EP4164690A1 - Small molecule ve-ptp inhibitors - Google Patents

Small molecule ve-ptp inhibitors

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Publication number
EP4164690A1
EP4164690A1 EP21825008.2A EP21825008A EP4164690A1 EP 4164690 A1 EP4164690 A1 EP 4164690A1 EP 21825008 A EP21825008 A EP 21825008A EP 4164690 A1 EP4164690 A1 EP 4164690A1
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European Patent Office
Prior art keywords
substituted
compound according
unsubstituted
previous
immediately previous
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EP21825008.2A
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German (de)
French (fr)
Inventor
Michael RYCZKO
Amy Ripka
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Mannin Research Inc
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Individual
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/22—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C07D277/28—Radicals substituted by nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • the present disclosure relates to compounds capable of inhibiting vascular endothelial protein tyrosine phosphatase (VE-PTP). These compounds are also capable of activating Tie2 receptor-mediated signaling. Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP.
  • the present disclosure also relates to pharmaceutically acceptable salts of said compounds, to pharmaceutical compositions comprising such compounds and/or pharmaceutically acceptable salts thereof, and to the use of such compounds, pharmaceutically acceptable salts thereof, and/or pharmaceutical compositions comprising the same in treating diseases and/or conditions mediated by VE- PTP signaling, such as those mediated by Angiopoietin/Tie2 signaling.
  • VE-PTP Vascular endothelial tyrosine phosphatase
  • ⁇ - ⁇ vascular endothelial tyrosine phosphatase beta
  • VE-PTP knockout mice have defective blood vessel remodeling (Dominguez, MG et al.
  • VE-PTP Vascular endothelial tyrosine phosphatase
  • Phosphotyrosine phosphatase R3 receptors Origin, evolution and structural diversification, PLoS ONE, 12(3), e0172887.; Matozaki, T et al. (2010 Dec) Expression, localization, and biological function of the R3 subtype of receptor-type protein tyrosine phosphatases in mammals, Cell Signal. 22(12):1811-7).
  • One of the substrates for VE-PTP is Tie2, an endothelial tyrosine kinase receptor that regulates angiogenesis and lymphangiogenesis and can support the integrity of endothelial junctions (Eklund, L et al.
  • VE-PTP Antibodies against the extracellular domain of VE-PTP dissociate the phosphatase from Tie2, leading to Tie2 phosphorylation and activated signaling (Winderlich, M et al. (2009 May) VE-PTP controls blood vessel development by balancing Tie-2 activity, J Cell Biol. 185(4):657-71). Pharmacological inhibition of VE-PTP activity also activates Tie2 and is capable of suppressing ocular neovascularization and VEGF-mediated vascular leakage (Goel, S et al. (2013 Aug) Effects of vascular-endothelial protein tyrosine phosphatase inhibition on breast cancer vasculature and metastatic progression.
  • the Angiopoietin-Tie2 signaling pathway is a major regulator of vascular development, vascular permeability, vessel remodeling, and post-natal angiogenesis, and altered expression of the Angiopoietin ligands or activity of the Tie2 receptor has been linked to a variety of lymphatic and blood vasculature pathologies including cancer, sepsis, diabetes, atherosclerosis, and ocular diseases (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci.
  • Angiopoietin2 In blood vascular endothelium, Angiopoietin2 (Angpt2 or Ang2) is reported to function as a competitive antagonist of Angiopoietinl /Tie2 signaling, inhibiting Angiopoietin 1 (Angptl or Angl>mediated phosphorylation of Tie2 (Thurston, G et al. (2012 Sep) The complex role of angiopoietin-2 in the angiopoietin-tie signaling pathway, Cold Spring Harb Perspect Med. 2(9):a006550.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov.
  • Tie2 receptor activators include, for example, Angptl recombinant proteins, vascular endothelial protein tyrosine phosphatase (VE-PTP) inhibitors, and Tie2-peptidomimetics (Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661.; Shen, J et al. (2014 Oct) Targeting VE-PTP activates TIE2 and stabilizes the ocular vasculature, J Clin Invest. 124(10):4564-76.; Souma, T et al.
  • angiopoietin 2 Context-dependent Junctions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115).
  • Angiopoietin- 1 decreases endothelial cell permeability and increases vascular stabilization via recruitment of pericytes and smooth muscle cells to growing blood vessels, while Angiopoietin-2 mediates angiogenic sprouting and vascular regression (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci.
  • angiopoietin 2 Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115).
  • Angpt2 deficiency in knockout mice resulted in widespread lymphatic dysfunction due to abnormal remodeling of the developing lymphatic vessels, resulting in lymphedema (Dellinger, M et al. (2008 Jul) Defective remodeling and maturation of the lymphatic vasculature in Angiopoietin-2 deficient mice, Dev Biol. 319(2):309-20.; Eklund, L et al.
  • Both Angptl/Angpt2 double knockout mice and Tie2 knockout mice have a defective Schlemm’s canal, leading to impaired drainage of aqueous humour, increased intraocular pressure (IOP) and glaucoma (Souma, T et al. (2016 Jul) Angiopoietin receptor TEK mutations underlie primary congenital glaucoma with variable expressivity, J Clin Invest. 126(7):2575-87.; Thomson, BR et al. (2014 Oct) A lymphatic defect causes ocular hypertension and glaucoma in mice, J Clin Invest. 124(10):4320-4.; Thomson, BR et al.
  • VE-PTP inhibitors particularly some that could have improved physicochemical properties, and are potent Tie2 activators.
  • Such therapeutic agents can be useful in the treatment of cancer, glaucoma, occlusive cardiovascular disease, vascular leaking syndromes, and other vascular-related diseases (Parikh, SM (2017 Jul) The Angiopoietin-Tie2 Signaling Axis in Systemic Inflammation, J Am Soc Nephrol. 28(7): 1973- 1982.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661.; Schmittnaegel, M et al. (2017 Dec) Reprogramming Tumor Blood Vessels for Enhancing Immunotherapy, Trends Cancer. 3(12):809-812).
  • the present disdosure relates to compounds and pharmaceutically acceptable salts thereof that are capable of inhibiting vascular endothelial protein tyrosine phosphatase (VE- PTP). These compounds are capable of activating the Tie-2 receptor.
  • VE- PTP vascular endothelial protein tyrosine phosphatase
  • the present disclosure also relates to pharmaceutical compositions comprising such compounds and/or pharmaceutically acceptable salts thereof.
  • the present disclosure also relates to the use of such compounds, pharmaceutically acceptable salts thereof, and/or pharmaceutical compositions in treating diseases and/or conditions mediated by VE-PTP activity, including those that are linked to Ang/Tie2-mediated signaling.
  • the present disclosure is directed to:
  • Ri is chosen from wherein
  • R 2 is chosen from substituted or unsubstituted heteroaiyl and substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more halogen atoms.
  • R 3 is chosen from substituted or unsubstituted heteroaiyl, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl; n is an integer chosen from 1, 2, 3, 4, 5, and 6;
  • R t is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted Ci-C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl;
  • R is chosen from H, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl; and each of Xi, X2, and X3 is independently chosen from CH and N, with the proviso that Xi, X2, and X3 are not simultaneously CH.
  • R2 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more halogen atoms.
  • R 2 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more fluorine atoms.
  • R 3 is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C3- Ce cyclic alkyl.
  • R 3 is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl.
  • R 3 is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl and substituted or unsubstituted C 3 -C 6 cyclic alkyl.
  • n is an integer chosen from 1 , 2, and 3.
  • n is an integer chosen from 1 and 2.
  • Re is substituted or unsubstituted heteroaryl, mis an integer chosen from 1, 2, 3, 4, 5, and 6;
  • R 7 is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 1 -C 6 cyclic alkyl;
  • Re is chosen from H, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 1 -C 6 cyclic alkyl; and each of Rg and Rio is independently selected from H, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 1 -C 6 cyclic alkyl.
  • Rn is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, substituted or unsubstituted C 3 -C 6 cyclic alkyl, and substituted or unsubstituted heterocyclyl; a is an integer chosen from 0, 1, 2, 3, 4, 5, and 6;
  • R12 is chosen from substituted or unsubstituted heteroaiyl and substituted or unsubstituted aiyl.
  • a method for inhibiting VE-PTP in a mammalian subject in need thereof comprising administering to the subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VE-PTP activity is inhibited.
  • a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof whereby VE-PTP activity is inhibited.
  • 151. The method of embodiment 150, wherein the subject suffers from one or more of cancer, ocular disorders, occlusive cardiovascular disease, a vascular leaking syndrome, or another vascular-related disease.
  • a method for reducing VE-PTP-mediated signaling in a mammalian subject in need thereof comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VEPTP-mediated signaling is reduced.
  • a method for increasing Tie2-mediated signaling in a mammalian subject in need thereof comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby Tie2-mediated signaling is increased.
  • FIG. 1. depicts Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
  • VEC Human Umbilical Vein Endothelial Cells
  • FIG. 2. depicts further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
  • VEC Human Umbilical Vein Endothelial Cells
  • FIG. 3. depicts still further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
  • HUVEC Human Umbilical Vein Endothelial Cells
  • FIG. 4. depicts yet further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
  • VEC Human Umbilical Vein Endothelial Cells
  • FIG. 5 depicts additional Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
  • HUVEC Human Umbilical Vein Endothelial Cells
  • FIG. 6. depicts results of dose response titration experiments with compounds of the disclosure having an IC50 for VE-PTP in the activity range of 0.1 to 5 nM over a range of 0.2 to 50 ⁇ and illustrates a clear dose response relationship.
  • Angiopoietin 1 is the cognate ligand for Tie2 receptor, used to activate the Tie2 signaling pathway.
  • numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions and results, and so forth, used to describe and claim certain embodiments of the presort disclosure are to be understood as being modified in some instances by the term “about.”
  • the term “about” is used to indicate that a value includes the standard deviation of the mean for tire device or method being employed to determine the value.
  • the numerical parameters set forth in the specification are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment.
  • the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the present disclosure are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the present disclosure may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
  • administering refers to the placement of a compound and/or a pharmaceutical composition comprising the compound into a mammalian tissue or a subject by a method or route that results in at least partial localization of the compound and/or composition at a desired site or tissue location.
  • Effectiveness can be established in vitro (biochemical and/or biological in cultured cells) and/or in vivo. Effectiveness in vitro may be used to extrapolate or predict some degree of effectiveness in vivo, in an animal or in a human subject. A reference or standard or comparison may be used.
  • the term “effective” at inhibiting or activating an enzyme (VE-PTP and/or Tie2, respectively), and/or signaling mediated by the enzyme in the context of this disclosure and claims means reducing/activating the activity of the enzyme and/or the activation and propagation of the signaling pathway in terms of activation of a downstream molecule or known biological effect by a detectable or measurable amount relative to the baseline activity.
  • the reduction or activation is measured in terms of percentage reduction or activation, relative to the activity in the absence of exposure to the compound of the disclosure, including, for example, at least 5%, at least 10%, 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or about 100%.
  • the activity might also fall within a range, e.g., about 5-10%, about 10-20%, and any other range interval between about 1% and 100%.
  • An amount is “effective” in vivo if it produces any benefit to the subject to which the compound is administered.
  • disease or “disorder” are used interchangeably herein and refer to any alteration in state of the body or of some of the organs, interrupting or disturbing the performance of the functions and/or causing symptoms such as discomfort, dysfunction, distress, or even death to the person afflicted or those in contact with a person.
  • a disease or disorder can also relate to a distemper, ailing, ailment, malady, sickness, illness, complaint, indisposition, or affection.
  • VE-PTP-mediated disorder or a “Tie-2 mediated disorder” is any disease or deleterious condition in which VE-PTP or Tie2 play a role. Non-limiting examples of such disorders are provided elsewhere in the disclosure.
  • phrases “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • Drug-approval agencies e.g., EMA, US-FDA
  • Examples are listed in Pharmacopeias.
  • phrases “pharmaceutically acceptable excipient” is employed herein to refer to a pharmaceutically acceptable material chosen from a solvent, dispersion media, diluent, dispersion, suspension aid, surface active agent, isotonic agent, thickening or emulsifying agent, preservative, polymer, peptide, protein, cell, hyaluronidase, and mixtures thereof.
  • the solvent is an aqueous solvent.
  • treatment refers to obtaining a desired pharmacologic and/or physiologic effect.
  • the effect can be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or can be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease.
  • subject is meant a mammal, including, but not limited to, a human or non- human mammal, such as a bovine, equine, canine, ovine, or feline.
  • Ranges provided herein are understood to be shorthand for all of the values within the range.
  • a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.
  • the same rule applies for any other ranges described herein, even if the values within the range are not specifically called out in this disclosure.
  • alkyl refers to a saturated monovalent hydrocarbon radical comprising one to twelve carbon atoms (C 1 -C 12 ).
  • Alkyl groups may be linear, branched, or cyclic. Alkyl groups may be unsubstituted, or they may be substituted as described elsewhere herein.
  • an alkyl group comprises one to eight carbon atoms (Ci-Cg). In some embodiments, an alkyl group comprises one to six carbon atoms (C 1 -C 6 ). In some embodiments, an alkyl group comprises one to four carbon atoms (Ci-C*).
  • a cyclic alkyl group comprises three to six carbon atoms (C 1 -C 6 ).
  • substituted and unsubstituted linear, branched, or cyclic alkyl groups include methyl, ethyl, «-propyl, iso-propyl, cyclopropyl, «-butyl, sec-butyl, iso-butyl, ieri-butyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl, chloromethyl, fluoromethyl, trifluoromethyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, dimethylaminomethyl, 2-dimethylaminoethyl, 3-dimethylaminopropyl, 4- dimethylaminobutyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, trifluoroethy
  • alkylene alkylene unit
  • alkylene group refers to a saturated divalent hydrocarbon radical comprising one to twelve carbon atoms (Ci-Cu).
  • Alkylene groups may be linear, branched, or cyclic. Alkylene groups may be unsubstituted, or they may be substituted as described elsewhere herein.
  • an alkylene group comprises one to eight carbon atoms (C 1 -C 6 ).
  • an alkylene group comprises one to six carbon atoms (C 1 -C 6 ).
  • an alkylene group comprises one to four carbon atoms (C1-C4).
  • Non-limiting examples of alkylene groups include methylene and ethylene.
  • alkenyl refers to a monovalent hydrocarbon radical comprising two to eight carbon atoms (C2- Cs) with at least one site of unsaturation (i.e., an sp 2 carbon-carbon double bond).
  • Alkenyl groups may be linear, branched, or cyclic. Alkenyl groups max' be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkenyl group comprises two to six carbon atoms (Cz-Ce). In some embodiments, an alkenyl group comprises two to four carbon atoms (C2-C4). Alkenyl groups max' have E or Z orientations. Non-limiting examples of alkenyl groups include ethenyl (also called vinyl), 1-propenyl, iso- propenyl, and 2-chloroethenyl.
  • alkenylene refers to a divalent hydrocarbon radical comprising two to eight carbon atoms (C2-C8) with at least one site of unsaturation (e.g., an sp 2 carbon-carbon double bond).
  • Alkenylene groups may be linear, branched, or cyclic. Alkenylene groups may be unsubstituted, or they may be substituted as described elsewhere herein.
  • an alkylene group comprises two to six carbon atoms (C2-Ce).
  • an alkylene group comprises two to four carbon atoms (C2-C4).
  • alkylene groups may have E ox Z orientations.
  • Non-limiting examples of alkenyl groups include ethenylene (also called vinylene).
  • alkynyl refers to a monovalent hydrocarbon radical comprising two to eight carbon atoms (C2- Cs) with at least one site of unsaturation ⁇ i.e., an sp carbon-carbon triple bond).
  • Alkynyl groups may be linear or branched. Alkynyl groups may be unsubstituted, or they may be substituted as described elsewhere herein.
  • an alkynyl group comprises two to six carbon atoms (Ca-Ce).
  • an alkynyl group comprises two to four carbon atoms (C2-C4) ⁇
  • Non-limiting examples of alkynyl groups include ethynyl.
  • alkynylene refers to a divalent hydrocarbon radical comprising two to eight carbon atoms (Ca-Cg) with at least one site of unsaturation (i.e., an sp carbon-carbon triple bond).
  • Alkynylene groups may be linear or branched. Alkynylene groups may be unsubstituted, or they may be substituted as described elsewhere herein.
  • an alkynylene group comprises two to six carbon atoms (C2-C6).
  • an alkynylene group comprises two to four carbon atoms (C 2 -C 4 ).
  • Non-limiting examples of alkynylene groups incl ude ethynylene.
  • aryl refers to a monovalent aromatic hydrocarbon radical comprising 6-20 carbon atoms (C6-C20) that is derived by removing a hydrogen atom from an aromatic ring.
  • Aryl groups can be unsubstituted, or they can substituted with one or more substituents as described elsewhere herein.
  • Non-limiting examples of unsubstituted and substituted aryl groups include phenyl, 2- fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-methylphenyl, 3-methylphenyl, 4- methylphenyl, 2-chlorophenyl, 3-chlorophenyl, 4-dilorophenyl, 2,6-dichlorophenyl, 3,4- difluorophenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2-methoxyphenyl, 3- methoxyphenyl, 4-methoxyphenyl, 2-phenoxyphenyl, 3-phenoxyphenyl, 4-phenoxyphenyl, 2- cyanophenyl, 3-cyanophenyl, 4-cyanophenyl, 2-dimethylaminophenyl, 3- dimethylaminophenyl, 4-dimethylaminophenyl, 3-methylsulfonylphenyl, 4- methylsulfonylphenyl
  • arylene refers to a divalent aromatic hydrocarbon radical comprising 6-20 carbon atoms (Ce- C20) that is derived by removing two hydrogen atoms from an aromatic ring.
  • Arylene groups can be unsubstituted, or they can be substituted with one or more substituents as described elsewhere herein.
  • Non-limiting examples of arylene groups include phenylene.
  • heterocycle refers to a saturated or partially unsaturated ring system comprising 3 to 20 atoms, wherein at least one of the ring atoms is a heteroatom chosen from nitrogen, oxygen, phosphorous, and sulfur.
  • a heterocyclic group may be unsubstituted or may be substituted with one or more substituents as described elsewhere herein.
  • a heterocyclic group comprises 3 to 10 atoms.
  • a heterocyclic group comprises 3 to 7 atoms.
  • a heterocyclic group is monocyclic.
  • a heterocyclic group is bicyclic.
  • a heterocyclic group comprises fused rings.
  • unsubstituted and substituted heterocyclic groups include pyrrolidinyl, JV-methylpyrrolidinyl, azetidinyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl, and 3- methoxypyrrolidinyl.
  • heteroaryl group refers to a monovalent aromatic radical comprising one or more 5-, 6-, or 7-membered rings and comprising one or more heteroatoms independently chosen from nitrogen, oxygen, phosphorous, and sulfur.
  • a heteroaryl group may be unsubstituted or may be substituted with one or more substituents as described elsewhere herein.
  • a heteroaryl group comprises 5 to 20 atoms.
  • a heteroaryl group comprises 5 to 9 atoms.
  • a heteroaryl group comprises 5 atoms.
  • a heteroaryl group comprises 6 atoms.
  • a heteroaryl group comprises 7 atoms. In some embodiments, a heteroaryl group is monocyclic. In some embodiments, a heteroaryl group is bicyclic. In some embodiments, a heteroaryl group comprises fused rings.
  • heteroaryl groups include pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, 2-thienyl, 3-thienyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl- 1,2,4- oxadiazolyl, 3-phenyl-l,2,4-oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, thiadiazolyl, furazanyl, benzofurazany
  • heteroaryl groups include:
  • substituted refers to the replacement of one or more hydrogen atoms on one or more of a hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaryl group with one or more substituents.
  • a substituted hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaryl group any number of hydrogen atoms may be replaced by substituents.
  • Non-limiting examples of substituents that replace a single hydrogen atom include halogen, hydroxyl, and amino.
  • Non-limiting examples of substituents that replace two hydrogen atoms include carbonyl.
  • Non-limiting examples of substituents that replace three hydrogen atoms include cyano.
  • Non-limiting examples of substituents that can substitute for hydrogen atoms on one or more of a hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaiyl group include:
  • Ci-C 6 linear, branched, or cyclic alkyl groups non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl sec-butyl, iso- butyl, tert- butyl, cyclobutyl, cyclopentyl, and cyclohexyl;
  • substituted or unsubstituted aryl groups non-limiting examples of which include phenyl, 2-fluorophenyl, 3-methylphenyl, 4-chlorophenyl, 2,6- dichlorophenyl, 3,4-difluorophenyl, 3-hydroxyphenyl, 4-cyanophenyl, 2- dimethylaminophenyl, 3-metiiylsulfonylphenyl, 4-trifluorometiiylphenyl, 3- isopropylphenyl, 1 -naphthyl, and 2-naphthyl;
  • substituted or unsubstituted heterocyclic groups include pyrrolidinyl, N-methylpyrrolidinyl, azetidinyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl, and 3- methoxypyrrolidinyl;
  • substituted or unsubstituted heteroaryl groups include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, frnyl, 2-thienyl, 3- thienyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl-l,2,4-oxadiazolyl, 3- phenyl-l,2,4-oxadiazolyl, indolyl, benzothiazolyl, and l//-pyrrolo[2,3- 6]pyridinyl;
  • -(CR a R b ) z S0 3 R c non-limiting examples of which include -S0 3 H, -S0 3 CH 3 , -CH 2 S0 3 H, -CH 2 S0 3 CH 3 , -S0 3 C 6 H 5 , and -CH 2 S0 3 C 6 H 5 ; wherein each of R a and R b is independently chosen from hydrogen and substituted or unsubstituted C 1 -C 6 linear, branched, or cyclic alkyl, each of R c and R d is independently chosen from hydrogen, substituted or unsubstituted Ci-C 6 linear, branched, or cyclic alkyl, and aryl, or wherein R c and R d together form a ring system comprising 3 to 7 atoms, and z is chosen from 0, 1, 2, 3, and 4.
  • Compounds of the disclosure may contain one or more chiral centers. Compounds of tiie disclosure thus may exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds described herein, including, as non-limiting examples, diastereomers, enantiomers, and mixtures thereof (including, as a non-limiting example, racemic mixtures) form parts of the disclosure.
  • a compound of the disclosure is a compound of Formula I Formula I wherein
  • Ri is chosen from wherein
  • R 2 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl;
  • R 3 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl; n is an integer chosen from 1, 2, 3, 4, 5, and 6;
  • R 4 is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl;
  • R is chosen from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl; and each of Xi, X 2 , and X 3 is independently chosen from CH and N, with the proviso that Xi, X 2 , and X 3 are not simultaneously CH.
  • R 2 is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R 2 is substituted or unsubstituted heteroaryl. In some embodiments, R 2 is substituted heteroaryl. In some embodiments, R 2 is unsubstituted heteroaryl. In some embodiments, R 2 is 2-thienyl. In some embodiments, R 2 is 3-thienyl.
  • R 2 is not substituted or unsubstituted heteroaryl. In some embodiments, R 2 is not substituted heteroaryl. In some embodiments, R 2 is not unsubstituted heteroaryl. In some embodiments, R 2 is not 2-thienyl. In some embodiments, R 2 is not 3- thienyl.
  • R 2 is substituted or unsubstituted linear alkyl. In some embodiments, R 2 is substituted or unsubstituted Ci-C 6 linear alkyl. In some embodiments, R 2 is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 2 is substituted C 1 -C 6 linear alkyl. In some embodiments, R 2 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more halogen atoms.
  • R 2 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more fluorine atoms.
  • R 2 is a trifluoroalkyl group. In some embodiments, R 2 is trifluoroethyl.
  • R 2 is not substituted or unsubstituted linear alkyl. In some embodiments, R 2 is not substituted or unsubstituted branched alkyl. In some embodiments, R 2 is not substituted or unsubstituted cyclic alkyl. In some embodiments, R 2 is not substituted or unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R2 is not unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R2 is not substituted C 1 -C 6 linear alkyl. In some embodiments, R 2 is not methyl. In some embodiments, R 2 is not ethyl. In some embodiments, R 2 is not isopropyl. In some embodiments, R 2 is not cyclopropyl.
  • R 3 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R 3 is substituted or unsubstituted heteroaryl. In some embodiments, R 3 is substituted heteroaryl. In some embodiments, R 3 is unsubstituted heteroaryl. In some embodiments, R 3 is 2-thienyl. In some embodiments, R 3 is 3-thienyl.
  • R 3 is not substituted or unsubstituted ary-l. In some embodiments, R 3 is not substituted aryl. In some embodiments, R 3 is not unsubstituted aryl. In some embodiments, R 3 is not phenyl. In some embodiments, R 3 is not substituted or unsubstituted heteroaryl. In some embodiments, R 3 is not substituted heteroaryl. In some embodiments, R 3 is not unsubstituted heteroaryl. In some embodiments, R 3 is not 2-thienyl. In some embodiments, R 3 is not 3-thienyl.
  • R 3 is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R 3 is chosen from substituted or unsubstituted Ci-C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, R 3 is chosen from substituted C 1 -C 6 linear alkyl, substituted C 1 -C 6 branched alkyl, and substituted C 3 -C 6 cyclic alkyl.
  • R 3 is chosen from unsubstituted C 1 -C 6 linear alkyl, unsubstituted C 1 -C 6 branched alkyl, and unsubstituted C 3 -C 6 cyclic alkyl.
  • R 3 is not substituted or unsubstituted linear alkyl. In some embodiments, R 3 is not substituted or unsubstituted branched alkyl. In some embodiments, R 3 is not substituted or unsubstituted cyclic alkyl. In some embodiments, R 3 is not substituted or unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 3 is not substituted C 1 -C 6 linear alkyl. In some embodiments, R 3 is not unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 3 is not methyl. In some embodiments, R 3 is not ethyl. In some embodiments, R 3 is not isopropyl. In some embodiments, R 3 is not cyclopropyl.
  • R 3 is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 3 is substituted C 1 -C 6 linear alkyl. In some embodiments, R 3 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with a C 5 -C 20 heteroaryl group. In some embodiments, R 3 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more halogen atoms.
  • R 3 is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more fluorine atoms.
  • R 3 is a trifluoroalkyl group.
  • R 3 is trifluoroethyl.
  • R 3 is trifluoropropyl.
  • R 3 is substituted or unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, R 3 is unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, R 3 is cyclopropyl.
  • R 3 is chosen from 2-thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl,
  • R 3 is 2-thienyl. In some embodiments, In some embodiments, R 3 is
  • R 4 is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 - C 6 cyclic alkyl. In some embodiments, R 4 is chosen from substituted Ci-C 6 linear alkyl, substituted C 1 -C 6 branched alkyl, and substituted C 3 -C 6 cyclic alkyl.
  • R 4 is chosen from unsubstituted C 1 -C 6 linear alkyl, unsubstituted C 1 -C 6 branched alkyl, and unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, R 4 is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 4 is methyl.
  • R 5 is H. hi some embodiments, R 5 is chosen from substituted or unsubstituted Ci-C 6 linear alkyl, substituted or unsubstituted C]-C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, R 5 is substituted C 1 -C 6 linear alkyl. In some embodiments, Rs is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, R 5 is methyl.
  • R 5 is not substituted C 1 -C 6 linear alkyl. In some embodiments, R 5 is not unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rs is not methyl. In some embodiments, R $ is not H.
  • Xi and X 2 are CH and X 3 is N. In some embodiments, Xi and X 3 are CH and X 2 is N. In some embodiments, X 2 and X 3 are CH and Xi is N. In some embodiments, Xi is CH and X 2 and X 3 are N. In some embodiments, X 2 is CH and Xi and X 3 are N. For the avoidance of doubt, it is meant to be understood that, in some embodiments, > > ,
  • n is an integer chosen from 1, 2, and 3. In some embodiments, n is an integer chosen from 1 and 2. In some embodiments, n is 1.
  • a compound of the disclosure is a compound of Formula ⁇
  • Re is substituted or unsubstituted heteroaryl, mis an integer chosen from 1, 2, 3, 4, 5, and 6;
  • R ? is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl;
  • Rg is chosen from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl; and each of Rg and Rio is independently selected from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl.
  • Re is substituted heteroaryl. In some embodiments, Re is unsubstituted heteroaryl. In some embodiments, Re is 2-thienyl. In some embodiments, Re is 3-thienyl.
  • R 7 is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C3- Ce cyclic alkyl. In some embodiments, R 7 is substituted C 1 -C 6 linear alkyl. In some embodiments, R 7 is unsubstituted Ci-C 6 linear alkyl. In some embodiments, R 7 is methyl.
  • Rg is H. In some embodiments, Rg is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted C 1 -C 6 branched alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, Rg is substituted C 1 -C 6 linear alkyl. In some embodiments, Rg is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rg is methyl.
  • each of Rg and Rio is independently selected from H and substituted or unsubstituted linear alkyl. In some embodiments, each of Rg and Rio is independently selected from H and substituted or unsubstituted linear C 1 -C 6 alkyl. In some embodiments, Rg is H. In some embodiments, Rio is H. In some embodiments, each of Rg and Rio is H. In some embodiments, each of Rg and Rio is independently substituted C 1 -C 6 linear alkyl. In some embodiments, each of Rg and Rio is independently unsubstituted Ci-C 6 linear alkyl. In some embodiments, Rg is methyl. In some embodiments, Rio is methyl. In some embodiments, each ofRg and Rio is methyl.
  • m is an integer chosen from 2, 3, 4, 5, and 6. In some embodiments, m is an integer chosen from 2, 3, 4, and 5. In some embodiments, m is an integer chosen from 2, 3, and 4. In some embodiments, m is an integer chosen from 3 and 4. In some embodiments, m is 3. In some embodiments, m is 4.
  • a compound of the disclosure is a compound of Formula III
  • Rn is chosen from substituted or unsubstituted aiyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl; a is an integer chosen from 0, 1 , 2, 3, 4, 5, and 6;
  • R12 is chosen from substituted or unsubstituted heteroaiyl and substituted or unsubstituted aryl.
  • Rn is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, Rn is chosen from substituted or unsubstituted C 1 -C 6 linear alkyl, substituted or unsubstituted branched C 1 -C 6 alkyl, and substituted or unsubstituted C 3 -C 6 cyclic alkyl.
  • Ru is substituted or unsubstituted heteroaryl. In some embodiments, Ru is substituted heteroaiyl. In some embodiments, Ru is unsubstituted heteroaryl. In some embodiments, Ru is 2-thienyl. In some embodiments, Ru is 3-thienyl.
  • Ru is not substituted or unsubstituted heteroaiyl. In some embodiments, Rn is not substituted heteroaryl. In some embodiments, Ru is not unsubstituted heteroaiyl. In some embodiments, Ru is not 2-thienyl. In some embodiments, Rn is not 3-thienyl.
  • Rn is not substituted aryl. In some embodiments, Rn is not unsubstituted aiyl. In some embodiments, Rn is not phenyl.
  • Rn is not substituted or unsubstituted linear alkyl. In some embodiments, Rn is not substituted or unsubstituted branched alkyl. In some embodiments, Rn is not substituted or unsubstituted cyclic alkyl. In some embodiments, Rn is not substituted or unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rn is not substituted C 1 -C 6 linear alkyl. In some embodiments, Rn is not unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rn is not methyl. In some embodiments, Rn is not ethyl. In some embodiments, Rn is not isopropyl. In some embodiments, Rn is not cyclopropyl.
  • Rn is substituted or unsubstituted linear alkyl. In some embodiments, Rn is substituted or unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rn is unsubstituted C 1 -C 6 linear alkyl. In some embodiments, Rn is substituted C 1 -C 6 linear alkyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more halogen atoms. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with one or more fluorine atoms. In some embodiments, Rn is a trifluoroalkyl group. In some embodiments, Rn is trifluoroethyl. In some embodiments, Rn is trifluoropropyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with a C 3 -C 6 cyclic alkyl group.
  • Rn is substituted Ci-C 6 linear alkyl, wherein said substituted Ci-C 6 linear alkyl is substituted with cyclopropyl group.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with cyclopentyl group.
  • Rn is cyclopropylmethyl.
  • Rn is cyclopentylmethyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted Ci-C 6 linear alkyl is substituted with a heteroaryl group. In some embodiments, Rn is substituted Ci-C 6 linear alkyl, wherein said substituted Ci-C 6 linear alkyl is substituted with an imidazolyl group.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with -N(R c )(R d ), wherein each of R c and R d are independently chosen from hydrogen, substituted or unsubstituted linear, branched, or cyclic alkyl, and aryl, or wherein R c and R d together form a ring system comprising 3 to 7 atoms.
  • Rn is substituted Ci-C 6 linear alkyl, wherein said substituted Ci-C 6 linear alkyl is substituted with -N(R c )(R d ), wherein each of R c and R d are independently chosen from H and substituted or unsubstituted C 1 -C 6 linear alkyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with -NfR ⁇ OR 11 ), wherein each of R c and R d are independently chosen from substituted or unsubstituted C 1 -C 6 linear alkyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -N(R c )(R d ), wherein each of R c and R d are independently chosen from unsubstituted C 1 -C 6 linear alkyl.
  • Rn is substituted C 1 -C 6 linear alkyl, wherein said substituted C 1 -C 6 linear alkyl is substituted with an amino group, a dimethylamino group, a diethylaminogroup, a di-n- propylamino group, or a di-n-butylamino group.
  • Rn is a dimethylaminomethyl group.
  • Rn is substituted or unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, Rn is substituted C 3 -C 6 cyclic alkyl. In some embodiments, Rn is unsubstituted C 3 -C 6 cyclic alkyl. In some embodiments, Rn is cyclopropyl.
  • Rn is substituted or unsubstituted heterocyclyl. In some embodiments, Rn is unsubstituted heterocyclyl. In some embodiments, Rn is substituted heterocyclyl. In some embodiments, Rn is chosen from pyrrolidinyl, 3-hydroxypyrrolidinyl, and 3-methoxypyrroIidinyl.
  • Rn is chosen from 2-thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl, cyclopropylmethyl, cyclopentylmethyl,
  • R12 is substituted or unsubstituted heteroaiyl.
  • Rn is substituted heteroaryl.
  • R12 is unsubstituted heteroaryl.
  • Rn is pyridinyl.
  • R12 is substituted or unsubstituted aryl.
  • R12 is substituted aryl.
  • Rn is monosubstituted aryl. In some embodiments, Rn is monosubstituted phenyl. In some embodiments, Rn is unsubstituted aryl. In some embodiments, Rn is phenyl. In some embodiments, Rn is chosen from 1-naphthyl and 2-naphthyl. In some embodiments, Rn is 1- naphthyl. In some embodiments, Rn is 2-naphthyl.
  • a is an integer chosen from 0, 1, 2, and 3. In some embodiments, a is an integer chosen from 0, 1, and 2. In some embodiments, a is an integer chosen from 0 and 1. In some embodiments, a is 2. In some embodiments, a is 1. In some embodiments, a is 0.
  • b is an integer chosen from 0, 1, 2, and 3. In some embodiments, b is an integer chosen from 0, 1, and 2. In some embodiments, b is an integer chosen from 0 and 1. In some embodiments, b is 2. In some embodiments, b is 1. In some embodiments, b is 0.
  • a compound of the disclosure is a compound of Formula III wherein Y1-Y2 is nothing and a and b are not simultaneously 0. It is thus to be understood that, in some embodiments, a compound of the disclosure is a compound having the structure . It is also thus to be understood that, in some embodiments, a compound of the disclosure is a compound having the structure
  • compounds of the disclosure are chosen from the compounds recited in Table I, below:
  • a compound of the disclosure is in the form of a
  • pharmaceutically acceptable salt refers to a salt that is pharmaceutically acceptable as defined herein and that has the desired pharmacological activity of the parent compound.
  • pharmaceutically acceptable salts include those derived from inorganic acids, non-limiting examples of which include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid, and those derived from organic acids, non-limiting examples of which include acetic acid, trifluoroacetic add, propionic acid, glycolic acid, pyruvic acid, oxalic acid, stearic acid, malic acid, maldc acid, malonic acid, salicylic add, succinic acid, fumaric add, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid,
  • Additional non-limiting examples of pharmaceutically acceptable salts include those formed when an acidic proton in a parent compound is replaced by a metal ion, non-limiting examples of which include an alkali metal ion and an alkaline earth metal ion, and those formed when an acidic proton present in a parent compound is replaced by a ammonium ion, a primary ammonium ion, a secondary ammonium ion, a tertiary ammonium ion, or a quaternary ammonium ion.
  • Non-limiting examples of alkali metals and alkaline earth metals include sodium, potassium, lithium, calcium, aluminum, magnesium, copper, zinc, iron, and manganese.
  • Additional non-limiting examples of pharmaceutically acceptable salts include those comprising one or more counterions and zwitterions.
  • compositions can be used therapeutically as pharmaceutical compositions.
  • pharmaceutical composition refers to a preparation that is in such form as to permit the biological activity of the active ingredient to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the composition would be administered.
  • such compositions may be sterile.
  • the components of the composition may be in solution, emulsions, or suspension (for example, incorporated into microparticles, nanoparticles, or liposomes). Colloidal dosage forms such as nanoparticles, nanomicelles, liposomes, and microemulsions can also be used.
  • an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render the formulation isotonic.
  • pharmaceutically acceptable carriers include, but are not limited to, saline, Ringer’s solution and dextrose solution.
  • the pH of the solution is preferably from about 5 to about 8, and more preferably from about 7 to about 7.5.
  • Pharmaceutical compositions may include carriers, thickeners, diluents, buffers, preservatives, and surface-active agents.
  • Further carriers include sustained release preparations such as semi-permeable matrices of solid hydrophobic polymers containing the compound of the disclosure, which matrices are in the form of shaped particles, e.g., films, liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of the composition being administered. Pharmaceutical compositions may also include one or more active ingredients such as antimicrobial agents, anti-inflammatory agents, and anesthetics.
  • compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments.
  • Said compounds, pharmaceutically acceptable salts, and/or pharmaceutical compositions can be administered in unit forms of administration to mammalian subjects, including human beings.
