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WO2021050688A1 - Composés et leurs utilisations - Google Patents

Composés et leurs utilisations Download PDF

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Publication number
WO2021050688A1
WO2021050688A1 PCT/US2020/050142 US2020050142W WO2021050688A1 WO 2021050688 A1 WO2021050688 A1 WO 2021050688A1 US 2020050142 W US2020050142 W US 2020050142W WO 2021050688 A1 WO2021050688 A1 WO 2021050688A1
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Prior art keywords
compound
alkyl
mmol
optionally substituted
stirred
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English (en)
Inventor
Esther C. Y. LEE
Andrew J. Mcriner
Christopher D. Hupp
Anna Kohlmann
Charles Joseph Eyermann
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X Chem Inc
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X Chem Inc
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    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline

Definitions

  • Oxalate is a metabolic end-product endogenously produced by the liver and filtered by the kidney to be excreted in urine. Oxalate tends to precipitate as insoluble calcium oxalate crystals, which is the main component of kidney stones and bladder stones.
  • Disorders e.g., primary hyperoxalurias (PHs)
  • PHs kidney stone disease
  • ESRD end stage renal disease
  • renal failure e.g., end stage renal disease (ESRD), and renal failure.
  • KSD kidney stone disease
  • ESRD end stage renal disease
  • therapies for disorders that lead to oxalate accumulation e.g., primary hyperoxalurias (PHs).
  • PHs Primary hyperoxalurias
  • PH1 primary hyperoxaluria type 1
  • AGT alanine-glyoxylate aminotransferase
  • Glyoxylate is produced via oxidation of glycolic acid (glycolate) by hydroxyacid oxidase 1 (HAO1), also known as glycolate oxidase (GO).
  • HAO1 hydroxyacid oxidase 1
  • LDH lactate dehydrogenase
  • the calcium salt of oxalate is insoluble and tends to precipitate as insoluble crystals in tissues. This potentially can result in kidney stones, kidney damage, kidney failure, hematuria, urinary tract infections, end stage renal disease (ESRD), systemic oxalosis, or injury to other organs.
  • HAO1 a life-threatening condition that prevents kidneys from effectively filtering fluids and waste products from the body.
  • Inhibition of HAO1 activity may reduce the production of glyoxylate thus reducing levels of toxic oxalate crystals in the kidneys of PH1 patients.
  • the present disclosure features compounds and methods useful for treating HAO1-associated disorders, for affecting the level and/or activity of HAO1, and/or for treating disorders (e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)) that lead to oxalate accumulation and the symptoms (e.g., end stage renal disease) that accompany such disorders.
  • disorders e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)
  • the symptoms e.g., end stage renal disease
  • the disclosure features a compound having the structure of Formula I: Formula I, where X 1 is N or CR 1 ; X 2 is N or CR 2 ; X 3 is N or CR 3 ; X 4 is N or C-COOH; X 5 is N or C-Het; each of R 1 , R 2 , and R 3 is, independently, H, cyano, hydroxyl, halogen, optionally substituted C 1 -C 6 alkyl, or optionally substituted C 1 -C 6 heteroalkyl; R 4 is H or optionally substituted C 1 -C 6 alkyl; R 5 is H, optionally substituted C 1 -C 6 alkyl, or optionally substituted C 1 -C 6 alkyl C 6 -C 10 aryl; each of R 6 and R 7 is, independently, H or optionally substituted C 1 -C 6 alkyl, or R 6 and R 7 combine with the carbon to which they are attached to form -C(O)-; each of R 8
  • X 1 is N. In some embodiments, X 1 is CR 1 . In some embodiments, R 1 is H, hydroxyl, halogen, optionally substituted C 1 -C 4 alkyl, or optionally substituted C 1 -C 4 heteroalkyl. In some embodiments, R 1 is H, hydroxyl, halogen, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, or –OR O1 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl. In some embodiments, R 1 is H, hydroxyl, . In some embodiments, R 1 is H, F, Cl, , or .
  • R 1 is H.
  • X 2 is N.
  • X 2 is CR 2 .
  • R 2 is H, hydroxyl, halogen, optionally substituted C 1 -C 4 alkyl, or optionally substituted C 1 -C 4 heteroalkyl.
  • R 2 is H, hydroxyl, halogen, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, or –OR O1 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl.
  • R 2 is H, hydroxyl, , or .
  • R 2 is H, F, Cl, , or . In some embodiments, R 2 is H. In some embodiments, X 3 is N. In some embodiments, X 3 is CR 3 . In some embodiments, R 3 is H, hydroxyl, halogen, optionally substituted C 1 -C 4 alkyl, or optionally substituted C 1 -C 4 heteroalkyl. In some embodiments, R 3 is H, hydroxyl, halogen, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, or –OR O1 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl.
  • R 3 is H, hydroxyl, F, Cl, , or . In some embodiments, R 3 is H, F, Cl, , or . In some embodiments, R 3 is H. In some embodiments, R 4 is H or optionally substituted C 1 -C 3 alkyl. In some embodiments, R 4 is H or C 1 -C 3 alkyl. In some embodiments, R 4 is H or . In some embodiments, R 4 is H. In some embodiments, R 5 is H or optionally substituted C 1 -C 3 alkyl. In some embodiments, R 5 is H or C 1 -C 3 alkyl. In some embodiments, R 5 is H, , or .
  • R 5 is optionally substituted C 1 -C 6 alkyl C 6 -C 10 aryl (e.g., phenethyl). In some embodiments, R 5 is H or . In some embodiments, R 5 is H. In some embodiments, R 5 is . In some embodiments, R 6 is H or optionally substituted C 1 -C 3 alkyl. In some embodiments, R 6 is H or C 1 -C 3 alkyl. In some embodiments, R 6 is H or . In some embodiments, R 6 is H. In some embodiments, R 7 is H or optionally substituted C 1 -C 3 alkyl. In some embodiments, R 7 is H or C 1 -C 3 alkyl.
  • R 7 is H or . In some embodiments, R 7 is H. In some embodiments, R 6 is H and R 7 is H. In some embodiments, R 6 and R 7 , together with the carbon to which they are attached, combine to form -C(O)-. In some embodiments, each of R 8 , R 9 , R 10 , and R 11 is, independently, H, cyano, hydroxyl, halogen, optionally substituted C 1 -C 4 alkyl, or optionally substituted C 1 -C 4 heteroalkyl, or R 8 and R 9 or R 9 and R 10 , together with the carbon atoms to which each is attached, combine to form an optionally substituted 5- or 6-membered aryl or heteroaryl.
  • each of R 8 , R 9 , R 10 , and R 11 is, independently, H, cyano, hydroxyl, halogen, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, or –OR O1 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl, or R 8 and R 9 or R 9 and R 10 , together with the carbon atoms to which each is attached, combine to form an optionally substituted 5- or 6-membered aryl or heteroaryl.
  • each of R 8 , R 9 , R 10 , and R 11 is, independently, H, cyano, hydroxyl, F, Cl, , , or , or R 8 and R 9 or R 9 and R 10 , together with the carbon atoms to which each is attached, combine to form an optionally substituted 5- or 6-membered aryl or heteroaryl.
  • each of R 8 , R 9 , R 10 , and R 11 is, independently, H, cyano, hydroxyl, F, Cl, , or , or R 8 and R 9 or R 9 and R 10 , together with the carbon atoms to which each is attached, combine to form an optionally substituted 5- or 6-membered aryl or heteroaryl.
  • R 8 is H.
  • R 9 is H, cyano, F, Cl, , or .
  • R 10 is H or F.
  • R 8 and R 9 together with the carbon atoms to which each is attached, combine to form a 5-membered heteroaryl.
  • R 8 and R 9 together with the carbon atoms to which each is attached, combine to form .
  • R 11 is H, OH, F, or Cl.
  • R 11 is OH.
  • X 5 is N.
  • the compound has the structure of Formula I-1: or a pharmaceutically acceptable salt thereof.
  • the compound has the structure of Formula I-2: , or a pharmaceutically acceptable salt thereof.
  • Het is optionally substituted 5-membered heteroaryl or optionally substituted 6-membered heteroaryl.
  • Het is 5-membered heteroaryl or 6- membered heteroaryl.
  • Het is optionally substituted 5-membered heteroaryl.
  • Het is 5-membered heteroaryl.
  • Het is , where R 12a is H, hydroxyl, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, or NR N1 R N2 ; R 12b is H, hydroxyl, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, or NR N1 R N2 ; R 12c is H or optionally substituted C 1 -C 6 alkyl, or optionally substituted C 3 -C 6 cycloalkyl; and each of R N1 and R N2 is, independently, H or optionally substituted C 1 -C 6 alkyl.
  • R 12a is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, or NR N1 R N2 .
  • R 12a is H, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, –OR O1 , or NR N1 R N2 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl.
  • each of R N1 and R N2 is, independently, H or C 1 -C 3 alkyl.
  • each of R N1 and R N2 is, independently, H or .
  • R 12a is H, , or . In some embodiments, R 12a is H or . In some embodiments, R 12a is H. In some embodiments, R 12b is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, or NR N1 R N2 . In some embodiments, R 12b is H, C 1 -C 4 alkyl, C 1 -C 4 fluoroalkyl, –OR O1 , or NR N1 R N2 , wherein R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl.
  • each of R N1 and R N2 is, independently, H or C 1 -C 3 alkyl. In some embodiments, each of R N1 and R N2 is, independently, H or .
  • R is H, , , , , , or .
  • R 12b is H.
  • R 12a is H and R 12b is H.
  • R 12c is H or C 1 -C 3 alkyl. In some embodiments, R 12c is H, methyl, iso- propyl, cyclopropyl, or cyclobutyl. In some embodiments, R 12c is H, , or .
  • R 12c is or 12 . In some embodiments, R c is . In some embodiments, Het is , where R 13 is H, hydroxyl, halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 3 -C 6 cycloalkyl, or –NR N1 R N2 , where each of R N1 and R N2 is, independently, H or optionally substituted C 1 -C 6 alkyl.
  • R 13 is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, optionally substituted C 3 -C 6 cycloalkyl, or NR N1 R N2 . In some embodiments, R 13 is optionally substituted C 3 -C 6 cycloalkyl. In some embodiments, R 13 is H, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 heteroalkyl, or NR N1 R N2 .
  • R 13 is H, C 1 -C 4 alkyl, C 1 -C 4 hydroxyalkyl, C 1 -C 4 fluoroalkyl, –OR O1 , C 3 -C 6 cycloalkyl, or –NR N1 R N2 , where R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl.
  • R 13 is H, C 1 -C 4 alkyl, C 1 -C 4 hydroxyalkyl, C 1 -C 4 fluoroalkyl, –OR O1 , or –NR N1 R N2 , where R O1 is C 1 -C 4 alkyl or C 1 -C 4 fluoroalkyl and each of R N1 and R N2 is, independently, H or optionally substituted C 1 -C 6 alkyl. In some embodiments, each of R N1 and R N2 is, independently, H or C 1 - C 3 alkyl. In some embodiments, each of R N1 and R N2 is, independently, H or . In some embodiments, R N1 is N2 and R is .
  • R 13 is H, , or . In some embodiments, R 13 is , , , , or In some embodiments, R 13 is or . In some embodimen 13 ts, R is . In some embodiments, R 13 is . In some embodiments, Het is , wherein X 6 is O, S, or NR 14b ; and each of R 14a and R 14b is, independently, H or optionally substituted C 1 -C 6 alkyl. In some embodiments, X 6 is O or S. In some embodiments, X 6 is O or NR 14b . In some embodiments, X 6 is O. In some embodiments, X 6 is S. In some embodiments, X 6 is NR 14b .
  • R 14b is H or C 1 -C 3 alkyl. In some embodiments, R 14b is H, methyl, or iso-propyl. In some embodiments, R 14b is methyl or iso-propyl. In some embodiments, R 14b is iso-propyl. In some embodiments, R 14a is H. In some embodiments, Het is , wherein X 7 is O or S; and each of R 15a and R 15b is, independently, H or optionally substituted C 1 -C 6 alkyl. In some embodiments, X 7 is O. In some embodiments, X 7 is S.
  • R 15a is H or C 1 -C 3 alkyl (e.g., methyl or iso-propyl). In some embodiments, R 15a is H. In some embodiments, R 15a is C 1 -C 3 alkyl (e.g., methyl or iso-propyl). In some embodiments, R 15b is H or C 1 C 3 alkyl (e.g., methyl or iso-propyl). In some embodiments R 15b is H. In some embodiments, R 15b is C 1 -C 3 alkyl (e.g., methyl or iso-propyl).
