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2C(0)0, 0CH2CH2CH2CH2CH2CH2C(0)0, 0CH2C(0)NH, 0CH2CH2C(0)NH, 0CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2C(0)NH, 0CH2CH2CH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2CH2C(0)0, NHCH2CH2CH2CH2C(0)NH, NHCH2CH2CH2CH2CH2C(0)NH, NHC(0)CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2CH2C(0)0, NHC(0)CH2CH2CH2C(0)NH, and NHC(0)CH2CH2CH2CH2C(0)NH;
R2 is a bond or a phosphate group;
R3 is H or CH2C02 ";
R4 is H;
R5a is H or methyl;
R5b is H or methyl; and
R5C is H or methyl.
34. The compound of claim 1, wherein the compound of Formula I is a compound of Formula IT:
Figure imgf000093_0003
II or a pharmaceutically acceptable salt thereof.
35. The compound of claim 1, wherein the compound of Formula I is a compound of Formula III:
Figure imgf000093_0004
III or a pharmaceutically acceptable salt thereof.
36. The compound of claim 1, wherein the compound of Formula I is a compound of Formula IV:
Figure imgf000093_0001
IV or a pharmaceutically acceptable salt thereof.
37. The compound of claim 1, wherein the compound of Formula I is a compound of Formula V:
Figure imgf000093_0002
V or a pharmaceutically acceptable salt thereof.
38. The compound of claim 1, wherein the compound of Formula I is selected from the group consisting of:
Figure imgf000094_0001
Figure imgf000095_0001
Figure imgf000096_0001
Figure imgf000097_0001
Figure imgf000098_0001
or a pharmaceutically acceptable salt thereof.
39. A pharmaceutical composition comprising a compound of any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.
40. A method of treating a disease or disorder selected from the group consisting of mitochondrial disorder, pain, a pain-related disease or disorder, a mood disease or disorder, a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, an optical disease or disorder, cancer, a gastrointestinal disease or disorder, a renal disease or disorder, a renal-related disease or disorder, a cardiovascular disease or disorder and a skin disease or disorder, comprising administering to a subject a therapeutic ally effective amount of a compound of any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 39.
41. The method of claim 40, wherein the disease or disorder is a mitochondrial disorder.
42. The method of claim 41, wherein the mitochondrial disorder is selected from the group consisting of Leber's hereditary optic neuropathy (LHON), Duchenne muscular dystrophy, mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS), mitochondrial myopathy, diabetes mellitus and deafness (DAD), Leigh syndrome, neuropathy, ataxia, and retinitis pigmentosa (NARP), ptosis, chronic progressive external ophthalmoplegia (CPEO), coenzyme Q10 deficiency, familial bilateral striatal necrosis, Friedreich ataxia, infantile-onset spinocerebellar ataxia (IOSC A), Keams-Sayre syndrome, mitochondrial DNA depletion syndrome (MDS) and Rett syndrome (RTT).
43. The method of claim 40, wherein the disease or disorder is pain or a pain- related disease or disorder.
44. The method of claim 43, wherein the pain or a pain-related disease or disorder is selected from the group consisting of acute pain, chronic pain, neuropathic pain, nociceptive pain, inflammatory pain, cancer pain, fibromyalgia, rheumatoid arthritis, osteoarthritis, surgery-related pain, and osteoporosis.
45. The method of claim 40, wherein the disease or disorder is a mood disease or disorder.
46. The method of claim 45, wherein the mood disease or disorder is selected from the group consisting of anxiety, depression, a sleeping disorder, an eating disorder, post-traumatic stress disorder, symptoms of drug or alcohol withdrawal or abuse, schizophrenia, obsessive-compulsive disorder, bipolar disorder, sexual dysfunction, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD).
47. The method of claim 40, wherein the disease or disorder is a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, or an optical disease or disorder.
48. The method of claim 47, wherein the disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, or an optical disease or disorder is selected from the group consisting of a demyelinating disease, glaucoma, age-related macular degeneration (AMD), amyotrophic lateral sclerosis (ALS), a cognitive disorder, Alzheimer’s disease, a movement disorder, Huntington’s chorea, Tourette’s syndrome, Niemann-Pick disease, Parkinson's disease, epilepsy, a cerebrovascular disorder, ischemic stroke, and brain injury.
49. The method of claim 48, wherein the demyelinating disease is selected from the group consisting of multiple sclerosis (MS), neuromyelitis optica (NMO), Devic’s disease, central nervous system neuropathy, central pontine myelinolysis, syphilitic myelopathy, leukoencephalopathies, leukodystrophies, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, anti -myelin-associated glycoprotein (MAG) peripheral neuropathy, Charcot-Marie-Tooth disease, peripheral neuropathy, myelopathy, optic neuropathy, progressive inflammatory neuropathy, optic neuritis, and transverse myelitis.
50. The method of statement 49, wherein the optic neuropathy is Leber’s hereditary optic neuropathy.
51. The method of any one of claims 40 to 50, further comprising administering to the subject one or more additional therapeutic agents.
52. One or more dosage forms, comprising a compound of any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent.
53. A method of treating a disease or disorder selected from the group consisting of mitochondrial disorder, pain, a pain-related disease or disorder, a mood disease or disorder, a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, an optical disease or disorder, cancer, a gastrointestinal disease or disorder, a renal disease or disorder, a renal-related disease or disorder, a cardiovascular disease or disorder and a skin disease or disorder, comprising administering to a subject one or more dosages forms of claim 52.
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