  • Suitable unit forms of administration include, as non-limiting examples, forms administered orally and forms administered via a parenteral/systemic route, non-limiting examples of which including inhalation, subcutaneous administration, intramuscular administration, intravenous administration, intradermal administration, intravitreal administration, as well as topical and local ocular (i.e, subconjunctival, intravitreal, retrobulbar, intracameral) modes of administration.
  • compositions for oral administration can be in tiie form of tablets, pills, powders, hard gelatine capsules, soft gelatine capsules, and/or granules.
  • a compound of the disclosure and/or a pharmaceutically acceptable salt of a compound of the disclosure is (or are) mixed with one or more inert diluents, non-limiting examples of which including starch, cellulose, sucrose, lactose, and silica.
  • such pharmaceutical compositions may further comprise one or more substances other than diluents, such as (as non-limiting examples), lubricants, coloring agents, coatings, or varnishes.
  • compositions for parenteral administration can be in the form of aqueous solutions, non-aqueous solutions, suspensions, emulsions, drops (including, as a non-limiting example, eye drops), or any combination(s) thereof.
  • such pharmaceutical compositions may comprise one or more of water, pharmaceutically acceptable glycol(s), pharmaceutically acceptable oil(s), pharmaceutically acceptable organic esters, or other pharmaceutically acceptable solvents.
  • a variety of vehicles suitable for administering compounds to the eye are known in the art. Specific nonlimiting examples are described in U.S. Pat. No. 6,261,547; U.S. Pat. No. 6,197,934; U.S. Pat. No. 6,056,950; U.S. Pat. No.
  • compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to inhibit VE-PTP. In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to reduce or activate VE-PTP-mediated signaling.
  • compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to increase Tie2-mediated signaling. In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to treat a Tie2-mediated disorder or a VEPTP-mediated disorder.
  • compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments for one or more of cancer, ocular disorders, occlusive cardiovascular disease, vascular leaking syndrome(s), and/or other vascular-related disease(s).
  • compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments for ocular disorders.
  • the mode (or modes) of administration, dosage (or dosages), and optimized pharmaceutical form (or forms) can be determined according to criteria generally considered during the establishment of a treatment of a patient, such as, by way of non-limiting examples, the potency of the compound(s) and/or pharmaceutically acceptable salts of the compound(s), the age of the patient, the body weight of the patient, the severity of the patient’s condition (or conditions), the patient’s tolerance to the treatment, and secondary effects observed in treatment. Determination of dosages effective to provide therapeutic benefit for specific modes and frequency of administration is within the capabilities of those skilled in the art.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 2,000 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 1,000 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 500 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 250 pg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 100 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 50 pg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 5,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 3,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 1 mg to about 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 500 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 50 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 50 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 100 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 10 mg to 50 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 250 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 25 mg to 50 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 100 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 100 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 2,000 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 500 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 1,000 mg.
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount of about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 10 pg, about 15 pg, about 20 pg, about 25 pg, about 30 pg, about 35 pg, about 40 pg, about 45 pg, about 50 pg, about 60 pg, about 70 pg, about 80 pg, about 90 pg, about 100 pg, about 125 pg, about 150 pg, about 175 pg, about 200 pg, about 225 pg, about 250 pg, about 300 pg, about 350 pg, about 400 pg, about 450 pg, about 500 pg, about 550 pg, about 600 pg, about 650 pg, about 700 pg, about 750 pg, about 800 pg, about 850
  • a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 1,000 mg, about 1,100 mg, about 1,200 mg, about 1,300 mg, about 1,400 mg, about 1,500 mg, about 1,600 mg, about
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mgZmL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 10 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 5 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 3 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 2 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 1 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 0.5 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 40 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 20 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 10 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 5 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 40 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 20 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 10 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 40 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 20 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 40 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 30 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 40 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 60 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 60 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 60 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 60 mg/mL to about 80 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 70 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 70 mg/mL to about 80 mg/mL.
  • a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of about 0.01 mg/mL, about 0.02 mg/mL, about 0.03 mg/mL, about 0.04 mg/mL, about 0.06 mg/mL, about 0.07 mg/mL, about 0.08 mg/mL, about 0.09 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 40 mg/mL, about 45 mg/mL, about 50 mg/mL, about 60 mg/mL, about 70 mg/mL, about 80 mg/mL, about 90 mg/mL, or about 100 mg/mL.
  • Effective amounts and dosages can be estimated initially from in vitro assays.
  • an initial dosage for use in animals can be formulated to achieve a circulating blood or serum concentration of active compound that is at or above an ICjo of the particular compound as measured in an in vitro assay. Calculating dosages to achieve such circulating blood or serum concentrations taking into account the bioavailability of the particular compound is well within the capabilities of skilled artisans. For guidance, the reader is referred to Fingl & Woodbury, "General Principles," In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, latest edition, Pergamagon Press, and the references cited therein, which methods are incorporated herein by reference in their entirety.
  • Initial dosages can also be estimated from in vivo data, such as animal models. Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases described in this disclosure are well-known in the art.
  • dosage amounts will be in the range of from about 0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but can be higher or lower, depending upon, among other factors, the activity of the compound, its bioavailability, the mode of administration and various factors discussed above. Dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s) which are sufficient to maintain therapeutic or prophylactic effect.
  • the compounds can be administered once per week, several times per week (e.g., every other day), once per day or multiple times per day, depending upon, among other things, the mode of administration, the specific indication being treated and the judgment of the prescribing physician.
  • the effective local concentration of active compound(s) may not be related to plasma concentration. Skilled artisans will be able to optimize effective local dosages without undue experimentation.
  • a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg,
  • a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg, 7.5 pg, 10 pg, 12.5 pg, 15 pg, 17.5 pg, 20 pg, 22.5 pg, 25 pg, 27.5 pg, or 30 pg.
  • a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg, 7.5 pg, 10 pg, 12.5 pg, 15 pg, 17.5 pg, 20 pg, 22.5 pg, 25 pg, 27.5 pg, or 30 pg once daily.
  • a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg, 7.5 pg, 10 pg,
  • a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg,
  • a therapeutic treatment comprises administration of a a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg, 7.5 mg, 10 mg,
  • a therapeutic treatment comprises administration of a a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg, 7.5 mg, 10 mg,
  • Phosphatase hydrolyzes phospho-substrate (DiFMUP) to release a fluorophore (DiFMU).
  • the fluorophore-DiFMU can be detected using FlexStation multi-mode microplate reader with excitation at 360 nm and emission at 460 nm.
  • the fluorescence intensity represents the reaction activity.
  • Buffer Preparation pH 7.4 buffer Potassium Phosphate, Monobasic 0.2 M. Dissolve 27.22 g of monobasic potassium phosphate (KH2PO4) in water, and dilute with water to 1000 mL. Place 50 mL of the monobasic potassium phosphate solution in a 200 mL volumetric flask, add 40 mL of sodium hydroxide solution, then add water to achieve desired volume and adjust pH to
  • HUVEC human umbilical vein endothelial cells
  • HUVEC AKT is a verified downstream signaling effector of Tie2 receptor tyrosine kinase, whose phosphorylation increases upon binding of Angiopoietin ligand to the receptor or by inhibiting VE-PTP, the negative inhibitor of the activated receptor (Souma, T et al. (2018 Jan)
  • Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115).
  • HUVEC-C Human Umbilical Vein/Vascular Endothelium cells
  • ATCC American Type Culture Collection
  • CRL-1730 Human Umbilical Vein/Vascular Endothelium cells
  • HUVEC ells were grown using EndoGro LS Complete Media Kit (Millipore, SCMEOOl) supplemented with Penicillin-Streptomycin cocktail (Coming, 30-002-CI) in an incubator at 37°C in 5%CC> 2 . Few days prior to the experiment, cells were counted using a hematocytometer and subcultured so as to reach 60-80% confluency on the day of the experiment.
  • DMSO dimethyl sulfoxide
  • the obtained whole cell lysate was spinned down for 5 minutes at 13,300 rpm, 4°C to remove cellular debris.
  • the supernatant was mixed with 4x Laemmli sample buffer (Bio-Rad, #161- 0747) to final lx concentration.
  • Subsequently samples were boiled at 95°C for 5 minutes and either used directly in polyacrylamide gel electrophoresis or stored at -20°C.
  • the PVDF membrane was washed once in lx TBS-T, followed by blocking with 5% BSA in TBS-T for 1 hour at room temperature with gentle shaking. The blocked membrane was cut according to the visible marker lanes to probe simultaneously but separately for different proteins of interest.
  • the resulting PVDF membrane stripes were incubated with either primary Phospho-AKT (Ser473) (D9E) XP® Rabbit mAb (Cell Signaling Technology', 4060) or with primary GAPDH (D16H11) XP® Rabbit mAb (Cell Signaling Technology, 5174) overnight, gently shaking at 4°C. Next day, membrane stripes were washed three times with lx TBS-T for 5 minutes, gently shaking at room temperature.
  • Radio immune precipitation assay buffer 50 mM Tris-Cl (pH 8.0)
  • Tween-20 Tris buffered saline-Tween-20 (TBS-T): 50 mM Tris pH 7.4,150 mM NaCl, and 0.01% (v/v) Tween-20
  • FIG. 1 to FIG. 6 depict Western blot analyses performed in accordance with the above- described protocol.
  • Compounds with IC50 for VE-PTP in the activity' range of 0.1 to 5 nM were dissolved in DMSO and used to treat Human Umbilical Vein Endothelial Cells
  • HUVEC cells treated for 10 minutes with indicated concentration of specific compound were used in whole cell lysate Western blot analysis to determine the effectiveness of compound at inhibiting VE-PTP and activating Tie2 signaling cascade, culminating in increase in downstream phosphorylation of serine/threonine-specific protein kinase AKT/PKB.
  • the glycolytic enzyme Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control.
  • Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control.
  • FIG. 1 to FIG. 5 show two biological replicates for each compound.
  • certain compounds are much more effective at inducing phosphorylation of AKT than others, despite being in the same ICso activity range for VE-PTP.
  • tire Western blots some compounds had tire same or slightly higher level of pAKT induction as DMSO at the concentrations tested, while others generated a more pronounced response.
  • FIG. 6 depicts dose response titration experiments with selected compounds over a range of 0.2 to 50 ⁇ and illustrates a clear dose response relationship.
  • Angiopoietin 1 600 ng/mL is the cognate ligand for Tie2 receptor, used to activate the Tie2 signaling pathway.
  • the ocular penetration study was performed to ascertain whether potent and soluble compounds 1, 2, 3 (diastereomer 1), 3 (diastereomer 2), 5, 16 and 17 administered topically as a single dose to the rat’s eye are present in the aqueous humor 30 minutes post-treatment.
  • Compounds 1, 2, 3, 5, 16 and 17 were dissolved in 50 mM PBS pH 7.4 at maximum formulatable concentration, ranging from 5 to 15 mg/mL, and dose volume delivered topically as 6 uL per eye.
  • Four rats per group were used, receiving bilateral doses to the central cornea using a calibrated micropipette, with a total of eight eyes per group collected post euthanasia. Rats were held such that the treatment remained in the eye for a period of few seconds before letting go of the animal. After receiving a single dose per eye the eyes and aqueous humour were collected 30 minutes post-dose and frozen immediately.
  • rats were euthanized by CO2 inhalation followed by thoracotomy performed in accordance with the accepted American Veterinary Medical Association guidelines. Immediately following confirmation of death the eye was washed with balanced salt solution. Aqueous humor was collected from both eyes of each animal, flash frozen in liquid nitrogen and stored in a -80 degree Celsius freezer. All tissues where then shipped on dry ice for bioanalysis. Mass spectrometry and chromatography methods were established to separate, detect and quantify each compound used in the study. Aqueous humor samples were analysed to detect and calculate the concentration of each analyte, which were then averaged and represented as ng/mL per group per compound.
  • the crude was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water(10mM NH4HCO3)- ACN];B%: 12%-42%,20min) to yield ammonium (4-((2S)-2-(2- methoxycarbonyl)(methyl)amino)-3-(pyridin-3-yl)propanamido)-2-(2-(thiophen-2- yl)thiazol-4-yl)ethyl)phenyl)sulfamate diastereomer 2 (101.50 mg, 34%) was obtained as a white solid.
  • reaction was bubbled withNHs (2.59 g, 152.24 mmol, 1.00 eq) at 0 °C, the mixture was stirred at 0 °C for 0.5 h.
  • the reaction was monitored by LCMS and TLC and when complete, the solution was washed with aq. HC1 (5%, 500 mL), HzO (2 x 400 mL) and aq. NaHCOs (5%, 500mL).
  • the combined organic layer was washed with brine (300 mL), dried with anhydrous NazSO*, filtered and concentrated in vacuo.
  • the mixture was stirred at 25 °C for 13 h.
  • the reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water 20 mL and extracted with ethyl acetate 30 mL (3 x 10 mL). The combined organic layer was washed with brine (30 mL), dried with Na 2 SC> 4 , filtered and concentrated under vacuo.
  • reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo, and the residue was diluted with water 20 mL and extracted with DCM 60 mL (3 x 20 mL). The combined organic layer was washed with brine (50 mL), dried with NaaSO*, filtered and concentrated in vacuo.
  • Diastereoisomer 1 To a solution of 18_diastereoisomer 1 (0.12 g, 232.70 umol, 1 eq) in pyridine (1 mL) andMeCN (1 mL) was cooled to 0 °C, then the sulfur trioxide pyridine complex (111.11 mg, 698.09 umol, 3 eq) was added at 0 °C and the reaction mixture was stirred at 0 °C for 0.17 h.
  • reaction was monitored by TLC and when complete, the reaction mixture was filtered, the filtrate was added diazomethane (12.75 g, 303.51 mmol, 7.09 eq) in ether (300 mL) and then the olution was stirred at 0 °C for 3 h.
  • the reaction was monitored by TLC and when complete, the eaction mixture was to yield intermediate 4 (14.31 g, crude) in THF (100 mL) was obtained as ellow liquid.
  • reaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured nto water (700 mL) and then the filter cake was dissolved into ethyl acetate (1000 mL) and washed with water (3 x 500 mL). Then the organic phase was washed with HC1 (5 %, 2 x 500 mL), water (2 x 400 mL) and NaHCCb (3 x 500 mL), then the organic phase was dried with nhydrous Na 2 SC> 4 and concentrated in vacuo.
  • reaction was monitored by TLC nd when complete, the reaction mixture was filtered, the filtrate was added ice-water (300 mL), nd then the mixture was extracted with ethyl acetate (3 x 300 mL), the organic phase was eutralized with aq. NaHCO .3 (5%, 3 x 200 mL). Then the combined organic phase was washed with brine (300 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo.
  • ntermediate IB (20 g, 212.51 mmol, 1 eq), intermediate 1C (33.83 g, 318.76 mmol, 32.22 mL, .5 eq) and ZnCh (11.59 g, 85.00 mmol, 0.4 eq) were stirred at 160 °C for 5 h.
  • the reaction was monitored by TLC and when complete, the reaction mixture was concentrated in vacuo.
  • ntermediate 2A (7 g, 38.41 mmol, 1 eq) was dissolved in dioxane (140 mL) and HiO (47 mL) was added OsC>4 (0.1 M, 5.88 mL, 1.53e '2 eq) and NaI04 (16.68 g, 77.98 mmol, 2.03 eq), and hen stirred at 25 °C for 6 h.
  • reaction was monitored y LCMS and when complete, the reaction mixture was diluted with water (30 mL) and xtracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with brine (30 mL), dried with anhydrous NazSC ⁇ , filtered and concentrated in vacuo to give a residue.

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Abstract

The present disclosure relates to compounds capable of inhibiting vascular endothelial protein tyrosine phosphatase (VE-PTP). These compounds are also capable of activating Tie2 receptor-mediated signaling. The present disclosure also relates to pharmaceutically acceptable salts of said compounds, to pharmaceutical compositions comprising such compounds and/or pharmaceutically acceptable salts thereof, and to the use of such compounds, pharmaceutically acceptable salts thereof, and/or pharmaceutical compositions comprising the same in treating diseases and/or conditions mediated by VE-PTP signaling, such as those mediated by Angiopoietm/Tie2 signaling.

Description

VE-PTP INHIBITORS
The present disclosure relates to compounds capable of inhibiting vascular endothelial protein tyrosine phosphatase (VE-PTP). These compounds are also capable of activating Tie2 receptor-mediated signaling. Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP. The present disclosure also relates to pharmaceutically acceptable salts of said compounds, to pharmaceutical compositions comprising such compounds and/or pharmaceutically acceptable salts thereof, and to the use of such compounds, pharmaceutically acceptable salts thereof, and/or pharmaceutical compositions comprising the same in treating diseases and/or conditions mediated by VE- PTP signaling, such as those mediated by Angiopoietin/Tie2 signaling.
Background of the Disclosure
Vascular endothelial tyrosine phosphatase (VE-PTP) is the mouse ortholog of human receptor-type tyrosine-protein phosphatase beta (ΗΡΤΡ-β), but the two terms are often used interchangeably (Matozaki, T et al. (2010 Dec) Expression, localization, and biological function of the R3 subtype of receptor-type protein tyrosine phosphatases in mammals , Cell Signal. 22(12): 1811-7). VE-PTP knockout mice have defective blood vessel remodeling (Dominguez, MG et al. (2007 Feb) Vascular endothelial tyrosine phosphatase (VE-PTP)-null mice undergo vasculogenesis but die embryonically because of defects in angiogenesis, Proc Natl Acad Sci U S A. 104(9):3243-8.; Winderlich, M et al. (2009 May) VE-PTP controls blood vessel development by balancing Tie-2 activity, J Cell Biol. 18;185(4):657-71). VE- PTP has an extracellular domain with multiple fibronectin type III repeats, a single transmembrane segment, and one cytoplasmic catalytic domain (Chicote, JU et al. (2017 Mar) Phosphotyrosine phosphatase R3 receptors: Origin, evolution and structural diversification, PLoS ONE, 12(3), e0172887.; Matozaki, T et al. (2010 Dec) Expression, localization, and biological function of the R3 subtype of receptor-type protein tyrosine phosphatases in mammals, Cell Signal. 22(12):1811-7). One of the substrates for VE-PTP is Tie2, an endothelial tyrosine kinase receptor that regulates angiogenesis and lymphangiogenesis and can support the integrity of endothelial junctions (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci. 131(1):87-103.; Frye, M et al. (2015 Dec) Interfering with VE-PTP stabilizes endothelial junctions in vivo via Tie-2 in the absence of VE-cadherin, J Exp Med. 212(13):2267-87.; Souma, T et al. (2018 Jan) Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115). Antibodies against the extracellular domain ofVE-PTP dissociate the phosphatase from Tie2, leading to Tie2 phosphorylation and activated signaling (Winderlich, M et al. (2009 May) VE-PTP controls blood vessel development by balancing Tie-2 activity, J Cell Biol. 185(4):657-71). Pharmacological inhibition of VE-PTP activity also activates Tie2 and is capable of suppressing ocular neovascularization and VEGF-mediated vascular leakage (Goel, S et al. (2013 Aug) Effects of vascular-endothelial protein tyrosine phosphatase inhibition on breast cancer vasculature and metastatic progression. J Natl Cancer Inst, 21 ;105(16): 1188-201.; Shen, J et al. (2014 Oct) Targeting VE-PTP activates TIE2 and stabilizes the ocular vasculature, J Clin Invest. 124(10):4564-76.; Souma, T et al. (2018 Jan) Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Nad Acad Sci U S A. pii: 1714446115). Indeed, stimulation of overall Tie2 activity via inhibition of VE-PTP can promote endothelial barrier function (Frye, M et al. (2015 Dec) Interfering with VE-PTP stabilizes endothelial junctions in vivo via Tie-2 in the absence of VE-cadherin, J Exp Med. 212(13):2267-87.; Gumik, S et al. (2016 May) Angiopoietin-2-induced blood-brain barrier compromise and increased stroke size are rescued by VE-PTP-dependent restoration ofT' ie2 signaling, Acta Neuropatiiol.131 (5):753- 73.; Milam, KE et al. (2015 Apr) The angiopoietin-Tie2 signaling axis in the vascular leakage of systemic inflammation , Tissue Barriers. 3(l-2):e957508).
The Angiopoietin-Tie2 signaling pathway is a major regulator of vascular development, vascular permeability, vessel remodeling, and post-natal angiogenesis, and altered expression of the Angiopoietin ligands or activity of the Tie2 receptor has been linked to a variety of lymphatic and blood vasculature pathologies including cancer, sepsis, diabetes, atherosclerosis, and ocular diseases (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci. 131(1):87-103.; Parikh, SM (2017 Jul) The Angiopoietin-Tie2 Signaling Axis in Systemic Inflammation, J Am Soc Nephrol. 28(7): 1973-1982.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin- TIE pathway, Nat Rev Drug Discov. 16(9):635-661). In blood vascular endothelium, Angiopoietin2 (Angpt2 or Ang2) is reported to function as a competitive antagonist of Angiopoietinl /Tie2 signaling, inhibiting Angiopoietin 1 (Angptl or Angl>mediated phosphorylation of Tie2 (Thurston, G et al. (2012 Sep) The complex role of angiopoietin-2 in the angiopoietin-tie signaling pathway, Cold Spring Harb Perspect Med. 2(9):a006550.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661). Tie2 receptor activators include, for example, Angptl recombinant proteins, vascular endothelial protein tyrosine phosphatase (VE-PTP) inhibitors, and Tie2-peptidomimetics (Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661.; Shen, J et al. (2014 Oct) Targeting VE-PTP activates TIE2 and stabilizes the ocular vasculature, J Clin Invest. 124(10):4564-76.; Souma, T et al. (2018 Jan) Context-dependent Junctions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115). Angiopoietin- 1 decreases endothelial cell permeability and increases vascular stabilization via recruitment of pericytes and smooth muscle cells to growing blood vessels, while Angiopoietin-2 mediates angiogenic sprouting and vascular regression (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci. 131(1):87-103.; Koh, GY (2013 Jan) Orchestral actions of angiopoietin-1 in vascular regeneration. Trends Mol Med. 19(l):31-9.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661). In addition, both Angptl and Angpt2 can promote lymphangiogenesis (Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci. 131(1):87-103.; Souma, T et al. (2018 Jan) Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115). Angpt2 deficiency in knockout mice resulted in widespread lymphatic dysfunction due to abnormal remodeling of the developing lymphatic vessels, resulting in lymphedema (Dellinger, M et al. (2008 Jul) Defective remodeling and maturation of the lymphatic vasculature in Angiopoietin-2 deficient mice, Dev Biol. 319(2):309-20.; Eklund, L et al. (2017 Jan) Angiopoietin-Tie signalling in the cardiovascular and lymphatic systems, Clin Sci. 131(1):87-103). The expression of Angl in the Ang2 genetic locus rescued the lymphatic abnormalities of Ang2 deficient mice, suggesting that Angl and Ang2 have overlapping functions in the lymphatic vasculature (Gale, NW et al. (2002 Sep) Angiopoietin- 2 is required for postnatal angiogenesis and lymphatic patterning, and only the latter role is rescued by Angiopoietin- 1, Dev Cell. 3(3):411-23).
Tie2 activation promotes formation of the Schlemm’s canal, a specialized hybrid lymphatic vessel formed around the eye (Souma, T et al. (2016 Jul) Angiopoietin receptor TEK mutations underlie primary congenital glaucoma with variable expressivity, J Clin Invest. 126(7):2575-87.; Thomson, BR et al. (2014 Oct) A lymphatic defect causes ocular hypertension and glaucoma in mice, J Clin Invest. 124(10): 4320-4.; Thomson, BR et al. (2017 Nov) Angiopoietin-1 is required for Schlemm’s canal development in mice and humans, J Clin Invest, pii: 95545). Both Angptl/Angpt2 double knockout mice and Tie2 knockout mice have a defective Schlemm’s canal, leading to impaired drainage of aqueous humour, increased intraocular pressure (IOP) and glaucoma (Souma, T et al. (2016 Jul) Angiopoietin receptor TEK mutations underlie primary congenital glaucoma with variable expressivity, J Clin Invest. 126(7):2575-87.; Thomson, BR et al. (2014 Oct) A lymphatic defect causes ocular hypertension and glaucoma in mice, J Clin Invest. 124(10):4320-4.; Thomson, BR et al. (2017 No \) Angiopoietin- 1 is required for Schlemm’s canal development in mice and humans, J Clin Invest, pii: 95545). However, knockout of VE-PTP rescues the Angptl/Angpt2 double knockout phenotype, as well as the Tie2 haploinsufficiency, demonstrating the power of VE-PTP inhibition in activating Tie2 signaling. (WO2017/190222). The introduction of VE-PTP null alleles into an Angptl/Angpt2 conditional knockout mouse eliminates phenotypic expression of high IOP and prevents pressure-related loss of retinal-ganglion cells leading to glaucoma. (WO2017/190222). There is thus a use for pharmacological VE-PTP inhibitors, particularly some that could have improved physicochemical properties, and are potent Tie2 activators. Such therapeutic agents can be useful in the treatment of cancer, glaucoma, occlusive cardiovascular disease, vascular leaking syndromes, and other vascular-related diseases (Parikh, SM (2017 Jul) The Angiopoietin-Tie2 Signaling Axis in Systemic Inflammation, J Am Soc Nephrol. 28(7): 1973- 1982.; Saharinen, P et al. (2017 Sep) Therapeutic targeting of the angiopoietin-TIE pathway, Nat Rev Drug Discov. 16(9):635-661.; Schmittnaegel, M et al. (2017 Dec) Reprogramming Tumor Blood Vessels for Enhancing Immunotherapy, Trends Cancer. 3(12):809-812).
Summary of the Disdosure
The present disdosure relates to compounds and pharmaceutically acceptable salts thereof that are capable of inhibiting vascular endothelial protein tyrosine phosphatase (VE- PTP). These compounds are capable of activating the Tie-2 receptor. The present disclosure also relates to pharmaceutical compositions comprising such compounds and/or pharmaceutically acceptable salts thereof. The present disclosure also relates to the use of such compounds, pharmaceutically acceptable salts thereof, and/or pharmaceutical compositions in treating diseases and/or conditions mediated by VE-PTP activity, including those that are linked to Ang/Tie2-mediated signaling.
Many modifications and other embodiments of the disclosures set forth herein will come to mind to one skilled in the art to which these disclosures pertain having the benefit of tire teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the disclosures are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
In some embodiments, the present disclosure is directed to:
1. A compound of Formula I
Formula 1 wherein
Ri is chosen from wherein
R2 is chosen from substituted or unsubstituted heteroaiyl and substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms.
R3 is chosen from substituted or unsubstituted heteroaiyl, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl; n is an integer chosen from 1, 2, 3, 4, 5, and 6;
Rt is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted Ci-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl;
Rs is chosen from H, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl; and each of Xi, X2, and X3 is independently chosen from CH and N, with the proviso that Xi, X2, and X3 are not simultaneously CH.
2. The compound according to embodiment 1, wherein Ri is
10
3. The compound according to embodiment 1, wherein Ri is
4. The compound according to embodiment 2, wherein R2 is substituted or unsubstituted heteroaryl.
5. The compound according to embodiment 2, wherein R2 is substituted heteroaiyl.
6. The compound according to embodiment 2, wherein R2 is unsubstituted heteroaiyl.
7. The compound according to embodiment 2, wherein R2 is 2-thienyl.
8. The compound according to embodiment 2, wherein R2 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. 9. The compound according to embodiment 2, wherein R2 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms.
10. The compound according to embodiment 2, wherein R2 is a trifluoroalkyl group.
11. The compound according to embodiment 2, wherein R2 is trifluoroethyl.
12. The compound according to embodiment 3, wherein R3 is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3- Ce cyclic alkyl.
13. The compound according to embodiment 3, wherein R3 is substituted or unsubstituted heteroaryl.
14. The compound according to embodiment 3, wherein R3 is substituted heteroaryl.
15. The compound according to embodiment 3, wherein R3 is unsubstituted heteroaryl.
16. The compound according to embodiment 3, wherein R3 is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl.
17. The compound according to embodiment 3, wherein R3 is chosen from substituted or unsubstituted C1-C6 linear alkyl and substituted or unsubstituted C3-C6 cyclic alkyl.
18. The compound according to embodiment 3, wherein R3 is substituted or unsubstituted C1-C6 linear alkyl. 19. The compound according to embodiment 3, wherein R3 is substituted C1-C6 linear alkyl.
20. The compound according to embodiment 3, wherein R3 is unsubstituted C1-C6 linear alkyl.
21. The compound according to embodiment 3, wherein R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms.
22. The compound according to embodiment 3, wherein R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms.
23. The compound according to embodiment 3, wherein R3 is a trifluoroalkyl group.
24. The compound according to embodiment 3, wherein R3 is trifluoroethyl.
25. The compound according to embodiment 3, wherein R3 is trifluoropropyl.
26. The compound according to embodiment 3, wherein R3 is substituted or unsubstituted C3-C6 cyclic alkyl.
27. The compound according to embodiment 3, wherein R3 is unsubstituted C3-C6 cyclic alkyl.
28. The compound according to embodiment 3, wherein R3 is cyclopropyl.
29. The compound according to embodiment 3, wherein R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a C5-C20 heteroaryl group. 30. The compound according to embodiment 3, wherein R3 is chosen from 2- thienyl, trifluoroethyl, trifluoropropyl,
31. The compound according to embodiment 3, wherein R3 is 2-thienyl.
32. The compound according to embodiment 3, wherein
N ll
33. The compound according to embodiment 3, wherein R3 is
34. The compound according to embodiment 1, wherein R4 is substituted or unsubstituted C1-C6 linear alkyl.
35. The compound according to embodiment 1, wherein R4 is unsubstituted Ci-C6 linear alkyl.
36. The compound according to embodiment 1, wherein R4 is methyl.
37. The compound according to embodiment 1, wherein R5 is chosen from H and substituted or unsubstituted Ci-C6 linear alkyl.
38. The compound according to embodiment 1, wherein is Rs is H.
39. The compound according to embodiment 1, wherein R5 is substituted C1-C6 linear alkyl.
40. The compound according to embodiment 1, wherein R5 is unsubstituted C1-C6 linear alkyl. 41. The compound according to embodiment 1, wherein R5 is methyl.
42. The compound according to embodiment 1, wherein Xi and X2 are CH and X3 isN.
43. The compound according to embodiment 1, wherein Xi and X3 are CH and X2 is N.
44. The compound according to embodiment 1, wherein X2 and X3 are CH and Xi isN.
45. The compound according to embodiment 1 , wherein Xi is CH and X2 and X3 areN.
46. The compound according to embodiment 1, wherein X2 is CH and Xi and X3 areN.
47. The compound according to embodiment 1, wherein n is an integer chosen from 1 , 2, and 3.
48. The compound according to embodiment 1, wherein n is an integer chosen from 1 and 2.
49. The compound according to embodiment 1, wherein n is 1.
50. A compound of Formula Π
wherein
Re is substituted or unsubstituted heteroaryl, mis an integer chosen from 1, 2, 3, 4, 5, and 6;
R7 is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C1-C6 cyclic alkyl;
Re is chosen from H, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C1-C6 cyclic alkyl; and each of Rg and Rio is independently selected from H, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C1-C6 cyclic alkyl.
51. The compound according to embodiment 50, wherein Re is substituted heteroaryl.
52. The compound according to embodiment 50, wherein Re is unsubstituted heteroaryl.
53. The compound according to embodiment 50, wherein Re is 2-thienyl.
54. The compound according to embodiment 50, wherein R? is substituted or unsubstituted Ct-Ce branched alkyl. 55. The compound according to embodiment 50, wherein R7 is substituted or unsubstituted C3-C6 cyclic alkyl.
56. The compound according to embodiment 50, wherein R7 is substituted or unsubstituted C1-C6 linear alkyl.
57. The compound according to embodiment 50, wherein R7 is substituted C1-C6 linear alkyl.
58. The compound according to embodiment 50, wherein R7 is unsubstituted Ci-
Ce linear alkyl.
59. The compound according to embodiment 50, wherein R7 is methyl.
60. The compound according to embodiment 50, wherein Rg is chosen from H and substituted or unsubstituted C1-C6 linear alkyl.
61. The compound according to embodiment 50, wherein is Rg is H.
62. The compound according to embodiment 50, wherein Rg is substituted or unsubstituted C1-C6 linear alkyl.
63. The compound according to embodiment 50, wherein Rg is substituted C1-C6 linear alkyl.
64. The compound according to embodiment 50, wherein Rg is unsubstituted Ci-
Ce linear alkyl.
65. The compound according to embodiment 50, wherein Rg is methyl.
66. The compound according to embodiment 50, wherein each of Rg and Rio is independently chosen from H and substituted or unsubstituted C1-C6 linear alkyl.
67. The compound according to embodiment 50, wherein each of R9 and Rio is H. 68. The compound according to embodiment 50, wherein each of Rg and Rio is independently substituted Ci-C6 linear alkyl.
69. The compound according to embodiment 50, wherein each of Rg and Rio is independently unsubstituted C1-C6 linear alkyl.
70. The compound according to embodiment 50, wherein each of Rg and Rio is methyl.
71. The compound according to embodiment 50, wherein m is an integer chosen from 1, 2, 3, and 4.
72. The compound according to embodiment 50, wherein m is an integer chosen from 2, 3, and 4.
73. The compound according to embodiment 50, wherein m is an integer chosen from 3 and 4.
74. The compound according to embodiment 50, wherein m is 3.
75. The compound according to embodiment 50, wherein m is 4.
76. A compound of Formula ΠΙ
Formula III wherein
Rn is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, substituted or unsubstituted C3-C6 cyclic alkyl, and substituted or unsubstituted heterocyclyl; a is an integer chosen from 0, 1, 2, 3, 4, 5, and 6;
Y1-Y2 is a group chosen from nothing, -CH2-CH2-, and -CH=CH-; b is an integer chosen from 0, 1, 2, 3, 4, 5, and 6; and
R12 is chosen from substituted or unsubstituted heteroaiyl and substituted or unsubstituted aiyl.
77. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted heteroaryl.
78. The compound according to embodiment 76, wherein Rn is substituted heteroaiyl.
79. The compound according to embodiment 76, wherein Rn is unsubstituted heteroaryl.
80. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted heterocyclyl.
81. The compound according to embodiment 76, wherein Rn is substituted heterocyclyl.
82. The compound according to embodiment 76, wherein Rn is unsubstituted heterocyclyl.
83. The compound according to embodiment 76, wherein Rn is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl. 84. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted C1-C6 linear alkyl.
85. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted C1-C6 branched alkyl.
86. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted Cs-Ce cyclic alkyl.
87. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl.
88. The compound according to embodiment 76, wherein Rn is substituted C1-C6 branched alkyl.
89. The compound according to embodiment 76, wherein Rn is substituted C3-C6 cyclic alkyl.
90. The compound according to embodiment 76, wherein Rn is unsubstituted Ci-
Ce linear alkyl.
91. The compound according to embodiment 76, wherein Rn is unsubstituted Ci- Ce branched alkyl.
92. The compound according to embodiment 76, wherein Rn is unsubstituted C3- Ce cyclic alkyl.
93. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms.
94. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. 95. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with a heteroaryl group.
96. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a C3-C6 cyclic alkyl group.
97. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with - (CRaRb)zN(Rc)(Rd), wherein each of Ra and Rb is independently chosen from hydrogen and substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl, each of Rc and Rd is independently chosen from hydrogen, substituted or unsubstituted Ci- Cft linear, branched, or cyclic alkyl, and aryl, or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms, and z is chosen from 0, 1, 2, 3, and 4.
98. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with NCR^fR0), wherein each of Rc and Rd is independently substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms.
99. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with wherein each of Rc and Rd is independently substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl.
100. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with NfR'OCR'1), wherein each of Rc and Rd is independently unsubstituted C1-C6 linear, branched, or cyclic alkyl. 101. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with N(R°)(Rd), wherein each of Rc and Rd is independently unsubstituted Ci-C6 linear alkyl.
102. The compound according to embodiment 76, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -Ν((Τ¼)2.
103. The compound according to embodiment 76, wherein Rn is chosen from 2- thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl, cyclopropylmethyl,
104. The compound according to embodiment 76, wherein Rn is 2-thienyl.
105. The compound according to embodiment 76, wherein Rn is a trifluoroalkyl group.
106. The compound according to embodiment 76, wherein Rn is trifluoroethyl.
107. The compound according to embodiment 76, wherein Rn is trifluoropropyl.
108. The compound according to embodiment 76, wherein Rn is cyclopropyl.
109. The compound according to embodiment 76, wherein Rn is cyclopropylmethyl.
110. The compound according to embodiment 76, wherein Rn is cyclopentylmethyl. 111. The compound according to embodiment 76, wherein Rn is +
112. The compound according to embodiment 76, wherein Rn is
113. The compound according to embodiment 76, wherein Rn is
114. The compound according to embodiment 76, wherein Rn is
115. The compound according to embodiment 76, wherein
N Ί,
116. The compound according to embodiment 76, wherein Rn is
117. The compound according to embodiment 76, wherein Rn is substituted or unsubstituted heteroaryl.
118. The compound according to embodiment 76, wherein Rn is substituted heteroaryl.
119. The compound according to embodiment 76, wherein Rn is unsubstituted heteroaryl.
120. The compound according to embodiment 76, wherein Rn is pyridinyl.
121. The compound according to embodiment 76, wherein Rn is substituted or
25 unsubstituted aryl. 122. The compound according to embodiment 76, wherein Rn is substituted aryl.