  • R 15a is C 1 -C 3 alkyl (e.g., methyl or iso-propyl) and R 15b is C 1 -C 3 alkyl (e.g., methyl or iso-propyl).
  • R 15a is H and R 15b is C 1 -C 6 alkyl (e.g., methyl or iso-propyl).
  • R 15a is C 1 -C 6 alkyl (e.g., methyl or iso-propyl) and R 15b is H.
  • R 15a is H and R 15b is H.
  • the compound of Formula I has the structure of Formula I-1a: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-1b: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1c: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1d: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1e: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1f: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1g: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-1h: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1i: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1j: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1k: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1m: or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula I has the structure of Formula I-1n: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-1o: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2a: , or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2b: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2c: et or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2d: , or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2e: or a pharmaceutically acceptable salt thereof.
  • the compound of Formula I has the structure of Formula I-2f: , or a pharmaceutically acceptable salt thereof.
  • the compound has the structure of any one of compounds 1-17 in Table 1A, or a pharmaceutically acceptable salt thereof.
  • the compound has the structure of any one of compounds 18-47 in Table 1B, or a pharmaceutically acceptable salt thereof.
  • the disclosure features a compound having the structure of any one of compounds 1-17 in Table 1A, or a pharmaceutically acceptable salt thereof.
  • the disclosure features a compound having the structure of any one of compounds 18-47 in Table 1B, or a pharmaceutically acceptable salt thereof.
  • Table 1A Compounds 1-17 of the Disclosure Table 1B.
  • the disclosure features a pharmaceutical composition including any of the foregoing compounds, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable excipient.
  • the disclosure features a method of reducing the level of HAO1 in a cell, where the method includes contacting the cell with an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disclosure features a method of inhibiting the activity of HAO1 in a cell, where the method includes contacting the cell with an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the cell is a liver cell.
  • the liver cell is a human liver cell.
  • the disclosure features a method of treating a disorder that leads to accumulation of oxalate in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disclosure features a method of treating an HAO1-associated disorder in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disorder is primary hyperoxaluria 1. In some embodiments, the disorder is primary hyperoxaluria 2.
  • the disclosure features a method of treating primary hyperoxaluria in a subject in need thereof (e.g., human in need thereof), where the method includes administering to the subject an effective amount of a compound of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the primary hyperoxaluria is primary hyperoxaluria 1.
  • the primary hyperoxaluria is primary hyperoxaluria 2.
  • the disclosure features a method of reducing the level of oxalate in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disclosure features a method of reducing the level of glyoxylate in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disclosure features a method of reducing the level of glycine in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the disclosure features a method of inhibiting the accumulation of oxalate in a subject in need thereof (e.g., human in need thereof), where the method comprises administering to the subject an effective amount of any of the foregoing compounds, or a pharmaceutically acceptable salt thereof, or any of the foregoing pharmaceutical compositions.
  • the subject is a human child, e.g., a human that is less than 18 years old (e.g., less than 15 years old, less than 10 years old, less than 9 years old, less than 8 years old, less than 7 years old, less than 6 years old, less than 5 years old, less than 4 years old, less than 3 years old, less than 2 years old, less than 1 years old).
  • the subject is a human adult, e.g., a human that is at least 18 years old (e.g., at least 19 years old, at least 20 years old, at least 30 years old, at least 40 years old, at least 50 years old).
  • a number following an atomic symbol indicates that total number of atoms of that element that are present in a particular chemical moiety.
  • other atoms such as hydrogen atoms, or substituent groups, as described herein, may be present, as necessary, to satisfy the valences of the atoms.
  • an unsubstituted C 2 alkyl group has the formula –CH 2 CH3.
  • a reference to the number of carbon atoms includes the divalent carbon in acetal and ketal groups but does not include the carbonyl carbon in acyl, ester, carbonate, or carbamate groups.
  • acyl represents a hydrogen or an alkyl group that is attached to a parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl.
  • exemplary unsubstituted acyl groups include from 1 to 6, from 1 to 11, or from 1 to 21 carbons.
  • alkyl refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms).
  • An alkylene is a divalent alkyl group.
  • alkenyl as used herein, alone or in combination with other groups, refers to a straight chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).
  • An alkenylene is a divalent alkenyl group.
  • alkynyl refers to a straight chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).
  • An alkynylene is a divalent alkynyl group.
  • amino represents –N(R N1 ) 2 , wherein each R N1 is, independently, H, OH, NO2, N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2 , an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), wherein each of these recited R N1 groups can be optionally substituted; or two R N1 combine to form an alkylene or heteroalkylene, and wherein each R N2 is, independently, H, alkyl, or aryl.
  • each R N1 is, independently, H, alkyl, or aryl.
  • the amino groups of the compounds described herein can be an unsubstituted amino (i.e., –NH 2 ) or a substituted amino (i.e., –N(R N1 )2).
  • aryl refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) radical of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H- indenyl.
  • arylalkyl represents an alkyl group substituted with an aryl group.
  • exemplary unsubstituted arylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C 1 -C 6 alkyl C 6 -C 10 aryl, C 1 -C 10 alkyl C 6 -C 10 aryl, or C 1 -C 20 alkyl C 6 -C 10 aryl), such as, benzyl and phenethyl.
  • azido represents a –N3 group.
  • cyano represents a —CN group.
  • Carbocyclyl refers to a monocarbocyclic or polycarbocyclic (e.g., bicyclic or tricyclic) radical of 3 to 12 carbon atoms in which no ring is aromatic.
  • Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals (e.g., cycloalkenyl and cycloalkynyl).
  • Examples of a carbocyclyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, norbornyl, adamantyl, and cyclooctynyl.
  • the term “cycloalkyl,” as used herein, refers to a saturated, non-aromatic, monovalent monocarbocyclic or polycarbocyclic radical of 3 to 12 carbon atoms. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.
  • halo or halogen, as used herein, means a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.
  • haloalkyl refers to an alkyl group that is substituted with one or more halogen atoms. Examples of haloalkyl include fluoroalkyl (e.g., difluoromethyl, trifluoroethyl, and perfluoroalkyl (e.g., trifluoromethyl and pentafluoroethyl)), chloroalkyl, bromoalkyl, and iodoalkyl.
  • heteroalkyl refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • heteroalkyl groups are an “alkoxy” which, as used herein, refers alkyl–O– (e.g., methoxy and ethoxy).
  • a heteroalkylene is a divalent heteroalkyl group.
  • heteroalkenyl refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • heteroalkenyl groups are an “alkenoxy” which, as used herein, refers alkenyl–O–.
  • a heteroalkenylene is a divalent heteroalkenyl group.
  • heteroalkynyl refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur.
  • heteroalkynyl groups are an “alkynoxy” which, as used herein, refers alkynyl–O–.
  • a heteroalkynylene is a divalent heteroalkynyl group.
  • heteroaryl refers to a monocyclic or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing at least 1 (e.g., 1, 2, 3, 4, or 5) ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon.
  • One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group.
  • heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl.
  • heteroarylalkyl represents an alkyl group substituted with a heteroaryl group.
  • exemplary unsubstituted heteroarylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C 1 -C 6 alkyl C 2 -C 9 heteroaryl, C 1 -C 10 alkyl C 2 -C 9 heteroaryl, or C 1 -C 20 alkyl C2-C9 heteroaryl).
  • heterocyclyl refers a monocyclic or polycyclic radical (e.g., bicyclic or tricyclic) having 3 to 12 atoms having at least one ring containing at least 1 (e.g., 1, 2, 3, 4, or 5) ring atoms selected from nitrogen, oxygen, or sulfur, where no ring is aromatic.
  • Heterocyclyl structures include heterocycloalkyl groups and unsaturated hetercyclyl radicals (e.g., heterocycloalkenyl and heterocycloalkynyl).
  • heterocyclyl groups are morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl.
  • heterocycloalkyl refers to a saturated, non-aromatic, monovalent monocyclic or polycyclic radical of 3 to 12 atoms having at least one ring containing at least 1 (e.g., 1, 2, 3, 4, or 5) ring atoms selected from nitrogen, oxygen, or sulfur, where no ring is aromatic.
  • hydroxyalkyl represents alkyl group substituted with an —OH group.
  • hydroxyl represents an —OH group.
  • N-protecting group represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, “Protective Groups in Organic Synthesis,” 3rd Edition (John Wiley & Sons, New York, 1999).
  • N-protecting groups include, but are not limited to, acyl, aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4- bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L, or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, and p-toluenesulfonyl; carbamate forming groups such as benzyl
  • N-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t- butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).
  • nitro represents an —NO2 group.
  • thiol represents an —SH group.
  • optionally substituted X e.g., optionally substituted alkyl
  • X optionally substituted
  • alkyl wherein said alkyl is optionally substituted
  • the functional groups described herein may be substituted or unsubstituted.
  • substituents there will generally be 1 to 6 (e.g., 1 to 5, 1 to 4, 1 to 3, or 1 to 2) substituents present, unless otherwise specified.
  • substituents include: alkyl (e.g., unsubstituted and substituted, where the substituents include any group described herein, e.g., aryl, halo, hydroxyl), aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluoro), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH 2 or mono- or dialkyl amino), azido, cyano, nitro, or thiol.
  • alkyl e.g., unsubstituted and substituted, where the substituents include any group described herein, e.g., aryl, halo, hydroxyl
  • Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted such as arylalkyl (e.g., substituted and unsubstituted benzyl)).
  • substituents include: (1) C 1-6 alkoxy; (2) C 1-6 alkylsulfinyl; (3) amino, as defined herein (e.g., unsubstituted amino (i.e., -NH 2 ) or a substituted amino (i.e., -N(R N1 )2, where R N1 is as defined for amino); (4) C 6-10 aryl-C 1-6 alkoxy; (5) azido; (6) halo; (7) (C 2-9 heterocyclyl)oxy; (8) hydroxyl, optionally substituted with an O-protecting group; (9) nitro; (10) oxo (e.g., carboxyaldehyde or acyl); (11) C 1 -7 spirocyclyl; (12) thioalkoxy; (13) thiol; (14) -CO2R A’ , optionally substituted with an O- protecting group and where R A’ is selected from the group consisting of (a) C 1-20 alkyl
  • each of these groups can be further substituted as described herein.
  • the alkylene group of a C 1 -alkaryl can be further substituted with an oxo group to afford the respective aryloyl substituent.
  • oxo group to afford the respective aryloyl substituent.
  • certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, tautomers) and/or isotopic (e.g., in which one or more atoms has been substituted with a different isotope of the atom, such as hydrogen substituted for deuterium) forms. Unless otherwise indicated or clear from context, a depicted structure can be understood to represent any such isomeric or isotopic form, individually or in combination.
  • one or more compounds depicted herein may exist in different tautomeric forms.
  • references to such compounds encompass all such tautomeric forms.
  • tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton.
  • a tautomeric form may be a prototropic tautomer, which is an isomeric protonation states having the same empirical formula and total charge as a reference form.
  • moieties with prototropic tautomeric forms are ketone - enol pairs, amide - imidic acid pairs, lactam - lactim pairs, amide - imidic acid pairs, enamine - imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H- and 3H- 1,2,3-triazole, 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.
  • the invention embraces all of these forms.
  • Compounds described herein can have one or more asymmetric carbon atoms and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, or mixtures of diastereoisomeric racemates.
  • the optically active forms can be obtained for example by resolution of the racemates, by asymmetric synthesis or asymmetric chromatography (chromatography with a chiral adsorbent or eluant). That is, certain of the disclosed compounds may exist in various stereoisomeric forms.
  • Stereoisomers are compounds that differ only in their spatial arrangement.
  • Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. "Enantiomer” means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon.
  • Racemate or “racemic mixture” means a compound containing two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light.
  • Geometric isomer means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system.
  • Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on 25 opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration.
  • "R,” “S,” “S*,” “R*,” “E,” “Z,” “cis,” and “trans,” indicate configurations relative to the core molecule.
  • Certain of the disclosed compounds may exist in atropisomeric forms.
  • Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers.
  • the compounds described herein may be prepared as individual isomers by either isomer-specific synthesis or resolved from an isomeric mixture.
  • Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide 35 of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods.
  • the stereochemistry of a disclosed compound is named or depicted by structure
  • the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers.
  • the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight optically pure.
  • the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight pure.
  • Percent optical purity is the ratio of the weight of the enantiomer or over the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or over the weight of all the diastereomers.
  • the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure relative to the other stereoisomers.
  • the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure.
  • the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by mole fraction pure.