123. The compound according to embodiment 76, wherein R12 is monosubstituted aryl.
124. The compound according to embodiment 76, wherein R12 is monosubstituted phenyl.
125. The compound according to embodiment 76, wherein is unsubstituted aryl.
126. The compound according to embodiment 76, wherein RM is phenyl.
127. The compound according to embodiment 76, wherein R12 is chosen from 1- naphthyl and 2-naphthyl.
128. The compound according to embodiment 76, wherein R12 is 1 -naphthyl.
129. The compound according to embodiment 76, wherein RI2 is 2-naphthyl.
130. The compound according to embodiment 76, wherein Y1-Y2 is nothing.
131. The compound according to embodiment 76, wherein Y1-Y2 is -CH2-CH2-.
132. The compound according to embodiment 76, wherein Y1-Y2 is -CH=CH
133. The compound according to embodiment 76, wherein a is an integer chosen fromO, 1, 2, and 3.
134. The compound according to embodiment 76, wherein a is an integer chosen from O, 1, and 2.
135. The compound according to embodiment 76, wherein a is an integer chosen from 0 and 1. 136. The compound according to embodiment 76, wherein a is 2.
137. The compound according to embodiment 76, wherein a is 1.
138. The compound according to embodiment 76, wherein a is 0.
139. The compound according to embodiment 76, wherein b is an integer chosen from 0, 1, 2, and 3.
140. The compound according to embodiment 76, wherein b is an integer chosen fromO, 1, and 2.
141. The compound according to embodiment 76, wherein b is an integer chosen from 0 and 1.
142. The compound according to embodiment 76, wherein b is 2.
143. The compound according to embodiment 76, wherein b is 1.
144. The compound according to embodiment 76, wherein b is 0.
145. The compound according to embodiment 76, wherein Yi-Y2 is nothing, a is 1, and b is 1.
146. The compound according to embodiment 76, wherein Yi-Y2 is nothing, a is 1, and b is 0.
147. The compound according to embodiment 76, wherein Yi-Y2 is -CH=CH-, a is
0, and b is 0.
148. The compound according to embodiment 76, with the proviso that, if Yi-Y2 is nothing, then a and b are not simultaneously 0. 149. A compound chosen from:
150. A method for inhibiting VE-PTP in a mammalian subject in need thereof, comprising administering to the subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VE-PTP activity is inhibited. 151. The method of embodiment 150, wherein the subject suffers from one or more of cancer, ocular disorders, occlusive cardiovascular disease, a vascular leaking syndrome, or another vascular-related disease.
152. A method for reducing VE-PTP-mediated signaling in a mammalian subject in need thereof, comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VEPTP-mediated signaling is reduced.
153. A method for increasing Tie2-mediated signaling in a mammalian subject in need thereof, comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of embodiments 1 to 149, or a pharmaceutically acceptable salt thereof, whereby Tie2-mediated signaling is increased.
154. A method for treating a Tie2-mediated disorder, or a VE-PTP-mediated disorder, in a mammalian subject in need thereof, comprising administering a therapeutically effective amount of a compound according to any one of embodiments 1 to 149, or pharmaceutically acceptable salt thereof, to the subject in need thereof.
155. The method of embodiment 154, wherein the subject suffers from one or more of cancer, ocular disorders, occlusive cardiovascular disease, a vascular leaking syndrome, or another vascular-related disease.
156. The method according to any one of embodiments 150 to 155, wherein the effective amount is determined in vitro by one or more biochemical and/or biological assays. Brief Description of the Drawings
FIG. 1. depicts Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
FIG. 2. depicts further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
FIG. 3. depicts still further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
FIG. 4. depicts yet further Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
FIG. 5. depicts additional Western blot results of Human Umbilical Vein Endothelial Cells (HUVEC) treated with the indicated concentrations of compounds of the disclosure that had an IC50 for VE-PTP in the activity range of 0.1 to 5 nM.
FIG. 6. depicts results of dose response titration experiments with compounds of the disclosure having an IC50 for VE-PTP in the activity range of 0.1 to 5 nM over a range of 0.2 to 50 μΜ and illustrates a clear dose response relationship. Angiopoietin 1 is the cognate ligand for Tie2 receptor, used to activate the Tie2 signaling pathway.
Detailed Description of the Disclosure
Reference is now made to certain embodiments of the disclosure. It will be understood that these references are not intended to limit the disclosure to those embodiments.
I. Definitions:
Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. Unless mentioned otherwise, the techniques employed or contemplated herein are standard methodologies well known to one of ordinary skill in the art. The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of microbiology, tissue culture, molecular biology, chemistry, biochemistry and recombinant DNA technology, which are within the skill of the art. The materials, methods and examples are illustrative only and not limiting.
In some embodiments, numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions and results, and so forth, used to describe and claim certain embodiments of the presort disclosure are to be understood as being modified in some instances by the term “about.” One of ordinary' skill in tire art would understand the meaning of the term “about” in the context of the value that it qualifies. In some embodiments, the term "about" is used to indicate that a value includes the standard deviation of the mean for tire device or method being employed to determine the value. In some embodiments, the numerical parameters set forth in the specification (into which the claims are incorporated in their entirety) are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the present disclosure are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the present disclosure may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
As used herein, the term “administering” refers to the placement of a compound and/or a pharmaceutical composition comprising the compound into a mammalian tissue or a subject by a method or route that results in at least partial localization of the compound and/or composition at a desired site or tissue location.
In this disclosure, “comprises,” “comprising,” “containing” and “having” and the like can have the meaning ascribed to them in U.S. Patent law and can mean “includes,” “including,” and the like; “consisting essentially of’ or “consists essentially” likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments. The “effectiveness” of a compound or composition of the disclosure can be assessed by any method known to one of ordinary skill in the art, including those described in the examples of this disclosure. Effectiveness can be established in vitro (biochemical and/or biological in cultured cells) and/or in vivo. Effectiveness in vitro may be used to extrapolate or predict some degree of effectiveness in vivo, in an animal or in a human subject. A reference or standard or comparison may be used. The term “effective” at inhibiting or activating an enzyme (VE-PTP and/or Tie2, respectively), and/or signaling mediated by the enzyme in the context of this disclosure and claims means reducing/activating the activity of the enzyme and/or the activation and propagation of the signaling pathway in terms of activation of a downstream molecule or known biological effect by a detectable or measurable amount relative to the baseline activity. This can be assessed in vitro or in vivo and, in some cases, extrapolated to what an activity or benefit in vivo might be by one of ordinary' skill in the art. In some embodiments, the reduction or activation is measured in terms of percentage reduction or activation, relative to the activity in the absence of exposure to the compound of the disclosure, including, for example, at least 5%, at least 10%, 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or about 100%. The activity might also fall within a range, e.g., about 5-10%, about 10-20%, and any other range interval between about 1% and 100%. An amount is “effective” in vivo if it produces any benefit to the subject to which the compound is administered.
The terms “disease” or “disorder” are used interchangeably herein and refer to any alteration in state of the body or of some of the organs, interrupting or disturbing the performance of the functions and/or causing symptoms such as discomfort, dysfunction, distress, or even death to the person afflicted or those in contact with a person. A disease or disorder can also relate to a distemper, ailing, ailment, malady, sickness, illness, complaint, indisposition, or affection.
A “VE-PTP-mediated disorder” or a “Tie-2 mediated disorder” is any disease or deleterious condition in which VE-PTP or Tie2 play a role. Non-limiting examples of such disorders are provided elsewhere in the disclosure.
The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Drug-approval agencies (e.g., EMA, US-FDA) provide guidance and approve pharmaceutically acceptable compounds, materials, compositions, and/or dosage forms. Examples are listed in Pharmacopeias.
The phrase “pharmaceutically acceptable excipient” is employed herein to refer to a pharmaceutically acceptable material chosen from a solvent, dispersion media, diluent, dispersion, suspension aid, surface active agent, isotonic agent, thickening or emulsifying agent, preservative, polymer, peptide, protein, cell, hyaluronidase, and mixtures thereof. In some embodiments, the solvent is an aqueous solvent.
As used herein, the terms "treatment," "treating," “effective,” “therapeutically effective” and the like, refer to obtaining a desired pharmacologic and/or physiologic effect. The effect can be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or can be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease.
By “subject” is meant a mammal, including, but not limited to, a human or non- human mammal, such as a bovine, equine, canine, ovine, or feline.
Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. The same rule applies for any other ranges described herein, even if the values within the range are not specifically called out in this disclosure.
The terms “alkyl,” “alkyl unit,” and “alkyl group” as used interchangeably herein refer to a saturated monovalent hydrocarbon radical comprising one to twelve carbon atoms (C1-C12). Alkyl groups may be linear, branched, or cyclic. Alkyl groups may be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkyl group comprises one to eight carbon atoms (Ci-Cg). In some embodiments, an alkyl group comprises one to six carbon atoms (C1-C6). In some embodiments, an alkyl group comprises one to four carbon atoms (Ci-C*). In some embodiments, a cyclic alkyl group comprises three to six carbon atoms (C1-C6). Non-limiting examples of substituted and unsubstituted linear, branched, or cyclic alkyl groups include methyl, ethyl, «-propyl, iso-propyl, cyclopropyl, «-butyl, sec-butyl, iso-butyl, ieri-butyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl, chloromethyl, fluoromethyl, trifluoromethyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, dimethylaminomethyl, 2-dimethylaminoethyl, 3-dimethylaminopropyl, 4- dimethylaminobutyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, trifluoroethyl, and trifluoropropyl.
The terms “alkylene,” “alkylene unit,” and “alkylene group” as used interchangeably herein refer to a saturated divalent hydrocarbon radical comprising one to twelve carbon atoms (Ci-Cu). Alkylene groups may be linear, branched, or cyclic. Alkylene groups may be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkylene group comprises one to eight carbon atoms (C1-C6). In some embodiments, an alkylene group comprises one to six carbon atoms (C1-C6). hi some embodiments, an alkylene group comprises one to four carbon atoms (C1-C4). Non-limiting examples of alkylene groups include methylene and ethylene.
The terms “alkenyl,” “alkenyl unit,” and “alkenyl group” as used interchangeably herein refer to a monovalent hydrocarbon radical comprising two to eight carbon atoms (C2- Cs) with at least one site of unsaturation (i.e., an sp2 carbon-carbon double bond). Alkenyl groups may be linear, branched, or cyclic. Alkenyl groups max' be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkenyl group comprises two to six carbon atoms (Cz-Ce). In some embodiments, an alkenyl group comprises two to four carbon atoms (C2-C4). Alkenyl groups max' have E or Z orientations. Non-limiting examples of alkenyl groups include ethenyl (also called vinyl), 1-propenyl, iso- propenyl, and 2-chloroethenyl.
The terms “alkenylene,” “alkenyl me unit,” and “alkenyl ene group” as used interchangeably herein refer to a divalent hydrocarbon radical comprising two to eight carbon atoms (C2-C8) with at least one site of unsaturation (e.g., an sp 2 carbon-carbon double bond). Alkenylene groups may be linear, branched, or cyclic. Alkenylene groups may be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkylene group comprises two to six carbon atoms (C2-Ce). In some embodiments, an alkylene group comprises two to four carbon atoms (C2-C4). alkylene groups may have E ox Z orientations. Non-limiting examples of alkenyl groups include ethenylene (also called vinylene).
The terms "alkynyl," “alkynyl unit,” and “alkynyl group” as used interchangeably herein refer to a monovalent hydrocarbon radical comprising two to eight carbon atoms (C2- Cs) with at least one site of unsaturation {i.e., an sp carbon-carbon triple bond). Alkynyl groups may be linear or branched. Alkynyl groups may be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkynyl group comprises two to six carbon atoms (Ca-Ce). In some embodiments, an alkynyl group comprises two to four carbon atoms (C2-C4)· Non-limiting examples of alkynyl groups include ethynyl.
The terms "alkynylene," “alkynylene unit,” and “alkynylene group” as used interchangeably herein refer to a divalent hydrocarbon radical comprising two to eight carbon atoms (Ca-Cg) with at least one site of unsaturation (i.e., an sp carbon-carbon triple bond). Alkynylene groups may be linear or branched. Alkynylene groups may be unsubstituted, or they may be substituted as described elsewhere herein. In some embodiments, an alkynylene group comprises two to six carbon atoms (C2-C6). In some embodiments, an alkynylene group comprises two to four carbon atoms (C2-C4). Non-limiting examples of alkynylene groups incl ude ethynylene.
The terms “aryl,” “aryl unit,” and “aryl group” as used interchangeably herein refer to a monovalent aromatic hydrocarbon radical comprising 6-20 carbon atoms (C6-C20) that is derived by removing a hydrogen atom from an aromatic ring. Aryl groups can be unsubstituted, or they can substituted with one or more substituents as described elsewhere herein. Non-limiting examples of unsubstituted and substituted aryl groups include phenyl, 2- fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-methylphenyl, 3-methylphenyl, 4- methylphenyl, 2-chlorophenyl, 3-chlorophenyl, 4-dilorophenyl, 2,6-dichlorophenyl, 3,4- difluorophenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2-methoxyphenyl, 3- methoxyphenyl, 4-methoxyphenyl, 2-phenoxyphenyl, 3-phenoxyphenyl, 4-phenoxyphenyl, 2- cyanophenyl, 3-cyanophenyl, 4-cyanophenyl, 2-dimethylaminophenyl, 3- dimethylaminophenyl, 4-dimethylaminophenyl, 3-methylsulfonylphenyl, 4- methylsulfonylphenyl, 3-aminophenyl, 3-methylaminophenyl, 3-(2-hydroxyethoxy)phenyl, 2- trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, 4-isopropylphenyl, 1 -naphthyl and 2-naphthyl.
The terms “arylene,” “arylene unit,” and “arylene group” as used interchangeably herein refer to a divalent aromatic hydrocarbon radical comprising 6-20 carbon atoms (Ce- C20) that is derived by removing two hydrogen atoms from an aromatic ring. Arylene groups can be unsubstituted, or they can be substituted with one or more substituents as described elsewhere herein. Non-limiting examples of arylene groups include phenylene.
The terms “heterocycle,” “heterocyclyl,” “heterocyclic unit,” and “heterocyclic group” as used interchangeably herein refer to a saturated or partially unsaturated ring system comprising 3 to 20 atoms, wherein at least one of the ring atoms is a heteroatom chosen from nitrogen, oxygen, phosphorous, and sulfur. A heterocyclic group may be unsubstituted or may be substituted with one or more substituents as described elsewhere herein. In some embodiments, a heterocyclic group comprises 3 to 10 atoms. In some embodiments, a heterocyclic group comprises 3 to 7 atoms. In some embodiments, a heterocyclic group is monocyclic. In some embodiments, a heterocyclic group is bicyclic. In some embodiments, a heterocyclic group comprises fused rings. Non-limiting examples of unsubstituted and substituted heterocyclic groups include pyrrolidinyl, JV-methylpyrrolidinyl, azetidinyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl, and 3- methoxypyrrolidinyl.
The terms ‘¾eteroaiyl,” “heteroaryl unit,” and “heteroaryl group” as used interchangeably herein refer to a monovalent aromatic radical comprising one or more 5-, 6-, or 7-membered rings and comprising one or more heteroatoms independently chosen from nitrogen, oxygen, phosphorous, and sulfur. A heteroaryl group may be unsubstituted or may be substituted with one or more substituents as described elsewhere herein. In some embodiments, a heteroaryl group comprises 5 to 20 atoms. In some embodiments, a heteroaryl group comprises 5 to 9 atoms. In some embodiments, a heteroaryl group comprises 5 atoms. In some embodiments, a heteroaryl group comprises 6 atoms. In some embodiments, a heteroaryl group comprises 7 atoms. In some embodiments, a heteroaryl group is monocyclic. In some embodiments, a heteroaryl group is bicyclic. In some embodiments, a heteroaryl group comprises fused rings. Non-limiting examples of heteroaryl groups include pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, 2-thienyl, 3-thienyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl- 1,2,4- oxadiazolyl, 3-phenyl-l,2,4-oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, and l//-pyrrolo[2,3-6]pyridinyl.
For the avoidance of doubt, it is meant to be understood that non-limiting examples of heteroaryl groups include:
The term “substituted” as used herein refers to the replacement of one or more hydrogen atoms on one or more of a hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaryl group with one or more substituents. On a substituted hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaryl group, any number of hydrogen atoms may be replaced by substituents. Non-limiting examples of substituents that replace a single hydrogen atom include halogen, hydroxyl, and amino. Non-limiting examples of substituents that replace two hydrogen atoms include carbonyl. Non-limiting examples of substituents that replace three hydrogen atoms include cyano.
Non-limiting examples of substituents that can substitute for hydrogen atoms on one or more of a hydrocarbon radical, alkyl group, alkylene group, alkenyl group, alkenylene group, alkynyl group, alkynylene group, aryl group, heterocyclic group, or heteroaiyl group include:
1. Ci-C6 linear, branched, or cyclic alkyl groups, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl sec-butyl, iso- butyl, tert- butyl, cyclobutyl, cyclopentyl, and cyclohexyl;
2. C2-C8 linear, branched, or cyclic alkenyl groups, non-limiting examples of which include ethenyl (also called vinyl), 1-propenyl, and iso-propenyl; 3. Cz-Cs linear or branched alkynyl groups, non-limiting examples of which include ethynyl;
4. substituted or unsubstituted aryl groups, non-limiting examples of which include phenyl, 2-fluorophenyl, 3-methylphenyl, 4-chlorophenyl, 2,6- dichlorophenyl, 3,4-difluorophenyl, 3-hydroxyphenyl, 4-cyanophenyl, 2- dimethylaminophenyl, 3-metiiylsulfonylphenyl, 4-trifluorometiiylphenyl, 3- isopropylphenyl, 1 -naphthyl, and 2-naphthyl;
5. substituted or unsubstituted heterocyclic groups, non-limiting examples of which include pyrrolidinyl, N-methylpyrrolidinyl, azetidinyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, 3-hydroxypyrrolidinyl, and 3- methoxypyrrolidinyl;
6. substituted or unsubstituted heteroaryl groups; non-limiting examples of which include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, frnyl, 2-thienyl, 3- thienyl, isoxazolyl, thiazolyl, oxadiazolyl, 3-methyl-l,2,4-oxadiazolyl, 3- phenyl-l,2,4-oxadiazolyl, indolyl, benzothiazolyl, and l//-pyrrolo[2,3- 6]pyridinyl;
7.
8.
9.
10.
11.
12.
13.
14.
15. 16. -(CRaRb),S02Rc; non-limiting examples of which include -S02H, -S02CH3, -CH2S02H, -CH2S02CH3, -S02C6H5, and -CH2S02C6H5; and
17. -(CRaRb)zS03Rc; non-limiting examples of which include -S03H, -S03CH3, -CH2S03H, -CH2S03CH3, -S03C6H5, and -CH2S03C6H5; wherein each of Ra and Rb is independently chosen from hydrogen and substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl, each of Rc and Rd is independently chosen from hydrogen, substituted or unsubstituted Ci-C6 linear, branched, or cyclic alkyl, and aryl, or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms, and z is chosen from 0, 1, 2, 3, and 4.
Compounds of the disclosure may contain one or more chiral centers. Compounds of tiie disclosure thus may exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds described herein, including, as non-limiting examples, diastereomers, enantiomers, and mixtures thereof (including, as a non-limiting example, racemic mixtures) form parts of the disclosure.
Various publications, articles and patents are cited or described in the background and throughout the specification; each of these references is herein incorporated by reference in its entirety. Discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is for the purpose of providing context for the invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any inventions disclosed or claimed.
II. Compounds
In some embodiments, a compound of the disclosure is a compound of Formula I Formula I wherein
Ri is chosen from wherein
R2 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl;
R3 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl; n is an integer chosen from 1, 2, 3, 4, 5, and 6;
R4 is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl;
Rs is chosen from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl; and each of Xi, X2, and X3 is independently chosen from CH and N, with the proviso that Xi, X2, and X3 are not simultaneously CH.
In some embodiments,
In some embodiments, In some embodiments, R2 is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R2 is substituted or unsubstituted heteroaryl. In some embodiments, R2 is substituted heteroaryl. In some embodiments, R2 is unsubstituted heteroaryl. In some embodiments, R2 is 2-thienyl. In some embodiments, R2 is 3-thienyl.
In some embodiments, R2 is not substituted or unsubstituted heteroaryl. In some embodiments, R2 is not substituted heteroaryl. In some embodiments, R2 is not unsubstituted heteroaryl. In some embodiments, R2 is not 2-thienyl. In some embodiments, R2 is not 3- thienyl.
In some embodiments, R2 is substituted or unsubstituted linear alkyl. In some embodiments, R2 is substituted or unsubstituted Ci-C6 linear alkyl. In some embodiments, R2 is unsubstituted C1-C6 linear alkyl. In some embodiments, R2 is substituted C1-C6 linear alkyl. In some embodiments, R2 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. In some embodiments, R2 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. In some embodiments, R2 is a trifluoroalkyl group. In some embodiments, R2 is trifluoroethyl.
In some embodiments, R2 is not substituted or unsubstituted linear alkyl. In some embodiments, R2 is not substituted or unsubstituted branched alkyl. In some embodiments, R2 is not substituted or unsubstituted cyclic alkyl. In some embodiments, R2 is not substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, R2 is not unsubstituted C1-C6 linear alkyl. In some embodiments, R2 is not substituted C1-C6 linear alkyl. In some embodiments, R2 is not methyl. In some embodiments, R2 is not ethyl. In some embodiments, R2 is not isopropyl. In some embodiments, R2 is not cyclopropyl.
In some embodiments, R3 is chosen from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R3 is substituted or unsubstituted heteroaryl. In some embodiments, R3 is substituted heteroaryl. In some embodiments, R3 is unsubstituted heteroaryl. In some embodiments, R3 is 2-thienyl. In some embodiments, R3 is 3-thienyl.
In some embodiments, R3 is not substituted or unsubstituted ary-l. In some embodiments, R3 is not substituted aryl. In some embodiments, R3 is not unsubstituted aryl. In some embodiments, R3 is not phenyl. In some embodiments, R3 is not substituted or unsubstituted heteroaryl. In some embodiments, R3 is not substituted heteroaryl. In some embodiments, R3 is not unsubstituted heteroaryl. In some embodiments, R3 is not 2-thienyl. In some embodiments, R3 is not 3-thienyl. In some embodiments, R3 is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, R3 is chosen from substituted or unsubstituted Ci-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl. In some embodiments, R3 is chosen from substituted C1-C6 linear alkyl, substituted C1-C6 branched alkyl, and substituted C3-C6 cyclic alkyl. In some embodiments, R3 is chosen from unsubstituted C1-C6 linear alkyl, unsubstituted C1-C6 branched alkyl, and unsubstituted C3-C6 cyclic alkyl.
In some embodiments, R3 is not substituted or unsubstituted linear alkyl. In some embodiments, R3 is not substituted or unsubstituted branched alkyl. In some embodiments, R3 is not substituted or unsubstituted cyclic alkyl. In some embodiments, R3 is not substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, R3 is not substituted C1-C6 linear alkyl. In some embodiments, R3 is not unsubstituted C1-C6 linear alkyl. In some embodiments, R3 is not methyl. In some embodiments, R3 is not ethyl. In some embodiments, R3 is not isopropyl. In some embodiments, R3 is not cyclopropyl.
In some embodiments, R3 is unsubstituted C1-C6 linear alkyl. In some embodiments, R3 is substituted C1-C6 linear alkyl. In some embodiments, R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a C5-C20 heteroaryl group. In some embodiments, R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. In some embodiments, R3 is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. In some embodiments, R3 is a trifluoroalkyl group. In some embodiments, R3 is trifluoroethyl. In some embodiments, R3 is trifluoropropyl.
In some embodiments, R3 is substituted or unsubstituted C3-C6 cyclic alkyl. In some embodiments, R3 is unsubstituted C3-C6 cyclic alkyl. In some embodiments, R3 is cyclopropyl.
In some embodiments, R3 is chosen from 2-thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl,
In some embodiments, R3 is 2-thienyl. In some embodiments, In some embodiments, R3 is
In some embodiments, R4 is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3- C6 cyclic alkyl. In some embodiments, R4 is chosen from substituted Ci-C6 linear alkyl, substituted C1-C6 branched alkyl, and substituted C3-C6 cyclic alkyl. In some embodiments, R4 is chosen from unsubstituted C1-C6 linear alkyl, unsubstituted C1-C6 branched alkyl, and unsubstituted C3-C6 cyclic alkyl. In some embodiments, R4 is unsubstituted C1-C6 linear alkyl. In some embodiments, R4 is methyl.
In some embodiments, R5 is H. hi some embodiments, R5 is chosen from substituted or unsubstituted Ci-C6 linear alkyl, substituted or unsubstituted C]-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl. In some embodiments, R5 is substituted C1-C6 linear alkyl. In some embodiments, Rs is unsubstituted C1-C6 linear alkyl. In some embodiments, R5 is methyl.
In some embodiments, R5 is not substituted C1-C6 linear alkyl. In some embodiments, R5 is not unsubstituted C1-C6 linear alkyl. In some embodiments, Rs is not methyl. In some embodiments, R$ is not H.
In some embodiments, Xi and X2 are CH and X3 is N. In some embodiments, Xi and X3 are CH and X2 is N. In some embodiments, X2 and X3 are CH and Xi is N. In some embodiments, Xi is CH and X2 and X3 are N. In some embodiments, X2 is CH and Xi and X3 are N. For the avoidance of doubt, it is meant to be understood that, in some embodiments, > > ,
In some embodiments, n is an integer chosen from 1, 2, and 3. In some embodiments, n is an integer chosen from 1 and 2. In some embodiments, n is 1.
In some embodiments, a compound of the disclosure is a compound of Formula Π
Formula II wherein
Re is substituted or unsubstituted heteroaryl, mis an integer chosen from 1, 2, 3, 4, 5, and 6;
R? is chosen from substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl;
Rg is chosen from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl; and each of Rg and Rio is independently selected from H, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl.
In some embodiments, Re is substituted heteroaryl. In some embodiments, Re is unsubstituted heteroaryl. In some embodiments, Re is 2-thienyl. In some embodiments, Re is 3-thienyl.
In some embodiments, R7 is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3- Ce cyclic alkyl. In some embodiments, R7 is substituted C1-C6 linear alkyl. In some embodiments, R7 is unsubstituted Ci-C6 linear alkyl. In some embodiments, R7 is methyl.
In some embodiments, Rg is H. In some embodiments, Rg is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl. In some embodiments, Rg is substituted C1-C6 linear alkyl. In some embodiments, Rg is unsubstituted C1-C6 linear alkyl. In some embodiments, Rg is methyl.
In some embodiments, each of Rg and Rio is independently selected from H and substituted or unsubstituted linear alkyl. In some embodiments, each of Rg and Rio is independently selected from H and substituted or unsubstituted linear C1-C6 alkyl. In some embodiments, Rg is H. In some embodiments, Rio is H. In some embodiments, each of Rg and Rio is H. In some embodiments, each of Rg and Rio is independently substituted C1-C6 linear alkyl. In some embodiments, each of Rg and Rio is independently unsubstituted Ci-C6 linear alkyl. In some embodiments, Rg is methyl. In some embodiments, Rio is methyl. In some embodiments, each ofRg and Rio is methyl.
In some embodiments, m is an integer chosen from 2, 3, 4, 5, and 6. In some embodiments, m is an integer chosen from 2, 3, 4, and 5. In some embodiments, m is an integer chosen from 2, 3, and 4. In some embodiments, m is an integer chosen from 3 and 4. In some embodiments, m is 3. In some embodiments, m is 4.
In some embodiments, a compound of the disclosure is a compound of Formula III
Formula ΠΙ wherein
Rn is chosen from substituted or unsubstituted aiyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, substituted or unsubstituted cyclic alkyl, substituted or unsubstituted linear alkenyl, substituted or unsubstituted branched alkenyl, substituted or unsubstituted cyclic alkenyl, substituted or unsubstituted linear alkynyl, and substituted or unsubstituted branched alkynyl; a is an integer chosen from 0, 1 , 2, 3, 4, 5, and 6;
Y1-Y2 is a group chosen from nothing, -CH2-CH2-, and -CH=CH-; b is an integer chosen from 0, 1, 2, 3, 4, 5, and 6; and
R12 is chosen from substituted or unsubstituted heteroaiyl and substituted or unsubstituted aryl.
In some embodiments, Rn is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted linear alkyl, substituted or unsubstituted branched alkyl, and substituted or unsubstituted cyclic alkyl. In some embodiments, Rn is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted branched C1-C6 alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl.
In some embodiments, Ru is substituted or unsubstituted heteroaryl. In some embodiments, Ru is substituted heteroaiyl. In some embodiments, Ru is unsubstituted heteroaryl. In some embodiments, Ru is 2-thienyl. In some embodiments, Ru is 3-thienyl.
In some embodiments, Ru is not substituted or unsubstituted heteroaiyl. In some embodiments, Rn is not substituted heteroaryl. In some embodiments, Ru is not unsubstituted heteroaiyl. In some embodiments, Ru is not 2-thienyl. In some embodiments, Rn is not 3-thienyl.
In some embodiments, Rn is not substituted aryl. In some embodiments, Rn is not unsubstituted aiyl. In some embodiments, Rn is not phenyl.
In some embodiments, Rn is not substituted or unsubstituted linear alkyl. In some embodiments, Rn is not substituted or unsubstituted branched alkyl. In some embodiments, Rn is not substituted or unsubstituted cyclic alkyl. In some embodiments, Rn is not substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is not substituted C1-C6 linear alkyl. In some embodiments, Rn is not unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is not methyl. In some embodiments, Rn is not ethyl. In some embodiments, Rn is not isopropyl. In some embodiments, Rn is not cyclopropyl.
In some embodiments, Rn is substituted or unsubstituted linear alkyl. In some embodiments, Rn is substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is substituted C1-C6 linear alkyl.
In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. In some embodiments, Rn is a trifluoroalkyl group. In some embodiments, Rn is trifluoroethyl. In some embodiments, Rn is trifluoropropyl.
In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a C3-C6 cyclic alkyl group. In some embodiments, Rn is substituted Ci-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with cyclopropyl group. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with cyclopentyl group. In some embodiments, Rn is cyclopropylmethyl. In some embodiments, Rn is cyclopentylmethyl. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with a heteroaryl group. In some embodiments, Rn is substituted Ci-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with an imidazolyl group.
In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -N(Rc)(Rd), wherein each of Rc and Rd are independently chosen from hydrogen, substituted or unsubstituted linear, branched, or cyclic alkyl, and aryl, or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms. In some embodiments, Rn is substituted Ci-C6 linear alkyl, wherein said substituted Ci-C6 linear alkyl is substituted with -N(Rc)(Rd), wherein each of Rc and Rd are independently chosen from H and substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -NfR^OR11), wherein each of Rc and Rd are independently chosen from substituted or unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -N(Rc)(Rd), wherein each of Rc and Rd are independently chosen from unsubstituted C1-C6 linear alkyl. In some embodiments, Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with an amino group, a dimethylamino group, a diethylaminogroup, a di-n- propylamino group, or a di-n-butylamino group. In some embodiments, Rn is a dimethylaminomethyl group.
In some embodiments, Rn is substituted or unsubstituted C3-C6 cyclic alkyl. In some embodiments, Rn is substituted C3-C6 cyclic alkyl. In some embodiments, Rn is unsubstituted C3-C6 cyclic alkyl. In some embodiments, Rn is cyclopropyl.
In some embodiments, Rn is substituted or unsubstituted heterocyclyl. In some embodiments, Rn is unsubstituted heterocyclyl. In some embodiments, Rn is substituted heterocyclyl. In some embodiments, Rn is chosen from pyrrolidinyl, 3-hydroxypyrrolidinyl, and 3-methoxypyrroIidinyl.
For the avoidance of doubt, it is to be understood that, in some embodiments, Rn is chosen from 2-thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl, cyclopropylmethyl, cyclopentylmethyl, In some embodiments, R12 is substituted or unsubstituted heteroaiyl. In some embodiments, Rn is substituted heteroaryl. In some embodiments, R12 is unsubstituted heteroaryl. In some embodiments, Rn is pyridinyl. In some embodiments, R12 is substituted or unsubstituted aryl. In some embodiments, R12 is substituted aryl. In some embodiments, Rn is monosubstituted aryl. In some embodiments, Rn is monosubstituted phenyl. In some embodiments, Rn is unsubstituted aryl. In some embodiments, Rn is phenyl. In some embodiments, Rn is chosen from 1-naphthyl and 2-naphthyl. In some embodiments, Rn is 1- naphthyl. In some embodiments, Rn is 2-naphthyl.
In some embodiments, Y1-Y2 is nothing. In some embodiments, Y1-Y2 is -CH2-CH2-. In some embodiments, Y1-Y2 is -CH=CH-.
In some embodiments, a is an integer chosen from 0, 1, 2, and 3. In some embodiments, a is an integer chosen from 0, 1, and 2. In some embodiments, a is an integer chosen from 0 and 1. In some embodiments, a is 2. In some embodiments, a is 1. In some embodiments, a is 0.
In some embodiments, b is an integer chosen from 0, 1, 2, and 3. In some embodiments, b is an integer chosen from 0, 1, and 2. In some embodiments, b is an integer chosen from 0 and 1. In some embodiments, b is 2. In some embodiments, b is 1. In some embodiments, b is 0.
In some embodiments, a compound of the disclosure is a compound of Formula ΙΠ wherein Y1-Y2 is nothing, a is 1, and b is 1. In some embodiments, a compound of the disclosure is a compound of Formula III wherein Y1-Y2 is nothing, a is 1, and b is 0. In some embodiments, a compound of the disclosure is a compound of Formula III wherein Y i-Y2 is - CH=CH-, a is 0, and b is 0. In some embodiments, a compound of the disclosure is a compound of Formula III wherein Y1-Y2 is -CH2-CH2-, a is 0, and b is 0. In some embodiments, a compound of the disclosure is a compound of Formula III wherein Y1-Y2 is nothing and a and b are not simultaneously 0. It is thus to be understood that, in some embodiments, a compound of the disclosure is a compound having the structure . It is also thus to be understood that, in some embodiments, a compound of the disclosure is a compound having the structure
In some embodiments, compounds of the disclosure are chosen from the compounds recited in Table I, below:
ΙΠ. Pharmaceutically Acceptable Salts
In some embodiments, a compound of the disclosure is in the form of a
5 pharmaceutically acceptable salt. As used herein, the term “pharmaceutically acceptable salt” refers to a salt that is pharmaceutically acceptable as defined herein and that has the desired pharmacological activity of the parent compound. Non-limiting examples of pharmaceutically acceptable salts include those derived from inorganic acids, non-limiting examples of which include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid, and those derived from organic acids, non-limiting examples of which include acetic acid, trifluoroacetic add, propionic acid, glycolic acid, pyruvic acid, oxalic acid, stearic acid, malic acid, maldc acid, malonic acid, salicylic add, succinic acid, fumaric add, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, and lactic acid.
Additional non-limiting examples of pharmaceutically acceptable salts include those formed when an acidic proton in a parent compound is replaced by a metal ion, non-limiting examples of which include an alkali metal ion and an alkaline earth metal ion, and those formed when an acidic proton present in a parent compound is replaced by a ammonium ion, a primary ammonium ion, a secondary ammonium ion, a tertiary ammonium ion, or a quaternary ammonium ion. Non-limiting examples of alkali metals and alkaline earth metals include sodium, potassium, lithium, calcium, aluminum, magnesium, copper, zinc, iron, and manganese. Additional non-limiting examples of pharmaceutically acceptable salts include those comprising one or more counterions and zwitterions.
IV. Pharmaceutical Compositions
The compounds of the disclosure can be used therapeutically as pharmaceutical compositions. The term “pharmaceutical composition” refers to a preparation that is in such form as to permit the biological activity of the active ingredient to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the composition would be administered. In some embodiments, such compositions may be sterile.
The components of the composition may be in solution, emulsions, or suspension (for example, incorporated into microparticles, nanoparticles, or liposomes). Colloidal dosage forms such as nanoparticles, nanomicelles, liposomes, and microemulsions can also be used. Typically, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render the formulation isotonic. Examples of pharmaceutically acceptable carriers include, but are not limited to, saline, Ringer’s solution and dextrose solution. The pH of the solution is preferably from about 5 to about 8, and more preferably from about 7 to about 7.5. Pharmaceutical compositions may include carriers, thickeners, diluents, buffers, preservatives, and surface-active agents. Further carriers include sustained release preparations such as semi-permeable matrices of solid hydrophobic polymers containing the compound of the disclosure, which matrices are in the form of shaped particles, e.g., films, liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of the composition being administered. Pharmaceutical compositions may also include one or more active ingredients such as antimicrobial agents, anti-inflammatory agents, and anesthetics.
Pharmaceutically acceptable carriers, excipients, vehicles, and diluents are well- known to persons having ordinary- skill in the art and are described in, as a non-limiting example, Remington: The Science and Practice of Pharmacy, 22nd Edition, Lippincott Williams & Wilkins, Philadelphia, Pa. (2013) and any other editions, which are hereby incorporated by reference.
V. Methods of Treatment and Administration
Compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments.
Said compounds, pharmaceutically acceptable salts, and/or pharmaceutical compositions can be administered in unit forms of administration to mammalian subjects, including human beings. Suitable unit forms of administration include, as non-limiting examples, forms administered orally and forms administered via a parenteral/systemic route, non-limiting examples of which including inhalation, subcutaneous administration, intramuscular administration, intravenous administration, intradermal administration, intravitreal administration, as well as topical and local ocular (i.e, subconjunctival, intravitreal, retrobulbar, intracameral) modes of administration.
In some embodiments, pharmaceutical compositions for oral administration can be in tiie form of tablets, pills, powders, hard gelatine capsules, soft gelatine capsules, and/or granules. In some embodiments of such pharmaceutical compositions, a compound of the disclosure and/or a pharmaceutically acceptable salt of a compound of the disclosure is (or are) mixed with one or more inert diluents, non-limiting examples of which including starch, cellulose, sucrose, lactose, and silica. In some embodiments, such pharmaceutical compositions may further comprise one or more substances other than diluents, such as (as non-limiting examples), lubricants, coloring agents, coatings, or varnishes. In some embodiments, pharmaceutical compositions for parenteral administration can be in the form of aqueous solutions, non-aqueous solutions, suspensions, emulsions, drops (including, as a non-limiting example, eye drops), or any combination(s) thereof. In some embodiments, such pharmaceutical compositions may comprise one or more of water, pharmaceutically acceptable glycol(s), pharmaceutically acceptable oil(s), pharmaceutically acceptable organic esters, or other pharmaceutically acceptable solvents. A variety of vehicles suitable for administering compounds to the eye are known in the art. Specific nonlimiting examples are described in U.S. Pat. No. 6,261,547; U.S. Pat. No. 6,197,934; U.S. Pat. No. 6,056,950; U.S. Pat. No. 5,800,807; U.S. Pat. No. 5,776,445; U.S. Pat. No. 5,698,219; U.S. Pat. No. 5,521,222; U.S. Pat. No. 5,403,841; U.S. Pat. No. 5,077,033; U.S.
Pat. No. 4,882,150; and U.S. Pat. No. 4,738,851, all of which are incorporate herein by reference in their entirety.
In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to inhibit VE-PTP. In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to reduce or activate VE-PTP-mediated signaling.
In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to increase Tie2-mediated signaling. In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered to a mammalian subject in need thereof to treat a Tie2-mediated disorder or a VEPTP-mediated disorder.
In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments for one or more of cancer, ocular disorders, occlusive cardiovascular disease, vascular leaking syndrome(s), and/or other vascular-related disease(s). In some embodiments, compounds of the disclosure, pharmaceutically acceptable salts of said compounds, and/or pharmaceutical compositions comprising said compounds and/or pharmaceutically acceptable salts thereof can be administered as therapeutic treatments for ocular disorders.
With regard to these therapeutic treatments, the mode (or modes) of administration, dosage (or dosages), and optimized pharmaceutical form (or forms) can be determined according to criteria generally considered during the establishment of a treatment of a patient, such as, by way of non-limiting examples, the potency of the compound(s) and/or pharmaceutically acceptable salts of the compound(s), the age of the patient, the body weight of the patient, the severity of the patient’s condition (or conditions), the patient’s tolerance to the treatment, and secondary effects observed in treatment. Determination of dosages effective to provide therapeutic benefit for specific modes and frequency of administration is within the capabilities of those skilled in the art.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 2,000 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 1,000 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 500 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 250 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 100 pg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 pg to 50 pg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 5,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 3,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 1 mg to about 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 1 mg to about 50 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 5 mg to 50 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 10 mg to 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 10 mg to 50 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 25 mg to 100 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 25 mg to 50 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 50 mg to 100 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 100 mg to 500 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is presort in a pharmaceutical composition in an amount from about 100 mg to 250 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 1,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 250 mg to 500 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 2,000 mg. In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount from about 500 mg to 1,000 mg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount of about 1 pg, about 2 pg, about 3 pg, about 4 pg, about 5 pg, about 10 pg, about 15 pg, about 20 pg, about 25 pg, about 30 pg, about 35 pg, about 40 pg, about 45 pg, about 50 pg, about 60 pg, about 70 pg, about 80 pg, about 90 pg, about 100 pg, about 125 pg, about 150 pg, about 175 pg, about 200 pg, about 225 pg, about 250 pg, about 300 pg, about 350 pg, about 400 pg, about 450 pg, about 500 pg, about 550 pg, about 600 pg, about 650 pg, about 700 pg, about 750 pg, about 800 pg, about 850 pg, about 900 pg, about 1,000 pg, about 1,100 pg, about 1,200 pg, about 1,300 pg, about 1,400 pg, about 1,500 pg, about 1,600 pg, about 1,700 pg, about 1,800 pg, about 1,900 pg, about 2,000 pg, about 2,100 pg, about 2,200 pg, about 2,300 pg, about
2,400 pg, about 2,500 pg, about 2,600 pg, about 2,700 pg, about 2,800 pg, about 2,900 pg, about 3,000 pg, about 3,100 pg, about 3,200 pg, about 3,300 pg, about 3,400 pg, about 3,500 pg, about 3,600 pg, about 3,700 pg, about 3,800 pg, about 3,900 pg, about 4,000 pg, about
4,100 pg, about 4,200 pg, about 4,300 pg, about 4,400 pg, about 4,500 pg, about 4,600 pg, about 4,700 pg, about 4,800 pg, about 4,900 pg, or about 5,000 pg.
In some embodiments, a compound of the disclosure or pharmaceutically acceptable salt thereof is present in a pharmaceutical composition in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 1,000 mg, about 1,100 mg, about 1,200 mg, about 1,300 mg, about 1,400 mg, about 1,500 mg, about 1,600 mg, about
1,700 mg, about 1,800 mg, about 1,900 mg, about 2,000 mg, about 2,100 mg, about 2,200 mg, about 2,300 mg, about 2,400 mg, about 2,500 mg, about 2,600 mg, about 2,700 mg, about 2,800 mg, about 2,900 mg, about 3,000 mg, about 3,100 mg, about 3,200 mg, about
3,300 mg, about 3,400 mg, about 3,500 mg, about 3,600 mg, about 3,700 mg, about 3,800 mg, about 3,900 mg, about 4,000 mg, about 4,100 mg, about 4,200 mg, about 4,300 mg, about 4,400 mg, about 4,500 mg, about 4,600 mg, about 4,700 mg, about 4,800 mg, about 4,900 mg, or about 5,000 mg.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mgZmL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 10 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 5 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 3 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 2 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 1 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 0.1 mg/mL to about 0.5 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 40 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 20 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 10 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 1 mg/mL to about 5 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 40 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 20 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 5 mg/mL to about 10 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 40 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 30 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 10 mg/mL to about 20 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 40 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 20 mg/mL to about 30 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 60 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 30 mg/mL to about 40 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 40 mg/mL to about 60 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 80 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 50 mg/mL to about 60 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 60 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 60 mg/mL to about 80 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 70 mg/mL to about 100 mg/mL. In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount from about 70 mg/mL to about 80 mg/mL.
In some embodiments, a pharmaceutical composition comprises a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of about 0.01 mg/mL, about 0.02 mg/mL, about 0.03 mg/mL, about 0.04 mg/mL, about 0.06 mg/mL, about 0.07 mg/mL, about 0.08 mg/mL, about 0.09 mg/mL, about 0.1 mg/mL, about 0.5 mg/mL, about 1 mg/mL, about 2 mg/mL, about 3 mg/mL, about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 40 mg/mL, about 45 mg/mL, about 50 mg/mL, about 60 mg/mL, about 70 mg/mL, about 80 mg/mL, about 90 mg/mL, or about 100 mg/mL.
Effective amounts and dosages can be estimated initially from in vitro assays. For example, an initial dosage for use in animals can be formulated to achieve a circulating blood or serum concentration of active compound that is at or above an ICjo of the particular compound as measured in an in vitro assay. Calculating dosages to achieve such circulating blood or serum concentrations taking into account the bioavailability of the particular compound is well within the capabilities of skilled artisans. For guidance, the reader is referred to Fingl & Woodbury, "General Principles," In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, latest edition, Pergamagon Press, and the references cited therein, which methods are incorporated herein by reference in their entirety. Initial dosages can also be estimated from in vivo data, such as animal models. Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases described in this disclosure are well-known in the art.
In one embodiment, dosage amounts will be in the range of from about 0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but can be higher or lower, depending upon, among other factors, the activity of the compound, its bioavailability, the mode of administration and various factors discussed above. Dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s) which are sufficient to maintain therapeutic or prophylactic effect. For example, the compounds can be administered once per week, several times per week (e.g., every other day), once per day or multiple times per day, depending upon, among other things, the mode of administration, the specific indication being treated and the judgment of the prescribing physician. In cases of local administration or selective uptake, such as local topical administration, the effective local concentration of active compound(s) may not be related to plasma concentration. Skilled artisans will be able to optimize effective local dosages without undue experimentation.
In some embodiments, a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg,
7.5 pg, 10 pg, 12.5 pg, 15 pg, 17.5 pg, 20 pg, 22.5 pg, 25 pg, 27.5 pg, or 30 pg. In some embodiments, a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg, 7.5 pg, 10 pg, 12.5 pg, 15 pg, 17.5 pg, 20 pg, 22.5 pg, 25 pg, 27.5 pg, or 30 pg once daily. In some embodiments, a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 pg, 7.5 pg, 10 pg,
12.5 pg, 15 pg, 17.5 pg, 20 pg, 22.5 pg, 25 pg, 27.5 pg, or 30 pg twice daily.
In some embodiments, a therapeutic treatment comprises administration of a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg,
7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, or 30 mg. In some embodiments, a therapeutic treatment comprises administration of a a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg, 7.5 mg, 10 mg,
12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, or 30 mg once daily. In some embodiments, a therapeutic treatment comprises administration of a a compound of the disclosure or pharmaceutically acceptable salt thereof in an amount of 5 mg, 7.5 mg, 10 mg,
12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, or 30 mg twice daily.
Other characteristics and advantages of the disclosure appear in the continuation of tiie description with the examples and the figures whose legends are represented below. The following non-limiting examples and data illustrate various aspects and features relating to the compounds and/or methods of the present invention, including the preparation of various VE-PTP inhibitor compounds, as are available through the synthetic methodologies described herein. In comparison with the prior art, in some embodiments, the present compounds and/or methods provide results and data which are surprising, unexpected and contrary thereto. While the utility of this invention is illustrated through the use of several compounds and moieties/groups which can be used therewith, it will be understood by those skilled in the art that comparable results are obtainable with various other compounds, moieties and/or groups, as are commensurate with the scope of this invention.
EXAMPLES
Examnle 1: PTP Inhibition in vitro
The potency and specificity of the compounds of the disclosure as inhibitors of three different phosphatases was determined by measuring IC$o values. The following methods, though written in present tense, were used in order to obtain the results described in this section.
Assay Principle:
Phosphatase hydrolyzes phospho-substrate (DiFMUP) to release a fluorophore (DiFMU). The fluorophore-DiFMU can be detected using FlexStation multi-mode microplate reader with excitation at 360 nm and emission at 460 nm. The fluorescence intensity represents the reaction activity.
Equipment
FlexStationMS (Molecular Devices) Bravo (Agilent)
Final assay conditions: lx reaction system: 25 mM HEPES pH 7.2, 50 mM NaCl, 5mM DTT, and 100 pg/mL
BSA
Procedure: Compound dilution:
Add 30 μΐ of compounds to undiluted well columns, and 20 pL DMSO to other column except 100% inhibition wells
Carry out 3-fold serial dilutions from lO mM/0.1 mM for 10 points by Bravo
3. Transfer 2 uL of compound/DMSO to 48 uL buffer except low control wells
4. Transfer 5 uL of cpd/DMSO solution to assay plate
5. Add 5 uL enzyme solution to assay plate, pre-incubate at 23°C for 15 min
6. Prepare vanadate:
6.1 Add 50 uL pre-activated vanadate to 140 uL 30% ¾(¾ 6.2 Add 20 uL vanadate and 40 uL DMSO to 940 uL buffer
6.3 Add 5 uL of vanadate solution to 100% inhibition wells
7. Add 10 ul of substrate solution to each well, centrifuge 1000 rpm for 10s, cover the plate, and incubate at 23°C for 120 min
8. Measure the signal on FlexStation with excitation at 360 nm and emission at 460 nm
30 Table 3 Activity Range
DEP-1 ICso (nM) 0.1 to 5 >5 to 100 >100 to 1,000
Compound No. 1, 2, 3, 17, 20, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 18, 10, 16, 19, 28, 56, 58, 61, 63, 21, 22, 23, 24, 25, 26, 27, 30, 31, 32, 29, 39, 50, 67, 65, 70, 71, 72, 33, 34, 35, 36, 37, 38, 40, 41, 42, 43, 69, 101, 105 76, 78, 81, 82, 44, 45, 46, 47, 48, 49, 51, 52, 53, 54, 83, 85, 86, 87, 55, 57, 59, 60, 62, 64, 66, 68, 73, 74, 90, 91, 92, 94, 75, 77, 79, 80, 84, 88, 89, 93, 96, 97, 95, 107, 108 98, 99, 100, 102, 103, 104, 106, 109,
110
Table 4
Activity Range
PTP1B ICso (nM) >1,000 to 100,000
Compound No. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,
42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,
99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110
Examole 2: Solubility Studies
Buffer Preparation pH 7.4 buffer: Potassium Phosphate, Monobasic 0.2 M. Dissolve 27.22 g of monobasic potassium phosphate (KH2PO4) in water, and dilute with water to 1000 mL. Place 50 mL of the monobasic potassium phosphate solution in a 200 mL volumetric flask, add 40 mL of sodium hydroxide solution, then add water to achieve desired volume and adjust pH to
7.4.
Procedure Around 1.5 mg of specific compound was weighed out into 2 vials and a volume of 150 pL of various buffers or vehicles was added for the target concentration of 10 mg/mL. All the formulations were stirred under room temperature for 24 hrs. After 24 hrs, pH was measured and appearance was observed. Supernatants were isolated by centrifugation at 14,000 rpm for 10 min. All the supernatants, diluted by 50% acetonitrile in water were measured with HPLC.
The solubility of compounds of the disclosure in pH 7.4 buffer is reported in Table 5.
Table 5
Example 3: Screening of Compounds in HUVEC
The ability of compounds of the disclosure to activate Tie2 and its downstream signaling (phosphorylation AKT kinase) in human umbilical vein endothelial cells (HUVEC) is reported below. HUVEC cells were chosen since they endogenously express Tie2/TEK receptor tyrosine kinase and its negative regulator VE-PTP (Souma, T et al. (2018 Jan) Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115). In HUVEC AKT is a verified downstream signaling effector of Tie2 receptor tyrosine kinase, whose phosphorylation increases upon binding of Angiopoietin ligand to the receptor or by inhibiting VE-PTP, the negative inhibitor of the activated receptor (Souma, T et al. (2018 Jan) Context-dependent functions of angiopoietin 2 are determined by the endothelial phosphatase VEPTP, Proc Natl Acad Sci U S A. pii: 1714446115). List of Abbreviations
BSA - Bovine Serum Albumin ECL - Enhanced Chemiluminescence EDTA - Ethylenediaminetetraacetic acid GAPDH - Glyceraldehyde 3-Phosphate Dehydrogenase HRP - Horse Horseradish Peroxidase HUVEC - Human Umbilical Vein Endothelial Cells pAKT - Phosphorylated Serine/Threonine Kinase 1 PVDF - Polyvinylidene fluoride RIP A - Radioimmunoprecipitation assay buffer SDS - Sodium dodecyl sulfate TBS - Tris Buffered Saline
1.0 Experimental procedure
1.1 Primary cell culture
Human Umbilical Vein/Vascular Endothelium cells (HUV-EC-C; ATCC; CRL-1730) were handled according to instructions provided by ATCC product sheet documentation. HUVEC ells were grown using EndoGro LS Complete Media Kit (Millipore, SCMEOOl) supplemented with Penicillin-Streptomycin cocktail (Coming, 30-002-CI) in an incubator at 37°C in 5%CC>2. Few days prior to the experiment, cells were counted using a hematocytometer and subcultured so as to reach 60-80% confluency on the day of the experiment.
1.2 Preparation of compounds
All compounds were diluted in 100% dimethyl sulfoxide (DMSO) (Amresco, N182), to a final 5 mM stock concentration.
1.3 Compound testing and samples preparation
Selected compounds were added directly to each well of a 6-well plate to achieve either 5 or 15 μΜ final concentration in 2 mL of complete HUVEC cell culture growth media. Equal volume of DMSO (Amresco, N182) was used as a vehicle control. Recombinant human Angiopoietinl protein (923-AN) was purchased from R&D Systems. Plates were gently swirled and incubated for 10 minutes at 37°C. Following the indicated treatment, the media was removed and each well was washed twice with ice-cold lx PBS (Coming, 21-031-CV). Cells were lysed using 100 pL of ice-cold RIPA buffer per well. The obtained whole cell lysate was spinned down for 5 minutes at 13,300 rpm, 4°C to remove cellular debris. The supernatant was mixed with 4x Laemmli sample buffer (Bio-Rad, #161- 0747) to final lx concentration. Subsequently samples were boiled at 95°C for 5 minutes and either used directly in polyacrylamide gel electrophoresis or stored at -20°C.
1.4 SDS polyacrylamide gel electrophoresis
Protein samples were separated on 4-12% Mini-Protean TGX Stain-Free Precast gels (Bio-Rad, #456-8086), along with Precision Plus Protein Dual Color Standards (Bio-Rad, #161-0374) using Mini-PROTEAN® Tetra Vertical Electrophoresis Cell (Bio-Rad, #165- 8005). Gels were run in lx Tris/Glycine/SDS running buffer (Bio-Rad, #161-0732) at 100 Volts for 10 minutes, followed by 25 minutes at 200 Volts.
1.5 Transfer to membrane
Transfer of proteins onto Immun-Blot® PVDF Membrane (Bio-Rad, #162-01777) was performed by either rapid wet-tank transfer for 1 hour at 100 Volts using Mini Trans- Blot Electrophoretic Transfer Cell (Bio-Rad, #170-3930), or by traditional semi-dry method for 30 minutes at 25 Volts using Trans-Blot® Turbo™ Transfer System (Bio-Rad, #170- 4150). As recommended by Bio-Rad transfer guidelines, gels and required amount of filter paper (Bio-Rad, #170-4085) were equilibrated in the Transfer Buffer for at least 15 minutes before assembling transfer sandwiches. Prior to use the PVDF membrane was pre-wetted in pure methanol for 20 seconds.
1.6 Western blotting
After transfer, the PVDF membrane was washed once in lx TBS-T, followed by blocking with 5% BSA in TBS-T for 1 hour at room temperature with gentle shaking. The blocked membrane was cut according to the visible marker lanes to probe simultaneously but separately for different proteins of interest. The resulting PVDF membrane stripes were incubated with either primary Phospho-AKT (Ser473) (D9E) XP® Rabbit mAb (Cell Signaling Technology', 4060) or with primary GAPDH (D16H11) XP® Rabbit mAb (Cell Signaling Technology, 5174) overnight, gently shaking at 4°C. Next day, membrane stripes were washed three times with lx TBS-T for 5 minutes, gently shaking at room temperature. Secondary anti-rabbit HRP-linked IgG (Cell Signaling Technology, 7074) antibody was applied for 1 hour, with gentle shaking at room temperature. Subsequently, the membrane was subjected to three 5 minute washes with shaking in lx TBS-T at room temperature. In addition, the phosphatase inhibitor cocktail was added to the wash buffer in 1:10,000 dilution during all washing steps in order to prevent the activity of residual phosphatases. For the final step, membranes were incubated with Clarity Western ECL substrate (Bio-Rad, #170-5060) and chemiluminescence signal was detected and digitally imaged using high-resolution Amersham Imager 600 (GE Healthcare).
2.0 Buffer compositions
All buffers were prepared using stock solutions directly ordered from the manufacturer.
Radio immune precipitation assay buffer (RIP A): 50 mM Tris-Cl (pH 8.0)
150 mM NaCl 1% IGEPAL CO-630 0.5 % Sodium Deoxycholate
0.1% SDS S mMEDTA lx Phosphatase inhibitor cocktail lx Protease inhibitor cocktail
Gel electrophoresis buffer: 25 mM Tris, 192 mM glycine and 0.1% SDS, pH ~8.6
Transfer buffer: 25 mM Tris, 192 mM glycine, 20% (V/V) methanol Tris buffered saline-Tween-20 (TBS-T): 50 mM Tris pH 7.4,150 mM NaCl, and 0.01% (v/v) Tween-20
Blocking buffer: 5% BSA (w/v) in lx TBS-T FIG. 1 to FIG. 6 depict Western blot analyses performed in accordance with the above- described protocol. Compounds with IC50 for VE-PTP in the activity' range of 0.1 to 5 nM were dissolved in DMSO and used to treat Human Umbilical Vein Endothelial Cells
(HUVEC).
HUVEC cells treated for 10 minutes with indicated concentration of specific compound were used in whole cell lysate Western blot analysis to determine the effectiveness of compound at inhibiting VE-PTP and activating Tie2 signaling cascade, culminating in increase in downstream phosphorylation of serine/threonine-specific protein kinase AKT/PKB. The glycolytic enzyme Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. The results depicted in FIG. 1 to FIG. 5 show two biological replicates for each compound. As depicted in the Western blot images, certain compounds are much more effective at inducing phosphorylation of AKT than others, despite being in the same ICso activity range for VE-PTP. As evidenced by tire Western blots, some compounds had tire same or slightly higher level of pAKT induction as DMSO at the concentrations tested, while others generated a more pronounced response.
FIG. 6 depicts dose response titration experiments with selected compounds over a range of 0.2 to 50 μΜ and illustrates a clear dose response relationship. Angiopoietin 1 (600 ng/mL) is the cognate ligand for Tie2 receptor, used to activate the Tie2 signaling pathway.
Example 4: Ocular Penetration
The ocular penetration study was performed to ascertain whether potent and soluble compounds 1, 2, 3 (diastereomer 1), 3 (diastereomer 2), 5, 16 and 17 administered topically as a single dose to the rat’s eye are present in the aqueous humor 30 minutes post-treatment.
All compounds delivered topically were detected in the aqueous humor post-dosing. The compounds varied in performance, with compound 16 performing the best - achieving 25.08 ng/mL in the aqueous humor. This compound also displayed the highest solubility of all the compounds tested. Finally, its IC50 was determined to be between 15 and 20 nM.
Protocol for ocular penetration and bioanalvsis
Fifteen week old brown Norway rats obtained from Charles River were used in the ocular penetration experiment. The Rats were fed ad libitum and housed in polycarbon cages with direct contact bedding, which was changed as often as necessary to keep animals dry and clean, and to minimize offensive odors. Environmental controls were set to maintain temperature, humidity and a 12-hour light-dark cycle. All animals were checked for general health by the animal facility stuff, and the study was performed in adherence to the protocol and in accordance with the Guide for the Care and Use of Laboratory Animals.
Compounds 1, 2, 3, 5, 16 and 17 were dissolved in 50 mM PBS pH 7.4 at maximum formulatable concentration, ranging from 5 to 15 mg/mL, and dose volume delivered topically as 6 uL per eye. Four rats per group were used, receiving bilateral doses to the central cornea using a calibrated micropipette, with a total of eight eyes per group collected post euthanasia. Rats were held such that the treatment remained in the eye for a period of few seconds before letting go of the animal. After receiving a single dose per eye the eyes and aqueous humour were collected 30 minutes post-dose and frozen immediately.
After treatment rats were euthanized by CO2 inhalation followed by thoracotomy performed in accordance with the accepted American Veterinary Medical Association guidelines. Immediately following confirmation of death the eye was washed with balanced salt solution. Aqueous humor was collected from both eyes of each animal, flash frozen in liquid nitrogen and stored in a -80 degree Celsius freezer. All tissues where then shipped on dry ice for bioanalysis. Mass spectrometry and chromatography methods were established to separate, detect and quantify each compound used in the study. Aqueous humor samples were analysed to detect and calculate the concentration of each analyte, which were then averaged and represented as ng/mL per group per compound.
Example 5: Synthesis of Exemplary Compounds of the Disclosure Synthesis of Compound 1
A mixture of (S)-2-amino-3-(pyridin-4-yl)propanoic add (4.50 g, 27.08 mmol, 1.00 eq) in MeOH (20.00 mL) was cooled to 0 °C, then sulfurous dichloride (86.44 g, 54.16 mmol, 3.93 mL, 2.00 eq) was added drop-wise, the mixture was then heated to 60 °C and stirred at 60 °C for 5 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield (S)-methyl 2-amino-3-(pyridin-4- yOpropanoaie (6.00 g, crude) was obtained as a white solid. LCMS: m/z = 181.1 (M+H+).
A mixture of (S)-methyl 2-amino-3-(pyridin-4-yl)propanoate (6.00 g, 23.70 mmol, 1.00 eq, 2HC1) in DCM (20.0 mL) was cooled to 0 °C, then diisopropylethylamine (12.25 g, 94.80 mmol, 16.55 mL, 4.00 eq) was added drop-wise, and then methyl carbonochloridate (2.24 g, 23.70 mmol, 1.84 mL, 1.00 eq) was added at 0 °C, the mixture was allowed to warm to 10 °C and stirred at 10 °C for 1 h. The reaction was monitored by TLC and when complete, the mixture was poured into water (50 mL) and extracted with DCM (3 x 50 mL), the organic phase was washed with brine (100 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to give the crude. The crude was purified by column chromatography (S1O2, Dichloromethane: Methanol = 10:1) to yield ( S)-methyl 2-((methoxycarbonyl)amino)-3-(pyridin-4-yl)propanoate (2.60 g, 46%) was obtained as a off-white oil.
To a solution of (S)-methyl 2-((methoxycarbonyl)amino)-3-(pyridin-4-yl)propanoate (1.30 g, 5.46 mmol, 1.00 eq) in DMF (10.00 mL) was added iodomethane (1.16 g, 8.19 mmol, 509.86 uL, 1.50 eq), the mixture was cooled to 0 °C, then sodium hydride (327.40 mg, 8.19 mmol, 60% purity, 1.50 eq) was added at 0 °C, the mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC and when complete, the solution was poured into sat. NH4CI (50 mL) and extracted with ethyl acetate (3 x 50 mL), the organic phase was washed with brine (100 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to give the crude. The crude was purified by column chromatography (S1O2, Dichloromethane: Methanol =100:1- 10:1) to yield (S)-methyl 2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-4-yl)propanoate (1.00 g, 33%) was obtained as a brown oil.
A solution of (S)-methyl 2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-4- yl)propanoate (1.00 g, 3.96 mmol, 1.00 eq) in THF (5.00 mL) and H2O (5.00 mL) was cooled to 0 °C, then lithium hydroxide hydrate (199.39 mg, 4.75 mmol, 1.20 eq) was added at 0 °C, the mixture was then allowed to warmed to 20 °C and stirred at 20 °C for 16 h The reaction was monitored by TLC and when complete, the solution was lyophilized to yield lithium ( S)-2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-4 - yl)propanoate (800.00 mg, crude) was obtained as a yellow solid. LCMS: m/z = 239.0 (M+H+).
To a solution of lithium (S)-2-((methoxycarbonyl)(methyi)amino)-3-(pyridin-4- yl)propanoate (700.00 mg, 2.87 mmol, 1.00 eq) and (S)-2-(4-nitrophenyl)~ l-(2- (thiophen-2-yI)thiazol~4-yl)ethanamine hydrobromide (591.03 mg, 1.43 mmol, 0.50 eq) in DMF (5.00 mL) was added li/-benzo|rf]|l,2,3|triazol-l-ol (387.37 mg, 2.87 mmol, 1.00 eq), then the mixture was cooled to 0 °C and diisopropylethylamine (741.02 mg, 5.73 mmol, 1.00 mL, 2.00 eq) was added drop-wise, and then JV1- ((ethylimino)methylene)-//3 r/V3-dimethylpropane- 1 ,3 -diamine hydrochloride (549.58 mg, 2.87 mmol, 1.00 eq) was added at 0 °C, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 5 h. The reaction was monitored by LCMS and TLC and when complete, the solution was poured into water (15 mL) and extracted with ethyl acetate (3 x 10 mL), the organic phase was washed with brine (20 mL), dried with anhydrous NaaSO*, filtered and concentrated in vacuo to give the crude. The crude was purified by column chromatography (S1O2, Dichloromethane: Methanol = 20:1) to yield methyl methyl((S)-l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2-yl)thiazol-4-yl)ethyl)amino)- l-oxo-3-(pyridin-4-yl)propan-2-yl)carbamate (600.00 mg, 38%) was obtained as a brown oil. LCMS: m/z = 552.2 (M+H+).
To a solution of methyl methyl((S)-l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2- yl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)carbamate (600.00 mg, 1.09 mmol, 1.00 eg) in EtOH (6.00 mL) and H2O (2 mL) was added NH4CI (290.90 mg, 5.44 mmol, 5.00 eg), the mixture was heated to 80 °C and Fe (303.73 mg, 5.44 mmol, 5.00 eq) was added at 80 °C, the mixture was stirred at 80 °C for 3 h. The reaction was monitored by TLC and when complete, the solution was filtered and the filtrate was concentrated in vacuo to give the crude. The crude was dissolved into water (10 mL) and extracted with DCM (3 x 10 mL), the organic phase was washed with brine (20 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield methyl ((S)- l-(((S)-2-(4-aminophenyl)-l-(2-(thiophen-2-yl)thiazol-4-yl)ethyl)amino)-l-oxo-3- (pyridin-4-yl)propan-2-yl)(methyl)carbamate (400.00 mg, crude) was obtained as a white solid. LCMS: m/z = 521.9 (M+H+)
A solution of methyl ((S)-l-(((S)-2-(4-aminophenyl)-l-(2-(thiophen-2-yl)thiazol-4- yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)caTbamate (400.00 mg, 766.80 umol, 1.00 eq) in pyridine (2.00 mL) and MeCN (2.00 mL) was cooled to 0 °C, then sulfur trioxide pyridine complex (366.13 mg, 2.30 mmol, 3.00 eq) was added in batches at 0 °C, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (5 mL) was added into the solution and the solution was stirred at 0 °C for 20 min, then the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini C18250*50mm*10 um;mobile phase: [water(10mM NH4HCO3)- ACN];B%: 20%-50%,23min) to yield ammonium (4-((S)-2-((S)-2-
((methoxycarbonyl)(methyi)amino)-3-(pyridin-4-yl)propanamido)-2-(2-(thiophen-2- yl)thiazol-4-yl)ethyi)phenyl)sulfamate (94.97 mg, 21%) was obtained as a white solid. LCMS: m/z = 602.1 (M+H+), 1H-NMR: (400 MHz, DMSO-i¾) δ = 8.33 - 8.49 (m, 3 H) 7.68 - 7.72 (m, 1 H) 7.60 - 7.64 (m, 1 H) 7.17 - 7.29 (m, 4 H) 7.1 - 7.17 (m, 1 H) 7.02 - 7.10 (m, 1 H) 6.93 - 6.99 (m, 1 H) 6.83 - 6.92 (m, 4 H) 5.05 - 5.20 (m, 1 H) 4.78 - 5.04 (m, 1 H) 3.34 - 3.55 (m, 3 H) 3.02 - 3.19 (m, 2H) 2.80 - 2.98 (m, 2 H) 2.57 (s, 3
H).
Synthesis of Compound 2 A solution of Methyl 2-acetamidomaIonate (14.00 g, 64.45 mmol, 1.00 eg) in DMF (50 mL) was cooled to 0 °C, then sodium hydride (7.73 g, 193.35 mmol, 60% purity, 3.00 eg) in DMF (150.00 mL) and 3-(bromomethyl)pyridine (16.30 g, 64.45 mmol, 1.00 eg) was added, the mixture was stirred at 0 °C for 2 h. The reaction was monitored by TLC and when complete, the solution was poured into water (400 mL) and extracted with ethyl acetate (3 x 400 mL), the organic phase was washed with H2O (3 x 400 mL), the combined organic phase was washed with brine (1000 mL), dried with anhydrous NaaSO-t, filtered and concentrated in vacuo to yield diethyl 2-acetamido-2-(pyridin-3- ylmethyl)malonate (11.86 g, crude) was obtained as a yellow solid.
A solution of diethyl 2-acetamido-2-(pyridin-3-ylmethyl)malonate (11.86 g, 38.47 mmol, 1.00 eg) in EtOH (230.82 mL) was cooled to 0 °C, then sodium hydroxide (6 M, 16.03 mL, 2.50 eg) was added, the mixture was then allowed to warm to 15 °C and stirred at 15 °C for 1.5 h. The reaction was monitored by TLC and when complete, N2 was bubbled into the solution to remove the solvent and the crude was dissolved into water (60 mL) and the solution was acidified by aq. HC1 (10 M, 9.62 mL). The solution was concentrated in vacuo to yield 2-acetamido-2-(pyridin-3-ylmethyl)malonlc add (11.86 g, crude) was obtained as a brown solid.
A solution of 2-acetamido-2-(pyridin-3-ylmethyl)malonic acid (11.00 g, 38.10 mmol, 1.00 eg) in H2O (120.00 mL) was heated to 110 °C and stirred at 110 °C for 15 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield 2-acetamido-3-(pyrldin-3-yl)propanoic acid (9.00 g, crude) was obtained as a brown solid. LCMS: m/z = 209.0 (M+H+).
A solution of 2-acetamido-3-(pyridin-3-yl)propanolc acid (9.00 g, 36.78 mmol, 1.00 eg) in hydrogen chloride (5 M, 140.00 mL) was heated to 100 °C and stirred at 100 °C for 4 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield 2-amino-3-(pyridin-3-yl)propanoic add (6.50 g, 87%) was obtained as a brown solid. LCMS: m/z = 167.0 (M+H+). A solution of 2-amino-3-(pyridin-3-yl)propanoic acid (6.20 g, 37.31 mmol, 1.00 eq) in MeOH (100.00 mL) was cooled to 0 °C, then sulfurous dichloride (13.32 g, 111.93 mmol, 8.12 mL, 3.00 eq) was added at 0 °C, the mixture was then heated to 60 °C and stirred at 60 °C for 10 h. The reaction was monitored by LCMS and when complete, tiie solution was concentrated in vacuo to yield methyl 2-amino-3-(pyridin-3- yOpropanoate (6.00 g, crude) was obtained as a brown solid. LCMS: m/z = 181.1 (M+H+).
A solution of methyl 2-amino-3-(pyruIin-3-yl)propanoate (6.00 g, 27.69 mmol, 1.00 eq) in DCM (150.00 mL) was cooled to 0 °C, then /V-ethyl-/V-isopropylpropan-2-amine (14.32 g, 110.77 mmol, 19.29 mL, 4.00 eq) was added, and then methyl carbonochloridate (2.62 g, 27.69 mmol, 2.14 mL, 1 eq) was added drop-wise, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 1 h. The reaction was monitored by LCMS and TLC and when complete, the solution was poured into water (100 mL) and extracted with DCM (3 x 100 mL), the combined organic layer was washed with brine (10 mL), dried with NazSO*, filtered and concentrated in vacuo to give the crude. The crude was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 1:1) to yield methyl 2-((methaxycarbonyl)amino)-3-(pyridin-3- yOpropanoate (2.90 g, 40%) was obtained as a brown oil. LCMS: m/z = 239.0 (M+H+).
To a solution of methyl 2-((methoxycarbonyl)amino)-3-(pyridin-3-yl)propanoate (2.90 g, 12.17 mmol, 1.00 eq) in DMF (20.00 mL) was added iodomethane (2.59 g, 18.26 mmol, 1.14 mL, 1.50 eq), then the mixture was cooled to 0 °C, and then sodium hydride (730.36 mg, 18.26 mmol, 60% purity, 1.50 eq) was added at 0 °C, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was poured into sat. NH*C1 (60 mL) and extracted with ethyl acetate (3 x 60 mL), the combined organic layer was washed with brine (100 mL), dried with NazSO*, filtered and concentrated in vacuo to yield methyl 2-((methaxycarbonyl)(methyl)amino)-3-(pyridin-3-yl)propanoate (1.68 g, 55%) was obtained as a brown oil. LCMS: m/z = 253.0 (M+H+).
A solution of methyl 2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-3- yl)propanoate (1.68 g, 6.66 mmol, 1.00 eq) in THF (10.00 mL) and H2O (3.00 mL) was cooled to 0 °C, then L1OH.H2O (335.33 mg, 7.99 mmol, 1.20 eq) was added at 0°C, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 16 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield lithium 2-((methoxycarbonyty(methyl)amino)-3-(pyridin-3-yfypropanoate (1.20 g, crude) was obtained as a brown solid. LCMS: m/z = 239.0 (M+H+).
To a solution of lithium 2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-3- yOpropanoate (1.20 g, 4.91 mmol, 1 eq) and (S)-2-(4-nitrophenyl)-l-(2-(thiophen-2- yl)thiazol-4-yl)ethanamine hydrobromide (1.01 g, 2.46 mmol, 0.5 eq) in DMF (10.0 mL) was added l//-benzo[i/][l,2,3]triazol-l-ol (664.06 mg, 4.91 mmol, 1.00 eq), the mixture was cooled to 0 °C, then /V-ethyl-N-isopropylpropan-2-amine (1.27 g, 9.83 mmol, 1.71 mL, 2.00 eq) and Arl-((ethylimino)methylene)-iV3,/V3-dimethylpropane- 1 ,3- diamine hydrochloride (942.12 mg, 4.91 mmol, 1.00 eq) was added at 0 °C, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 5 h. The reaction was monitored by LCMS and TLC and when complete, the solution was poured into water (50 mL) and extracted with ethyl acetate (3 x 50 mL), the combined organic layer was washed with brine (80 mL), dried with NazSO*, filtered and concentrated in vacuo to give the crude. The crude was purified by column chromatography (S1O2, Dichloromethane: Methanol = 20:1) to yield methyl methy!(l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2- yl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-3-yl)propan-2-yl)carbamate (1.00 g, 37%) was obtained as a brown oil. LCMS: m/z = 552.0 (M+H+).
To a solution of methyl methyl(l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2-yl)thiazol- 4-yl)ethyl)amino)-l-oxi>-3-(pyridin-3-yl)propan-2-yl)carbamate (l.OO g, 1.81 mmol, 1 eq) in EtOH (9.0 mL) and H2O (3.0 mL) was added NH4CI (484.83 mg, 9.06 mmol, 5 eq), then the mixture was heated to 80 °C, and Fe (506.22 mg, 9.06 mmol, 5 eq) was added, the mixture was stirred at 80 °C for 3 h. The reaction was monitored by LCMS and TLC and when complete, the solution was filtered and the filtrate was concentrated in vacuo to give the crude. The crude was purified by prep-TLC (Dichloromethane: Methanol = 15:1) and by prep_HPLC (column: DuraShell 150*25mm*5um;mobile phase: [water(0.1%TFA)-ACN];B%: l%-40%,8min) to yield 14_diastereomer 1 (0.12 g, 13%) and 14_diastereomer 2 (0.08 g, 8%) was obtained as a brown oil. LCMS: m/z = 522.1 (M+H+).