  • Percent purity by mole fraction is the ratio of the moles of the enantiomer or over the moles of the enantiomer plus the moles of its optical isomer.
  • percent purity by moles fraction is the ratio of the moles of the diastereomer or over the moles of the diastereomer plus the moles of its isomer.
  • the term “a” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and/or”; and (iii) the terms “including” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps.
  • the terms “about” and “approximately” refer to a value that is within 10% above or below the value being described. For example, the term “about 5 nM” indicates a range of from 4.5 to 5.5 nM.
  • the term “accumulation of oxalate” refers to an increased level of oxalate (e.g., urinary oxalate level) in a subject (e.g., human), as compared to a reference (e.g., the level of oxalate in a healthy subject or the level of oxalate prior to administration of a compound of the invention).
  • a reference e.g., the level of oxalate in a healthy subject or the level of oxalate prior to administration of a compound of the invention.
  • the term “inhibiting accumulation of oxalate” refers to reducing the rate of accumulation of oxalate in a subject compared to the rate of accumulation of oxalate in the subject prior to administration of a compound of the invention (e.g., a decrease in the rate of accumulation by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease in the rate of accumulation of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease in the rate of accumulation by more than about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.
  • level is meant a level of, e.g., oxalate, glyoxylate, or glycine, as compared to a reference.
  • the reference can be any useful reference, as defined herein.
  • a “decreased level” or an “increased level” of oxalate is meant a decrease or increase in, e.g., oxalate level (e.g., urinary oxalate level or blood oxalate level), glyoxylate level, or glycine level, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an
  • a level of, e.g., oxalate e.g., urinary oxalate level or blood oxalate level
  • glyoxylate, or glycine may be expressed in mass/vol (e.g., g/dL, mg/mL, ⁇ g/mL, or ng/mL), in mass/vol/time (e.g., g/dL/hours, mg/mL/hours, ⁇ g/mL/hours, or ng/mL/hours), or percentage relative to total oxalate in a sample (e.g., a urine sample or blood sample).
  • administration refers to the administration of a composition (e.g., a compound or a preparation that includes a compound as described herein) to a subject or system.
  • Administration to an animal subject may be by any appropriate route.
  • administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, and vitreal.
  • a “combination therapy” or “administered in combination” means that two (or more) different agents or treatments are administered to a subject as part of a defined treatment regimen for a particular disease or condition.
  • the treatment regimen defines the doses and periodicity of administration of each agent such that the effects of the separate agents on the subject overlap.
  • the delivery of the two or more agents is simultaneous or concurrent and the agents may be co-formulated.
  • the two or more agents are not co-formulated and are administered in a sequential manner as part of a prescribed regimen.
  • administration of two or more agents or treatments in combination is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one agent or treatment delivered alone or in the absence of the other.
  • the effect of the two treatments can be partially additive, wholly additive, or more than additive (e.g., synergistic).
  • each therapeutic agent can be effected by any appropriate route including, but not limited to, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucous membrane tissues.
  • the therapeutic agents can be administered by the same route or by different routes.
  • a first therapeutic agent of the combination may be administered by intravenous injection while a second therapeutic agent of the combination may be administered orally.
  • the terms “effective amount,” “therapeutically effective amount,” and “a “sufficient amount” of an agent refer to a quantity of an agent described herein sufficient to, when administered to the subject, including a human, effect beneficial or desired results, including clinical results, and, as such, an “effective amount” or synonym thereto depends on the context in which it is being applied.
  • the agent that reduces the level and/or activity of HAO1 or reduces the level of oxalate sufficient to achieve a treatment response as compared to the response obtained without administration of the agent that reduces the level and/or activity of HAO1 or reduces the level of oxalate.
  • a given agent that reduces the level and/or activity of HAO1 or reduces the level of oxalate described herein that will correspond to such an amount will vary depending upon various factors, such as the given agent, the pharmaceutical formulation, the route of administration, the type of disease or disorder, the identity of the subject (e.g., age, sex, and/or weight) or host being treated, and the like, but can nevertheless be routinely determined by one of skill in the art.
  • a “therapeutically effective amount” of an agent is an amount which results in a beneficial or desired result in a subject as compared to a control.
  • a therapeutically effective amount of an agent that reduces the level and/or activity of HAO1, reduces the level of oxalate, or treats an HAO1-associated disorder may be readily determined by one of ordinary skill by routine methods known in the art. Dosage regimen may be adjusted to provide the optimum therapeutic response.
  • HAO1-associated disorder refers to a disorder in which an activity related to HAO1, or a downstream effect (e.g., increased level of oxalate or increased level of glyoxylate) is amplified compared to a reference.
  • agents described herein are useful for the treatment of an HAO1-associated disorder and/or alleviation of a symptom associated therewith.
  • pharmaceutically acceptable excipient refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient.
  • Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, and waters of hydration.
  • antiadherents antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, and waters of hydration.
  • excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C,
  • pharmaceutically acceptable salt means any pharmaceutically acceptable salt of a compound described herein.
  • pharmaceutically acceptable salts of any of the compounds described herein include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008.
  • the salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable organic acid.
  • the compounds described herein may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts.
  • These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds described herein, be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparation of the appropriate salts are well-known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases.
  • Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pe
  • alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.
  • pharmaceutical composition represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal.
  • compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.
  • PFS progression-free survival
  • progression-free survival refers to the length of time during and after treatment during which a disease being treated (e.g., HAO1-associated disorder) does not get worse.
  • Progression-free survival may include the amount of time patients have experienced a complete response or a partial response, as well as the amount of time patients have experienced stable disease.
  • PFS may be defined as the time from randomization or the beginning of treatment to the first documented disease progression.
  • reducing the level of glycine is meant decreasing the level of glycine in a subject (e.g., human).
  • the level of glycine may be measured using any method known in the art.
  • reducing the level of glyoxylate is meant decreasing the level of glyoxylate in a subject (e.g., human).
  • the level of glyoxylate may be measured using any method known in the art.
  • reducing the level of oxalate is meant decreasing the level of oxalate in a subject (e.g., human).
  • the level of oxalate may be measured using any method known in the art.
  • reducing the level of HAO1 is meant decreasing the level of HAO1 in a cell or a subject (e.g., a human cell or a human subject).
  • the level of HAO1 is determined by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample) to obtain the physical entity or value.
  • “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third-party laboratory that directly acquired the physical entity or value).
  • Methods to measure HAO1 level generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescent polarization, phosphorescence, immunohistochemical analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS), and flow cytometry, as well as assays based on a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners.
  • ELISA enzyme-linked immunosorbent assay
  • the level of HAO1 is decreased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; decreased by more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; or decreased by more than about 1.2-fold, about 1.4- fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, 25 about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more).
  • inhibiting the activity of HAO1 is meant decreasing the level of an activity related to HAO1, or a related downstream effect (e.g., increased level of oxalate or increased level of glyoxylate).
  • the activity level of HAO1 may be measured using any method known in the art, e.g., a horseradish peroxidase- based assay as described herein.
  • an agent that inhibits the activity of HAO1 is a compound as described herein.
  • the activity of HAO1 is decreased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; decreased by more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; or decreased by more than about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0- fold, 25 about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100- fold, about 1000-fold, or more).
  • the term “subject” refers to any organism to which a composition in accordance with the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.
  • animal e.g., mammals such as mice, rats, rabbits, non-human primates, and humans.
  • a subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.
  • treat means both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results.
  • Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease.
  • Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.
  • the present disclosure features compounds and methods useful for treating HAO1-associated disorders and/or disorders (e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)) that lead to oxalate accumulation and its symptoms (e.g., end stage renal disease).
  • HAO1-associated disorders and/or disorders e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)
  • oxalate accumulation and its symptoms e.g., end stage renal disease
  • the disclosure further features compositions and methods useful for reduction of the level and/or activity of HAO1, e.g., for the treatment of disorders that lead to oxalate accumulation and its symptoms in a subject in need thereof (e.g., human).
  • Glyoxylate Enzymes Alanine-glyoxylate aminotransferase (AGT), glyoxylate reductase-hydroxypyruvate reductase (GRHPR), and 4-hydroxy-2-oxoglutarate aldolase (HOGA1) are involved in the breakdown and processing of protein building blocks (e.g., amino acids) and other compounds.
  • AGT is involved in the breakdown of L-alanine and glyoxylate.
  • GRHPR is involved in the breakdown of glyoxylate and hydroxypyruvate.
  • HOGA1 is involved in the breakdown of amino acids, which results in the formation of glyoxylate.
  • HAO1 oxidizes glycolic acid (glycolate) to glyoxylate.
  • the liver-specific peroxisomal enzymes alanine-glyoxylate aminotransferase (AGT) and glyoxylate reductase-hydroxypyruvate reductase (GRHPR) catalyze the detoxification (transamination) of glyoxylate to glycine.
  • AGT alanine-glyoxylate aminotransferase
  • GRHPR glyoxylate reductase-hydroxypyruvate reductase
  • LDH lactate dehydrogenase
  • Oxalate tends to precipitate as insoluble calcium oxalate crystals in tissues, leading to harmful kidney stones and bladder stones. Renal damage is caused by a combination of toxicity from oxalate (e.g., death of tubular epithelial cells of the renal tubules of the kidneys), nephrocalcinosis, and renal obstruction by the stones.
  • oxalate e.g., death of tubular epithelial cells of the renal tubules of the kidneys
  • nephrocalcinosis e.g., renal obstruction by the stones.
  • reduction of HAO1 activity should reduce endogenous oxalate production, e.g., oxidation of glyoxylate by LDH, by reducing the production of glyoxylate from glycolic acid. Ultimately, this should lead to a reduction of oxalate and therefore calcium oxalate.
  • Primary Hyperoxaluria Metabolic disorders can be caused by mutations in genes that encode enzymes of a metabolic pathway. Mutation can result in a harmful accumulation of a compound that is normally metabolized by the enzyme.
  • Primary hyperoxalurias are rare autosomal recessive disorders affecting the glyoxylate or hydroxyproline pathways that result in an overproduction or accumulation of oxalate.
  • Primary hyperoxaluria is estimated to affect 1 in 58,000 individuals worldwide.
  • PH primary hyperoxaluria
  • PH1 primary hyperoxaluria type 1
  • ESRD end stage renal disease
  • PH2 primary hyperoxaluria type 2
  • PH3 primary hyperoxaluria type 3
  • PH1 accounts for approximately 80% of primary hyperoxaluria cases and eventually leads to renal failure after several years.
  • PH2 and PH3 each account for about 10% of primary hyperoxaluria cases and have a less severe course.
  • Kidney stones can be treated, e.g., by surgical removal of the stones, dietary changes to increase fluid intake, dietary changes to restrict oxalate intake, urine alkalization, diuretics, and crystallization inhibitors (e.g., citrate, bicarbonate, and magnesium).
  • Compounds described herein reduce the level and/or activity of HAO1 and/or reduce the level of oxalate, glyoxylate, and/or glycine for the treatment of disorders (e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)) that lead to oxalate accumulation and its symptoms.
  • disorders e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1)
  • Exemplary compounds described herein have the structure according to Formula I.
  • Formula I is shown below: where X 1 is N or CR 1 ; X 2 is N or CR 2 ; X 3 is N or CR 3 ; X 4 is N or C-COOH; X 5 is N or C-Het; each of R 1 , R 2 , and R 3 is, independently, H, cyano, hydroxyl, halogen, optionally substituted C 1 -C 6 alkyl, or optionally substituted C 1 -C 6 heteroalkyl; R 4 is H or optionally substituted C 1 -C 6 alkyl; R 5 is H, optionally substituted C 1 -C 6 alkyl, or optionally substituted C 1 -C 6 alkyl C 6 -C 1 0 aryl; each of R 6 and R 7 is, independently, H or optionally substituted C 1 -C 6 alkyl, or R 6 and R 7 combine with the carbon to which they are attached to form -C(O)-; each of R 8 , R 9 , R 10 , and
  • the compounds described herein are useful in the methods of the invention and, while not bound by theory, are believed to exert their desirable effects through their ability to modulate the level of oxalate, glyoxylate, and/or glycine, e.g., by modulating the level, status, and/or activity of HAO1 in a cell in a subject (e.g., mammal (e.g., human)).
  • An aspect of the present invention relates to methods of treating disorders such as primary hyperoxalurias (e.g., primary hyperoxaluria 1 or primary hyperoxaluria 2) related to oxalate levels and/or HAO1 levels and/or activity in a subject in need thereof.