Diastereoisomer 1:
A solution of 14_diastereomer 1 (0.30 g, 575.09 umol, 1.00 eq) in pyridine (2.0 mL) and CH3CN (2.0 mL) was cooled to 0 °C, then sulfur trioxide pyridine complex (274.60 mg, 1.73 mmol, 3.00 eq) was added at 0 °C, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (6 mL) was added drop-wise, then the mixture was dried by blowing N2 to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 12%-42%,20min) to yield ammonium (4-((2S)-2-(2-methoxycarbonyl)(methyl)amino)-3-(pyridin-3- yl)propanamido)-2-(2-(thiophen-2-yl)thiazol-4-yl)ethyl)phenyl)sulfamate diastereomer 1 (165.20 mg, 45%) was obtained as a white solid. LCMS: m/z = 602.0 (M+Hf), 'H-NMR: (400 MHz, DMSO-<¾) δ = 8.37 - 8.58 (m, 3 H) 7.80 - 7.90 (m, 1 H) 7.68 - 7.77 (m, 1 H) 7.60 - 7.67 (m, 1 H) 7.4 - 7.50 (m, 1 H) 7.24 - 7.29 (m, 1 H) 7.22 (s, 1 H) 7.17 (dd, .7=4.89, 3.79 Hz, 1 H) 7.09 (s, 1 H) 6.96 (s, 1 H) 6.91 (s, 3 H) 5.07 - 5.25 (m, 1 H) 4.75 - 5.03 (m, 1 H) 3.50 (s, 2 H) 3.34 (s, 1 H) 3.14 - 3.22 (m, 1 H) 3.05 - 3.13 (m, 1 H) 2.86 - 2.99 (m, 2 H) 2.57 (s, 3 H).
Diastereoisomer 2:
A solution of 14_diasteretmer 2 (240.00 mg, 460.08 umol, 1.00 eq) in pyridine (2.0 mL) and MeCN (2.0 mL) was cooled to 0 °C, then sulfur trioxide pyridine complex (219.68 mg, 1.38 mmol, 3.00 eq) was added at 0°C, the mixture was stirred at 0 °C for 5 min. 7% ammonium hydroxide (4 mL) was added drop-wise, then the mixture was dried by blowing N2 to give a residue. The crude was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water(10mM NH4HCO3)- ACN];B%: 12%-42%,20min) to yield ammonium (4-((2S)-2-(2- methoxycarbonyl)(methyl)amino)-3-(pyridin-3-yl)propanamido)-2-(2-(thiophen-2- yl)thiazol-4-yl)ethyl)phenyl)sulfamate diastereomer 2 (101.50 mg, 34%) was obtained as a white solid. LCMS: m/z = 602.0 (M+H+), 1H-NMR: (400 MHz, Methanol^) δ = 8.50 (s, 1 H) 8.41 (s, 1 H) 8.30 - 8.38 (m, 1 H) 7.80 - 8.00 (m, 1 H) 7.57 - 7.61 (m, 1 H) 7.53 - 7.57 (m, 2 H) 7.16 (s, 1 H) 7.13 (dd, .7=5.01, 3.79 Hz, 1 H) 6.99 - 7.07 (m, 4 H) 5.23 - 5.37 (m, 1 H) 4.70 - 4.83 (m, 1 H) 3.45 - 3.66 (m, 3 H) 3.15 - 3.28 (m, 2 H) 2.86 - 3.03 (m, 2 H) 2.78 (s, 3 H).
Synthesis of Compound 3
A solution of diethyl 2-acetamidopropanedioate (16.08 g, 74.02 mmol, 1.04 eq) in DMF (4.0 mL) was cooled to 0 °C, then sodium hydride (8.54 g, 213.51 mmol, 60% purity, 3.00 eq) in DMF (160 mL) and 2-(bromomethyl)pyridine (18.00 g, 71.17 mmol, 1.00 eq, HBr) was added, the mixture was stirred at 0 °C for 2 h. The reaction was monitored by TLC and when complete, the solution was poured into sat. NH*C1 (500 mL) and extracted with ethyl acetate (3 x 500 mL), the combined organic phase was washed with brine (1000 mL), dried with anhydrous NazSO*, filtered and concentrated in vacuo. The residue was washed with petroleum ether (150 mL) and filtered the filter cake was dried in vacuo to yield diethyl 2-acetamido-2-(pyridin-2-ylmethyl)malonate (12.95 g, crude) was obtained as a yellow solid.
A solution of diethyl 2-acetamido-2-(pyridin-2-ylmethyl)malonate (12.95 g, 42.00 mmol, 1.00 eq) in EtOH (150 mL) was cooled to 0 °C, then sodium hydroxide (6 M, 17.50 mL, 2.50 eq) was added, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 1.5 h. The reaction was monitored by LCMS and when complete, the solution was dried by blowing Nz. The residue was dissolved into water (80 mL) and acidified with aq. hydrogen chloride (10 M, 10.5 mL). The solution was concentrated in vacuo to yield 2-acetamido-2-(pyridin-2-ylmethyl)malonic acid (12.00 g, crude) was obtained as a brown solid.
A solution of 2-acetamido-2-(pyridin-2-yhnethyl)malonic add (12.00 g, 41.57 mmol, 1.00 eq, HC1) in HzO (120 mL) was heated to 100 °C and stirred at 100 °C for 15 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield 2-acetamido-3-(pyridin-2-yl)propanoic add (9.00 g, crude) was obtained as a brown solid. LCMS: m/z = 209.0 (M+H4).
A solution of 2-acetamido-3-(pyridin-2-yl)propanoic add (9.00 g, 36.78 mmol, 1.00 eq, HC1) in hydrogen chloride (5 M, 140.00 mL) was heated to 100 °C and stirred at 100 °C for 4 h. The reaction was monitored by TLC and when complete, the solution was concentrated in vacuo to yield 2-amino-3-(pyridin-2-yl)propanoic add (7.00 g, crude) was obtained as a brown solid. Asolution of2-amino-3-(pyridin-2-yl)propanoicadd (6.8 g, 33.56 mmol, 1.00 eq, HC1) in MeOH (5.00 mL) was cooled to 0 °C, then thionyl chloride (7.98 g, 67.11 mmol, 4.87 mL, 2.00 eq) was added, the mixture was then heated to 60 °C and stirred at 60 °C for 10 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield methyl 2-amino-3-(pyridin-2-yl)propanoate (7 g, crude) was obtained as a yellow solid. LCMS: m/z = 180.9 (M+H+).
A solution of methyl 2-amino-3-(pyridin-2-yl)propanoate (7 g, 32.31 mmol, 1.00 eq, HC1) in DCM (20 mL) was cooled to 0 °C, then diisopropylethylamine (16.70 g, 129.23 mmol, 22.57 mL, 4.00 eq) and methyl carbonochloridate (3.05 g, 32.31 mmol, 2.50 mL, 1.00 eq) was added drop-wise at 0 °C, the mixture was allowed to warm to 15 °C and stirred at 15 °C for 1 h. The reaction was monitored by TLC and when complete, the solution was poured into water (50 mL) and extracted with DCM (3 x 50 mL), the combined organic phase was washed with brine (100 mL), dried with anhydrous NazSO*, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (SiCh, Dichloromethane: Methanol =100:1 to 20:1) to yield methyl 2-((methoxycarbonyl)amino)-3-(pyridin-2-yl)propanoate (3.5 g, 44%) was obtained as a brown solid.
To a solution of methyl 2-((methoxycarbonyl)amino)-3-(pyridin-2-yl)propanoate (3.5 g, 14.69 mmol, 1.00 eq) in DMF (20 mL) was added iodomethane (3.13 g, 22.04 mmol, 1.37 mL, 1.50 eq), then the mixture was cooled to 0 °C, and sodium hydride (881.47 mg, 22.04 mmol, 60% purity, 1.50 eq) was added, tire mixture was then allowed to warm to 15 °C and stirred at 15 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was poured into sat. NH4CI (50 mL) and extracted with ethyl acetate (3 x 50 mL), the combined organic phase was washed with brine (100 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield methyl 2- ((methoxycarbonyl)(methyl)amino)-3-(pyridin-2-yl)propanoate (2.5 g, crude) was obtained as a brown solid. LCMS: m/z = 253.2 (M+H+). A solution of methyl 2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-2- yl)propanoate (2.5 g, 9.91 mmol, 1.00 eq) in THF (10.0 mL) and ¾€) (10.0 mL) was cooledto 0 °C, then lithium hydroxide hydrate (499.00 mg, 11.89 mmol, 1.20 eq) was added at 0 °C, the mixture was then allowed to warm to 15 °C and stirred at 15 °C for 16 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield lithium 2-( (methoxycarbonyl)(methyl)amino)-3- (pyridin-2-y0propanoate (1.8 g, crude) was obtained as a brown solid. LCMS: m/z = 239.2 (M+H+).
To a solution of lithium 2-((methaxycarbonyl)(methyl)amino)-3-(pyridin-2- yOpropanoate (1.8 g, 7.37 mmol, 1.00 eq) and (S)-2-(4-nitrophenyl)-l-(2-(thiophen- 2-yl)thiazol-4-yl)etkanamine hydrobromide (1.52 g, 3.69 mmol, 0.50 eq) in DMF (20 mL) was added 17/-benzo[i/][l,2,3]triazol-l-ol (996.08 mg, 7.37 mmol, 1.00 eq), then the mixture was cooled to 0 °C and diisopropylethylamine (1.91 g, 14.74 mmol, 2.57 mL, 2.00 eq) and jV1-((ethylimino)methylene) dimethylpropane-l, 3-diamine hydrochloride (1.41 g, 7.37 mmol, 1.00 eq) was added drop-wise at 0 °C, the mixture was then allowed to warm to 15 °C and stirred at 15 °C for 5 h. The reaction was monitored by LCMS and TLC and when complete, the solution was poured into water (60 mL) and extracted with ethyl acetate (3 x 40 mL), the organic phase was washed with water (2 x 40 mL), tire combined organic phase was washed with brine (100 mL), dried with anhydrous NaaSO*, filtered and concentrated in vacuo. The residue was purified by column chromatohraphy (Si02, Dichloromethane: Methanol = 100:1 to 20:1) to yield methyl methyl(l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2-yl)thiazol-4- yl)ethyl)amino)-l-oxo-3-(pyridin-2-yl)propan-2-yl)carbamate (1.40 g, crude) was obtained as a brown oil. LCMS: m/z = 552.1 (M+H+).
To a solution of methyl methyl(l-(((S)-2-(4-nitrophenyl)-l-(2-(thiophen-2-yl)thiazol- 4-yl)ethyl)amino)-l-oxo-3-(pyridin-2-yl)propan-2-yl)carbamate (1.40 g, 2.54 mmol, 1 eq) in EtOH (10.0 mL) and H2O (3.0 mL) was added NH4CI (135.75 mg, 2.54 mmol, 1 eq), then the mixture was heated to 80 °C, and then Fe (141.74 mg, 2.54 mmol, 1 eq) was added, the mixture was stirred at 80 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was filtered and the filtrate was concentrated in vacuo. The residue was dissolved into DCM (10 mL) and filtered, the filtrate was concentrated in vacuo. The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 0:1) and by prep-HPLC (column: Luna C18 100*305u; mobile phase: [water (0.1 %TFA)- ACN] ;B% : 10%-30%,10min) to yield lOJOasteremsomer 1 (0.15 g, 18%) was obtained as a brown oil LCMS: m/z = 522.3 (M+H4) and 1 O diastereoisomer 2 (0.1 g, 18%) which was obtained as a brown oil. LCMS: m/z = 522.3 (M+H+).
Diastereoisomer 1:
A solution of JO_diastereoisomer 1 (200.00 mg, 383.40 umol, 1.00 eq) in pyridine (2.0 mL) and MeCN (2.0 mL) was cooled to 0 °C, and then sulfiir trioxide pyridine complex (183.06 mg, 1.15 mmol, 3.00 eq) was added in batches, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (4 mL) was added slowly, then the mixture was dried by blowing Nz. The residue was purified by prep_HPLC (column: Phenomenex Gemini Cl 8 250*50 10u;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 15%-45%,20min) to yield ammonium (4-((2S)-2-(2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-2- yl)prop<mamido)-2-(2-(thiophen-2-yl)thiazol-4-y[)ethyl)phenyl)sulfamate diastereoisomer 1 (0.074 g, 31%) was obtained as a white solid. LCMS: m/z = 602.1 (M+H+), Ή-ΝΜΚ: (400 MHz, METHANOL-i¾) δ = 8.46 (br d, .7=4.85 Hz, 1 H) 7.86 (br s, 1 H) 7.57 (dd, J= 11.14, 4.30 Hz, 2 H) 7.24 - 7.48 (m, 2 H) 6.98 - 7.17 (m, 6 H) 5.32 (br dd, J= 9.37, 5.84 Hz, 1 H) 4.98 - 5.20 (m, 1 H) 3.45 - 3.69 (m, 3 H) 3.39 (br d, .7=13.89 Hz, 1 H) 3.24 (br d, J= 9.26 Hz, 1 H) 2.95 - 3.15 (m, 2 H) 2.54 - 2.74 (m, 3 H).
Diastereoisomer 2:
A solution of lOjMastereoisomer 2 (180.00 mg, 345.06 umol, 1.00 eq) in pyridine (2.00 mL) and CHBCN (2.00 mL) was cooled to 0 °C, then sulfiir trioxide pyridine complex (164.76 mg, 1.04 mmol, 3.00 eq) was added in batches, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (4 mL) was added slowly, then the mixture was dried by blowing Nz to to give a residue. The residue was purified by prep_HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 12%- 42%,20min) to yield ammonium (4-((2S)-2-(2-((methoxycarbonyl)(methyl)amino)-3- (pyridin-2-yl)propanamido)-2-(2-(thiophen-2-yl)thiazol-4-yl)ethyl)phenyl)sulfamate (70.70 mg, 34%) was obtained as a white solid. LCMS: m/z = 602.1 (M+H+), 1H-NMR: (400 MHz, METHANOLS) δ = 8.52 (d, J= 4.77 Hz, 1 H) 7.93 (br s, 1 H) 7.54 - 7.64 (m, 2 H) 7.41 (br s, 2 H) 7.15 (dd, J=5.01, 3.79 Hz, 2 H) 6.97 - 7.11 (m, 4 H) 5.32 (dd, J=8.93, 6.11 Hz, 1 H) 5.05 (br s, 1 H) 3.45 - 3.68 (m, 3 H) 3.37 (br s, 1 H) 3.17 - 3.29 (m, 1 H) 2.93 - 3.16 (m, 2 H) 2.80 (s, 3 H).
Carbonyl di-imidazole (20.64 g, 127.29 mmol, 1 eq) in DCM (50 mL) was poured into 3,3,3-trifluoropropanoic acid (16.3 g, 127.29 mmol, 11.24 mL, 1 eq) in DCM (60 mL) at 25 °C, then the mixture was heated to 40 °C and refluxed at 40 °C for 0.5 h. Then the mixture was cooled to 25 °C and ammonia (2.17 g, 127.29 mmol, 2.13 mL, 1 eq) was added and stirred at 25 °C for another 0.5 h. The reaction was monitored by LCMS and when complete, the reaction mixture was poured into aq. HC1 (2N, 100 mL) and extracted with ethyl acetate (2 x 60 mL). The combined organic phase was washed with brine (100 mL), dried with anhydrous NazSO^ filtered and concentrated in vacuo to yield 3,3,3-trifluoropropanamide (10 g, crude) was obtain as a white solid. LCMS: m/z = 128.0 (M+H+), Ή-ΝΜΚ: (400 MHz, DMSO^e) δ = 7.63 (s, 1 H) 7.27 (s, 1 H) 3.19 (q, 7=11.40 Hz, 2 H).
A mixture of 3,3,3-trifluoropropanamide (5 g, 39.35 mmol, 1 eq), Lawesson's reagent (19.10 g, 47.22 mmol, 1.2 eq) and THF (50 mL) was degassed and purged with Na for 3 times, and then the mixture was stirred at 25 °C for 13 h under Na atmosphere. The reaction was monitored by TLC and when complete, the reaction mixture was diluted with saturated NaHCCh (150 mL) and extracted with ethyl acetate (3 x 60 mL). The combined organic phase was washed with brine (150 mL), dried with anhydrous NaaSO*, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiOa, Petroleum ether/Ethyl acetate = 20:1 to 5:1) to yield 3,3,3- trifluoropropanethioamide (2.7 g, 48%) was obtained as a yellow solid. ¾ NMR: (400 MHz, CDCb) δ = 7.97 (hr s, 1 H) 7.27 (hr s, 1 H) 3.60 (q, 7=10.36 Hz, 2 H).
A mixture of (S)-tert-buiyl (4-bromo-l-(4-nitropheny[)-3-axobutan-2-yl)carbamate (2 g, 5.16 mmol, 1 eq) and 3,3,3-trifluoropropanethioamide (739.26 mg, 5.16 mmol, 1 eq) in MeCN (10 mL) was heated to 80 °C and stirred at 80 °C for 10 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to give the crude. The crude was washed with Petroleum ether: Ethyl acetate (1:1, 5 mL) and filtrated the solid to yield (S)-2-(4-nitrophenyl)-l-(2-(2,2,2-trifluoroethyl)thiazol- 4-yl)ethanamine hydrobromide (1.6 g, crude) was obtained as a brown solid. LCMS: m/z = 332.0 (M+H+).
To a solution of ( S)-2-(4-nitrophenyl)-l-(2-(2,2,2-trifluoroethyl)thiazol-4 - yl)ethanamine hydrobromide (1.95 g, 4.73 mmol, 1 eq) and (S)-2- ((methoxycarbonylXmethyl)amino)-3-(pyridin-4-yl)propanoic acid (1.88 g, 4.73 mmol, 1 eg) in DMF (30 mL) was added triethylamine (2.87 g, 28.38 mmol, 3.95 mL, 6 eg), then the mixture was cooled to 0 °C, and 2,4,6-tripropyl-l,3,5,2,4,6- trioxatriphosphinane 2,4,6-trioxide (6.02 g, 9.46 mmol, 5.63 mL, 50% purity, 2 eg) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The reaction was monitored by LCMS and TLC and when complete, the reaction mixture was poured into ice/water (150 mL) slowly, and extracted with ethyl acetate (3 x 50 mL), the organic phase was washed with brine (150 mL), dried with anhydrous NazSO*, concentrated in vacuo to give a residue. The residue was purified by prep-TLC (Petroleum ether/Ethyl acetate=l :2) to yield methyl metkyl( (S)-l-(((S)-2-(4- nitrophenyl)-l-(2-(2,2,2-trifluoroethyI)thiazol-4-yl)ethyl)amino)-]-oxo-3-(pyridin-4- yl)propan-2-yl)carbamate (1.5 g, 57%) was obtained as a yellow oil. LCMS: m/z = 552.2 (M+H+).
To a solution of methyl methyl((S)-l-(((S)-2-(4-nitrophenyl)-l-(2-(2,2,2- trifluoroethyl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2- yl)carbamate (1.4 g, 2.54 mmol, 1 eg) in EtOH (12 mL) and H2O (4 mL) was added NH4CI (678.88 mg, 12.69 mmol, 5 eg), then the mixture was heated to 90 °C and then Fe (708.84 mg, 12.69 mmol, 5 eg) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 250*50mm*10 um;mobile phase: [water(0.1%TFA)-ACN];B%: l%-25%,20min) to yield methyl ((S)-l-(((S)-2- (4-aminophenyl)-l-(2-(2,2,2-trifluoroethyl)thiazol-4-yl)ethyl)amino)-l-oxo-3- (pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.5 g, 38%) was obtained as a yellow solid. LCMS: m/z = 522.2 (M+H4).
A mixture of methyl ((S)-l-(((S)-2-(4-aminophenyl)-l-(2-(2,2,2- trifluoroethyl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2- yl)(methyl)carbamate (0.3 g, 575.20 umol, 1 eg) in MeCN (0.5 mL) and pyridine (0.5 mL) was cooled to 0 °C, then SOs-pyridine (274.65 mg, 1.73 mmol, 3 eq) was added in batches. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, then the mixture was dried by blowing N2 to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN];B%: 5%-30%,20min) to yield ammonium (4- ((S)-2-((S)-2-((methoxycarbonyI)(methyl)amino)-3-(pyridin-4-yt)propanamido)-2- (2-(2,2,2-trifluoroethyI)thiazol-4-yl)ethyl)phenyl)sulfamate (75.57 mg, 21%) was obtained as a white solid. LCMS: m/z = 602.1 (M+Hf), Ή-NMR: (400 MHz, D20) δ = 8.29 (d, .7=5.87 Hz, 2 H) 7.21 (s, 2 H) 6.97 - 7.13 (m, 5 H) 5.23 (dd, .7=10.03, 5.50 Hz, 1 H) 4.80 - 4.91 (m, 1 H) 3.86 - 4.01 (m, 2 H) 3.37 - 3.63 (m, 3 H) 3.05 - 3.25 (m, 2 H) 2.82 - 2.98 (m, 2 H) 2.40 - 2.58 (m, 3 H).
To a solution of 4,4,4-trifiuorobutanoic add (20.00 g, 140.77 mmol, 1 eq) in DCM (20.0 mL) was added a solution of carbonyl diimidazole (23.08 g, 142.32 mmol, 1.01 eq) in DCM (300.00 mL), then warmed to 40 °C, the mixture was stirred at 40 °C for 0.5 h, and then cooled to 15 °C, then NH3 (gas) was bubbled into the solution, the mixture was stirred at 15 °C for 10 h. The reaction was monitored by LCMS and when complete, the solution was washed with aq. HC1 (10%, 2 x 200.00 mL), concentrated in vacuo to yield 4,4,4-trifluorobutanamide (10 g, crude) was obtained as a white solid. LCMS: m/z = 142.1 (M+H+).
To a solution of 4,4,4-trifluorobutanamide (6 g, 42.53 mmol, 1 eq) in THF (100 mL) was added 2,4-bis(4-methoxyphenyi)-2,4-dithioxo-l,3,2,4dithiadiphosphetane (20.64 g, 51.03 mmol, 1.2 eq), the mixture was stirred at 15 °C for 10 h. The reaction was monitored by TLC and when complete, the mixture was diluted with NaHCOs (aq, 2 x 100.0 mL) and extracted by ethyl acetate (100.0 mL), dried with NaaSO* and the solution was filtered and concentrated in vacuo. The residue was purified by column chromatography (Dichloromethane: Methanol = 20:1) to yield 4,4,4- trifluorobutanethioamide (4 g, 60%) was obtained as a white solid.
A solution of (S)-tert-butyl (4-bromo-l-(4-nitrophenyi)-3-oxobutan-2-yl)carbamate (2 g, 5.16 mmol, 1 eq) and 4,4,4-trifluorobutanethioamide (811.71 mg, 5.16 mmol, 1 eq) in MeCN (10 mL) was warmed to 80 °C and stirred at 80 °C for 10 h. The reaction was monitored by LCMS and when complete, the solution was washed with petroleum ether: ethyl acetate = 1:1 (8 mL), the filterer cake was dried in vacuo to yield (S)-2-(4- nitrophenyl)-l-(2-(3,3,3-trifluoropropyl)thiazol-4-yl)ethanamine hydrobromide (1.50 g, crude) was obtained as a brown solid. LCMS: m/z = 346.0 (M+H+).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-(3,3,3-trifluoropropyl)thiazoi-4- yl)ethanamine hydrobromide (0.80 g, 1.88 mmol, 1 eq) lithium (S)-2- ((methaxycarbonyl)(methyl)amino)-3-(pyrUUn-4-yl)propanoate (705.03 mg, 1.88 mmol, 1 eq) in DMF (10 mL) was added triethylamine (1.14 g, 11.26 mmol, 1.57 mL, 6 eq), then cooled to 0 °C, and then 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (1.79 g, 2.82 mmol, 1.67 mL, 50% purity, 1.5 eq) was added drop-wise at 0 °C under N2, the reaction was stirred at 20 °C for 16 h. The reaction was monitored by LCMS and when complete, the reaction mixture was washed with H2O (15 mL) and extracted with acethyl acetate (3 x 15 mL), the solution was filtered and concentrated in vacuo. The residue was purified by prep-TLC (Dichloromethane: Methanol = 15:1) to yield methyl methyl((S)-l-(((S)-2-(4-nitrophenyQ-l-(2-(3,3,3- trifluoropropyl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2- yQcarbamate (0.5 g, 47%) was obtained as a brown oil. LCMS: m/z = 566.1 (M+H+).
A solution of methyl methyl((S)-l-(((S)-2-(4-nitrophenyt)-l-(2-(3,3,3- trifluoropropyl)thiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2- yQcarbamate (0.5 g, 884.07 umol, 1 eq) and NH4CI (236.45 mg, 4.42 mmol, 5 eq) in EtOH (4 mL) and H2O (2 mL) was heated to 90 °C, then Fe (246.88 mg, 4.42 mmol, 5 eq) was added, the mixture was stirred at 900 C for 1 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(0.1%TFA)-ACN];B%: 1%-30%,10 min) to yield methyl ((S)-l-(((S)-2-(4- aminophenyl)-l-(2-(3,3,3-trifluoropropyl)thiazol-4-yl)ethyI)amino)-l-oxo-3- (pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.3 g, 61%) was obtained as a brown oil. LCMS: m/z = 536.2 (Μ+Η').
To a solution of methyl ((S)-l-(((S)-2-(4-aminophenyl)-]-(2-(3,3,3- trifluoropropyl)thiazjol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2- yl)(methyl)carbamate (0.27 g, 504.13 umol, 1.00 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, added SOs.pyridine (240.71 mg, 1.51 mmol, 3.00 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% ammonium hydroxide (4 mL) was added drop-wise, and then the mixture was dried by blowing N2, and the residue was purified by prep-HPLC (column: Phenomenex Gemini Cl 8 250*50 10u;mobile phase: |water(10mM NH4HCO3)- ACN];B%: 10%-40%, 20min) to yield ammonium (4-((S)-2-((S)-2-
((methoxycarbonyl)(methyl)amino)-3-(pyridin-4-yl)propanamido)-2-(2-(3,3,3- trifluoropropyl)thiazpl-4-yl)ethyl)phenyl)sulfamate (0.1 g, 31%) was obtained as a white solid. LCMS: m/z = 616.1 (M+H+), 1H-NMR: (400 MHz, D20) δ = 8.32 (d, J= 5.99 Hz, 2 H) 7.28 (br s, 2 H) 6.87 - 7.11 (m, 5 H) 5.18 (dd, J= 9.96, 5.44 Hz, 1 H) 4.83 (br s, 1 H) 3.32 - 3.65 (m, 3 H) 3.05 - 3.26 (m, 4 H) 2.78 - 2.99 (m, 2 H) 2.53 - 2.69 (m, 2 H) 2.47 (br s, 3 H).
Synthesis of Compound 6
To a mixture of (S)-tert-hutyl (4-bromo-l-(4-nitrophenyl)-3-oxobutan-2-yl)carbamate (2 g, 5.16 mmol, 1 eq) in MeCN (60 mL) was added cyclopropanecarbothioamide (522.54 mg, 5.16 mmol, 1 eq), then warmed to 90 °C and stirred for 16 h at 90 °C. The reaction was monitored by LCMS and when complete the solvent was removed in vacuo. The residue was purified by column chromatography (Si02, a little triethylamine, DCMiMeOH = 80:0 to 80:1) to yield (S)-l-(2-cyclopropylthiazol-4-yl)- 2-(4-nitrophenyl)ethanamine hydrobromide (1.2 g, 63%). LCMS: m/z = 289.9 (M+H+). To a solution of (S)-l-(2-cyclopropylthiazol-4-yl)-2-(4-nitrophenyl)ethanamlne hydrobromide (0.7 g, 1.51 mmol, 1 eq) and (S)-2-((methoxycarbonylXmethyl)amino)- 3-(pyridin-4-yl)propanoic acid (615.49 mg, 1.51 mmol, 1 eq) in DMF (10 mL) was added triethylamine (918.26 mg, 9.07 mmol, 1.26 mL, 6 eq), then cooled to 0 °C, and tiien 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (1.44 g, 2.27 mmol, 1.35 mL, 50% purity, 1.5 eq) was added drop-wise at 0 °C under N2, the reaction was stirred at 25 °C for 16 h. The reaction was monitored by LCMS and when complete, the reaction mixture was washed with H2O (15 mL) and extracted with acethyl acetate (5 mL x 3), dried with saturated brines, the solution was filtered and concentrated in vacuo and purified by prep-HPLC(column: Phenomenex luna(2) Cl 8 250*50 10u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 26%-56%, 20min) to yield methyl ((S)-l-(((S)-l-(2-cyclopropylthiazol-4-yl)-2-(4-nUrophenyt)ethyl)amino)-l- oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate as a off-white solid (0.48 g, 62%). LCMS: m/z = 510.1 (M+H+).
A solution of methyl ((S)-l-(((S)-l-(2-cyclopropylthiazol-4-yl)-2-(4- nitrophenyl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.34 g, 667.22 umol, 1 eq) and NH4CI (178.45 mg, 3.34 mmol, 116.64 uL, 5 eq) in EtOH (10 mL) and H2O (3 mL) was heated to 90 °C, then Fe (186.30 mg, 3.34 mmol, 5 eq) was added, the mixture was stirred at 900 C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo and was purified by prep-HPLC (column: Phenomenex Gemini Cl 8250*50 lOu; mobile phase: [ water(0.1 %TF A)- ACN] ;B%: 5%-30%,20min) to yield methyl ((S)-l-(((S)-2-(4- aminophenyl)-l-(2-cyclopropylthiazol-4-yl)ethyl)amino)-l-oxo-3-(pyridin-4- yl)propan-2-yl)(methyl)carbamate (0.34 g, 86%, TFA) was obtained as a yellow oil. LCMS: m/z = 480.1 (M+H+).
To a solution of methyl ((S)-l-(((S)-2-(4-aminophenyl)-l-(2-cyclopropylthiazfll-4- yl)ethyl)amlno)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.32 g, 539.07 umol, 1 eq, TFA) in pyridine (3 mL) and MeCN (3 mL) was cooled to 0 °C, added SCb.pyridine (257.40 mg, 1.62 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and HPLC when complete, 7% ammonium hydroxide (1 niL) was added dropwise, and then the mixture was dried by- flowing N2, and the residue was purified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 10%- 40%,20min) to yield ammonium (4-( (S)-2-(2-cyclopropylthiazol-4-yl)-2-( (S)-2- ((methoxycarbonyl)(methyl)amino)-3-(pyridin-4- yl)propanamuL})ethyl)phenyl)sulfamate (0.09 g, 29%) was obtained as a white solid. LCMS: m/z = 560.1 (M+H+), 1H-NMR: (400 MHz, DMSO-dfe) δ = 8.47 (d, J=5.95 Hz, 2 H) 8.22 - 8.37 (m, 1 H) 7.28 - 7.36 (m, 2 H) 7.22 (s, 1 H) 7.05 - 7.13 (m, 1 H) 6.96 (s, 2 H) 6.80 - 6.93 (m, 4 H) 4.77 - 5.14 (m, 2 H) 3.35 - 3.58 (m, 3 H) 3.09 - 3.21 (m, 1 H) 2.95 - 3.05 (m, 1 H) 2.87 (td, .7=14.28, 10.03 Hz, 2 H) 2.57 (s, 3 H) 2.38 (ddd, J= 12.95, 8.32, 4.96 Hz, 1 H) 1.05 - 1.17 (m, 2 H) 0.85 - 0.98 (m, 2 H).
Synthesis of Compound 7
To a solution of (2S)-2-amino-3-(4-nitrophenyl)propanolc acid (32.00 g, 152.24 mmol, 1.00 eq) and 4-methylmorpholine (15.40 g, 152.24 mmol, 16.74 mL, 1.00 eq) in DMF (300.00 mL) was added Iso-butyl-chloroformate (20.79 g, 152.24 mmol, 19.99 mL, 1.00 eq) drop- wised at 0 °C, then the mixture was stirred at 0 °C for 2 h. then the reaction was bubbled withNHs (2.59 g, 152.24 mmol, 1.00 eq) at 0 °C, the mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by LCMS and TLC and when complete, the solution was washed with aq. HC1 (5%, 500 mL), HzO (2 x 400 mL) and aq. NaHCOs (5%, 500mL). The combined organic layer was washed with brine (300 mL), dried with anhydrous NazSO*, filtered and concentrated in vacuo. The residue was washed with Petroleum ether/Ethyl acetate (3:1) to yield tert-butyl N-f(lS)-2-amino- l-[(4-nitrophenyl)methyl]-2-oxo-ethyl]carbamate (22.00 g, 47%) was obtained as a white solid. LCMS: m/z = 254.0 (M-55).
A solution of tert-butyl N-[(lS)-2-amino-]-[(4-nitrophenyI)methyl]-2-oxo- ethyl/carbamate (10.00 g, 32.33 mmol, 1.00 eq ) and 2,4-bis(4-methoxyphenyl)-2,4- dithioxo- 1 ,3,2,4dithiadiphosphetane (32.69 g, 80.82 mmol, 2.50 eq) in THF (100.00 mL) was stirred at 15 °C for 3 h. The reaction was monitored by TLC and when complete, the solution was washed with aq. NaaCOs (500 mL). The combined organic layer was washed with brine (100 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by re-crystallized from Petroleum ether (300 ml) to yield tert-butyl N-[(lS)-2-amino-l-[(4-nitrophenyl)methyl]-2- th ioxo-ethy!] carbamate (8.00 g, 76%) was obtained as a white solid.
A solution of (S)-tert-butyl (l-amino-3-(4-nitrophenyl)-l-thioxopropan-2- yl)carbamate (2.00 g, 6.15 mmol, 1.00 eq) and 2-bromo- 1 -(thiophen-2-y l)ethanone (1.26 g, 6.15 mmol, 1.00 eq) in MeCN (3.00 mL) was stirred at 80 °C for 12 h. The reaction was monitored by LCMS and when complete, the mixture was stirred at 25 °C for 1 h, then filtered and the filter cake dried in vacuo to yield (S)-2-(4-nitrophenyl)-l- (4-(thiophen-2-yl)thiazol-2-yl)ethanamine hydrobromide (1.70 g, 83%) was obtained as a yellow solid. LCMS: m/z = 332.0 (M+H+).
To a solution of (S)-2-(4-nitrophenyl)-l-(4-(thiophen-2-yl)thiazol-2-yl)ethanamine hydrobromide (0.6 g, 1.46 mmol, 1 eq, HBr) and (S)-2-
((methoxycarbonyl)(methyl)amino)-3-(pyridin-4-yl)propanotc acid (577.80 mg, 1.46 mmol, 1 eq) in DMF (10 mL) was added 2,4,6-tripropyl-l ,3,5,2,4,6- trioxatriphosphinane 2,4,6-trioxide (1.85 g, 2.91 mmol, 1.73 mL, 50% purity, 2 eq) and triethylamine (883.49 mg, 8.73 mmol, 1.22 mL, 6 eg) at 0 °C, then the reaction was stirred at 25 °C for 13 h under N2. The reaction was monitored by LCMS and when complete, the reaction mixture was poured into ice/water (100 mL) slowly, and extracted with ethyl acetate (3 x 30 mL), the organic phase was washed with brine (100 mL), dried withNa2S04, filtered and concentrated in vacuo to yield methyl methyl((S)- l-(((S)-2-(4-nitrophenyl)-l-(4-(thiophen-2-yl)thiazol-2-yl)ethyl)amino)-l-oxo-3- (pyridin-4-yl)propan-2-yl)carbamate (0.8 g, crude) was obtained as a yellow solid. LCMS: m/z = 552.1 (M+H+).
To a solution of methyl methyl((S)-l-(((S)-2-(4-nitrophenyl)-l-(4-(thiophen-2- yI)thiazol-2-yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)carbamate (0.25 g, 453.20 umol, 1 eq) in EtOH (2 mL) and HaO (2 mL) was added Fe (126.56 mg, 2.27 mmol, 5 eq) andNH+Cl (121.21 mg, 2.27 mmol, 5 eq) at 90 °C. The mixture was stirred at 90 °C for 2 h under N2. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated under vacuo. The residue was purified by prep-TLC (Si02, dichloromethane: methyl alcohol = 10:1) to yield methyl ((S)-l-(((S)-2-(4-aminophenyl)-l-(4-(thiophen-2-yl)thiazol-2-yl)ethyl)ammo)-l-oxo- 3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.25 g, 74%) was obtained as a yellow solid. LCMS: m/z = 522.1 (M+H+). methyl ((S)-J-(((S)-2-(4-aminophenyl)-J-(4-(thiophen-2-yl)thiazol-2- yl)ethyl)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (0.18 g, 345.06 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, then SOs.pyridine (164.76 mg, 1.04 mmol, 3 eq) was added and stirred at 0 °C for 0.17 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, 7% (2 mL) of ammonium hydroxide was added to the reaction mixture drop- wise, then the mixture was dried by blowing N2. The residue was purified by prep-HPLC (column: Phenomenex Gemini €18 250*50 10u;mobile phase: |water(10mM NH4HC03)- ACN];B%: 10%-40%,20min) to yield ammonium (4-((S)-2-((S)-2-
((methoxycarbonyl)(methyi)amino)-3-(pyridin-4-yl)propanamido)-2-(4-(thiophen-2- yl)thiazol-2-yl)ethyi)phenyl)sulfamate (45.55 mg, 22%) was obtained as a white solid. LCMS: m/z = 602.1 (M+H+). ‘H-NMR: (400 MHz, DMSO-dfe) δ = 8.77 - 8.95 (m, 1 H) 8.47 - 8.67 (m, 2 H) 7.84 (s, 1 H) 7.41 - 7.61 (m, 4 H) 7.23 (s, 1 H) 6.87 - 7.15 (m, 6 H) 5.20 - 5.37 (m, 1 H) 4.78 - 5.10 (m, 1 H) 3.51 (hr s, 3 H) 3.23 - 3.30 (m, 2 H) 2.88 - 3.13 (m, 2 H) 2.52 - 2.56 (m, 3 H).