  • the compound is administered in an amount and for a time effective to result in one of (or more, e.g., two or more, three or more, four or more of): (a) reduced level of oxalate production (e.g., reduced urinary oxalate level); (b) reduced oxalate accumulation; (c) decreased frequency and/or size of kidney stones; (d) delayed onset of end stage renal disease; (e) increased survival of a subject (e.g., mammal (e.g., human)); and (f) increased progression free survival of a subject (e.g., human).
  • the methods of the invention result in an increase in the length of time during and after treatment that the subject lives with an HAO1-associated disorder, but the disorder does not get worse (e.g., the amount of renal and/or bladder stones are stable or decreased and/or the amount of kidney and/or bladder infections are stable or decreased).
  • Administration of compounds of the invention may result in reduced levels of oxalate in a subject or cell.
  • the level of oxalate is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • administration of compounds of the invention may reduce further accumulation of oxalate in a subject or cell.
  • oxalate is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • Administration of compounds of the invention may result in reduced levels of glyoxylate in a subject or cell.
  • the level of glyoxylate is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • Administration of compounds of the invention may result in reduced levels of glycine in a subject or cell.
  • the level of glycine is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • Administration of compounds of the invention may result in a decrease in number of kidney stones in a subject.
  • kidney stone number is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the number prior to treatment.
  • Administration of compounds of the invention may result in a reduced frequency of new kidney stones in a subject.
  • the frequency of new kidney stones is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • Administration of compounds of the invention may result in decreased overall mass of kidney stones in a subject.
  • the overall mass of kidney stones is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the level prior to treatment.
  • Administration of compounds of the invention may result in delayed onset of end stage renal disease in a subject.
  • the onset of end stage renal disease is delayed by more than 30 days (more than 60 days, 90 days, or 120 days) or by more than 1 year (e.g., more than 2 years, more than 5 years, more than 10 years, more than 15 years, more than 20 years, more than 30 years, more than 40 years, or more than 50 years) in a subject.
  • An increase in average onset of end stage renal disease of a population may be measured by any reproducible means.
  • an increase in average onset of end stage renal disease of a population may be measured by calculating for a population the average length of time following initiation of treatment with a compound described herein before onset of end stage renal disease.
  • An increase in average length of time before onset of end stage renal stage disease may also be measured, for example, by calculating for a population the average length of time before onset of end stage renal stage disease following completion of a first round of treatment with a pharmaceutically acceptable salt of a compound described herein.
  • Administration of compounds of the invention can result in an increase in average survival time of subjects treated according to the present invention in comparison to a population of untreated subjects.
  • the average survival time is increased by more than 30 days (more than 60 days, 90 days, or 120 days) or by more than 1 year (more than 2 years, more than 5 years, more than 10 years, or more than 20 years).
  • An increase in average survival time of a population may be measured by any reproducible means.
  • an increase in average survival time of a population may be measured by calculating for a population the average length of survival following initiation of treatment with the compound described herein.
  • An increase in average survival time of a population may also be measured, for example, by calculating for a population the average length of survival following completion of a first round of treatment with a pharmaceutically acceptable salt of a compound described herein.
  • Administration of compounds of the invention can also result in a decrease in the mortality rate of subjects treated in comparison to an untreated population. For example, the mortality rate is decreased by more than 2% (e.g., more than 5%, 10%, or 25%).
  • a decrease in the mortality rate of a population of treated subjects may be measured by any reproducible means by calculating for a population the average number of disease-related deaths per unit time following initiation of treatment with a pharmaceutically acceptable salt of a compound described herein.
  • a decrease in the mortality rate of a population may also be measured, for example, by calculating for a population the average number of disease-related deaths per unit time following completion of a first round of treatment with a pharmaceutically acceptable salt of a compound described herein.
  • a method of the invention can be used alone or in combination with additional therapy (e.g., a compound (e.g., vitamin B6)) that treats HAO1-associated disorders and/or disorders that lead to oxalate accumulation (e.g., primary hyperoxalurias (e.g., primary hyperoxaluria 1 or primary hyperoxaluria 2)) or symptoms associated therewith.
  • additional therapy e.g., a compound (e.g., vitamin B6)
  • the dosages of one or more of the additional therapies e.g., a compound (e.g., vitamin B6)
  • doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)). In this case, dosages of the compounds when combined should provide a therapeutic effect.
  • the first and second therapy are administered simultaneously or sequentially, in either order.
  • the first therapeutic agent may be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to, 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to hours 16, up to 17 hours, up 18 hours, up to 19 hours up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours up to 24 hours or up to 1-7, 1-14, 1-21 or 1-30 days or up to 1-5 years, 5-10 years, or 10-20 years before or after the second therapeutic agent.
  • Additional therapy that treats HAO1-associated disorders and/or disorders that lead to oxalate accumulation can also be a non-drug treatment.
  • the additional therapy may be shock wave lithotripsy, cystoscopy and ureteroscopy, or percutaneous nephrolithotomy to remove kidney stones, or an oxalate-limited diet.
  • two or more additional therapies are administered.
  • an additional therapy e.g., a compound (e.g., vitamin B6) is administered with an oxalate- limited diet.
  • the additional therapy e.g., a compound (e.g., vitamin B6)
  • a compound e.g., vitamin B6
  • the additional therapy is administered without an oxalate-limited diet.
  • Pharmaceutical Compositions The pharmaceutical compositions described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo.
  • the compounds described herein may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the methods described herein.
  • the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art.
  • the compounds described herein may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, intratumoral, or transdermal administration and the pharmaceutical compositions formulated accordingly.
  • Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration.
  • Parenteral administration may be by continuous infusion over a selected period of time.
  • a compound described herein may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard- or soft-shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet.
  • a compound described herein may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers.
  • a compound described herein may also be administered parenterally.
  • Solutions of a compound described herein can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
  • compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders.
  • Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device.
  • the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use.
  • the dosage form includes an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon.
  • the aerosol dosage forms can also take the form of a pump-atomizer.
  • compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerine.
  • Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter.
  • a compound described herein may be administered intratumorally, for example, as an intratumoral injection. Intratumoral injection is injection directly into the tumor vasculature and is specifically contemplated for discrete, solid, accessible tumors. Local, regional, or systemic administration also may be appropriate.
  • a compound described herein may advantageously be contacted by administering an injection or multiple injections to the tumor, spaced for example, at approximately, 1 cm intervals.
  • the present invention may be used preoperatively, such as to render an inoperable tumor subject to resection.
  • Continuous administration also may be applied where appropriate, for example, by implanting a catheter into a tumor or into tumor vasculature.
  • the compounds described herein may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.
  • the dosage of the compounds described herein, and/or compositions including a compound described herein can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated.
  • One of skill in the art can determine the appropriate dosage based on the above factors.
  • the compounds described herein may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, satisfactory results may be obtained when the compounds described herein are administered to a human at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as the solid form).
  • Dose ranges include, for example, between 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered. Alternatively, the dosage amount can be calculated using the body weight of the patient. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may range from 0.1-50 mg/kg (e.g., 0.25-25 mg/kg).
  • the dose may range from 0.5-5.0 mg/kg (e.g., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg/kg) or from 5.0-20 mg/kg (e.g., 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg/kg).
  • 0.5-5.0 mg/kg e.g., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg/kg
  • 5.0-20 mg/kg e.g., 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg/kg.
  • kits including (a) a pharmaceutical composition including an agent (e.g., a compound of Formula I) that reduces the level of oxalate, glyoxylate, and/or glycine, and/or reduces the level and/or activity of HAO1 in a cell or subject described herein, and (b) a package insert with instructions to perform any of the methods described herein.
  • an agent e.g., a compound of Formula I
  • the kit includes (a) a pharmaceutical composition including an agent (e.g., a compound of Formula I) that reduces the level of oxalate, glyoxylate, and/or glycine, and/or reduces the level and/or activity of HAO1 in a cell or subject described herein, (b) an additional therapy (e.g., vitamin B6 or an oxalate-limited diet), and (c) a package insert with instructions to perform any of the methods described herein.
  • an agent e.g., a compound of Formula I
  • an additional therapy e.g., vitamin B6 or an oxalate-limited diet
  • Example 1 Synthesis of 5-((2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl) benzyl) amino)-1H-indazole-3- carboxylic acid (compound 1)
  • Step 1 Synthesis of 2-methoxy-3-(1-methyl-1H-pyrazol-3-yl) benzaldehyde
  • a solution of 3-bromo-1-methyl-1H-pyrazole (320 mg, 2 mmol), (3-formyl-2- methoxyphenyl)boronic acid (360 mg, 2 mmol), Pd(P(Ph3))4 (116 mg, 0.1 mmol) and K2CO3 (276 mg, 2 mmol) in dimethylformamide (DMF)/ H 2 O 4: 1 (5 mL) was stirred at 80 °C under the protection of nitrogen for 6 hours (h).
  • Step 2 Synthesis of 2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl) benzaldehyde
  • BBr3 2-methoxy-3-(1-methyl-1H-pyrazol-3-yl) benzaldehyde
  • LCMS showed the reaction was completed.
  • the reaction mixture was concentration under vacuum to give 2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl) benzaldehyde (290 mg, crude) as a yellow oil.
  • Step 3 Synthesis of 5-((2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl) benzyl) amino)-1H-indazole-3-carboxylic acid
  • 2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl) benzaldehyde 290 mg, ⁇ 1 mmol
  • 5- amino-1H-indazole-3-carboxylic acid 178 mg, 1 mmol
  • InCl 3 (293 mg, 1 mmol)
  • triethylsilane 232 mg, 2 mmol
  • Step 2 Synthesis of 5-((2-hydroxy-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 2-hydroxy-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)benzaldehyde 200 mg, 0.86 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 177 mg, 1 mmol
  • acetic acid (AcOH) 0.1 mL
  • 10 mL MeOH was stirred at rt for 0.5 h, then NaBH3CN (124 mg, 2 mmol) was added, then the reaction was stirred at rt overnight.
  • Example 3 Synthesis of 5-[([6-[3-(dimethylamino)-1,2,4-oxadiazol-5-yl]-1H-indazol-4- yl]methyl)amino]-1H-indazole-3-carboxylic acid (Compound 3) Step 1: Synthesis of (6-bromo-1H-indazol-4-yl)methanol A solution of methyl 6-bromo-1H-indazole-4-carboxylate (2.00 g, 7.84 mmol) and LiAlH 4 (889.8 mg, 23.4 mmol) in tetrahydrofuran (THF) (25 mL) was stirred at room temperature for 4 h under nitrogen atmosphere.
  • THF tetrahydrofuran
  • Step 2 Synthesis of 6-bromo-1H-indazole-4-carbaldehyde MnO 2 (2.87 g, 33.02 mmol) were added (6-bromo-1H-indazol-4-yl)methanol (1.50 g, 6.61 mmol) in DCM (20 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was filtered, and the filter cake was washed with DCM. The filtrate was concentrated under reduced pressure.
  • Step 3 Synthesis of methyl 4-formyl-1H-indazole-6-carboxylate
  • 6-bromo-1H-indazole-4-carbaldehyde 730 mg, 3.24 mmol
  • MeOH MeOH
  • Pd(dppf)Cl 2 474.7 mg, 0.65 mmol
  • Et 3 N 984.7 mg, 9.73 mmol
  • the resulting mixture was stirred at 90 oC under CO (5 atm) overnight.
  • the resulting mixture was purified using C18 flash chromatography with the following conditions (Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% to 100% in 30 min).
  • Step 5 Synthesis of 4-formyl-N-[N'-hydroxy-N,N-dimethylcarbamimidoyl]-1H-indazole-6-carboxamide
  • DMF dimethyl sulfoxide
  • Step 6 Synthesis of 6-[3-(dimethylamino)-1,2,4-oxadiazol-5-yl]-1H-indazole-4-carbaldehyde
  • 4-formyl-N-[N'-hydroxy-N,N-dimethylcarbamimidoyl]-1H-indazole-6- carboxamide 220 mg, 0.79 mmol
  • THF 6 mL
  • the resulting mixture was stirred at 60 oC for 1 h.
  • the resulting mixture was concentrated under vacuum.