Synthesis of Compound 8
A mixture of 4,4,4-trifluorobutan-2-one (3 g, 23.80 mmol, 1 eq) in ethyl acetate (15 mL) and CHCb (7 mL) was added CuBra (6.75 g, 30.22 mmol, 1.42 mL, 1.27 eq), then the mixture was heated to 80 °C and stirred at 80 °C for 16 h. The reaction was monitored by TLC and when complete, the solution was concentrated in vacuo to yield 1 -bromo-4, 4, 4-trifluorobutan-2-one (4.88 g crude) was obtained as a white solid.
A mixture of (S)-tert-butyl (l-amino-3-(4-nitrophenyl)-l-thioxopropan-2- yl)carbamate (2 g, 6.15 mmol, 1 eq) and 4, 4, 4-trifluorobutan-2-one (1.26 g, 6.15 mmol, 1 eq) in MeCN (80 mL) was heated to 80 °C and stirred at 80 °C for 16 h. The reaction was monitored by TLC and when complete, the solution was concentrated in vacuo to give the crude. The crude was purified by column chromatography (SiOz, Petroleum ether/Ethyl acetate=2/l to 0/1) to yield (S)-2-(4-nitropheny0-l-(4-(2,2,2- trifluoroethyl)thiazol-2-yl)ethanamine hydrobromide (1.5 g, 41% ) was obtained as a yellow solid.
To a solution of (lS)-2-(4-nitrophenyl)-l-/4-(2,2,2-trifluoroethyl)thiazot-2- yljethanamine hydrobromide (0.62 g, 601.61 umol, 1 eq) and f(2S)-2- [methoxycarbonyl(methyl)aminoJ-3-(4-pyridyl)propanoyl]oxylithium (176.28 mg, 721.94 umol, 1.2 eq) inDMF (15 mL) was added triethylamine (365.26 mg, 3.61 mmol, 502.43 uL, 6 eq), then then the mixture was cooled to 0 °C and 2,4,6-tripropyl- 1 ,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (574.26 mg, 902.42 umol, 536.70 uL, 50% purity, 1.5 eq) was added at 0 °C, the mixture was allowed to warm to 25 °C and stirred at 25 °C for 16 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (15 mL) and extracted with ethyl acetate (3 x 10 mL), the organic phase was washed with brine (20 mL), dried with anhydrous NazSCU, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiOz, Petroleum ether/Ethyl acetate=2/l to 0:1) to yield methyl methyl(l-(((S)-2-(4-nitrophenyl)-l-(4-(2,2,2-trifluoroethyl)thiazol-2- yl)ethyI)amino)-l-oxo-3-(pyridin-4-yl)propan-2-yl)carbamate (400 mg, crude) was obtained as a yellow solid. LCMS: m/z = 552.3 (M+H+).
To a solution of methyl methyl(l-(((S)-2-(4-nitrophenyl)-l-(4-(2,2,2- trifluoroethyl)thiazo!-2-yI)ethyl)amino)-l-oxo-3-(pyridin-4-yI)propan-2- yl)carbamate (400.00 mg, 725.25 umol, 1.00 eq) in EtOH (9.00 mL) and H2O (3 mL) was added NH4CI (193.97 mg, 3.63 mmol, 126.78 uL, 5 eq), the mixture was heated to 90 °C and Fe (202.52 mg, 3.63 mmol, 5 eq) was added at 90 °C, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the solution was filtered and the filtrate was concentrated in vacuo to give the crude. The crude was dissolved into water (10 mL) and extracted with DCM (3 x 10 mL), the organic phase was washed with brine (20 mL), dried with anhydrous NazSO* and concentrated in vacuo to yield methyl (l-(((S)-2-(4-aminophenyl)-l-(4-(2,2,2-trifluoroethyl)thiazol-2- yl)ethyl)amlno)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (400.00 mg, crude) was obtained as a yellow solid. LCMS: m/z = 522.3 (M+H+). methyl (l-(((S)-2-(4-aminophenyI)-l-(4-(2,2,2-trifluoroethyl)thlazol-2- yl)ethyl)amlno)-l-oxo-3-(pyridin-4-yl)propan-2-yl)(methyl)carbamate (400.00 mg, 766.94 umol, 1 eq) was purified by prep-TLC (S1O2, DCM: MeOH = 20:1, 1% NH3.H2O) to yield 8_diastereoisomer 1 (100 mg, 25%) and 8_diastereoisomer 2 (75 mg, 18%) was obtained as a yellow solid. LCMS: m/z = 522.3 (M+H+).
Diastereoisomer 1:
A solution of 8_diastereoisomer 1 (80 mg, 153.39 umol, 1 eq) in pyridine (1.00 mL) and MeCN (1.00 mL) was cooled to 0 °C, then sulfur trioxide pyridine complex (73.24 mg, 460.16 umol, 3 eq) was added in batches at 0 °C, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (5 mL) was added into the solution and the solution was stirred at 0 °C for 20 min, then the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 5%-30%,llmin) to yield ammonium (4-((2S)- 2-(2-((methoxycarbonyl)(methyl)amino)-3-(pyridin-4-yl)propanamido)-2-(4-(2,2,2- trifluoroethyl)thiazol-2-yl)ethyl)phenyl)sulfamate diastereomer 1 (41 mg, 39%) was obtained as a white solid. LCMS: m/z = 602.2 (M+H+), Ή-NMR: (400 MHz, D20) δ = 8.51 (hr d, J=5.99 Hz, 2 H) 7.53 (hr s, 2 H) 7.47 (s, 1 H) 7.00 - 7.17 (m, 4 H) 5.42 (hr d, J=3.67 Hz, 1 H) 4.83 - 4.90 (m, 1 H) 3.69 (q, 1=10.84 Hz, 2 H) 3.49 - 3.64 (m, 3 H) 3.36 (hr dd, J=13.88, 5.07 Hz, 1 H) 3.40 - 3.22 (m, 1 H) 2.95 - 3.10 (m, 2 H) 2.73 (s, 3
H).
Diastereoisomer 2:
A solution of 8_diastereoisomer 2 (70 mg, 134.21 umol, 1 eq) in pyridine (1.00 mL) and MeCN (1.00 mL) was cooled to 0 °C, then sulfur trioxide pyridine complex (64.08 mg, 402.64 umol, 3 eq) was added in batches at 0 °C, the mixture was stirred at 0 °C for 5 min. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (5 mL) was added into the solution and the solution was stirred at 0 °C for 20 min, then the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: YMC- Actus Triart C18 100*30mm*5um;mobile phase:[water(0.04%NH3H2O)-ACN];B%: 20%-40%,12min) to yield ammonium (4- ((2S)-2-(2-((methoxycarbonyl)(methyl)amino)-3-{pyridin-4-yl)propanamido)-2-(4- (2,2,2-trifluoroethyl)thiazol-2-yl)ethyl)phenyi)sulfamate diastereomer 2 (23 mg, 28%) was obtained as a white solid. LCMS: m/z = 602.2 (M+H+), ‘H-NMR: (400 MHz, D2O) δ = 8.51 (hr d, J=5.99 Hz, 2 H) 7.53 (hr s, 2 H) 7.47 (s, 1 H) 7.00 - 7.17 (m, 4 H) 5.42 (hr d, J=3.67 Hz, 1 H) 4.83 - 4.90 (m, 1 H) 3.69 (q, J=10.84 Hz, 2 H) 3.49 - 3.64 (m, 3 H) 3.36 (hr dd, J=13.88, 5.07 Hz, 1 H) 3.40 - 3.22 (m, 1 H) 2.95 - 3.10 (m, 2 H) 2.73 (s, 3 H).
Synthesis of Compound 9
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4- nitrophenyl)ethanamine hydrochloride (0.5 g, 1.37 mmol, 1 eq) and [(2S)-2- [methoxycarbonyl(methyl)amino]-3-(4-pyridyl)propanoyl]oxylithium (333.72 mg, 1.37 mmol, 1 eq) in DMF (7 mL) was added triethylamine (829.79 mg, 8.20 mmol, 1.14 mL, 6 eq), then cooled to 0 °C, and then 2,4,6-tripropyl-l ,3,5,2,4,6- trioxatriphosphinane 2,4,6-trioxide (1.74 g, 2.73 mmol, 1.63 mL, 50% purity, 2 eq) was added drop- wise at 0°C under N2, the reaction was stirred at 25 °C for 16 h. The reaction was monitored by LCMS and when complete, the reaction mixture was washed with H2O (30 mL) and extracted with acethyl acetate (3 x 15 mL), dried with saturated brine (45 mL), the solution was filtered and concentrated in vacuo and the residue was purified by column chromatography (Si02, Ethyl acetate/Methanol = 5/1) to yield methyl N-[(lS)-2-[[(lS)-l-[2-(lH-imidazol-S-ylmethyl)thiazol-4-yl]-2-(4- nitrophenyl)ethylJamino]-2-oxo-l-(4-pyridylmethyl)ethyl]-N-methyl-carbamate was obtained as a yellow solid (0.3 g, 32%). LCMS: m/z = 550.3 (M+H+).
A solution of methyl N-[(lS)-2-[l(lS)-l-l2-(lH-imidazol-5-ylmethyl)thiazol-4-yl]-2- (4-nitrophenyl)ethyl]amino]-2-oxo-l-(4-pyridylmethyl)ethyl]-N-methyl-carbamate
(0.3 g, 545.85 umol, 1 eq) and NH4CI (145.99 mg, 2.73 mmol, 5 eq) in EtOH (1.5 mL) and H2O (0.5 mL) was heated to 90 °C, then Fe (152.43 mg, 2.73 mmol, 5 eq) was added, the mixture was stirred at 900 C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo. The residue was purified by prep-TLC (Si02, ethyl acetate : ethyl alcohol = 2:1) to yield methyl N- [(lS)-2-[[(lS)-2-(4-aminophenyl)-l-[2-(lH-imidazol-5-ylmethyl)thiazol-4- ylJethylJaminoJ-2-oxo-l-(4-pyridylmethyl)ethylJ-N-methyl-carbamate (0.1 g, 32%) was obtained as a yellow oil. LCMS: m/z = 520.3 (M+H+).
To a solution of methyl N-[(lS)-2-[[(lS)-2-(4-aminophenyl)-l-[2-(lH-imidazol-5- ylmethyl)thiazol-4-yl]ethylJamino]-2-oxo-l-(4-pyriifylmethyl)ethyl]-N-methyl- carbamate (0.2 g, 373.35 umoL, 1 eq) in pyridine (1.5 mL) and MeCN (1.5 mL) was cooled to 0 °C, added SO?. pyridine (178.27 mg, 1.12 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% ammonium hydroxide (2.5 mL) was added drop-wise, and then the mixture was dried by blowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HC03>-ACN]; B%: 5%- 30%,12min) to yield (4-((S)-2-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-((S)-2- ((methoxycarbonyl)(methyI)amino)-3-(pyridin-4- yl)propanamido)ethyl)phenyl)sulfamate (0.07 g, 30%) was obtained as a white solid. LCMS: m/z = 598.1 (M+H+), 1H-NMR: (400 MHz, D20) δ = 8.27 (hr s, 2 H) 7.66 (hr s, 1 H) 7.15 (hr s, 2 H) 7.04 (hr d, Λ=4.40 Hz, 6 H) 5.08 - 5.32 (m, 1 H) 4.82 - 4.92 (m, 1 H) 4.07 - 4.29 (m, 2 H) 3.28 - 3.75 (m, 3 H) 2.99 - 3.24 (m, 2 H) 2.75 - 2.94 (m, 2 H) 2.52 (hr s, 3 H).
Synthesis of Compound 10
To a solution of (JS)-l-[2-[(3-methylimidazol-4-yl)methylJthUizol-4-ylJ-2-(4- nitrophenyl)ethanamlne;hydrobromide (868.88 mg, 2.05 mmol, 1 eq) and f(2S)-2- [methoxycarbonyl(methyl)amino]-3-(4-pyridyl)propanoyl]oxylithium (0.5 g, 2.05 mmol, 1 eq) in DMF (5 mL) was added triethylamine (1.24 g, 12.29 mmol, 1.71 mL, 6 eq), and cooled to 0 °C, then 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphinane 2,4,6- trioxide (260.62 g, 4.10 mmol, 243.57 mL, 0.5% purity, 2 eq) was added at 0 °C. The mixture was stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water 20 mL and extracted with ethyl acetate 30 mL (3 x 10 mL). The combined organic layer was washed with brine (30 mL), dried with Na2SC>4, filtered and concentrated under vacuo. The residue was purified by column chromatography (Si02, Ethyl acetate to Ethyl acetate/ methyl alcohol = 5:1) to yield methyl N-methyl-N-[(lS)-2-[[(lS)-l-[2-[(3-methylimidazol-4- yl)methylJthlazol-4-yl]-2-(4-nitrophenyl)ethyl]amino]-2-oxo-l-(4- pyrUfylmethyl)ethyl/carbamate (0.3 g, 22%) was obtained as a yellow oil. LCMS: m/z = 564.1 (M+H+).
To a solution of methyl N-methyl-N-[(lS)-2-[[(lS)-l-[2-[(3-methylimidazol-4- yl)methylfthiazol-4-yl]-2-(4-nitrophenyl)ethyl]amino]-2-oxo-l-(4- pyridyhnethyl)ethyllcarbamate (0.33 g, 585.49 umol, 1 eq) in EtOH (6 mL) and HzO (2 mL) , then Fe (163.50 mg, 2.93 mmol, 5 eq) and NH4CI (156.59 mg, 2.93 mmol, 5 eq) was added at 90 °C. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated in vacuo. The residue was purified by prep-TLC (Si02, dichloromethane: methyl alcohol = 7:1) to give methyl N-[(lS)-2-[[(lS)-2-(4-aminophenyl)-l-l2-[(3- methylimidazol-4-yl)methyl]thiazol-4-yl]ethyl]amino]-2-oxo-l-(4- pyridylmethyl)ethyl]-N-methyl-carbamate (0.12 g 38%) was obtained as a yellow oil. LCMS: m/z = 534.3 (M+H+).
To a solution of msihy\N-[(lS)-2-[[(lS)-2-(4-aminophenyI)-l-[2-[(3-methylimidazol- 4-yl)methyl]thiazol-4-yl]ethyl]amino]-2-oxo-l-(4-pyridylmethyl)ethyl]-N-methyl- carbamate (0.1 g, 187.39 umol, 1 eq ) in pyridine (1 mL) and MeCN (1 mL) was added SOS.pyridine (89.48 mg, 562.17 umol, 3 eq). The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (3 mL) was added and was dried by blowing Nz. The residue was purified by prep-HPLC (column: YMC-Actus Triart C18 100*30mm*5um;mobile phase: [water(10mM NH4HC03)- ACN] ;B%: 10%-30%,12min) to yield ammonium (4-((S)- 2-((S)-2-((methoxycarbonyI)(methyl)amino)-3-(pyridin-4-yl)propanamido)-2-(2-((l- methyl-lH-imidazol-5-yl)methyl)thiazol-4-yl)ethyl)phenyI)sulfamate (0.03810 g, 33%) was obtained as a white solid. LCMS: m/z = 614.2 (M+Hf). 1HNMR(400 MHz, D20) δ = 8.38 (s, 1 H) 8.30 (d, J= 6.11 Hz, 2 H) 7.21 (br s, 2 H) 7.14 (s, 1 H) 6.97 - 7.08 (m, 5 H) 5.22 (dd, ^=8.99, 6.66 Hz, 1 H) 4.85 - 4.93 (m, 1 H) 4.40 (s, 2 H) 3.40 - 3.66 (m, 6 H) 3.05 - 3.21 (m, 2 H) 2.84 - 2.99 (m, 2 H) 2.57 (br s, 3 H).
Synthesis of Compound 11
A solution of (S)-5-(benzyloxy)-4-((tert-butaxycarbonyl)amino)-5-oxopentanoic acid (50.00 g, 148.21 mmol, 1 eq) in ethyl acetate (150.00 mL) and HCl/EtOAc (4 M, 800.00 mL, 21.59 eq) was stirred at 0 °C for 1 h. The mixture was filtered and the filter cake was dried in vacuo to yield ( S)-4-amino-5-(benzylaxy)-5-oxopentanoic acid (32.00 g crude) was obtained as a white solid.
To a solution of ( S)-4-amino-5-(benzyloxy)-5-oxopentanoic acid (10.3 g, 37.63 mmol, 1 eq, HC1) in DCM (1200.00 mL) was added diisopropylethylamine (19.45 g, 150.52 mmol, 26.22 mL, 4 eq) at 0 °C, then methyl carbonochloridate (3.56 g, 37.63 mmol, 2.91 mL, 1 eq) was added at 0 °C, the mixture was stirred at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (2500 mL) and adjust pH= 2 - 3 with HC1 (1 M), and extracted with DCM (3 x 2500 mL), the organic phase was washed with brine (50 mL), dried with anhydrous NazSO*, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (SiOz, Dichloromethane: Methanol = 15:1) to yield (S)-S-(benzyloxy)- 4-((methoxycarbonyl)amino)-5-oxopentanoic add (25.0 g, 63%) as a colorless oil. LCMS: m/z = 296.0 (M+H+).
To a solution of (S)-5-benzytoxy-4-(methaxycarbonylamino)-5-oxo-pentanoic acid (10 g, 33.87 mmol, 1 eq) in DME (40 mL) was added 4-methylmorpholine (3.43 g, 33.87 mmol, 3.72 mL, 1 eq) at -15 °C, then isobutyl chloroformate (4.63 g, 33.87 mmol, 4.45 mL, 1 eq) was added drop-wise, the mixture was stirred at -15 °C for 0.5 h. Then NaBH* (1.92 g, 50.80 mmol, 1.5 eq) was added at 0 *C, and then HzO (20 mL) was added at 0 *C, the mixture was stirred at 25 *C for 10 h. The reaction was monitored by TLC and when complete, the solution was poured into water (100 mL), and then extracted with ethyl acetate (3 x 50 mL), the organic phase was washed with brine (50 mL), dried with anhydrous NaaSO^ filtered and and concentrated to give a residue. The residue was purified with column (SiOz, Petroleum ether: Ethyl acetate^ 1 : 1) to yield benzyl (2S)- 5-hydroxy-2-(methoxycarbonylamino)pentanoate (5.91 g, 62%) was obtained as a colorless oil.
To a solution of benzyl (2S)-5-hydroxy-2-(methoxycarbonylamino)pentanoate (12.5 g, 44.44 mmol, 1 eq) in DCM (300 mL) wras added imidazole (7.56 g, 111.09 mmol, 2.5 eq) and tert-butylchlorodimethylsilane (8.04 g, 53.32 mmol, 6.53 mL, 1.2 eq) at 0 °C. The mixture was allowed warm to 25 °C and stirred for 13 h. The reaction was monitored by TLC and when complete, the solution was poured into HzO (50 ml), and then extracted with DCM (3 x 50 mL), washed with brine (50 mL), dried over NazS04, and then concentrated in vacuo. The residue was purified by column (S1O2, Petroleum ether: Ethyl acetate= 5: 1) to yield benzyl (2S)-5-[tert-butyl(dimethyl)sUyl]oxy-2- (methoxycarbonylamino)pentanoate (17.24 g, 98% ) was obtained as a colorless oil.
To a solution of benzyl (2S)-5-ltert-butyl(dimethyl)silyl]axy-2- (methoxycarbonylamino)pentanoate (10 g, 25.28 mmol, 1 eg) in DMF (50 mL) was added iodomethane (5.38 g, 37.92 mmol, 2.36 mL, 1.5 eq) at -20 °C, then sodium hydride (1.52 g, 37.92 mmol, 60% purity, 1.5 eq) was added at -20 °C, the mixture was stirred at -20 °C for 1 h. The reaction was monitored by TLC and when complete, the solution was poured into HaO (50 mL), and then extracted with ethyl acetate (3 x 50 mL), then concentrated in vacuo. The residue was purified by column (S1O2, Petroleum ether: Ethyl acetate = 5: 1) to yield benzyl (2S)-5-[tert-butyl(dimethyl)silylJoxy-2- [methoxycarbonyl(methyl)aminoJpentanoaie (7 g, 67%) was obtained as a colorless oil.
To a soltion of benzyl ( 2S)-5-[tert-butyl(dimethyl)silyl]oxy-2 - lmethoxycarbonyl(methyl)amino]pentanoate (0.5 g, 1.22 mmol, 1 eq) in THF (10 mL) was added aq. HC1 (1 M, 3.66 mL, 3 eg) at 0 °C, the mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by LCMS and when complete, the solution was dried by blowing N2. The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate =2: 3) to yield benzyl (2S)-5-hydroxy-2-[metkaxycarbonyl(methyl)amino]pentanoate (0.26 g, 72%) was obtained as colorless oil. LCMS: m/z = 296.1 (M+H+).
To a solution of benzyl (2S)-5-hydraxy-2-
[methoxycarbonyl(methyl)amino]pentanoate ( 0.7 g, 2.37 mmol, 1 eq) in DCM (12 mL) was added Dess-Martin (1.11 g, 2.61 mmol, 807.18 uL, 1.1 eq) at 0 °C, the reaction was stirred at 25 °C for 2 h under N2. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition NaS20330 mL at 25 °C, and extracted with DCM 50 mL (3 x 25 mL). The combined organic layer was washed with saturated NaHC03 (50 mL) and washed with brine (50 mL), dried with Na2S04, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiOa, Petroleum ether: Ethyl acetate=5: 1 to 1 : 1) to yield benzyl ( 2S 2-fmethoxycarbonyl(methyl)aminoJ-5-oxo-pentanoaie (0.6 g, 77%) was obtained as a white solid, LCMS: m/z = 294.1 (M+H+).
A solution of benzyl (2S)-2-/methoxycarbonyl(meihyl)aminoJ-5-oxo-pentanoate (0.55 g, 1.88 mmol, 1 eq) and N-methylmethanamine (764.53 mg, 9.38 mmol, 4.69 mL, 5 eq, HC1) in MeOH (10 mL) was treated withNaBH3CN (235.67 mg, 3.75 mmol, 2 eq) at 0 °C, then the reaction was allowed warm to 25 °C and stirred for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo, and the residue was diluted with water 20 mL and extracted with DCM 60 mL (3 x 20 mL). The combined organic layer was washed with brine (50 mL), dried with NaaSO*, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiOa, Ethyl acetate/methyl alcohol = 50:1 to 10:1) to yield benzyl (2S)-5-(diniethylamino)-2-[methoxycarbonyl(methyl)aminoJpentanoate (0.6 g, 84%) was obtained as a colorless oil, LCMS: m/z = 323.1 (M+H+).
To a solution of benzyl (2S)-5-(dimethylamino)-2- [methoxycarbonyl(methyl)amino]pentanoate (0.56 g, 1.74 mmol, 1 eq) in MeOH (5 mL) was added Pd/C (1.74 mmol, 10% purity, 1 eq), the reaction was stirred at 25 °C for 2 h under Ha (3.51 mg, 1.74 mmol, 1 eq) (15 psi). The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated in vacuo to yield (2S)-5-(dimethylamino)-2-[methoxycarbonyl(methyl)amino]pentanoic acid (0.4 g crude) was obtained as a off white solid, LCMS: m/z = 233.1 (M+H+).
To a solution of (lS)-2-(4-niirophenyl)-l-f2-(2-thienyQtkiazol-4- yl]ethanamine;hydrobromide (710.06 mg, 1.72 mmol, 1 eq) and S-(dimethylamino)- 2-[methoxycarbonyl(methyl)amino]pentanoic acid (0.4 g, 1.72 mmol, 1 eq) in DMF (5 mL) was added triethylamine (1.05 g, 10.33 mmol, 1.44 mL, 6 eq), then the mixture was cooled to 0 °C and 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (2.19 g, 3.44 mmol, 2.05 mL, 50% purity, 2 eq) was added at 0 °C, the mixture was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was dried by blowing N2. The resiue was purified by column chromatography (S1O2, Ethyl acetate: methyl alcohol: ammonium hydroxide = 3: 1: 0.05) to yield methyl N-[4-(dimethylamino)-l-[[(lS)-2- (4-nitrophenyl)-l-[2-(2-thienyl)thiazol-4-yl]ethylJcarbamoylJbutylJ-N-methyl- carbamate (0.3 g, 28%) was obtained as a yellow oil. LCMS: m/z = 546.1 (M+H1).
To a solution of methyl N-[4-(dimethylamino)-l-ff(lS)-2-(4-nitrophenyl)-l-[2-{2- thienyl)thiazol-4-yl]ethylJcarbamoylJbutylJ-N-methyl-carbamate (0.3 g, 549.78 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (147.04 mg, 2.75 mmol, 5 eq), the mixture was heated to 90 °C and Fe (153.53 mg, 2.75 mmol, 5 eq) was added at 90 °C, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated under reduced pressure to give methyl N-[l-[[(lS)-2-(4-aminophenyl)-l-[2-(2-thienyl)thiazol-4- yl]ethyl]carbamoyl]-4-(dimethylamino)butyl]-N-methyl-carbamate (0.25 g, 88%) was obtained as a yellow oil.
LCMS: m/z = 516.1 (M+H+).
A mixture of methyl N-fl-[[(lS)-2-(4-aminophenyl)-l-f2-(2-thienyl)thiazol-4- yl]ethyl]carbamoyl]-4-(dimethylamino)butyl]-N-methyl-carbamate (0.25 g, 484.79 umol, 1 eq) was separated by SFC to yield 18_Diastereoisomer 1 (0.13 g, 46%) was obtained as a yellow oil. and 18_Diastereoisomer 2 (0.09 g, 32%) was obtained as a yellow oil. LCMS: m/z = 516.1 (M+H+).
Diastereoisomer 1: To a solution of 18_diastereoisomer 1 (0.12 g, 232.70 umol, 1 eq) in pyridine (1 mL) andMeCN (1 mL) was cooled to 0 °C, then the sulfur trioxide pyridine complex (111.11 mg, 698.09 umol, 3 eq) was added at 0 °C and the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by blowing Nz, and the residue was purified by prep-HPLC (column: Agela DuraShell 150mm_25mm_5um;mobile phase: [water(10mM NH4HC03)- ACN] ;B%: 20%-
50%,10min) to yield ammonium (4-((2S)-2-(S-(dimethylamino)-2- ((methoxycarbonyl)(methyl)amino)pentanamido)-2-(2-(thiophen-2-yl)thiazol-4- ytyethyQpkenyQsulfamate diastereoisomer 1 (1.9 mg, 2%) was obtained as a white solid. LCMS: m/z = 594.1 (M-H+). ¾-NMR: (400 MHz, D20) δ = 7.34 - 7.59 (m, 2 H) 7.07 - 7.15 (m, 6 H) 5.17 - 5.38 (m, 1 H) 4.28 - 4.50 (m, 1 H) 3.57 (br s, 3 H) 3.14 - 3.34 (m, 1 H) 2.82 - 3.06 (m, 3 H) 2.76 (s, 6 H) 2.68 (br s, 3 H) 1.18 - 1.63 (m, 4 H).
Diastereolsomer 2:
To a solution of 18_diastereoisomer 2 (0.08 g, 155.13 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, then the sulfur trioxide pyridine complex (74.07 mg, 465.40 umol, 3 eq) was added at 0 °C and the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing Nz, and the residue was purified by prep-HPLC (column: Agela DuraShell 150mm_25mm_5um;mobile phase: [water(10mM NH4HCO3)- ACN] ;B%: 20%-
50%,10min) to yield ammonium (4-((2S)-2-(S-(dimethylamino)-2- ((methoxycarbonyl)(methyl)amino)pentanamUio)-2-(2-(thiophen-2-yl)thiazol-4- yi)ethyl)phenyl)sulfamate diastereoisomer 2 (1.9 mg, 2%) was obtained as a white solid. LCMS: m/z = 594.1 (M-H+). ‘H-NMR: (400 MHz, DzO) δ = 7.34 - 7.59 (m, 2 H) 7.07 - 7.15 (m, 6 H) 5.17 - 5.38 (m, 1 H) 4.28 - 4.50 (m, 1 H) 3.57 (br s, 3 H) 3.14 - 3.34 (m, 1 H) 2.82 - 3.06 (m, 3 H) 2.76 (s, 6 H) 2.68 (br s, 3 H) 1.18 - 1.63 (m, 4 H).
Synthesis of Compound 12
To a solution of (S)-6-amino-2-(((benzylaxy)carbonyt)amino)hexanoic add (10 g, 35.67 mmol, 1 eq) inMeOH (100 mL), then cooled to 0°C , sulfurous dichloride (14.77 g, 124.14 mmol, 9.01 mL, 3.48 eq) was added at 0°C, then warmed to 25 °C and stirred for 13 h at 25 °C under Nz . The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with saturated NaHCO? (100 mL) and extracted with dichloromethane (100 mL). The combined organic layers were washed with brine (100 mL), dried with NazSO^ filtered and concentrated in vacuo to yield (S)-methyl 6- amino-2-(((benzyloxy)carbonyl)amino)hexanoate (9.8 g, crude) was obtained as a yellow oil. LCMS: m/z = 295.2 (M+Hf). To a solution of phthalic acid (9.93 g, 59.79 mmol, 1 eq) in MeCN (200 mL) was added carbonyl diimidazole (20.94 g, 129.15 mmol, 2.16 eq) under Na and (S)-methyl 6- amino-2-(((benzytoxy)carbonyl)amino)hexanoate (22 g, 59.79 mmol, 1 eq) was added and the reaction was stirred at 25 °C for 13 h The reaction was monitored by TLC and when complete, the solution was concentrated in vacuo and purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=10:l to 2:1) to yield (S)-methyl 2-(((benzyloxy)carbonyl)amino)-6-(l,3-dioxoisoindolin-2-yl)hexanoate (9 g, 28%) was obtained as a yellow oil.
A solution of ( Symethy I 2-(((benzyloxy)carbonyl)amino)-6-(l,3-dioxoisoindolin-2- yQhexanoate (9.00 g, 21.20 mmol, 1 eq) in DMF (100 mL) was cooled to 0 °C , then added sodium hydride (1.10 g, 27.57 mmol, 60% purity, 1.5 eq) at 0°C, the mixture was stirred at 0°C for 0.5h, then warmed to 25 °C , then added potassium iodide (24.08 g, 169.63 mmol, 10.56 mL, 8 eq) was added, the mixture was stirred at 25 °C for 1.5 h. The reaction was monitored by LCMS and when complete, the solution was poured into ice water (300 mL) and extracted with ethyl acetate (3 x 200 mL), the solution was concentrated in vacuo and was purified by silica gel column chromatography (Petroleum ether: Ethyl acetate = 10:1-5:1)) to yield methyl 2-
(((benzyloxy)carbonyl)(methyl)amino)-6-(l,3-dioxolsoindolin-2-yl)hexanoate (4.3 g, 26%) was obtained as a colorless oil. LCMS: m/z = 439.0 (Μ+Η').
To a solution of methyl 2-(((benzyloxy)carbonyl)(methyl)amlno)-6-(l,3- dloxoisoindolin-2-yl)hexanoate (4.30 g, 9.81 mmol, 1 eq) in ethyl acetate (150 mL) was added Pd/C (6.00 g, 9.81 mmol, 10% purity, 1.00 eq) under Ha, the mixture was stirred at 20°C under Ha at 15 psi for 10 h The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to yield methyl 6-(l,3- dioxoisoindolin-2-yl)-2-(methylamino)hexanoate (2.7 g, crude) was obtained as a colorless oil. LCMS: m/z = 305.0 (M+H+). A solution of methyl 6-(l,3-dioxoisoindolin-2-yl)-2-(methylamino)hexanoate (2.70 g, 8.87 mmol, 1 eq) in dichloromethane (40.0 mL) was cooled to 0 °C, then added diisopropylethylamine (3.44 g, 26.61 mmol, 4.64 mL, 3 eq) and methyl carbonochloridate (838.34 mg, 8.87 mmol, 687.17 uL, 1 eq) at 0 °C, then warmed to 15 °C , the mixture was stirred at 15°C for 10 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (40 mL) and extracted with dichloromethane (3 x 20 mL), the solution was concentrated in vacuo and purified by silica gel column chromatography (Petroleum ether: Ethyl acetate = 25:1 - 3:1) to yield (S)-methyl6-(l,3-dioxoisoindolin-2-yl)-2-
((methoxycarbonyl)(methyl)amino)hexanoate (2.1 g, 52% ) was obtained as colorless oil. LCMS: m/z = 363.0 (M+H4).
To a solution of (S)-methyl6-( l,3-dioxoisoindolin-2-yl)-2- ((methoxycarbonyl)(methyl)amino)hexanoate (1.3 g, 3.59 mmol, 1 eq) in ethyl alcohol (20 mL) was added hydrazine hydrate (633.84 mg, 10.76 mmol, 615.37 uL, 85% purity, 3 eq), then warmed to 80 °C, the mixture was stirred at 80 °C for 10 h. The reaction was monitored by LCMS and when complete, the solution was concentrated in vacuo and purified by column chromatography (SiOz, dichloromethane: Methanol= 2:1) to yield methyl 6-amino-2-[methoxycarbonyl(methyl)amino]hexanoate (0.56 g, 67%) was obtained as a colorless oil. LCMS: m/z = 233.2.0 (M+H+).
To a solution of methyl 6-amino-2-fmethoxycarbonyl(methyl)aminoJhexanoate (0.6 g, 2.58 mmol, 1 eq) in MeOH (20 mL) was added Pd/C (0.1 g, 2.58 mmol, 50% purity,
1 eq) and formaldehyde (1.05 g, 12.97 mmol, 965.42 uL, 5.02 eq). The mixture was stirred at 25 °C for 13 h under Hz (2.58 mmol, 1 eq) at 50 psi. The reaction w'as monitored by LCMS and w'hen complete, the solution was concentrated in vacuo to yield methyl 6-(dimethylamino)-2-((methoxycarbonyl)(methyl)amino)hexanoate (0.7 g, crude) was obtained as a colorless oil. LCMS: m/z = 260.9 (M+H4).
To a solution of methyl 6-(dimethylamino)-2-
((methoxycarbonyl)(methyl)amino)hexanoate (0.75 g, 1.73 mmol, 1 eq) in THF (9 mL) and H2O (3 mL) was added lithium hydroxide hydrate (49.68 mg, 2.07 mmol, 1.2 eq) slowly. The mixture was stirred at 25 °C for 0.5 h. The mixture was dried by lyophilization to yield 6-(dimethylamino)-2-
((methoxycarbonyl)(methyl)amino)hexanoic acid (0.55 g, crude) was obtained as a white solid. LCMS: m/z = 247.2(M+H+).
To a solution of 6-(dimethylamino)-2-((methoxycarbonyl)(methyl)amino)hexanoic acid (0.55 g, 2.23 mmol, 1 eq) and (lS)-2-(4-nilrophenyl)-l-(2-(thiophen-2-yl)thiaz€d- 4-yl)ethanamine hydrobromide (920.73 mg, 2.23 mmol, 1 eq)) in DMF (10 mL) was added triethylamine (1.36 g, 13.40 mmol, 1.86 mL, 6 eq), then cooled to 0 °C, and then 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (2.84 g, 4.47 mmol, 2.66 mL, 50% purity, 2 eq) was added drop-wise at 0°C under N2, the reaction was stirred at 25 °C for 13 h. The reaction was monitored by LCMS when complete, the reaction mixture was diluted with ice water (50 mL) and extracted with ethyl acetate 60 mL (2 x 30 mL) and washed with brine (60 mL) , dried over NaiSO-i, filtered and concentratedthe in vacuo and purified by column chromatography (Si02, Ethyl acetate/ Methanol = 10/1 to 2:1) to yield methyl (6-(dimethylamino)-l-(((S)-2-(4-nitrophenyl)- l-(2-(thiophen-2-yl)thiazol-4-yl)ethyl)amino)-l-oxohexan-2-yl)(methyl)carbamate (800 mg, 64%) was obtained as a yellow oil. LCMS: m/z =560.3(M+H+).
A solution of methyl (6-(dimeihylamtno)-l-(((S)-2-(4-nitrophenyl)-l-(2-(thwphen-2- yl)thiazol-4-yl)ethyl)amino)-l-oxohexan-2-yl)(methyl)carbamate (0.1 g, 178.67 umol, 1 eq) and NH4CI (47.78 mg, 893.33 umol, 31.23 uL, 5 eq) in ethyl alcohol (3 mL) and H2O (1 mL) was heated to 90 °C, then Fe (359.13 mg, 6.43 mmol, 10 eq) was added, the mixture was stirred at 90 0 C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to yield methyl (1-(((S)~ 2-(4-aminophenyl)-l-(2-(thiophen-2-yQthiazol-4-yl)ethyQamino)-6- (dimethylamino)-J-axohexan-2-yl)(methyl)carbamate (100 mg, crude) was obtained as a white solid. LCMS: m/z = 530.0 (M+H1). A mixture of methyl N-[l-[[(lS)-2-(4-aminophenyl)-l-f2-(2-thienyl)thiazol-4- yl]ethyl]carbamoyl]-5-(dimethylamino)pentylJ-N-methyl-carbamate (0.4 g, 755.12 umol, 1 eq) was separated by SFC. The residue was separated by SFC (column: AD(250mm*30mm,5um);mobile phase: [0.1%NH3H2O ETOH];B%: 48%- 48%,12min). Compound diastereomer 1 (0.22 g, 42%) and diastereomer 2 (0.16 g, 34%) were obtained as yellow solids.
5 diastereoisomer 1:
To a solution of diastereomer 1 (0.2 g, 377.56 umol, 1 eq) in pyridine (4 mL) and MeCN (1.3 mL) was cooled to 0 °C, added SO3. pyridine (180.28 mg, 1.13 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then tiie mixture was dried by blowing N2, and the residue was purified by prep-HPLC (column: Gemini 200*30 ΙΟμ; mobile phase: [water(10mM NH4HC03)-ACN];B%:
10%-60%, 12min) to yield (4-((2S)-2-(6-dimethylamino)-2- ((methoxycarbonyI)(methyl)amino)hexanamido)-2-(2-(thiophen-2-yl)thiazol-4- yl)ethy!)phenyl)sulfamic acid S diastereomer 1 (52.31 mg, 21%) was obtained as a white solid. LCMS: m/z = 610.2 (M+H+), Ή-ΝΜΚ: (400 MHz, DMSO-rfe) δ = 8.46 (br s, 1 H) 7.92 (s, 1 H) 7.72 (dd, 7=5.07, 0.88 Hz, 1 H) 7.65 (dd, J= 3.75, 1.10 Hz, 1 H) 7.32 (br s, 1 H) 7.17 (dd, >4.96, 3.64 Hz, 1 H) 7.00 - 7.07 (m, 2 H) 6.93 - 7.00 (m, 2 H) 5.14 (brt, J=8.05 Hz, 1 H) 4.19 - 4.50 (m, 1 H) 3.57 (s, 3 H) 3.14 (dd, >13.89, 3.75 Hz, 1 H) 2.70 - 2.92 (m, 13 H) 1.38 - 1.56 (m, 3 H) 1.30 (br d, >10.58 Hz, 1 H) 0.96 (br s, 1 H) 0.76 (br s, 1 H)
5 diastereoisomer 2:
To a solution of diastereomer 2 (148.87 mg, 281.03 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SCh.pyridine (134.19 mg, 843.09 umol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% ammonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by blowing N2, and the residue was purified by prep-HPLC (column: Gemini 200*30 10p;mobile phase: [water(10mM NH4HC03)- ACN] ;B%:
15%-45%,12min) to yield (4-((2S)-2-(6-dimethylamino)-2- ((methoxycarbonyl)(metkyl)amino)hexanamido)-2-(2-(tkiophen-2-yl)thiazol~i- yl)ethyl)phenyl)sulfamic add S diastereoisomer 2 (51.8 mg, 29%) was obtained as a white solid. LCMS: m/z = 610.2 (M+H+), 1H-NMR: (400 MHz, DMSO-cfe) δ = 8.98 - 9.35 (m, 1 H) 8.12 - 8.26 (m, 1 H) 7.68 - 7.76 (m, 2H) 7.65 (dd, J= 3.67, 1.10 Hz, 1 H) 7.30 (s, 1 H) 7.18 (dd, .7=5.01, 3.79 Hz, 1 H) 6.85 - 6.98 (m, 4 H) 5.11 (td, ^8.74, 5.26 Hz, 1 H) 4.30 - 4.59 (m, 1 H) 3.54 - 3.75 (m, 3 H) 3.06 - 3.16 (m, 1 H) 2.85 - 3.03 (m, 3 H) 2.72 (s, 6 H) 2.62 (br s, 3 H) 1.67 - 1.83 (m, 1 H) 1.41 - 1.65 (m, 3 H) 1.01 - 1.18 (m, 2H).
ynthesis of (S)-tert-butyI-(4-bromo-l-(4-nitrophenyl)-3-axobutan-2-yI)-carbamate
To a solution of intermediate 2 (13.28 g, 42.80 mmol, 1 eq) and NMM (4.76 g, 47.08 mmol, .17 mL, 1.1 eq) in THF (100 mL) was added intermediate 3 (6.43 g, 47.08 mmol, 6.18 mL, 1.1 q) drop-wise at -15 °C under Nz. And the temperature was maintained below -15 °C for 0.5 h. The reaction was monitored by TLC and when complete, the reaction mixture was filtered, the filtrate was added diazomethane (12.75 g, 303.51 mmol, 7.09 eq) in ether (300 mL) and then the olution was stirred at 0 °C for 3 h. The reaction was monitored by TLC and when complete, the eaction mixture was to yield intermediate 4 (14.31 g, crude) in THF (100 mL) was obtained as ellow liquid.
To a solution of intermediate 4 (14.3 g, 42.77 mmol, 1 eq) in THF (100 mL) was added aqueous HBr (17.42 mL, 128.32 mmol, 40 % purity, 3 eq) drop-wise at 0 °C under N2. And then the eaction mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC and when omplete, the reaction mixture was poured into aqueous NaaCCb (150 mL). The mixture was xtracted with ethyl acetate (2 x 100 mL), and then the combined organic phase was washed with rine (100 mL), dried with anhydrous Na2S04 and concentrated in vacuo. The residue was riturated with TBME (70 mL) and filtered and then the filter cake was dried in vacuo to yield S)-tert-butyl-(4-bromo-l-(4-nitropheny1)-3-axobutan-2-y1)-carbamate (12 g, 72%) was btained as a white solid. !H-NMR: (400 MHz, DMSO -φ>) 5 = 8.18 (d, >8.33 Hz, 2 H) 7.37 (d, >8.33 Hz, 2 H) 4.96 - 5.18 (m, 1 H) 4.71 - 4.90 (m, 1 H) 4.02 (s, 2 H) 3.27 - 3.40 (m, 1 H) 2.97 3.12 (m, 1 H) 1.40 (s, 9 H).
Synthesis of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yI)thiazol-4-yl)ethanamine hydrobromide salt
To a solution of (S)-tert-butyl-(4-bromo-l-(4-nitrophenyI)-3-oxobutan-2-yl)-carbamate (20 g, 1.65 mmol, 1 eq) in MeCN (200 mL) was added intermediate S (7.40 g, 51.65 mmol, 1 eq), hen the reaction mixture was stirred at 90 °C for 16 h. The reaction was monitored by TLC and when complete, the reaction mixture was filtered, and the filter cake was dried in vacuo to yield S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (19.3 g, rude) was obtained as a white solid. 1H-NMR: (400 MHz, DMSO-de) δ = 8.64 (s, 3 H) 8.13 (d, >8.77 Hz, 2 H) 7.76 (d, >4.82 Hz, 1 H) 7.69 (d, >3.07 Hz, 1 H) 7.64 (s, 1 H) 7.46 (br d, >8.77 Hz, 2 H) 7.18 (t, >4.39 Hz, 1 H) 4.76 - 4.96 (m, 1 H) 3.35 - 3.50 (m, 2 H).
Synthesis of (S)-tert-butyl-(l-amino-3-(4-nitrophenyl)-l-thioxopropan-2-yl)-carbamate
To a solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-nitrophenyl)propanoic acid (50.00 g, 61.13 mmol, 1.00 eq) and NMM (16.30 g, 161.13 mmol, 17.72 mL, 1.00 eq) in DMF (250 mL) was added isobutyl chloroformate (22.01 g, 161.13 mmol, 21.16 mL, 1.00 eq) drop-wise at 0 C, the mixture was stirred at 0 °C for 2 h. Then N¾ (gas) was bubbled at 0 °C for 0.5 h. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured nto water (700 mL) and then the filter cake was dissolved into ethyl acetate (1000 mL) and washed with water (3 x 500 mL). Then the organic phase was washed with HC1 (5 %, 2 x 500 mL), water (2 x 400 mL) and NaHCCb (3 x 500 mL), then the organic phase was dried with nhydrous Na2SC>4 and concentrated in vacuo. The residue was triturated by PE: EA = 6:1 (100 mL) filtered and the filter cake dried in vacuo to yield (S)-tert-butyl-(l-amino-3-(4- itrophenyI)-l-oxopropan-2-yl)-carbamate (31.00 g, 62 %) as a white solid. LCMS: m/z = 54.2 (M-55). 1H-NMR: (400 MHz, DMSO-tfc) δ = 8.16 (d, >8.60 Hz, 2 H) 7.54 (d, >8.60 Hz, 2 H) 7.40 - 7.46 (m, 1 H) 7.07 (s, 1 H) 6.93 (d, >8.82 Hz, 1 H) 4.09 - 4.22 (m, 1 H) 3.03 - 3.15 (m, 1 H) 2.80 - 2.92 (m, 1 H) 1.27 (s, 9 H). To a solution of (S)-tert-butyl-(l-amino-3-(4-nitrophenyl)-l-axopropan-2-yl)-carbamate (25.3 , 81.79 mmol, 1.00 eq) in THF (200 mL) was added lawesson reagent (49.62 g, 122.69 mmol, .50 eq) at 0 °C, the mixture was stirred at 15 °C for 10 h. The reaction was monitored by TLC nd when complete, the reaction mixture was filtered, the filtrate was added ice-water (300 mL), nd then the mixture was extracted with ethyl acetate (3 x 300 mL), the organic phase was eutralized with aq. NaHCO.3 (5%, 3 x 200 mL). Then the combined organic phase was washed with brine (300 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo. The residue was riturated by petroleum ether: ethyl acetate = 80:1 (200 mL,), filtered and the filter cake dried in acuo to yield (S)-tert-butyl-(l-amino-3-(4-nitrophenyl)-l-thioxopropan-2-y1)-carbamate (18 g, 7%) was obtained as a off-white solid. Ή-NMR: (400 MHz, DMSO-<¾) δ = 9.65 (s, 1 H) 9.28 s, 1 H) 8.16 (d, .7=8.44 Hz, 2 H) 7.58 (d, .7=8.44 Hz, 2 H) 6.93 - 6.96 (m, 1 H) 4.46 - 4.51 (m, 1 H) 2.86 - 3.17 (m, 2 H) 1.28 (s, 9 H).
Synthesis of Lithium (S)-2-((methaxycarbonyl)(methyl)amino)-3-(pyridine-4-yl)propanoate
To a solution of (S)-2-amino-3-(pyridine-4-yl)propanoic acid (30 g, 180.53 mmol, 1 eq) in MeOH (250 mL) was added SOClz (42.96 g, 361.06 mmol, 26.19 mL, 2 eq) dropwise at 0°C, hen the reaction was heated to 60 °C and stirred at 60 °C for 13 h under N2 atmosphere. The eaction was monitored by LCMS and HPLC and when complete, the reaction was mixture was oncentrated in vacuo to yield (S)-methyl-2-amino-3-(pyridine-4-yl)propanoate (30 g, 27%) was btained as a white solid. LCMS: m/z = 181.1 (M+fT).
To a solution of (S)-methyl-2-amino-3-(pyridine-4-yl)propanoate (30 g, 138.46 mmol, 1 eq, HC1) in DCM (250 mL) was added DIPEA (71.58 g, 553.85 mmol, 96.47 mL, 4 eq) drop-wise at 0°C, after stirring for 10 min, methyl chloroformate (13.08 g, 138.46 mmol, 10.72 mL, 1 eq) was added dropwise at 0 °C, then the reaction was stirred at 25 °C for 50 min under N2. The eaction was monitored by TLC and HPLC and when complete, the reaction mixture was oncentrated in vacuo. The residue was purified by column chromatography (S1O2,
Dichlorom ethane: Methanol=l :0 to 30:1) to yield (S)-methyl-2-((methaxycarbonyt)amino)-3-pyridine-4-yt)propanoate (7 g, 21%) was obtained as a yellow oil. 1H-NMR: (400 MHz, CDCI3) δ = 8.45 - 8.56 (m, 2 H) 7.02 - 7.11 (m, 2 H) 5.38 - 5.47 (m, 1 H) 4.63 - 4.76 (m, 1 H) .72 (s, 3 H) 3.65 (s, 3 H) 2.94 - 3.24 (m, 2 H).
To a solution of (S)-methyl-2-((methaxycarbonyl)amino)-3-(pyridine-4-yl)propanoate (2 g, .39 mmol, 1 eq) and iodomethane (1.79 g, 12.59 mmol, 783.92 uL, 1.5 eq) in DMF (80 mL) was added NaH (503.70 mg, 12.59 mmol, 60% purity, 1.5 eq) slowly at -15 °C, then the reaction was stirred at -15°C for lh under N2. The reaction was monitored by LCMS and HPLC and when complete, the solution was quenched by sat. NH4CI (200 mL). The mixture was extracted with ethyl acetate (3 x 100 mL), then the combined organic phase was washed with brine (200 mL), dried with anhydrous NazS04 and concentrated in vacuo to yield (S)-methyl~2- (methoxycarbonyl) ( methyl) amino) -3- (pyridine-4-yl)propan oaie (1.8 g, 83%) was obtained as a ellow oil. LCMS: m/z = 253.2 (M+H4).
To a solution of (S)-methyl-2-((methaxycarbonyl)(methyl)amino)-3-(pyridine-4-yI)propanoate 1.8 g, 7.14 mmol, 1 eq) in THF (10 mL) and H20 (20 mL) was added LiOH (205.07 mg, 8.56 mmol, 1.2 eq) slowly at 0 °C, then the reaction was stirred at 25 °C for 2 h under N2. The eaction was monitored by LCMS and HPLC and when complete, the reaction mixture was dried y flowing N2 to yield Lithium (S)-2-((methaxycarbonyl)(methyI)amino)-3-(pyridine-4- l)propanoate (2 g, 97%) was obtained as a green oil. LCMS: m/z = 239.1 (M+H4).
Synthesis of (S)-l-(2-((lH-imidazol-5-yl)methyI)thiazol-4-yl)-2-(4-nitrophenyI)ethanamine
To a solution of 2-(lH-imidazol-5-yl)acetonitrile (5 g, 46.68 mmol, 1 eq) in DMF (30 mL) was dded thioacetamide (7.01 g, 93.36 mmol, 2 eq), then the mixture was stirred at 100 °C for 2 h. The reaction was monitored by TLC and when complete, HCl/EtOAc (4 M, 300 mL) was added nd stirred at 25 °C for 1 h. Then the solvent was evaporated in vacuo, and the residue was riturated with acetone to yield 2-(lH-imidazol-5-yl)ethanethioamide (16.2 g, 76%) as a light ellow solid. JH-NMR: (400 MHz, DMSO -ck) δ = 14.45 (br s, 1 H) 9.42 - 10.15 (m, 2 H) 9.02 s, 1 H) 7.49 (s, 1 H) 3.97 (s, 2 H).
To a solution of 2-(lH-imidazol-5-yl)ethanethioamide (7 g, 49.58 mmol, 1 eq) in EtOH (100 mL) was added (S)-tert-butyl-(4-bromo-l-(4-nitrophenyl)-3-oxobutun-2-yl)-carbumate (16 g, 1.32 mmol, 8.33e-1 eq), the reaction mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to ield (S)-tert-butyl-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4- itrophenyI)ethyl)carbamate (20 g, crude) was obtained as a white solid. LCMS: m/z = 329.9 M-100). S)-tert-butyl-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethyl)carbamate 10 g, 23.28 mmol, 1 eq) in HCl/EtOAc (50 mL) was stirred at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated in acuo to yield (S)-l-(2-((lH-imidazol-S-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine hydrochloride salt (11 g, crude) was obtained as a yellow solid. LCMS: m/z = 351.9 (M+Na+). H-NMR: (400 MHz, Methanol -<&) δ = 8.95 (d, >1.32 Hz, 1 H) 8.09 - 8.16 (m, 2 H) 7.57 (s, 1 H) 7.42 (s, 1 H) 7.37 (d, >8.82 Hz, 2 H) 4.76 - 4.86 (m, 1 H) 4.52 - 4.66 (m, 2 H) 3.45 - 3.51 m, 2 H).
Synthesis of Compound 13
To a solution of (S)-l -(2-((lH-imidazol-S-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl) ethanamine ydrobromide salt (0.45 g, 1.23 mmol, 1 eq, HC1) and 3-phenylpropanoic acid (184.72 mg, .23 mmol, 172.64 uL, 1 eq) in DMF (10 mL) was added EfcN (746.82 mg, 7.38 mmol, 1.03 mL, eq) at 25 °C, and then the solution was cooled to 0 °C, T3P (1.17 g, 1.85 mmol, 1.10 mL, 50 % urity, 1.5 eq) was added dropwise, the mixture was stirred at 25 °C for 10 h. The reaction was monitored by LCMS and TLC and when complete, the solution was poured into water (10 mL) nd extracted with ethyl acetate (3 x 10 mL), and then comined organic phase was washed with rine (20 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo. The residue was urified by silica gel column chromatography (S1O2, Dichloromethane: Methanol = 15:1) to ield (S)-N-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethyl)-3- henylpropanamide (0.35 g, crude) was obtained as brown oil. LCMS: m/z = 462.0 (M+H+).
To a solution of (S)-N-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethyl)-3- henylpropanamide (0.35 g, 758.34 umol, 1 eq) in EtOH (3 mL) and H2O (1 mL) was added NH4CI (202.82 mg, 3.79 mmol, 132.56 uL, 5 eq) at 20 °C, then Fe (211.77 mg, 3.79 mmol, 5 eq) was added at 90 °C, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and HPLC and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo. The residue was purified by prep-HPLC (column: YMC-Actus Triart C18 00*30mm*5um;mobile phase: [water(10mMNH4HCO3)-ACN]; B%: 15%-38%, 10 min) to ield (S)-N-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-aminophenyi)ethyl)-3- henylpropanamide (0.1 g, 26%) was obtained as brown oil. LCMS: m/z = 432.3 (M+H4).
To a solution of (S)-N-(l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-aminophenyl)ethyl)~ -phenylpropanamide (0.2 g, 463.44 umol, 1 eq) in C¾CN (2 mL) and pyridine (2 mL) was dded SOs-pyridine (221.29 mg, 1.39 mmol, 3 eq) at 0 °C, the reaction was stirred at 0 °C for .17 h. The reaction was monitored by LCMS and HPLC and when complete, the solution was ried by flowing N2. And the residue was purified by prep-HPLC (column: Gemini 200*30 10μ; mobile phase: [water (lOmM NH4HCO3)- ACN] ; B%: 5%-35%, 12 min) to yield Compound 13 0.09 g, 35%) was obtained as a white solid. LCMS: m/z = 512.1 (M+H4). Ή-NMR: (400 MHz, DMSO-ife) δ = 8.18 - 8.34 (m, 2 H) 7.72 (br s, 1 H) 7.21 - 7.29 (m, 2 H) 7.13 - 7.20 (m, 3 H) .08 (s, 1 H) 7.00 (s, 1 H) 6.87 - 6.93 (m, 2 H) 6.79 - 6.84 (m, 2 H) 5.02 - 5.16 (m, 1 H) 4.30 (s, H) 2.95 - 3.00 (m, 1 H) 2.74 - 2.83 (m, 3 H) 2.36 - 2.45 (m, 2 H).
Synthesis of Compound 14 To a solution of (S)-l-(2-((lH-imidazoI-5-yl)methyI)thiazol~4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (200 mg, 546.69 umol, 1 eq, HC1) and intermediate 2B (82.64 mg, 546.69 mol, 1 eq) in DMF (6 mL) was added EtsN (331.92 mg, 3.28 mmol, 456.56 uL, 6 eq), and then he solution was cooled to 0 °C and T3P (521.84 mg, 820.04 umol, 487.70 uL, 50 % purity, 1.5 q) was added drop-wise, and then the solution was allowed to warm to 25 °C and stirred for 16 . The reaction was monitored by LCMS and when complete, the reaction mixture was poured nto ice-water (20 mL) and the mixture was extracted with ethyl acetate (3 x 20 mL), then the ombined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4 and oncentrated in vacuo to yield intermediate 8 (424 mg, crude) was obtained as yellow oil. LCMS: m/z = 463.3 (M+H+).
To the solution of intermediate 8 (0.3 g, 648.62 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (173.48 mg, 3.24 mmol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (181.11 mg, 3.24 mmol, 5 eq) was added, and then the solution was stirred for 2 h at 90 °C. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 9 (450 mg, crude) was obtained as a yellow olid. LCMS: m/z = 433.0 (M+I T").
To a solution of intermediate 9 (210 mg, 485.50 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was added SOa-pyridine (231.82 mg, 1.46 mmol, 3 eq) slowly at 0 °C, then stirred at 0 °C or 10 min. The reaction was monitored by LCMS and HPLC and when complete, NH3.H2O 7%, 1 mL) was added drop-wise to above mixture at 0 °C, and stirred at 0 °C for 5 min. and hen the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Gemini 200*30 10μ; mobile phase: [water(10mM NHjHCCbJ-ACN]; B%: 5%-30%, 12 min) to ield Compound 14 (42 mg, 16%) was obtained as a white solid. LCMS: m/z = 513.2 (M+H+), H-NMR: (400 MHz, D20) δ =8.27 (d, >6.24 Hz, 2 H) 8.12 (s, 1 H) 7.16 (d, J=6.11 Hz, 2 H) .08 (s, 1 H) 6.90 - 6.99 (m, 4 H) 6.87 (s, 1 H) 5.03 - 5.13 (m, 1 H) 4.30 (s, 2 H) 2.78 - 3.03 (m, H) 2.52 - 2.59 (m, 2 H).
Synthesis of Compound IS
ntermediate IB (20 g, 212.51 mmol, 1 eq), intermediate 1C (33.83 g, 318.76 mmol, 32.22 mL, .5 eq) and ZnCh (11.59 g, 85.00 mmol, 0.4 eq) were stirred at 160 °C for 5 h. The reaction was monitored by TLC and when complete, the reaction mixture was concentrated in vacuo. The esidue was purified by column (S1O2, petroleum ether: ethyl acetate: ethyl alcohol = 4: 3: 1) to ield intermediate 2Λ (14 g, 36%) was obtained as a light yellow solid. 1H-NMR: (400 MHz, Methanol-t/4) δ = 9.37 (dd, >2.08, 1.10 Hz, 1 H) 9.07 - 9.09 (m, 1 H) 7.79 - 7.87 (m, 1 H) 7.64 - .76 (m, 3 H) 7.34 - 7.43 (m, 3 H) 7.19 (d, >16.51 Hz, 1 H). ntermediate 2A (7 g, 38.41 mmol, 1 eq) was dissolved in dioxane (140 mL) and HiO (47 mL) was added OsC>4 (0.1 M, 5.88 mL, 1.53e'2 eq) and NaI04 (16.68 g, 77.98 mmol, 2.03 eq), and hen stirred at 25 °C for 6 h. The reaction was monitored by TLC and when complete, the eaction mixture was diluted with dioxane (20 mL), and filtered, and then concentrated in vacuo, he residue was Purified by column (S1O2, petroleum ether: ethyl acetate =1: 1) to yield ntermediate 1 (3 g, 72%) was obtained as a yellow solid. 1 H-NMR: (400 MHz, Methanol-^) 5 = 9.22 - 9.25 (m, 1 H) 9.15 - 9.18 (m, 1 H) 7.73 - 7.83 (m, 1 H). To a suspension of NaH (1.66 g, 41.63 mmol, 60 % purity, 1.5 eq) in THF (20 mL) was added rop-wise methyl intermediate 1A (5.83 g, 27.75 mmol, 1 eq) at 0 °C. After the addition was ompleted, the mixture was stuffed for 30 minutes. Then intermediate 1 (3 g, 27.75 mmol, 1 eq) n THF (10 mL) was added. The reaction mixture was allowed to warm to 25 °C for 2 h. The eaction was monitored by TLC and when complete, the reaction mixture was neutralized with at. NH4CI (50 mL), The mixture was extracted with ethyl acetate (3 x 50 mL), then combined rganic phase was washed with brine (3 x 25 mL), dried with anhydrous Na2SC>4 and oncentrated in vacuo. The residue was purified by column (S1O2, petroleum ether: ethyl acetate 3: 2) to yield intermediate 2 (1.5 g, 30%) was obtained as a yellow solid. LCMS: m/z = 165.1 M+H+).
To a solution of intermediate 2 (1.3 g, 7.30 mmol, 1 eq) in ethyl acetate (10 mL) was added d/C (700 mg, 7.30 mmol, 10 % purity, 1.00 eq) under N2. The suspension was degassed under acuum and purged with H2 several times. Then the mixture was stirred under Hz (15 psi) at 25 C for 15 min. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 3 (0.693 g, 53%) was btained as a colorless oil. LCMS: m/z = 167.1 (M+H+). ntermediate 3 (230.54 mg, 1.39 mmol, 1 eq) in THF (6 mL) and H2O (12 mL) was added LiOH.H20 (69.86 mg, 1.66 mmol, 1.2 eq) at 25 °C, and then the solution was stirred at 25 °C for h. The reaction was monitored by LCMS and when complete, the reaction mixture was oncentrated to yield intermediate 2B (0.25 g, crude) was obtained as a colorless oil. LCMS: m/z = 153.1 (M+H4).
To a solution of intermediate 2C (480.89 mg, 1.31 mmol, 1 eq) and intermediate 2B (200 mg, .31 mmol, 1 eq) in DMF (6 mL) was added EtsN (798.08 mg, 7.89 mmol, 1.10 mL, 6 eq), and hen the solution was cooled to 0 °C, then T3P (1.25 g, 1.97 mmol, 1.17 mL, 50 % purity, 1.5 eq) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 18 h. The reaction was monitored by LCMS and TLC and when complete, the reaction mixture was dried by flowing N2 o give a residue. The residue was purified by prep-TLC (S1O2, dichloromethane: methyl alcohol: mmonium hydroxide = 20:2:1) to yield intermediate 10 (150 mg, 23%) was obtained as a yellow solid. LCMS: m/z = 464.0 (M+H+). To a solution of intermediate 10 (130 mg, 280.47 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (75.01 mg, 1.40 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (78.31 mg, 1.40 mmol, 5 eq) was added, then the mixture was stirred t 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 11 (150 mg, crude) was obtained as yellow solid. LCMS: m/z = 434.1 (M+H4).
To a solution of intermediate 11 (100 mg, 230.67 umol, 1 eq) in pyridine (0.5 mL) and MeCN 0.5 mL) was cooled to 0 °C, then the SCb-pyridine (110.14 mg, 692.00 umol, 3 eq) was added at °C and the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (4 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 36%-66%,10min) o yield Compound IS (12 mg, 10%) was obtained as white solid. LCMS: m/z = 514.1 (M+H+). H-NMR: (400 MHz, DMSO-de) δ = 8.98 - 9.14 (m, 2 H) 8.68 (s, 1 H) 8.32 (d, J= 7.72 Hz, 1 H) .58 - 7.91 (m, 1 H) 7.37 (s, 1 H) 7.07 - 7.20 (m, 3 H) 6.73 - 7.00 (m, 5 H) 5.08 - 5.10 (m, , 1 H) .36 (s, 2 H) 2.91 - 3.00 (m, 2 H) 2.78 - 2.82 (m, 4 H).
A mixture of intermediate 1 (3 g, 7.75 mmol, 1 eq) and intermediate 2 (915.75 mg, 5.92 mmol, 7.64 e'1 eq, HC1) in MeCN (20 mL) were degassed and purged with N2 for 3 times, and then the mixture was stirred at 90 °C for 13 h under N2 atmosphere. The reaction was monitored by LCMS nd HPLC and when complete, the solution was concentrated in vacuo. The residue was purified y prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water (0.05% mmonia hydroxide v/v)-ACN];B%: 5%-48%,20min) to yield intermediate 3 (0.7 g, 23%) was btained as a yellow oil. LCMS: m/z = 307.2 (M+H÷).
To a solution of intermediate 3 (0.7 g, 1.81 mmol, 1 eq) and intermediate 4 (327.85 mg, 2.17 mmol, 1.2 eq) in DMF (6 mL) was added TEA (1.10 g, 10.84 mmol, 1.51 mL, 6 eq), and then the olution was cooled to 0 °C and then T3P (862.63 mg, 2.71 mmol, 806.19 uL, 1.5 eq) was added rop-wise, then the mixture was stirred at 25 °C for 13 hr. The reaction was monitored by LCMS when complete, the solution was dried by flowing N2 to yield intermediate 5 (0.794 g, 90%) was btained as a light yellow oil. LCMS: m/z = 440.3 (M+FT).
To a solution of intermediate 5 (0.794 g, 1.81 mmol, 1 eq) in EtOH (4 mL) and H2O (1 mL) was dded NH4CI (483.16 mg, 9.03 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm o 90 °C, Fe (504.41 mg, 9.03 mmol, 5 eq) was added, then the solution was heated to 90 °C for h. The reaction was monitored by LCMS and HPLC and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC column: Waters Xbridge 150*25 5u;mobile phase: [water(10MmNH4HCO3)-ACN];B%: 5%- 5%,10min) to yield intermediate 6 (600 mg, 81%) was obtained as a colorless oil. LCMS: m/z 410.2 (M+H+).
To a solution of intermediate 6 (0.6 g, 1.47 mmol, 1 eq) in pyridine (5 mL) and MeCN (5 mL) was added SCb-pyridine (699.54 mg, 4.40 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for .17 hr. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Phenomenex Gemini C18 250*50 10u;mobile phase: water(0.04%NH3H20)-ACN];B%: 0%-30%,21min) to yield Compound 16 (222.23 mg, 30%) was obtained as a white solid. LCMS: m/z = 490.2 (M+FT), 1H-NMR: (400 MHz, DMSO-fd6) δ 8.44 (d, J=5.99 Hz, 2 H) 8.33 (d, J=8.44 Hz, 1 H) 7.53 - 7.92 (m, 1 H) 7.37 (s, 1 H) 7.20 (d, J=5.87 Hz, 2 H) 6.89 (d, J=8.44 Hz, 2 H) 6.80 (d, J=8.44 Hz, 2 H) 5.10 - 5.22 (m, 1 H) 4.48 (s, 2 H) 2.95 - 3.07 (m, 1 H) 2.78 - 2.90 (m, 3 H) 2.70 (s, 6 H) 2.44 - 2.49 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide sah (200 mg, 603.48 umol, 1 eq) and intermediate 2 (100.35 mg, 663.83 umol, .1 eq) in DMF (4 mL) was added EtgN (366.40 mg, 3.62 mmol, 503.98 uL, 6 eq), then cooled to °C, and then T3P (288.02 mg, 905.22 umol, 269.18 uL, 1.5 eq) was added dropwise at 0 °C nder N2, the reaction was stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition ice water (40 mL) at 25 °C, and xtracted with ethyl acetate (3 x 30 mL). The combined organic layer was washed with brine (30 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo, the residue was purified y column chromatography silica gel (S1O2, Petroleum ether: Ethyl acetate = 1: 1 to Dichlorom ethane: Methanol = SO: 1) to yield intermediate 3 (172 mg, 55%) was obtained as a ellow solid. LCMS: m/z = 465.2 (M+H+). IH-NMR: (400 MHz, DMSO-de) δ = 8.45 (d, >8.68 Hz, 1 H) 8.28 - 8.38 (m, 2 H) 8.09 (d, >8.56 Hz, 2 H) 7.71 (d, >5.01 Hz, 1 H) 7.64 - 7.67 (m, 1 H) 7.41 - 7.52 (m, 3 H) 7.27 (s, 1 H) 7.13 - 7.23 (m, 2 H) 5.23 - 5.31 (m, 1 H) 3.28 - 3.32 (m, 3 H) 3.03 - 3.15 (m, 1 H) 2.69 - 2.80 (m, 2 H) 2.42 (td, >7.37, 3.61 Hz, 2 H).
A solution of intermediate 3 (258 mg, 555.37 umol, 1 eq) and NH4CI (148.73 mg,2.78 mmol, 5 q) in EtOH (6 mL) was heated to 90 °C, then Fe (155.07 mg, 2.78 mmol, 5 eq) was added, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the olution was filtered and concentrated in vacuo to yield intermediate 4 (400 mg, crude) was btained as a yellow solid. LCMS: m/z = 435.1 (Μ+ΙΓ).
To a solution of intermediate 4 (255 mg, 586.78 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, added SO3. pyridine (280.18 mg, 1.76 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: YMC- Actus Triart Cl 8 00*30mm*5um;mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 10%-40%,12 min) to ield Compound 17 (74.5 mg, 24%) was obtained as a white solid. LCMS: m/z = 512.9 (M-fT), H-NMR: (400 MHz, DMSO-<¾) δ = 8.35 - 8.45 (m, 2 H) 8.31 (d, >8.56 Hz, 1 H) 7.71 (d, >5.01 Hz, 1 H) 7.57 - 7.66 (m, 2 H) 7.31 (dd, >7.76, 4.95 Hz, 1 H) 7.21 (br s, 1 H) 7.14 - 7.19 m, 2 H) 7.08 (br s, 1 H) 6.96 (br s, 1 H) 6.84 - 6.93 (m, 4 H) 5.02 - 5.18 (m, 1 H) 3.03 (dd, >13.63, 5.69 Hz, 1 H) 2.72 - 2.87 (m, 3 H) 2.40 - 2.47 (m, 2 H).
Synthesis of Compound 18
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (200 mg, 603.48 umol, 1 eq) and intermediate 2 (95.78 mg, 633.65 umol, .05 eq) in DMF (4 mL) was added EtsN (366.40 mg, 3.62 mmol, 503.98 uL, 6 eq), then cooled o 0 °C, and then T3P (288.02 mg, 905.22 umol, 269.18 uL, 1.5 eq) was added dropwise at 0 °C under N2, the reaction was stirred at 25 °C for 13 h. The reaction was monitored by TLC when omplete; the reaction mixture was quenched by addition ice water (30 mL) at 25 °C, extracted with ethyl acetate (3 x 10 mL). The combined organic layer was washed with brine (10 mL), ried with anhydrous NaaSO-t, filtered and concentrated in vacuo, the residue was purified by olumn chromatography silica gel (S1O2, Dichloromethane: Methanol = 100: 1 to 50: 1) to yield ntermediate 3 (154 mg, 49%) was obtained as a white solid. 'H-NMR: (400 MHz, DMSO-de) δ 8.49 (d, >8.56 Hz, 1 H) 8.30 - 8.38 (m, 2 H) 8.11 (d, >8.68 Hz, 2 H) 7.72 (dd, >5.01, 0.98 Hz, 1 H) 7.65 (dd, >3.67, 1.10 Hz, 1 H) 7.48 (d, >8.80 Hz, 2 H) 7.31 (s, 1 H) 7.17 (dd, >5.07, .73 Hz, 1 H) 7.11 (d, >5.87 Hz, 2 H) 5.27 (td, >8.86, 5.62 Hz, 1 H) 3.33 - 3.38 (m, 1 H) 3.14 dd, >13.63, 9.35 Hz, 1 H) 2.75 (t, >7.46 Hz, 2 H) 2.38 - 2.46 (m, 2 H).
A solution of intermediate 3 (228 mg, 490.79 umol, 1 eq) and NH4CI (131.075 mg, 2.78 mmol, 5 q) in EtOH (6 mL) was heated to 90 °C, then Fe (137.04 mg, 2.45 mmol, 5 eq) was added, the mixture was stirred at 90 0 C for 2 h. The reaction was monitored by LCMS when complete, the olution was filtered and concentrated in vacuo to yield intermediate 4 (240 mg, crude) was btained as a yellow solid. LCMS: m/z = 435.2 (M+FT).
To a solution of intermediate 4 (240 mg, 552.26 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, added SCb.pyridine (263.70 mg, 1.66 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: YMC-Actus Triart Cl 8 00*30mm*5um;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 15%-35%,12min) to yield Compound 18 (42.3 mg, 14%) was obtained as a white solid. LCMS: m/z = 515.0 (M+H÷), 1H- NMR: (400 MHz, DMSCMO δ = 8.39 (br d, >5.95 Hz, 2 H) 7.52 - 7.63 (m, 2 H) 7.41 (d, >5.95 Hz, 2 H) 7.08 - 7.16 (m, 2 H) 7.00 (d, >8.16 Hz, 2 H) 6.74 (d, >8.16 Hz, 2 H) 5.04 (dd, >9.15, 5.84 Hz, 1 H) 3.06 (br dd, >13.89, 5.73 Hz, 1 H) 2.82 - 2.92 (m, 2 H) 2.78 (br dd, >13.78, 9.37 Hz, 1 H) 2.42 - 2.44 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide sah { 400 mg, 970.11 umol, 1 eq, HBr) and intermediate 2 (159.16 mg, 1.07 mmol, .1 eq) in DMF (6 mL) was added Et?N (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq), then cooled to °C, and then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 1.5 eq) was added drop-wise t 0 °C under N2, the reaction was stirred at 25 °C for 13 h. The reaction was monitored by LCMS when complete, the reaction mixture was quenched by addition ice water (30 mL) at 25 °C, and hen diluted with ice water (60 mL) and extracted with ethyl acetate (3 x 20 mL). The combined rganic layers were washed with brine (2 x 20 mL), dried with anhydrous NazSO^ filtered and oncentrated in vacuo. The residue was purified by column chromatography silica gel (S1O2, etroleum ether: Ethyl acetate = 1: 1 to Dichloromethane : Methanol = 100: 1) to yield intermediate (383 mg, 85%) was obtained as a yellow solid. LCMS: m/z = 463.1 (M+H+), 1H-NMR: (400 MHz, DMSO-£&) δ = 8.89 (br d, >8.56 Hz, 1 H) 8.60 (d, >5.87 Hz, 2 H) 8.13 (d, >8.56 Hz, 2 H) 7.63 - 7.74 (m, 2 H) 7.44 - 7.57 (m, 5 H) 7.36 (d, >15.89 Hz, 1 H) 7.17 (dd, >4.89, 3.91 Hz, H) 6.89 (d, >15.89 Hz, 1 H) 5.37 - 5.50 (m, 1 H) 3.42 (br dd, >13.57, 5.99 Hz, 1 H) 3.24 - 3.31 m, 1 H).
A solution of intermediate 3 (393 mg, 849.65 umol, 1 eq) and NH4CI (227.375 mg, 4.25 mmol, 5 q) in EtOH (6 mL) was heated to 90 °C, then Fe (237.24 mg, 4.25 mmol, 5 eq) was added, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the olution was filtered and concentrated in vacuo to yield intermediate 4 (371 mg, crude) was btained as a yellow solid. LCMS: m/z = 433.2 (Μ+ΙΓ).