  • Step 7 Synthesis of 5-[([6-[3-(dimethylamino)-1,2,4-oxadiazol-5-yl]-1H-indazol-4-yl]methyl)amino]-1H- indazole-3-carboxylic acid
  • 6-[3-(dimethylamino)-1,2,4-oxadiazol-5-yl]-1H-indazole-4-carbaldehyde 190 mg, 0.74 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 157 mg, 0.89 mmol
  • Step 2 Synthesis of 5-((2-chloro-3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 2-chloro-3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde 150 mg, 0.60 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 106 mg, 0.60 mmol
  • MeOH MeOH
  • InCl 3 398 mg, 1.80 mmol
  • TES triethylsilane
  • Example 5 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H- pyrazolo[3,4-b]pyridine-3-carboxylic acid (Compound 5)
  • Step 1 Synthesis of 5-amino-1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid
  • Pd/C 10%, 100 mg
  • Step 2 Synthesis of 3-hydroxy-1,1-dimethylguanidine To a solution of hydroxylamine (500 mg, 7.25 mmol) and NaHCO 3 (1.2 g, 14.50 mmol) in EtOH (10 mL) was added N,N-dimethylcyanamide (508 mg, 7.25 mmol). The reaction was stirred at rt for 2 h.
  • Step 3 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde
  • EDCI 126 mg, 6.60 mmol
  • HOBT 89 mg, 6.60 mmol
  • DIEA 851 mg, 6.60 mmol
  • Step 4 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-pyrazolo[3,4-b]pyridine-3- carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde (434 mg, 2.00 mmol)
  • 5-amino-1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid 356 mg, 2.00 mmol) in MeOH (10 mL) was added InCl3 (1.3 g, 6.00 mmol) followed by TES (0.7 mL, 6.00 mmol).
  • Example 6 Synthesis of 5-((3-chloro-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H- indazole-3-carboxylic acid (Compound 6)
  • Step 1 Synthesis of 3-chloro-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde
  • EDCI 516 mg, 2.70 mmol
  • HOBT 365 mg, 2.70 mmol
  • DIEA 348 mg, 2.70 mmol
  • Step 2 Synthesis of 5-((3-chloro-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-chloro-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde 230 mg, 0.91 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 161 mg, 0.91 mmol
  • MeOH MeOH
  • InCl3 603 mg, 2.73 mmol
  • TES 317 mL, 2.73 mmol
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxy-5-methylbenzyl)amino)-1H- indazole-3-carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxy-5-methylbenzaldehyde 140 mg, 0.56 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 187 mg, 1.00 mmol
  • one drop of acetic acid in MeOH (2 mL) was stirred at rt overnight.
  • Example 8 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-methoxybenzyl)amino)- 1H-indazole-3-carboxylic acid (Compound 8) Step 1: Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-methoxybenzaldehyde To a solution of 3-formyl-5-methoxybenzoic acid (500 mg, 2.91 mmol) and 3-hydroxy-1,1- dimethylguanidine (300 mg, 2.91 mmol) in DMF (5 mL) was added HATU (1.3 g, 3.49 mmol) followed by DIEA (1.5 mL, 8.73 mmol).
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-methoxybenzyl)amino)-1H-indazole- 3-carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-methoxybenzaldehyde 100 mg, 0.40 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 71 mg, 0.40 mmol
  • one drop of acetic acid in MeOH (3 mL) was stirred at rt overnight.
  • Example 9 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-fluorobenzyl)amino)-1H- indazole-3-carboxylic acid (Compound 9)
  • Step 1 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-fluorobenzaldehyde
  • 2-fluoro-3-formylbenzoic acid 300 mg, 1.79 mmol
  • 3-hydroxy-1,1- dimethylguanidine 202 mg, 1.79 mmol
  • EDCI 514 mg, 2.68 mmol
  • HOBT 359 mg, 2.68 mmol
  • DIEA (691 mg, 5.36 mmol) in DMF (5 mL) stirred at room temperature for 16 h.
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-fluorobenzyl)amino)-1H-indazole-3- carboxylic acid
  • a solution of product 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-fluorobenzaldehyde 100 mg, 0.43 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 75 mg, 0.43 mmol
  • InCl 3 188 mg, 0.85 mmol
  • TES 148 mg, 1.28 mmol
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-fluorobenzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-fluorobenzaldehyde 150 mg, 0.65 mmol
  • 5-amino-1H-indazole-3-carboxylic acid (112 mg, 0.65 mmol)
  • AcOH 0.1 mL
  • Example 11 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-4-fluorobenzyl)amino)- 1H-indazole-3-carboxylic acid (Compound 11)
  • Step 1 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-4-fluorobenzaldehyde A solution of 2-fluoro-5-formylbenzoic acid (500 mg, 2.98 mmol), 3-hydroxy-1,1- dimethylguanidine (337 mg, 2.98 mmol), EDCI (857 mg, 4.46 mmol), HOBT (603 mg, 4.46 mmol), DIEA (1152 mg, 8.93 mmol) in DMF (10 mL) stirred at room temperature for 16 h.
  • 2-fluoro-5-formylbenzoic acid 500 mg, 2.98 mmol
  • 3-hydroxy-1,1- dimethylguanidine 337
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-4-fluorobenzyl)amino)-1H-indazole-3- carboxylic acid
  • a solution of product 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-4-fluorobenzaldehyde 50 mg, 0.21 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 41 mg, 0.21 mmol
  • InCl 3 94 mg, 0.43 mmol
  • TES 74 mg, 1.64 mmol
  • Step 2 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde A solution of 3-hydroxy-1,1-dimethylguanidine (1.72 g, 16.66 mmol), 3-formylbenzoic acid (1.00 g, 6.67 mmol), HATU (3.80 g, 10 mmol) and NaHCO3 (1.68 g, 10 mmol) in DMF (20 mL) was stirred at rt overnight.
  • Step 3 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde 75 mg, 0.35 mmol
  • 5- amino-1H-indazole-3-carboxylic acid 92 mg, 0.52 mmol
  • NaBH3CN 26 mg, 0.42 mmol
  • Example 13 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzyl)(methyl)amino)-1H-indazole-3-carboxylic acid (Compound 13)
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde 100 mg, 0.43 mmol
  • Step 2 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde
  • 3-hydroxy-1,1-dimethylguanidine 3.0 g, crude
  • 3-formyl-2-hydroxybenzoic acid 1.3 g, 7.3 mmol
  • HATU 5.5 g, 15 mmol
  • NaHCO 3 1.8 g, 21.85 mmol
  • the reaction mixture was diluted with H 2 O (30 mL) and extracted with ethyl acetate (30 mL x 3), then dried over with anhydrous Na2SO4.
  • Step 3 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole- 3-carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde 120 mg, 0.50 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 180 mg, 0.7 mmol
  • Example 15 Synthesis of 5-((3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-hydroxybenzyl)amino)-1H- indazole-3-carboxylic acid (Compound 15)
  • Step 1 Synthesis of 3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-methoxybenzaldehyde
  • 3-bromo-1,5-dimethyl-1H-pyrazole 346 mg, 2 mmol
  • Pd(dppf)Cl 2 100 mg, 0.1 mmol
  • K 3 PO 4 (424 mg, 4 mmol
  • H 2 O 2 mL
  • Step 2 Synthesis of 3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-hydroxybenzaldehyde
  • a solution of 3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-methoxybenzaldehyde (220 mg, 0.95 mmol) in CH 2 Cl 2 (5 mL) was added BBr 3 (0.5 mL) at 0 °C and stirred at room temperature for 2 h. Then the mixture was evaporated under vacuum to give 3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-hydroxybenzaldehyde (250 mg, crude) as a yellow oil.
  • LC-MS (M+H) + 216.
  • Step 3 Synthesis of 5-((3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-hydroxybenzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(1,5-dimethyl-1H-pyrazol-3-yl)-2-hydroxybenzaldehyde 250 mg, ⁇ 0.95 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 177 mg, 1 mmol
  • AcOH 60 mg, 1 mmol
  • NaBH3CN 335 mg, 5 mmol
  • Step 2 Synthesis of 3-[4-(hydroxymethyl)-1-methylpyrazol-3-yl]-2-methoxybenzaldehyde
  • 3-formyl-2-methoxyphenylboronic acid (294.90 mg, 1.60 mmol, 1.50 equiv)
  • NaHCO3 275.30 mg, 3.30 mmol, 3.00 equiv
  • Pd(dppf)Cl2 (30.00 mg, 0.04 mmol, 0.04 equiv) and H 2 O (2.00 mL).
  • Step 3 Synthesis of 2-hydroxy-3-[4-(hydroxymethyl)-1-methylpyrazol-3-yl]benzaldehyde
  • 3-[4-(hydroxymethyl)-1-methylpyrazol-3-yl]-2-methoxybenzaldehyde (200.00 mg, 0.80 mmol, 1.00 equiv) in DCM (10.00 mL) was added aluminum chloride (649.70 mg, 4.90 mmol, 6.00 equiv) at -5 °C.
  • the resulting mixture was stirred at -5 °C for 8 h, and then, was stirred at room temperature for another 28 h.
  • the solvent was removed under vacuum.
  • Step 4 Synthesis of 5-[([2-hydroxy-3-[4-(hydroxymethyl)-1-methylpyrazol-3-yl] phenyl] methyl) amino]-1H- indazole-3-carboxylic acid
  • 2-hydroxy-3-[4-(hydroxymethyl)-1-methylpyrazol-3-yl]benzaldehyde (120.00 mg, 0.50 mmol, 1.00 equiv) in MeOH (8.00 mL) was added 5-amino-1H-indazole-3-carboxylic acid (137.30 mg, 0.80 mmol, 1.50 equiv), AcOH (0.05 mL) and NaBH3CN (97.40 mg, 1.50 mmol, 3.00 equiv).
  • Step 2 Synthesis of 3-bromo-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzonitrile 3-hydroxy-1,1-dimethylguanidine (413 mg, 4 mmol), 3-bromo-5-cyanobenzoic acid (900 mg, 4 mmol), HOBT (1080 mg, 8 mmol), EDCI (1520 mg, 8 mmol) and DIEA (1040 mg, 8 mmol) were dissolved in 10 mL DMF and the mixture was stirred at 60 °C for overnight. The completion of the reaction was monitored by TLC.
  • Step 3 Synthesis of methyl 3-cyano-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzoate 3-bromo-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzonitrile (295 mg, 1 mmol), xantphos (58 mg, 0.1 mmol), Pd(OAc)2 (23 mg, 0.1 mmol) and triethylamine (1 mL) were dissolved in MeOH (10 mL) and DMF (1 mL), and the mixture was stirred at 70 °C for overnight under carbon monoxide atmosphere. The completion of the reaction was monitored by TLC.
  • Step 4 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-(hydroxymethyl)benzonitrile Methyl 3-cyano-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzoate (55 mg, 0.2 mmol) was dissolved in MeOH (10 mL) and then LiBH4 (9 mg, 0.4 mmol) was added. The mixture stirred at room temperature for 4 h and monitored by TLC.
  • Step 5 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-formylbenzonitrile 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-(hydroxymethyl)benzonitrile (36 mg, 0.15 mmol) was dissolved in dichloromethane (10 ml). PCC (65 mg, 0.3 mmol) was added, and stirred at room temperature for overnight.
  • Step 6 Synthesis of 5-((3-cyano-5-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-5-formylbenzonitrile 32 mg, 0.13 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 24 mg, 0.13 mmol
  • AcOH 0.1 ml
  • Example 18 Synthesis of 5-((2-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H- indazole-3-carboxylic acid (Compound 18) Step 1: Synthesis of 2-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)benzaldehyde To a solution of 3-formyl-2-hydroxybenzoic acid (200 mg, 1.88 mmol) in 10 mL CH 2 Cl 2 was added dropwise Oxalyl chloride (1 mL) at rt, then accelerated by catalytic amount of DMF, then the reaction was stirred at rt for 2h.
  • Step 2 Synthesis of 5-((2-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • a solution of 2-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)benzaldehyde (60 mg, 0.29 mmol), 5- amino-1H-indazole-3-carboxylic acid (52 mg, 0.49 mmol), InCl 3 (221 mg, 1 mmol) and Et 3 SiH (2 mL) in 5 mL DMF was stirred at 80oC for 4 h, the solution was concentrated and purified by prep-HPLC to give 5- ((2-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-indazole-3-carboxylic acid (13 mg) as a brown solid.
  • Example 19 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzyl)(isopropyl)amino)-1H-indazole-3-carboxylic acid (Compound 19)
  • Step 1 Synthesis of 5-(isopropylamino)-1H-indazole-3-carboxylic acid
  • Acetone (1 mL) in MeOH (20 mL) was added Acetic acid (0.5 mL) and the mixture was stirred at room temperature for 1 h.
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)(isopropyl)amino)- 1H-indazole-3-carboxylic acid
  • Example 20 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzyl)(phenethyl)amino)-1H-indazole-3-carboxylic acid (Compound 20)
  • 2-phenylacetaldehyde 24 mg, 0.2 mmol
  • 5-((3-(3-(dimethylamino)-1,2,4- oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole-3-carboxylic acid 50 mg, 0.127 mmol
  • MeOH MeOH
  • Acetic acid 0.1 mL
  • Example 21 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzyl)(ethyl)amino)-1H-indazole-3-carboxylic acid (Compound 21)
  • Step 1 Synthesis of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde
  • a solution of 3-formyl-2-hydroxybenzoic acid 600 mg, 3.6 mmol
  • compound 3-hydroxy-1,1- dimethylguanidine 412 mg, 4 mmol
  • HOBT 540 mg, 4 mmol
  • EDCI 760 mg, 4 mmol
  • DIEA 516 mg, 4 mmol
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole- 3-carboxylic acid
  • Step 3 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)(ethyl)amino)-1H- indazole-3-carboxylic acid
  • a solution of acetaldehyde (9 mg, 0.2 mmol) compound 5-((3-(3-(dimethylamino)-1,2,4- oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole-3-carboxylic acid (29 mg, 0.073 mmol), InCl 3 (45 mg, 0.2 mmol), HSiEt3 (24 mg, 0.2 mmol) in DMF (5 mL) was stirred at 80 ⁇ C for 2 h.
  • Example 22 Synthesis of 5-((2-hydroxy-3-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)benzyl)amino)- 1H-indazole-3-carboxylic acid (Compound 22) Step 1: Synthesis of 2-hydroxy-3-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)benzaldehyde To a solution of 3-formyl-2-hydroxybenzoic acid (200 mg, 1.88 mmol) in 10 mL CH 2 Cl2 was added Oxalyl chloride (1 mL) at rt, then accelerated by catalytic amount of DMF, then the reaction was stirred at rt for 2h.
  • Step 2 Synthesis of 5-((2-hydroxy-3-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)benzyl)amino)- 1H-indazole-3-carboxylic acid
  • 2-hydroxy-3-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)benzaldehyde 95 mg, 0.37 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 65 mg, 0.37 mmol
  • AcOH 0.1 mL
  • 10 mL MeOH was stirred at rt for 0.5 h
  • NaBH 3 CN(62 mg, 1 mmol) was added, then the reaction was stirred at rt for overnight, the solution was concentrated and purified by Prep-HPLC to give 5-((2-hydroxy-3-(3- (trifluoromethyl)-1,2,4-oxadiazol-5-yl)benzyl)amino)-1H-in
  • Step 2 Synthesis of 3-acetyl-5-bromo-2-hydroxybenzoic acid A mixture of 2-acetoxy-5-bromobenzoic acid (100 g, 386.0 mmol) and AlCl3 (154.41 g, 1158.03 mmol) were heated at 160°C for 3 h. The reaction mixture was allowed to cool to room temperature, poured into ice cold 2N aq.
  • Step 3 Synthesis of 3-acetyl-2-hydroxybenzoic acid
  • MeOH MeOH
  • 10% Pd/C 0.64 gm.
  • the reaction mixture was filtered over a Celite® pad, washed with MeOH (30 mL X 3) and the filtrate was concentrated to afford 3-acetyl-2-hydroxybenzoic acid (2.8 g, crude) as a brown solid. This material was directly used for next step.
  • Step 4 Synthesis of methyl 3-acetyl-2-methoxybenzoate To a stirred solution of 3-acetyl-2-hydroxybenzoic acid (2.8 g, 15.54 mmol) in DMF (28 mL) were added K2CO3 (10.74 g, 77.7 mmol) and methyl iodide (6.617 g, 46.62 mmol) at room temperature. The reaction mixture was stirred at 40°C for 16 h and then quenched with water (25 mL) and extracted with EtOAc (30 mL X 3).
  • Step 5 Synthesis of methyl (E)-3-(1-(hydroxyimino) ethyl)-2-methoxybenzoate
  • 3-acetyl-2-methoxybenzoate 1.5 g, 7.204 mmol
  • EtOH:H2O 8:2
  • NaOAc 1.77 g, 21.61 mmol
  • NH 2 OH•HCl 0.551 g, 7.924 mmol
  • Step 6 Synthesis of methyl 3-(2-isopropylthiazol-4-yl)-2-methoxybenzoate
  • methyl (E)-3-(1-(hydroxyimino)ethyl)-2-methoxybenzoate 1.7 g, 7.61 mmol
  • Isobutyric anhydride 3.614 g, 22.84 mmol
  • CuI 0.725 gm, 3.81 mmol
  • KSCN (1.110 g, 11.42 mmol
  • Step 7 Synthesis of (3-(2-isopropylthiazol-4-yl)-2-methoxyphenyl) methanol
  • methyl 3-(2-isopropylthiazol-4-yl)-2-methoxybenzoate (0.34 g, 1.167 mmol) in THF (8.5 mL) was added was added LAH (2.4M solution in THF, 0.1 mL, 2.33 mmol) at 0 °C.
  • the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 6 h.
  • the reaction mixture was cooled to 0 °C, quenched with EtOAc (5 mL), and saturated aq. Na 2 SO 4 solution (3 mL).
  • Step 8 Synthesis of 3-(2-isopropylthiazol-4-yl)-2-methoxybenzaldehyde To a stirred solution of (3-(2-isopropylthiazol-4-yl)-2-methoxyphenyl) methanol (0.25 g, 0.949 mmol) in CHCl3 (5 mL) was added MnO2 (0.825 g, 9.49 mmol) at room temperature and the reaction mixture was stirred at room temperature for 16 h.
  • Step 9 Synthesis of 2-hydroxy-3-(2-isopropylthiazol-4-yl)benzaldehyde
  • DCM 4-methylethyl-sulfonate
  • BBr3 1-methylethyl-sulfonyl-sulfonyl-sulfonyl-sulfonyl-sulfonyl-sulfonyl-sulfonyl-sulfonate
  • the reaction mixture was allowed to cool to room temperature and stirred at room temperature for 16 h.
  • the reaction mixture was diluted with DCM (10 mL) and quenched with ice cold water (10 mL).
  • Step 10 Synthesis of methyl 5-((2-hydroxy-3-(2-isopropylthiazol-4-yl)benzyl)amino)-1H-indazole-3- carboxylate
  • 2-hydroxy-3-(2-isopropylthiazol-4-yl)benzaldehyde 100 mg, 0.404 mmol
  • MeOH MeOH
  • methyl-5-amino-1H-indazole-3-carboxylate 93 mg, 0.485 mmol
  • AcOH 0.05 mL
  • NaCNBH 3 38 mg, 0.606 mmol
  • Step 11 Synthesis of 5-((2-hydroxy-3-(2-isopropylthiazol-4-yl)benzyl)amino)-1H-indazole-3-carboxylic acid
  • methyl 5-((2-hydroxy-3-(2-isopropylthiazol-4-yl)benzyl)amino)-1H- indazole-3-carboxylate 80 mg, 0.189 mmol
  • MeOH 0.8 mL
  • THF 0.5 mL
  • water 0.3 mL
  • LiOH.H 2 O 24 mg, 0.568 mmol
  • Step 2 Synthesis of Methyl 3-(4-isopropyl-1,3-oxazol-2-yl)-2-methoxybenzoate
  • a solution of methyl 3-carbamoyl-2-methoxybenzoate (500 mg, 2.39 mmol, 1.0 equiv), 1-bromo- 3-methylbutan-2-one (493 mg, 3.00 mmol, 1.25 equiv) and AgOTf (768 mg, 3.00 mmol, 1.3 equiv) in EtOAc (5 mL) was stirred at 60oC for 2 hours. The resulting mixture was filtered. The filter cake was washed with EtOAc (3 x 10 mL). The filtrate was concentrated under reduced pressure.
  • Step 3 Synthesis of [3-(4-isopropyl-1,3-oxazol-2-yl)-2-methoxyphenyl]methanol
  • a solution of methyl 3-(4-isopropyl-1,3-oxazol-2-yl)-2-methoxybenzoate (480 mg, 1.74 mmol, 1.0 equiv) and LiAlH 4 (5 mL, 10.00 mmol, 5.7 equiv, 2M in THF) in THF (5 mL) was stirred at 0 oC for 1 h. The reaction was quenched with Na2SO4 . 10H 2 O at room temperature. The resulting mixture was diluted with EtOAc (50 mL). The resulting mixture was filtered.
  • Step 4 Synthesis of 3-(4-isopropyl-1,3-oxazol-2-yl)-2-methoxybenzaldehyde
  • a mixture of [3-(4-isopropyl-1,3-oxazol-2-yl)-2-methoxyphenyl]methanol (430 mg, 1.74 mmol, 1.0 equiv) and MnO 2 (1.51 g, 17.39 mmol, 10.0 equiv) in CH 2 Cl 2 (10 mL) was stirred at room temperature overnight. The resulting mixture was filtered. The filter cake was washed with CH 2 Cl 2 (3 x 10 mL).
  • Step 5 Synthesis of 2-hydroxy-3-(4-isopropyl-1,3-oxazol-2-yl)benzaldehyde
  • the residue was purified using flash chromatography with the following conditions (Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 50 mL/min; Gradient: 0% B to 100% B in 40 min; 254; 220 nm).
  • the resulting mixture was purified by Prep-HPLC (Column: XBridge Shield RP18 OBD Column 30*150mm,5um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 20% B to 40% B in 8 min; 220 nm; Rt: 6.85) to afford 5-([[2-hydroxy-3-(4-isopropyl-1,3- oxazol-2-yl)phenyl]methyl]amino)-1H-indazole-3-carboxylic acid (25.4 mg, 12.5%) as a yellow solid.
  • Prep-HPLC Column 30*150mm,5um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 20% B to 40% B in 8 min; 220 nm; Rt: 6.85
  • the resulting mixture was stirred at 60oC for 1 h under nitrogen atmosphere.
  • the resulting mixture was diluted with water (30 mL).
  • the resulting mixture was extracted with EtOAc (3 x 20 mL).
  • the combined organic layers were washed with brine (2x30 mL), dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure.
  • the reaction mixture was purified using flash chromatography with the following conditions (Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 50% B in 50 min; 254/220 nm to afford 3-(4-isopropyl-1,3- oxazol-2-yl)benzaldehyde (320.00 mg, 30.7%) as a light yellow oil.
  • Mobile Phase A Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 50% B in 50 min; 254/220 nm to afford 3-(4-isopropyl-1,3- oxazol-2-yl)benzaldehyde (320.00 mg, 30.7%) as a light yellow oil.
  • [M+H] + 216.
  • Step 2 Synthesis of 5-([[3-(4-isopropyl-1,3-oxazol-2-yl)phenyl]methyl](methyl)amino)-1H-pyrazolo[3,4- b]pyridine-3-carboxylic acid
  • 3-(4-isopropyl-1,3-oxazol-2-yl)benzaldehyde (300.00 mg, 1.39 mmol, 1.00 equiv)
  • 5-amino-1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid (248.29 mg, 1.39 mmol, 1.00 equiv) in MeOH (2.00 mL) were added NaBH 3 CN (350.34 mg, 5.58 mmol, 4.00 equiv).
  • the resulting mixure was purified using Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30 ⁇ 150 mm 5 ⁇ m; Mobile Phase A:Water (10 mmol/L NH4HCO3), Mobile Phase B:ACN; Flow rate:60 mL/min; Gradient:10% B to 30% B in 9 min; 254/220 nm; RT1:9.33;) to afford 5-([[3-(4-isopropyl-1,3-oxazol-2-yl)phenyl]methyl](methyl)amino)-1H-pyrazolo[3,4- b]pyridine-3-carboxylic acid (14.70 mg, 2.7%) as a yellow solid.
  • Example 26 Synthesis of 5-(methyl((2-propoxypyridin-3-yl)methyl)amino)-1H-indazole-3- carboxylic acid (Compound 26)
  • 2-propoxynicotinaldehyde 500 mg, 3.03 mmol
  • 5-amino-1H-indazole-3- carboxylic acid 532 mg, 3.03 mmol
  • NaBH3CN 381 mg, 6.06 mmol
  • paraformaldehyde 136.4 mg, 4.55 mmol
  • Step 2 Synthesis of 2-hydroxy-3-(1-methyl-1H-imidazol-4-yl) benzaldehyde To a solution of 2-methoxy-3-(1-methyl-1H-imidazol-4-yl) benzaldehyde (310mg,1.43mmol) in CH 2 Cl2 (5 mL) was added BBr3 (1.5 g, 7.15 mmol) dropwise at 0 degree. Then the reaction mixture was stirred at rt for 1h.
  • Step 2 Synthesis of 3-((tert-butyldiphenylsilyl)oxy)-N'-hydroxypropanimidamide
  • a solution of 3-[(tert-butyldiphenylsilyl)oxy]propanenitrile (2, 2.10 g, 6.79 mmol, 1.00 equiv) and NH 2 OH (50% in water) (493.07 mg, 7.46 mmol, 1.10 equiv) in EtOH (10.00 mL) was stirred at room temperature for 1 h under nitrogen atmosphere. The resulting mixture was concentrated under vacuum. This resulted in 3-[(tert-butyldiphenylsilyl)oxy]-N-hydroxypropanimidamide (3.5 g, crude) as a white solid.
  • Step 4 Synthesis of 3-(3-(2-((tert-butyldiphenylsilyl)oxy)ethyl)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzaldehyde
  • a solution of N-(3-((tert-butyldiphenylsilyl)oxy)-1-(hydroxyimino)propyl)-3-formyl-2- hydroxybenzamide (500.00 mg, 1.02 mmol, 1.00 equiv) in THF (5.00 mL) was stirred at 80 oC for overnight under nitrogen atmosphere. The resulting mixture was concentrated under vacuum.
  • Step 5 Synthesis of 5-((3-(3-(2-((tert-butyldiphenylsilyl)oxy)ethyl)-1,2,4-oxadiazol-5-yl)-2- hydroxybenzyl)amino)-1H-indazole-3-carboxylic acid
  • 3-(3-[2-[(tert-butyldiphenylsilyl)oxy]ethyl]-1,2,4-oxadiazol-5-yl)-2- hydroxybenzaldehyde 100.00 mg, 0.21 mmol, 1.00 equiv
  • 5-amino-1H-indazole-3-carboxylic acid 37.49 mg, 0.21 mmol, 1.00 equiv
  • NaBH3CN 26.59 mg, 0.42 mmol, 2.00 equiv
  • Step 6 Synthesis of tert-butyl (1-(4-(5-(((1-carbamoylpiperidin-3-yl)methyl)(methyl)amino)-1,3,4- oxadiazol-2-yl)phenyl)cyclobutyl)carbamate
  • Step 2 Synthesis of 2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl)benzaldehyde
  • BBr3 250 mg, 1 mmol
  • the reaction mixture was concentrated under vacuum to give the crude 2-hydroxy-3-(1-methyl-1H-pyrazol-3- yl)benzaldehyde (215 mg, crude) as a red oil.
  • LC-MS (M+H) + 203.
  • Step 3 Synthesis of 5-((2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl)benzyl)(methyl)amino)-1H-indazole-3- carboxylic acid
  • 2-hydroxy-3-(1-methyl-1H-pyrazol-3-yl)benzaldehyde 215 mg, ⁇ 0.8 mmol
  • 5- amino-1H-indazole-3-carboxylic acid 179 mg, 1 mmol
  • AcOH 1 drop
  • NaBH3CN 124 mg, 2 mmol
  • the reaction mixture was degassed three times with nitrogen, and then heated at 100 °C for 3 h.
  • the reaction mixture was concentrated under vacuum.
  • the residue was diluted with ethyl acetate and H 2 O (30 mL), filtered with kieselguhr, and extracted with ethyl acetate (20 mLx3).
  • the combined organic layer was washed with brine (15 mLx3), then dried over with anhydrous Na 2 SO 4 .
  • Step 2 Synthesis of 2-hydroxy-3-(1-isopropyl-1H-pyrazol-3-yl)benzaldehyde To a solution of 3-(1-isopropyl-1H-pyrazol-3-yl)-2-methoxybenzaldehyde (250 mg, 1.03 mmol) in CH 2 Cl2 (3 mL) was added dropwise BBr3 (1 mL, 10.25 mmol) and stirred at rt overnight. The reaction mixture was concentrated under vacuum. The residue was diluted with H 2 O and CH 2 Cl 2 (20 mL), then extracted with CH 2 Cl 2 (15 mLx3). The combined organic layer was washed with brine (15 mLx3), then dried over with anhydrous Na2SO4.
  • Step 3 Synthesis of 5-((2-hydroxy-3-(1-isopropyl-1H-pyrazol-3-yl)benzyl)amino)-1H-indazole-3- carboxylic acid
  • 2-hydroxy-3-(1-isopropyl-1H-pyrazol-3-yl)benzaldehyde 100 mg, 0.44 mmol
  • 5- amino-1H-indazole-3-carboxylic acid 92 mg, 0.52 mmol
  • InCl3 192 mg, 0.87 mmol
  • Et 3 SiH 151 mg, 1.31 mmol
  • Step 2 Synthesis of 3-formyl-2-hydroxy-N-(1-(hydroxyimino)-2-((tetrahydro-2H-pyran-2- yl)oxy)ethyl)benzamide
  • N-hydroxy-2-(oxan-2-yloxy)ethanimidamide (2, 500.00 mg, 2.87 mmol, 1.00 equiv)
  • 3-formylsalicylic acid (476.84 mg, 2.87 mmol, 1.00 equiv) in DMF(6.00 mL) were added HOBT(581.76 mg, 4.31 mmol, 1.50 equiv) and EDCI (825.35 mg, 4.31 mmol, 1.50 equiv).
  • the resulting mixture was stirred at room temperature for 2 h.
  • the resulting mixture was purified using reverse phase flash with the following conditions (Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 100% B in 25 min; 254/220 nm) to afford 3-formyl-2-hydroxy-N-[1-(hydroxyimino)-2- (oxan-2-yloxy)ethyl]benzamide (120 mg, 12.97%) as a yellow solid.
  • the resulting solution was purified using reverse phase flash with the following conditions (Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 100% B in 25 min; 254/220 nm) to afford 2-hydroxy-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]benzaldehyde (50 mg, 60.99%) as a yellow solid.
  • the resulting mixture was stirred at room temperature for 2 h.
  • the resulting mixture was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30*150 mm 5 ⁇ m; Mobile Phase A: Water (0.05 % TFA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 30% B in 7 min; 220 nm; RT1: 6.69 min).
  • Step 2 Synthesis of 2-hydroxy-3-(4-isopropylthiazol-2-yl)benzaldehyde
  • BBr 3 1-M solution in CH 2 Cl 2 , 19.15 mL, 19.15 mmol
  • the reaction mixture was allowed to warm to rt and stirred at rt for 16 h.
  • the reaction mixture was diluted with CH 2 Cl2 (30 mL) and quenched with ice cold water (30 mL).
  • Step 3 Synthesis of methyl 5-((2-hydroxy-3-(4-isopropylthiazol-2-yl)benzyl)amino)-1H-indazole-3-carboxylate
  • 2-hydroxy-3-(4-isopropylthiazol-2-yl)benzaldehyde 400 mg, 1.62 mmol
  • MeOH MeOH
  • methyl 5-amino-1H-indazole-3-carboxylate 309 mg, 1.62 mmol
  • AcOH 0.4 mL
  • Step 4 Synthesis of 5-((2-hydroxy-3-(4-isopropylthiazol-2-yl)benzyl)amino)-1H-indazole-3-carboxylic acid
  • MeOH MeOH
  • THF trifluoroethyl
  • water 5 mL
  • LiOH LiOH
  • Example 33 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)(methyl)amino)-1H- pyr- azolo[3,4-b]pyridine-3-carboxylic acid (Compound 33)
  • Step 1 Synthesis of 5-amino-1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid
  • Pd/C 40 mg
  • Step 2 Synthesis of 5-((3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzyl)(methyl)amino)-1H- pyrazolo[3,4-b]pyridine-3-carboxylic acid
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)benzaldehyde 100 mg, 0.461 mmol
  • 5-amino-1H-pyrazolo[3,4-b]pyridine-3-carboxylic acid 82 mg, 0.461 mmol
  • InCl3 204 mg, 0.922 mmol
  • HSiEt3 160 mg, 1.383 mmol
  • Step 2 Synthesis of 2-hydroxy-3-(2-isopropyl-1,3-oxazol-4-yl)benzoic acid
  • a solution of methyl 3-(2-bromoacetyl)-2-hydroxybenzoate (650.00 mg, 2.38 mmol, 1.00 equiv) and isobutyramide (829.48 mg, 9.52 mmol, 4.00 equiv) in NMP (0.50 mL) was stirred at 110oC for 3 h.
  • Step 3 Synthesis of 2-(hydroxymethyl)-6-(2-isopropyl-1,3-oxazol-4-yl)phenol
  • 2-hydroxy-3-(2-isopropyl-1,3-oxazol-4-yl)benzoic acid (180.00 mg, 0.73 mmol, 1.00 equiv) in THF (7.00 mL)
  • BH 3 -THF (1M in THF, 4.37 mL, 4.37 mmol, 6.00 equiv
  • the resulting solution was stirred at 0oC for 6 h.
  • the reaction was quenched by MeOH (10.00 mL) at 0oC.
  • the resulting solution was concentrated under vacuum.
  • Step 4 Synthesis of 2-hydroxy-3-(2-isopropyl-1,3-oxazol-4-yl)benzaldehyde
  • a solution of 2-(hydroxymethyl)-6-(2-isopropyl-1,3-oxazol-4-yl)phenol (180.00 mg, 0.77 mmol, 1.00 equiv) and MnO 2 (2.01 g, 23.15 mmol, 30.00 equiv) in DCM (360.00 mL) was stirred at room temperature for 6 h.
  • the reaction solution was filtrated and washed with MeOH (20.00 mL). The filtrate was concentrated under vacuum.
  • Step 5 Synthesis of 5-([[2-hydroxy-3-(2-isopropyl-1,3-oxazol-4-yl)phenyl]methyl]amino)-1H-indazole-3- carboxylic acid
  • 2-hydroxy-3-(2-isopropyl-1,3-oxazol-4-yl)benzaldehyde 50.00 mg, 0.22 mmol, 1.00 equiv
  • 5-amino-1H-indazole-3-carboxylic acid 153.22 mg, 0.86 mmol, 4.00 equiv
  • AcOH (12.98 mg, 0.22 mmol, 1.00 equiv
  • Example 35 Synthesis of 5-((2-hydroxy-3-(3-isopropyl-1,2,4-oxadiazol-5- yl)benzyl)(methyl)amino)-1H-indazole-3-carboxylic acid (Compound 35)
  • a solution of Compound 2 50 mg, 0.13 mmol
  • Formalin 0.5 mL
  • AcOH 0.1 mL
  • NaBH3CN 62 mg, 1 mmol
  • the reaction was stirred at rt for overnight, the solution was concentrated and purified by Prep-HPLC to give 5-((2-hydroxy- 3-(3-isopropyl-1,2,4-oxadiazol-5-yl)benzyl)(methyl)amino)-1H-indazole-3-carboxylic acid (16 mg) as a yellow solid.
  • Example 36 Synthesis of 5-((3-(3-(tert-butyl)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H- indazole -3-carboxylic acid (Compound 36)
  • Step 1 Synthesis of N-hydroxypivalimidamide
  • NH 2 OH HCl 1.247 g, 18.072 mmol
  • K 2 CO 3 4.988 g, 36.144 mmol
  • Step 2 Synthesis of 3-(3-(tert-butyl)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde To a solution of 3-formyl-2-hydroxybenzoic acid (200 mg, 1.205 mmol) and (COCl)2 (1391 mg, 10.950 mmol) in CH 2 Cl 2 (5 mL) was added DMF (Catalytic amount) at rt. The reaction mixture was stirred at rt for 1 h. It was concentrated under vacuum.
  • Step 3 Synthesis of 5-((3-(3-(tert-butyl)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole -3- carboxylic acid
  • 3-(3-(tert-butyl)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde 70 mg, 0.285 mmol
  • 5-amino-1H-indazole-3-carboxylic acid 61 mg, 0.342 mmol
  • MeOH 8 mL
  • InCl 3 126 mg, 0.57 mmol
  • HSiEt3 165 mg, 1.425 mmol
  • Step 2 Synthesis of 5-((3-(3-cyclopropyl-1,2,4-oxadiazol-5-yl)-2-hydroxybenzyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(3-cyclopropyl-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde 120 mg, 0.52 mmol
  • InCl 3 231 mg, 1.04 mmol
  • Et 3 SiH 121 mg, 1.04 mmol
  • Step 2 Synthesis of 3-(4,5-dimethyl-1,3-thiazol-2-yl)-2-hydroxybenzaldehyde
  • BBr3 1-M, 3.23 mL, 3.23 mmol, 2.0 equiv
  • Step 3 Synthesis of 5-([[3-(4,5-dimethyl-1,3-thiazol-2-yl)-2-hydroxyphenyl]methyl]amino)-1H-indazole-3- carboxylic acid
  • 3-(4,5-dimethyl-1,3-thiazol-2-yl)-2-hydroxybenzaldehyde (288 mg, 1.24 mmol, 1 equiv)
  • 5-amino-1H-indazole-3-carboxylic acid (219 mg, 1.24 mmol, 1.0 equiv) in DMSO (2 mL) were added CH 3 COOH (74 mg, 1.24 mmol, 1.0 equiv) and NaBH 3 CN (310 mg, 4.94 mmol, 4.0 equiv).
  • the resulting mixture was stirred at room temperature for 1 h under nitrogen atmosphere.
  • the resulting mixture was purified using Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30*150mm 5um, n; Mobile Phase A:Water (0.1% FA), Mobile Phase B:ACN; Flow rate:60 mL/min; Gradient:56% B to 77% B in 8 min; 254 nm; RT1:8.12;) to afford 5-([[3-(4,5-dimethyl-1,3-thiazol- 2-yl)-2-hydroxyphenyl]methyl]amino)-1H-indazole-3-carboxylic acid (25.00 mg, 5.1%) as a yellow solid.
  • Step 2 Synthesis of 5-(methyl(3-(4-methylthiazol-2-yl)benzyl)amino)-1H-indazole-3-carboxylic acid
  • 3-(4-methyl-1,3-thiazol-2-yl)benzaldehyde 340 mg, 1.67 mmol, 1.0 equiv
  • 5-amino-1H-indazole-3-carboxylic acid 296 mg, 1.67 mmol, 1.0 equiv
  • AcOH 0.2 mL
  • the resulting mixture was stirred at room temperature for 30 min.
  • NaBH 3 CN 420 mg, 6.69 mmol, 4.0 equiv).
  • Example 40 Synthesis of 5-((3-(3-isopropyl-1,2,4-oxadiazol-5-yl)benzyl)(methyl)amino)-1H- indazole-3- carboxylic acid (Compound 40)
  • 3-(3-isopropyl-1,2,4-oxadiazol-5-yl)benzaldehyde 200 mg, 0.93 mmol
  • 5- amino-1H-indazole-3-carboxylic acid 197 mg, 1.11 mmol
  • MeOH 5 mL
  • Step 2 Synthesis of 2-hydroxy-3-(4-isopropyl-1,3-thiazol-2-yl)benzaldehyde
  • BBr 3 3-(4-isopropyl-1,3-thiazol-2-yl)-2-methoxybenzaldehyde
  • reaction solution was purified using Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30*150mm 5um, n; Mobile Phase A:Water (0.1% FA), Mobile Phase B:ACN; Flow rate:60 mL/min; Gradient: 33% B to 77% B in 8 min; 254 nm; RT1:7.49;) to afford 5-([[2-hydroxy-3-(4-isopropyl- 1,3-thiazol-2-yl)phenyl]methyl](methyl)amino)-1H-indazole-3-carboxylic acid (28.60 mg, 7.6%) as a yellow solid.
  • Step 2 Synthesis of 5-([[3-(4-isopropyl-1,3-thiazol-2-yl)phenyl]methyl](methyl)amino)-1H-indazole-3- carboxylic acid
  • 3-(4-isopropyl-1,3-thiazol-2-yl)benzaldehyde 100.00 mg, 0.43 mmol, 1.00 equiv
  • 5-amino-1H-indazole-3-carboxylic acid 76.59 mg, 0.43 mmol, 1.00 equiv
  • MeOH 1.00 mL
  • CH3COOH 25.96 mg, 0.43 mmol, 1.00 equiv
  • NaBH3CN 108.67 mg, 1.73 mmol, 4.00 equiv).
  • the resulting mixture was purified using Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 30*150mm, 5um ; Mobile Phase A:Water (0.1%FA), Mobile Phase B:ACN; Flow rate:60 mL/min; Gradient:20% B to 45% B in 8 min; 220/254 nm; RT1:6.57 min ) to afford 5-([[3-(4-isopropyl-1,3-thiazol-2- yl)phenyl]methyl](methyl)amino)-1H-indazole-3-carboxylic acid (25.10 mg, 13.8%) as a yellow solid.
  • Step 2 Synthesis of 2-hydroxy-3-(4-methylthiazol-2-yl)benzaldehyde To a stirred solution of 2-methoxy-3-(4-methyl-1,3-thiazol-2-yl)benzaldehyde (590 mg, 2.53 mmol, 1.00 equiv) in CH 2 Cl 2 (6 mL) was added BBr 3 (1M, 5.06 mL, 5.06 mmol, 2.00 equiv) at 0 oC. The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by the addition of water (3 mL) at 0 oC. The resulting mixture was extracted with CH 2 Cl2, dried over anhydrous Na2SO4.
  • Step 2 Synthesis of 2-hydroxy-3-(1,3-thiazol-2-yl)benzaldehyde To a stirred solution of 2-methoxy-3-(1,3-thiazol-2-yl)benzaldehyde (195.00 mg, 0.89 mmol, 1.00 equiv) in DCM (3.00 mL) was added BBr3 (445.60 mg, 1.78 mmol, 2.00 equiv) in portions at 0oC. The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched with H 2 O at 0oC. The organic layer was collected and solvent was removed under vacuum.
  • the resulting mixture was stirred at room temperature for 2 h.
  • the resulted solution was purified using C18 flash chromatography with the following conditions (Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 100% B in 30 min; 254/220 nm).
  • the crude was further purified by prep-HPLC with following conditions: Column: Xselect CSH OBD Column 30*150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 46% B to 68% B in 8 min; 254 nm; RT: 7.77 min.
  • Example 45 Synthesis of 5-((3-(4,5-dimethylthiazol-2-yl)benzyl)(methyl)amino)-1H-indazole-3- carboxylic acid (Compound 45)
  • Step 1 Synthesis of 3-(4,5-dimethylthiazol-2-yl)benzaldehyde
  • 2-bromo-4,5-dimethyl-1,3-thiazole 500 mg, 2.60 mmol, 1.0 equiv
  • 3- formylphenylboronic acid (468 mg, 3.12 mmol, 1.2 equiv) in 1,4-dioxane (6 mL) and H 2 O (2 mL) were added C s2 CO3 (2.54 g, 7.81 mmol, 3.0 equiv) and Pd(dppf)Cl2 (190 mg, 0.26 mmol, 0.1 equiv).
  • Step 2 Synthesis of 5-((3-(4,5-dimethylthiazol-2-yl)benzyl)(methyl)amino)-1H-indazole-3-carboxylic acid
  • 3-(4,5-dimethyl-1,3-thiazol-2-yl)benzaldehyde 200 mg, 0.92 mmol, 1 equiv
  • 5-amino-1H-indazole-3-carboxylic acid 163 mg, 0.92 mmol, 1.0 equiv
  • MeOH MeOH
  • AcOH 0.2 mL
  • Step 2 Synthesis of 1H-[1,2,3]triazolo[4,5-b]pyridin-6-amine To a stirred solution of 6-nitro-1H-[1,2,3]triazolo[4,5-b]pyridine (500 mg, 3.03 mmol) in THF: MeOH (1:1) (10 mL) was added Pd/C (100 mg, 20%w/w) at room temperature and stirred at hydrogen atmosphere for 16 h. The reaction was monitored by TLC.
  • Step 3 Synthesis of 2-(((1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)amino)methyl)-6-(3-(dimethylamino)-1,2,4- oxadiazol-5-yl)phenol
  • 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde (220 mg, 0.94 mmol)
  • K2CO3 195 mg, 1.41 mmol
  • 1H- [1,2,3]triazolo[4,5-b]pyridin-6-amine (191.1 mg, 1.41 mmol) and stirred the reaction mixture at 65 °C for 16 h.
  • reaction mixture cooled to room temperature, NaCNBH 3 (58.5 mg, 0.94 mmol) was added to reaction mixture and stirred at RT for 2 h. After completion of reaction (by TLC), the reaction mixture was filtered and filtrate was concentrated under reduced pressure. Resulting crude was purified by SFC to afford 2-(((1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)amino)methyl)-6-(3-(dimethylamino)-1,2,4-oxadiazol-5- yl)phenol (41 mg, 12%) as off white solid.
  • Step 3 Synthesis of 2-(((3H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)(methyl)amino)methyl)-6-(3- (dimethylamino)-1,2,4-oxadiazol-5-yl)phenol
  • a solution of 3-(3-(dimethylamino)-1,2,4-oxadiazol-5-yl)-2-hydroxybenzaldehyde (100mg, 0.43mmol), 1H-[1,2,3]triazolo[4,5-b]pyridin-6-amine (58 mg, 0.43mmol) and AcOH (0.1 mL) in 5 mL MeOH was stirred at 45 °C for 4 h, then NaBH3CN (62 mg, 1 mmol) was added at rt, the reaction mixture was stirred at rt for 3 h, then Formalin(0.2 mL) was added at room temperature, the reaction mixture was stirred at rt for 2 h, then NaBH3CN
  • Recombinant HAO1 at a concentration of 10 nM was added to test compounds dissolved in DMSO or DMSO alone as a control that had been dispensed into 384 well assay plates. After a 30-minute preincubation of HAO1 with compound, hydrogen peroxide detection reagents were added such that the final concentrations were 50 mM Amplex Red and 0.5 U/ml horseradish peroxidase. The reaction was initiated by the addition of glycolic acid to a final concentration of 50 mM. The rate of fluorescence increase was read kinetically over 15 minutes using a Molecular Devices SpectraMax Paradigm Microplate Reader (ex.563 nm, em.587 nm).
  • CHO cells stably transformed with vector expressing glycolate oxidase are cultured in Ham’s F-12 /Glutamax media supplemented with 10% fetal bovine serum (FBS) and 100 units/mL penicillin/100 ⁇ g/mL streptomycin with additional supplementation of 400 ⁇ g/ml zeocin.
  • CHO- WT cells are maintained in the same media conditions without the addition of zeocin.
  • Cells are plated in 96-well plate (2000 cells/well) in standard medium (Ham’s F12 Glutamax F-12 with 10% FBS and antibiotics) and incubated with compound at different concentrations for 1 hour followed by addition of glycolic acid (final concentration 125 or 250 ⁇ M; Sigma).
  • the final DMSO concentration in the assay is 0.5 %. Viability is monitored using live cells imager (IncuCyte Zoom) for up to 96 hours. CHO-WT cells are used as a control. Data Analysis. Data is analyzed using customized software developed internally. EC50 values (half maximum effective concentration) are calculated from dose-response curves using Prism software (Graph pad Software). At each compound concentration the % rescue is calculated as the % confluence of CHO-GO cells treated with the compound in presence of glycolic acid divided by the % confluence of CHO-GO cells treated with the compound in the absence of glycolic acid.

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Abstract

La présente invention concerne des compositions et des procédés pour le traitement de troubles associés à HA01, tels que l'hyperoxalurie primaire 1.
PCT/US2020/050142 2019-09-10 2020-09-10 Composés et leurs utilisations Ceased WO2021050688A1 (fr)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11427558B1 (en) 2019-07-11 2022-08-30 ESCAPE Bio, Inc. Indazoles and azaindazoles as LRRK2 inhibitors

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Publication number Priority date Publication date Assignee Title
WO2006091737A1 (fr) * 2005-02-24 2006-08-31 Kemia, Inc. Modulateurs de l'activite de gsk-3
US20070049607A1 (en) * 2001-01-10 2007-03-01 Vernalis Research Limited Triazolo [4,5-d] pyrimidine derivatives and their use as purinergic receptor antagonists
WO2010114726A1 (fr) * 2009-03-31 2010-10-07 Merck Sharp & Dohme Corp. Dérivés d'aminobenzotriazole

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070049607A1 (en) * 2001-01-10 2007-03-01 Vernalis Research Limited Triazolo [4,5-d] pyrimidine derivatives and their use as purinergic receptor antagonists
WO2006091737A1 (fr) * 2005-02-24 2006-08-31 Kemia, Inc. Modulateurs de l'activite de gsk-3
WO2010114726A1 (fr) * 2009-03-31 2010-10-07 Merck Sharp & Dohme Corp. Dérivés d'aminobenzotriazole

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DATABASE PubChem Compound 12 January 2012 (2012-01-12), ANONYMOUS: "Compound Summary for CID 70217172", XP055805064, retrieved from NCBI Database accession no. CID 70217172 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11427558B1 (en) 2019-07-11 2022-08-30 ESCAPE Bio, Inc. Indazoles and azaindazoles as LRRK2 inhibitors

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