To a solution of intermediate 4 (255 mg, 589.51 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, added SCh.pyridine (281.49 mg, 1.77 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: YMC- Actus Triart Cl 8 00*30mm*5um; mobile phase: [ water( 1 OmM NH4HC O3 )- ACN] ; B%: 15%-35%, 12min) to ield Compound 19 (17 mg, 5%) was obtained as a yellow solid. LCMS: m/z = 511.0 (M-H+), H-NMR: (400 MHz, D20) δ = 8.33 (d, J=4.65 Hz, 2 H) 7.39 (d, J=4.52 Hz, 2 H) 7.14 - 7.26 (m, H) 7.01 (m, 5 H) 6.69 - 6.86 (m, 2 H) 5.32 (m, J=6.42 Hz, 1 H) 3.02 - 3.18 (m, 1 H) 2.85 - 2.98 m, 1 H).
Synthesis of Compound 20
4
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (200 mg, 603.48 umol, 1 eq) and intermediate 2 {99.69 mg, 663.83 umol, 3.17 uL, 1.1 eq) in DMF (4 mL) was added EtsN (366.40 mg, 3.62 mmol, 503.98 uL, 6 eq), then cooled to 0 °C, and then T3P (288.02 mg, 905.22 umol, 269.18 uL, 1.5 eq) was added ropwise at 0 °C under N2, the reaction was stirred at 25 °C for 13 h. The reaction was monitored y LCMS when complete, the reaction mixture was quenched by addition ice water (30 mL) at 5°C, and extracted with ethyl acetate (3 x 10 mL). The combined organic layer was washed with brine (40 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo, the residue was purified by column chromatography silica gel (S1O2, Petroleum ether: Ethyl acetate = 5: 1 to : 1) to yield intermediate 3 (158 mg, 50%) was obtained as a yellow solid. LCMS: m/z = 464.2 M+H*), ¾-NMR: (400 MHz, DMSO^) δ = 8.42 (d, >8.68 Hz, 1 H) 8.10 (d, >8.68 Hz, 2 H) .69 - 7.75 (m, 1 H) 7.61 - 7.67 (m, 1 H) 7.47 (d, >8.68 Hz, 2 H) 7.06 - 7.26 (m, 7 H) 5.13 - .36 (m, 1 H) 3.37 - 3.40 (m, 1 H) 3.13 (dd, >13.63, 9.35 Hz, 1 H) 2.73 (t, >7.52 Hz, 2 H) 2.35 2.43 (m, 2 H).
A solution of intermediate 3 (237 mg, 511.25 umol, 1 eq) and NH4CI (136.96 mg, 2.78 mmol, 5 q) in EtOH (8 mL) and H2O (3 mL) was heated to 90 °C, then Fe (142.75 mg, 2.56 mmol, 5 eq) was added, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when omplete, the solution was filtered and concentrated in vacuo to yield intermediate 4 (312 mg, rude) was obtained as a yellow solid. LCMS: m/z = 434.1 (M+FT).
To a solution of intermediate 4 (312 mg, 719.58 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, added SCb.pyridine (343.59 mg, 2.16 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell Cl 8 150*25 5u; mobile phase: [water(10mM NH4HCC>3)-ACN]; B%: 22%-52%,10min) to yield Compound 20 62 mg, 16%) was obtained as a white solid. LCMS: m/z = 514.0 (M+H+), ‘H-NMR: (400 MHz, DMSO-<fc) δ = 8.26 (d, >8.44 Hz, 1 H) 7.70 (dd, >5.07, 1.04 Hz, 1 H) 7.63 (dd, >3.67, .98 Hz, 2 H) 6.97 - 7.36 (m, 11 H) 6.85 - 6.91(m, 4 H) 5.06 - 5.11 (m, 1 H) 3.02 - 3.07 (m, 1 H) .81 - 2.86 (m, 1 H) 2.75 - 2.78 (m, 2 H) 2.38 - 2.42 (m, 2 H).
Synthesis of Compound 21
4
To a solution of intermediate 1 (0.3 g, 727.76 umol, 1 eq) and intermediate 2 (109.29 mg, 27.76 umol, 102.14 uL, 1 eq) in DMF (3 mL) was added EtsN (441.85 mg, 4.37 mmol, 607.77 L, 6 eq), then cooled to 0 °C, and then T3P (926.24 mg, 1.46 mmol, 865.64 uL, 50% purity, 2 q) was added dropwise at 0 °C under N2, the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS when complete, the reaction mixture was quenched by addition ice water (30 mL) at 25 °C, and then diluted with ice water (20 mL) and extracted with ethyl acetate 3 x 40 mL). The combined organic layers were washed with brine (2 x 30 mL), dried with nhydrous NazSO-i, filtered and concentrated in vacuo to yield intermediate 3 (360 mg, crude) was obtained as a brown solid. LCMS: m/z =464.2 (M+H+). 1H-NMR: (400 MHz, Methanol-^) = 8.06 (d, >8.68 Hz, 2 H) 7.33 (d, >8.68 Hz, 2 H) 7.18 - 7.23 (m, 2 H) 7.10 - 7.16 (m, 3 H) .03 (s, 1 H) 5.34 - 5.41 (m, 1 H) 3.95 - 4.08 (m, 2 H) 3.37 (dd, >13.57, 6.11 Hz, 2 H) 3.11 (dd, >13.69, 8.80 Hz, 1 H) 2.79 - 2.86 (m, 2 H) 2.43 - 2.52 (m, 2 H).
A solution of intermediate 3 (0.35 g, 755.17 umol, 1 eq) in ethyl acetate (3.5 mL) was heated to 0 °C, then SnCl2.2H20 (852.01 mg, 3.78 mmol, 5 eq) was added, the mixture was stirred at 50 C for 2 h. The reaction was monitored by LCMS when complete, potassium sodium tartrate (15 mL) was added into the solution and was filtered and concentrated in vacuo to yield intermediate 4 (150 mg, crude) was obtained as a brown solid. LCMS: m/z = 434.2 (M+H+), *H NMR (400 MHz, DMSO-ifc) 5 = 8.31 (d, >8.68 Hz, 1 H) 7.05 - 7.32 (m, 8 H) 6.97 (d, >8.19 Hz, 2 H) 6.73 br d, >8.07 Hz, 2 H) 4.99 - 5.19 (m, 1 H) 4.15 - 4.31 (m, 2 H) 3.06 (dd, >13.82, 5.75 Hz, 1 H) .78 - 2.87 (m, 1 H) 2.71 - 2.75 (m, 2 H) 2.34 - 2.41 (m, 2 H).
To a solution of intermediate 4 (150 mg, 346.03 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SCb.pyridine (165.23 mg, 1.04 mmol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20)-ACN];B%: l%-40%,10min) to yield Compound 21 (60.3 mg, 3% yield) was obtained as a white solid. LCMS: m/z = 514.1 (M+H4), 1H-NMR: (400 MHz, DMSO-de) δ = 8.22 (d, >8.44 Hz, 1 H) 7.61 (s, 1 H) 7.20 - 7.28 (m, 2 H) 7.13 - 7.19 (m, 4 H) .06 (br s, 4 H) 6.85 - 6.91 (m, 2 H) 6.75 - 6.81 (m, 2 H) 4.98 - 5.16 (m, 1 H) 4.10 - 4.29 (m, 2 H) 3.00 (dd, >13.82, 5.99 Hz, 1 H) 2.72 - 2.84 (m, 3 H) 2.36 - 2.43 (m, 2 H).
Synthesis of Compound 22
A mixture of intermediate 1 (500 mg, 1.29 mmol, 1 eq) and intermediate 2 (130.63 mg, 1.29 mmol, 1 eq) in MeCN (15 mL) was heated to 90 °C, and then the reaction was stirred at 90 °C or 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was oncentrated in vacuo to yield intermediate 3 (500 mg, crude) was obtained as a light yellow olid. LCMS: m/z = 390.1 (M+H+).
A mixture of intermediate 3 (502 mg, 1.29 mmol, 1 eq) in HCl/EtOAc (4 M, 10 mL) was stirred t 25 °C for 1 h. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to yield intermediate 4 (250 mg, crude) was obtained as a light ellow solid. LCMS: m/z = 290.1 (M+H+), ‘H-NMR: (400MHz, Methanol^») δ = 8.14 (d, >8.56 Hz, 2 H) 7.36 (d, >8.68 Hz, 2 H) 7.17 (s, 1 H) 3.54 - 3.67 (m, 1 H) 3.43 - 3.44 (m, 2 H) .37 - 2.47 (m, 1 H) 1.17 - 1.22 (m, 2 H) 1.08 - 1.13 (m, 2 H).
To a solution of intermediate 4 (250 mg, 767.31 umol, 1 eq, HC1) and intermediate S (115.23 mg, 767.31 umol, 107.69 uL, 1 eq) in DMF (3 mL) was added EtsN (465.87 mg, 4.60 mmol, 40.80 uL, 6 eq) and then the mixture was cooled to 0 °C, T3P (976.57 mg, 1.53 mmol, 912.68 L, 50% purity, 2 eq) was added at 0 °C, then the mixture was allowed to warm to 25 °C and tirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ¾0 (20 mL), and extracted with ethyl acetate (2 x 10 mL), the ombined organic layer was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered nd concentrated in vacuo to yield intermediate 6 (200 mg, crude) was obtained as a light yellow olid. LCMS: m/z = 422.2 (M+H+).
To a solution of intermediate 6 (150 mg, 355.86 umol, 1 eq) in EtOAc (10 mL) was added nCh.2H20 (401.50 mg, 1.78 mmol, 5 eq), then the mixture was heated to 50 °C and stirred for h. The reaction was monitored by LCMS and when complete, the reaction mixture was added at. seignette salt (20 mL), then filtered and the filtrate was extracted with ethyl acetate (2 x 10 mL), the combined oiganic layer was washed with brine (5 mL), then dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield intermediate 7 (90 mg, crude) was obtained s a light yellow solid. LCMS: m/z = 392.2 (M+H1").
A mixture of intermediate 7 (0.09 g, 229.87 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SCb-pyridine (109.76 mg, 689.61 umol, 3 eq) was added. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, 7% ammonium hydroxide (1 mL) was added dropwise, then the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 u;mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 15%-45%,10min) to yield Compound 2 (32.59 mg, 30%) was obtained as a white solid. LCMS: m/z = 470.0 (M-H+), 1H-NMR: (400 MHz, DMSO-ifc) 5= 8.14 (d, >8.56 Hz, 1 H) 7.20 - 7.27 (m, 3 H) 7.12 - 7.19 (m, 3 H) 7.08 (s, 1 H) 6.96 (s, 1 H) 6.86 - 6.91 (m, 2 H) 6.76 - 6.83 (m, 3 H) 4.94 - 5.08 (m, 1 H) 2.94 - 2.99 (m, 1 H) 2.69 - 2.85 (m, 3 H) 2.34 - 2.43 (m, 3 H) 1.05 - 1.17 (m, 2 H) 0.87 - 1.00 (m, 2 H).
Synthesis of Compound 23
Preparation of Intermediate 2
CDI (1.78 g, 10.99 mmol, 1.1 eg) in DCM (10 mL) was poured into a solution of intermediate 7 1 g, 9.99 mmol, 1 eq) in DCM (2 mL) at 0 °C, then the mixture was stirred at 25 °C for 0.5 h, hen N¾ (gas) was bubbled to the mixture for 1 h at 0 °C. The reaction was monitored by TLC nd when complete, the reaction mixture neutralized with aq. HC1 (10%, 20 mL), extracted with thyl acetate (3 x 20 mL), and then combined organic phase was washed with brine (10 mL), ried with anhydrous Na2$04 and concentrated in vacuo to yield intermediate 8 (0.35 g, crude) was obtained as yellow solid.. Ή-NMR: (400 MHz, DMSO-i&) δ = 7.15 (s, 1 H) 6.71 (s, 1 H) 1.93 (d, >7.09 Hz, 2 H) 0.88 - 1.01 (m, 1 H) 0.37 - 0.50 (m, 2 H) 0.00 - 0.19 (m, 2 H). A mixture of intermediate 8 (0.34 g, 3.43 mmol, 1 eq) and lawesson’s reagent (1.39 g, 3.43 mmol, eq) in THF (3.5 mL) was degassed and purged with Nz for 3 times, and then the mixture was tirred at 40 °C for 16 h. The reaction was monitored by LCMS and when complete, the reaction mixture neutralized with sat. NaHCCb (10 mL) extracted with ethyl acetate (2 x 10 mL), and then ombined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered nd concentrated in vacuo to yield intermediate 2 (0.16 g, crude) was obtained as yellow solid. Ή-ΝΜΚ: (400 MHz, DMSO-ifc) δ = 9.34 (s, 1 H) 9.05 (s, 1 H) 2.37 (d, J= 7.09 Hz, 2 H) 1.07 - .14 (m, 1 H) 0.36 - 0.52 (m, 2 H) 0.15 - 0.19 (m, 2 H).
To a solution of intermediate 1 (0.15 g, 387.37 umol, 1 eq) and intermediate 2 (44.62 mg, 87.37 umol, 1 eq) in MeCN (5 mL) was stirred at 90 °C for 13 h. The reaction was monitored y LCMS and when complete, the reaction mixture was concentrated in vacuo to yield ntermediate 3 (0.17 g, crude) as a yellow solid. LCMS: m/z = 304.0 (M+H+).
To a solution of intermediate 3 (0.16 g, 416.35 umol, 1 eq) and intermediate 4 (62.53 mg, 16.35 umol, 58.44 uL, 1 e) in DMF (3 mL) was added EtsN (252.78 mg, 2.50 mmol, 347.71 uL, eq), and then the solution was cooled to 0 °C, then T3P (529.90 mg, 832.70 umol, 495.23 uL, 0% purity, 2 eq) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was dried with flowing N2 to yield intermediate S (0.25 g, crude) was obtained as a yellow solid, LCMS: m/z = 436.1 M+ET). ntermediate 5 (0.24 g, 551.04 umol, 1 eq) was dissolved in EtOAc (3 mL), and then the nCh.2H20 (746.05 mg, 3.31 mmol, 6 eq) was added and stirred at 50 °C for 13 h. The reaction was monitored by LCMS and when complete, sat. Potassium Sodium tartrate solution (20 mL) was poured into the reaction mixture, then the mixture was filtered and the aqueous phase was xtracted with ethyl acetate (2 x 10 mL), the combined organic phase was washed with brine (20 mL) and dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield intermediate (85 mg, crude) was obtained as a yellow solid. LCMS: m/z = 406.3 (Μ+ΕΓ).
To a solution of intermediate 6 (0.08 g, 197.26 umol, 1 eq) in pyridine (0.5 mL) and MeCN (0.5 mL) was cooled to 0 °C, then the SCb-pyridine (94.19 mg, 591.78 umol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7 % ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H2O+ 1 OmM NH4HCO3)- ACN ] to yield Compound 23 (20 mg, 21%) was obtained as light yellow solid. LCMS: m/z = 486.1 (M+H+). H-NMR: (400 MHz, DMSO-<fc) δ = 8.18 (d, >8.68 Hz, 1 H) 7.19 - 7.27 (m, 3 H) 7.12 - 7.18 m, 3 H) 7.08 (s, 1 H) 6.91 - 6.98 (m, 2 H) 6.85 - 6.90 (m, 2 H) 6.78 - 6.84 (m, 2 H) 4.99 - 5.11 m, 1 H) 2.95 - 3.04 (m, 1 H) 2.87 (d, >6.97 Hz, 2 H) 2.72 - 2.81 (m, 3 H) 2.35 - 2.42 (m, 2 H) .03 - 1.17 (m, 1 H) 0.51 - 0.63 (m, 2 H) 0.26 - 0.33 (m, 2 H).
To a mixture of intermediate 1 (2 g, 15.60 mmol, 1.96 mL, 1 eq) in DCM (20 mL) was added OCI2 (6.31 g, 53.02 mmol, 3.85 mL, 3.40 eq) at 0 °C, then warmed to 25 °C and stirred for 2 h at 5 °C under N2 atmosphere. The reaction mixture was concentrated in vacuo to yield intermediate (2.5 g, crude) was obtained as a yellow oil.
To a mixture of intermediate 2 (2.5 g, 17.05 mmol, 1.96 mL, 1 eq) in DCM (20 mL) was added NH3 (290.39 mg, 17.05 mmol, 1 eq) at 0 °C, then warmed to 25 °C and stirred for 20 min at 25 °C under N2 atmosphere. The mixture was poured into water (50 mL) and then the mixture was extracted with DCM (2 x 10 mL). The combined organic phase was washed with HC1 solution (1 N, 10 mL), sat. NaHCOs solution (10 mL) and brine (10 mL), dried with anhydrous NaaSO*, filtered and concentrated in vacuo to yield intermediate 3 (1 g, 46%) was obtained as a yellow olid. 1H-NMR: (400 MHz, DMSO-tife) 5 = 7.21 (s, 1 H) 6.66 (s, 1 H) 2.07 - 2.20 (m, 1 H) 1.95 - .03 (m, 2 H) 1.66 - 1.75 (m, 2 H) 1.40 - 1.64 (m, 4 H) 1.03 - 1.30 (m, 2 H).
To a mixture of intermediate 3 (1 g, 7.86 mmol, 1.96 mL, 1 eq) in THF (10 mL) was added Lawesson’s reagent (3.18 g, 7.86 mmol, 1 eq) at 25 °C and stirred at 25 °C for 16 h under N2 tmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured nto water (50 mL) and then the mixture was extracted with dichloromethane (2 x 10 mL). The ombined organic phase was washed with sat. NaHCOs solution (10 mL) and brine (10 mL), ried with anhydrous Na2S04, filtered and concentrated in vacuo to yield intermediate 4 (1.5 g, rude) was obtained as a yellow oil. LCMS: m/z = 144.1 (M+H+).
A mixture of (S)-tert-butyl-(4-bromo-l-(4-nitrophenyl)-3-oxobutan-2-yl)-carbamate (2.97 g, .68 mmol, 1.00 eq) and intermediate 4 (1.1 g, 7.68 mmol, 1 eq) in MeCN (50 mL) was heated o 90 °C and stirred for 16 h at 90 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to yield intermediate (3.2 g, crude, HBr salt) was obtained as a black brown oil. LCMS: m/z = 332.2 (M+fF).
To a mixture of intermediate 6 (400 mg, 970.06 umol, 1 eq, HBr) and intermediate 7 (145.68 mg, 70.06 umol, 1 eq) inDMF (5 mL) was dropwise added EtgN (588.96mg, 5.82 mmol, 810.13 uL, eq) and cooled to 0 °C, then T3P (925.97 mg, 1.46 mmol, 865.39 uL, 50% purity, 1.5 eq) was dded at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 hours at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete, the mixture was poured nto ice-water (25 mL) and stirred for 10 minutes. Then the mixture was extracted with ethyl cetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with nhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by column hromatography (S1O2, Petroleum ether: Ethyl acetate = 10: 1 to 2: 1) to yield intermediate 8 (370 mg, 82%) was obtained as a yellow solid. LCMS: m/z = 464.2 (M+H+), ^-NMR: (400 MHz, CDCh) δ = 8.04 - 8.06 (m, 1 H) 7.18 - 7.37 (m, 6 H) 7.10 (d, >8.68 Hz, 2 H) 6.63 (s, 1 H) 6.49 d, >8.19 Hz, 1 H) 5.26 - 5.39 (m, 1 H) 3.25 - 3.32 (m, 1 H) 3.11 - 3.19 (m, 1 H) 2.95 - 3.04 (m, H) 2.48 - 2.59 (m, 2 H) 2.22 - 2.33 (m, 1 H) 1.76 - 1.88 (m, 2 H) 1.67 - 1.75 (m, 2 H) 1.58 - 1.66 (m, 2 H) 1.25 - 1.34 (m, 2 H). A solution of intermediate 8 (370 mg, 798.12 umol, 1 eq) and NH4CI (213.46 mg, 3.99 mmol, 5 q) in EtOH (12 mL) and Η20 (4 mL) was heated to 90 °C and then Fe (222.85 mg, 3.99 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 9 (345 mg, crude) was obtained as a yellow solid. LCMS: m/z = 434.3 (M+H4).
A solution of intermediate 3 (345 mg, 795.65 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCh-pyridine (379.91 mg, 2.39 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop- wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10MmNH4HC03)-ACN]; B%: 15%-50%,10min) to yield Compound 24 (122.02 mg, 28%) was obtained as a light yellow solid. LCMS: m/z = 512.1 (M- H4), 1H-NMR: (400 MHz, DMSO-dk) δ = 8.18 (d, >8.56 Hz, 1 H) 7.21 - 7.27 (m, 3 H) 7.12 - .19 (m, 3 H) 7.08 (s, 1 H) 6.96 (s, 1 H) 6.90 (s, 1 H) 6.87 (d, >8.44 Hz, 2 H) 6.77 (d, >8.31 Hz, 2 H) 4.98 - 5.10 (m, 1 H) 2.93 - 3.02 (m, 3 H) 2.72 - 2.81 (m, 3 H) 2.36 - 2.42 (m, 2 H) 2.19 2.27 (m, 1 H) 1.70 - 1.80 (m, 2 H) 1.48 - 1.65 (m, 4 H) 1.20 - 1.30 (m, 2 H).
Synthesis of Compound 25
Preparation of intermediate 2
To a solution of intermediate 2A (1 g, 14.06 mmol, 1.17 mL, 1 eq) in CH3CI (10 mL) was added solution of isothiocyanato(trimethyl)silane (2.40 g, 18.28 mmol, 1.3 eq) drop-wise under N2. Then the reaction mixture was stirred at 25 °C for 16 h. MeOH (50 mL) was added and stirred or 1 hat 25 °C. Then the mixture was concentrated in vacuo to yield intermediate 2 (0.65 g, rude) was obtained as a yellow solid. LCMS: m/z = 131.0 (M+H+).
A mixture of intermediate 1 (500.86 mg, 1.23 mmol, 1 eq) and intermediate 2 (0.2 g, 1.23 mmol, 117.37 uL, 1 eq) in EtOH (10 mL) was heated to 85 °C and stirred for 2 h.
The reaction was monitored by TLC and LCMS and when complete, the mixture was cooled and oncentrated in vacuo to yield intermediate 5 (1.0 g, crude) was obtained as a yellow solid. LCMS: m/z = 419.1 (M+H+). The solution of intermediate 3 (1.0 g, 1.15 mmol, 1 eq) in HCl/EtOAc (4 mL) was stirred at 25 C for 5 min. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered and concentrated in vacuo, the solid was washed with ethyl acetate (5 mL) o yield intermediate 4 (0.4 g, 81%) was obtained as a white solid. LCMS: m/z = 319.2 (M+H+), H-NMR: (400 MHz, DMSO-ffc) 6 = 8.58 (s, 2 H) 8.15 (d, >8.559 Hz, 2 H) 7.42 - 7.52 (m, 2 H) 6.69 (s, 1 H) 4.50 - 4.60 (m, 1 H) 3.39 - 3.43 (m, 3 H) 3.02 - 3.13 (m, 3 H) 1.99 - 2.02 (m, 2 H) 1.80 - 1.84 (m, 2 H).
To a solution of intermediate 4 (0.4 g, 1.03 mmol, 1 eq, HC1) and intermediate 5 (201.12 mg, .34 mmol, 1.3 eq) in DMF (2 mL) was added EtsN (625.46 mg, 6.18 mmol, 860.33 uL, 6 eq), nd the mixture was cooled to 0 °C, then T3P (983.34 mg, 1.55 mmol, 919.01 uL, 50% purity, .5 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for h. The reaction was monitored by LCMS and when complete, the reaction mixture was uenched with H2O (20 mL) and then filtered, the filtrate was concentrated in vacuo. The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 1 : 1) to yield intermediate 6 (0.45 g, 8%) was obtained as a yellow solid. LCMS: m/z = 451.2 (M+fT).
To a solution of intermediate 6 (462.37 mg, 513.11 umol, 1 eq) in EtOH (5 mL) and H20 (1 mL) was added NH4CI (164.68 mg, 3.08 mmol, 107.63 uL, 6 eq), and then the solution was heated to 0 °C, Fe (171.94 mg, 3.08 mmol, 6 eq) was added, then the reaction mixture was stirred at 90 C for 2 h. The reaction was monitored by TLC and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo. The residue was purified by prep-TLC (Petroleum ther: Ethyl acetate = 0: 1) to yield intermediate 7 (0.17 g, 71%) was obtained as a yellow solid.
A solution of intermediate 7 (200 mg, 475.55 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SOj-pyridine (227.07 mg, 1.43 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell Cl 8 150*25 u;mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 20%-50%,10min) to yield Compound 5 (70.66 mg, 29%) was obtained as a white solid. LCMS: m/z = 499.1 (M-fT), 1H-NMR: (400 MHz, DMSO-i¾) δ = 7.19 - 7.26 (m, 2 H) 7.09 - 7.17 (m, 3 H) 6.85 - 6.95 (m, 4 H) 6.18 (s, 1 H) .80 - 4.90 (m, 1 H) 3.30 - 3.40 (m, 4 H) 2.95 (dd, >13.82, 5.38 Hz, 1 H) 2.63 - 2.75 (m, 3 H) .31 - 2.42 (m, 2 H) 1.90 - 2.05 (m, 4 H).
Synthesis of Compound 26
A mixture of (S)-tert-butyl-(4-bromo-l-(4-nitrophenyl)-3-oxobutan-2-yI)-carbamate (2 g, 5.16 mmol, 1 eg) and intermediate 2 (471.79 mg, 6.20 mmol, 117.37 uL, 1.2 eg) in EtOH (10 mL) was heated to 85 °C and stirred for 2 h. The reaction was monitored by TLC and LCMS and when complete, the mixture was cooled and concentrated in vacuo to yield intermediate 3 (1.8 g, 4%) was obtained as a white solid. LCMS: m/z = 365.0 (M+H+). The solution of intermediate 3 (1.7 g, 3.31 mmol, 1 eq) in HCl/EtOAc (4 mL) was stirred at 25 C for 2 h. The reaction was monitored by TLC and LCMS and when complete, the reaction was ltered and the filter cake was washed with ethyl acetate (5 mL) and then dried in vacuo to yield ntermediate 4 (0.85 g, 85%, HC1 salt) was obtained as a white solid. LCMS: m/z = 264.9 M+H^), ‘H-NMR: (400 MHz, DMSO-dfe) 5 = 8.73 - 8.95 (m, 3 H) 8.17 (d, >8.56 Hz, 2 H) .49 (d, >8.56 Hz, 2 H) 6.84 (s, 1 H) 4.53 - 4.69 (m, 1 H) 3.28 - 3.50 (m, 2 H).
To a solution of intermediate 4A (439.39 mg, 2.93 mmol, 410.65 uL, 1.1 eq ) and intermediate 4 0.8 g, 2.66 mmol, 1 eq, HC1) in DMF (10 mL) was added a mixture ofEtgN (565.22 mg, 5.59 mmol, 777.48 uL, 2.1 eq) and HATU (1.21 g, 3.19 mmol, 1.2 eq) drop-wise at 0 °C under Nz tmosphere. During which the temperature was maintained below 0 °C. Then the reaction mixture was warmed to 25 °C and stirred at 25 °C for 2 h. The reaction was monitored by TLC nd LCMS when complete, the reaction mixture was quenched by H2O (20 mL) and then ltered. The filter cake was dried in vacuo. The residue was purified by prep-TLC (Petroleum ther: Ethyl acetate = 1 : 1) to yield intermediate 5 (0.89 g, 42%) was obtained as a yellow solid. LCMS: m/z = 397.2 (M+H+).
To a mixture of intermediate 5 (0.3 g, 756.69 umol, 1 eq) and O1CI2 (162.78 mg, 1.21 mmol, 1.6 q) in MeCN (10 mL) was added tert-butyl nitrite (117.05 mg, 1.14 mmol, 135.00 uL, 1.5 eq) rop-wise at 0 °C under N2. The mixture was stirred for 2 h at 0 °C. The reaction was monitored y TLC and LCMS when complete, the reaction was quenched by H2O (50 mL) and then xtracted with ethyl acetate (2 x 10 mL). The combined organic phase was washed with brine 10 mL), dried with anhydrous NazSO,*, filtered and concentrated in vacuo. The residue was urified by prep-TLC (Petroleum ether: Ethyl acetate = 3: 1) to yield intermediate 7 (0.11 g, 8%) was obtained as a yellow solid. LCMS: m/z = 416.2 (M+H+), 1H-NMR: (400 MHz, CDCb) δ = 8.06 - 8.08 (m, 2 H) 7.19 - 7.26 (m, 3 H) 7.12 - 7.18 (m, 4 H) 6.65 (s, 1 H) 6.02 (br d, >8.56 Hz, 1 H) 5.24 - 5.30 (m, 1 H) 3.19 - 3.25 (m, 1 H) 3.01 - 3.13 (m, 1 H) 2.91 - 2.96 (m, 2 H) 2.44 - 2.53 (m, 2 H).
To a solution of intermediate 8 (35.40 mg, 406.36 umol, 32.78 uL, 1.3 eq) and intermediate 7 0.13 g, 312.58 umol, 1 eq) in DMSO (4 mL) was added a solution of KI (5.19 mg, 31.26 umol, .1 eq) and DBU (95.17 mg, 625.16 umol, 94.23 uL, 2 eq) drop wise under N2 atmosphere. The reaction mixture was heated to 130 °C and stirred at 130 °C for 1 h under microwave. The eaction was monitored by TLC and LCMS when complete, the reaction was quenched by water 20 mL) and then extracted with ethyl acetate (2 x 5 mL). The combined organic phase was washed with brine (3 mL), dried with anhydrous NaaSCU, filtered and concentrated in vacuo. The esidue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 0: 1) to yield intermediate 9 0.11 g, 72%) was obtained as a yellow solid. LCMS: m/z = 467.2 (M+H+). ‘H-NMR: (400 MHz, CDC13) δ = 8.03 - 8.05 (m, 2 H) 7.27 - 7.29 (m, 1 H) 7.24 - 7.26 (m, 1 H) 7.17 - 7.23 (m, 3 H) 7.11 - 7.17 (m, 2 H) 6.26 (br d, >7.95 Hz, 1 H) 5.86 - 5.90 (m, 1 H) 4.99 - 5.11 (m, 1 H) 4.60 4.71 (m, 1 H) 3.58 - 3.68 (m, 2 H) 3.48 - 3.56 (m, 2 H) 3.43 - 3.46 (m, 1 H) 3.18 - 3.22 (m, 1 H) .05 - 3.10 (m, 1 H) 2.94 - 3.01 (m, 2 H) 2.48 - 2.52 (m, 2 H) 2.18 - 2.24 (m, 1 H) 2.08 - 2.16 (m, H).
To a solution of intermediate 9 (0.11 g, 223.98 umol, 1 eq) in EtOH (5 mL) and H2O (1 mL) was dded NH4CI (71.89 mg, 1.34 mmol, 46.98 uL, 6 eq) and heated to 90 °C, then Fe (75.06 mg, .34 mmol, 6 eq) was added. The reaction mixture was stirred at 90 °C for 2 h. The reaction was monitored by TLC and when complete, the reaction was filtered and concentrated in vacuum.
The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 0:1) to yield ntermediate 10 (0.08 g, 79%) was obtained as a yellow oil.
To a solution of intermediate 5 (80 mg, 183.25 umol, 1 eq) in pyridine (1 mL) and CH3CN (1 mL) was cooled to 0 °C, then the S03-pyridine (87.50 mg, 549.74 umol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7% ammonium hydroxide (2 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbiidge 50*25 5u;mobile phase: [water(0.04%NH3H20+10niM NH4HC03)-ACN]; B%: l%-30%, Omin) to yield Compound 26 (25.3 mg, 26%) was obtained as a light yellow solid. LCMS: m/z 515.1 (M-lF). Ή-NMR: (400 MHz, DMSO-tfc) δ = 7.05 - 7.28 (m, 6 H) 6.97 (d, >8.19 Hz, 1 H) 6.83 - 6.92 (m, 1 H) 6.66 (d, >8.19 Hz, 1 H) 6.00 - 6.07 (m, 1 H) 4.76 - 4.87 (m, 1 H) 4.39 br s, 1 H) 3.35 - 3.50 (m, 3 H) 3.20 - 3.22 (m, 1 H) 2.92 - 3.03 (m, 1 H) 2.60 - 2.74 (m, 3 H) .29 - 2.37 (m, 2 H) 2.00 - 2.09 (m, 1 H) 1.87 - 1.91 (m, 1 H). Synthesis of Compound 27 .1 Preparation of intermediate 2
To a solution of intermediate 2A (0.2 g, 1.45 mmol, 1.17 mL, 1 eq, HC1) in THF (4 mL) was dded a solution of di(imidazol- 1 -yl)methanethione (336.72 mg, 1.89 mmol, 1.3 eq) drop-wise nder N2. The reaction mixture was stirred at 25 °C for 16 h. The reaction was monitored by LCMS and when complete, MeOH (50 mL) was added and stirred for 1 h at 25 °C. Then the mixture was concentrated in vacuum. The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 0: 1) to yield intermediate 2 (70 mg, 15%) was obtained as a yellow solid. LCMS: m/z = 161.1 (Μ+ΙΓ).
A mixture of ( S)-tert-butyl-(4-bromo-l-(4-nitrophenyl)-3-oxobutan-2-yl)-carbamate (457.88 mg, 1.12 mmol, 1 eq) and intermediate 2 (0.2 g, 1.12 mmol, 117.37 uL, 1 eq) in EtOH (10 mL) was heated to 85 °C and stirred for 2 h. The reaction was monitored by LCMS and when complete, the mixture was cooled and concentrated in vacuo. The residue was triturated with thyl acetate (5 mL) and dried in vacuo to yield intermediate 3 (0.7 g, 83%) was obtained as a ellow solid. LCMS: m/z = 449.3 (M+H+)
The solution of intermediate 3 (0.8 g, 1.07 mmol, 1 eq) in HCl/EtOAc (4 mL) was stirred at 25 C for 5 min. The reaction was monitored by LCMS and when complete, the reaction was filtered and concentrated in vacuo. The residue was washed with ethyl acetate (5 mL) and dried n vacuo to yield intermediate 3 (0.6 g, 73%, HC1 salt) was obtained as a yellow solid. LCMS: m/z = 349.1 (M+H+)
To a solution of intermediate 4A (254.47 mg, 1.69 mmol, 237.82 uL, 1.2 eq) and intermediate 3 0.6 g, 1.41 mmol, 1 eq, HC1) in DMF (6 mL) was added Et3N (857.34 mg, 8.47 mmol, 1.18 mL, eq), then cooled to 0 °C and T3P (1.35 g, 2.12 mmol, 1.26 mL, 50% purity, 1.5 eq) drop-wise t 0°C under N2. Then the reaction mixture was warmed to 25 °C and stirred at 25 °C for 2 h.
The reaction was monitored by LCMS and when complete, the reaction mixture was quenched y H2O (20 mL) and then filtered, the filtrate was dried in vacuo. The residue was purified by rep-TLC (Petroleum ether: Ethyl acetate = 1 : 1) to yield intermediate 5 (0.6 g, crude) was btained as a black brown oil. LCMS: m/z = 481.3 (M+fL).
To a solution of intermediate 5 (0.6 g, 873.95 umol, 1 eq) in EtOH (5 mL) and H2O (1 mL) was dded NH4CI (280.48 mg, 5.24 mmol, 183.32 uL, 6 eq) and heated to 90 °C, then Fe (292.86 mg, .24 mmol, 6 eq) was added and stirred at 90 °C for 2 h. The reaction was monitored by TLC nd when complete, the reaction was filtered and concentrated in vacuum. The residue was urified by prep-TLC (Petroleum ether/Ethyl acetate = 0:1) to yield intermediate 6 (0.2 g, 48%) was obtained as a yellow solid. LCMS: m/z = 481.3 (M+FT).
To a solution of intermediate 6 (0.25 g, 554.82 umol, 1 eq) in pyridine (2 mL) and CH3CN (2 mL) was cooled to 0 °C, then the SOs-pyridine (264.92 mg, 1.66 mmol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7 % ammonium hydroxide (2 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM N¾HC03)-ACN];B%: 5%- 5%,10min) to yield Compound 27 (183.3 mg, 61%) was obtained as white solid. LCMS: m/z = 29.0 (M-H+). !H-NMR: (400 MHz, DMSO-de) δ =7.16 - 7.24 (m, 2 H) 7.04 - 7.16 (m, 4 H) .80 - 6.96 (m, 4 H) 6.00 (d, >5.14 Hz, 1 H) 4.76 - 4.87 (m, 1 H) 4.07 - 4.08 (m, 1 H) 3.27 - .50 (m, 4 H) 3.22 (s, 3 H) 2.91 - 3.07 (m, 1 H) 2.65 - 2.74 (m, 2 H) 2.56 - 2.65 (m, 1 H) 2.29 - .40 (m, 2 H) 2.03 - 2.09 (m, 2 H).
Synthesis of Compound 28
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (0.1 g, 243.73 umol, 1 eq) and intermediate 2 (33.18 mg, 243.73 umol, 30.73 L, 1 eq) in DMF (3 mL) was added TEA (147.98 mg, 1.46 mmol, 203.55 uL, 6 eq), and then the olution was cooled to 0 °C, and then T3P (23.27 g, 365.60 umol, 21.74 mL, 50% purity, 1.5 eq) was added drop-wise, the mixture was stirred at 20 °C for 13 hr. The reaction was monitored by LCMS, and when complete, the solution was poured into cold-water (20 mL) and extracted with thyl acetate (3 x 10 mL), and then combined organic phase was washed with brine (2 x 10 mL), ried with anhydrous NaiSOd and concentrated in vacuo. The residue was purified by prep-TLC Dichloromethane: Methanol = 10: 1) to yield intermediate 3 (80 mg, 73%) was obtained as a white solid. LCMS: m/z = 448.2 (M+H+).
To a solution of intermediate 3 (0.08 g, 178.77 umol, 1 eq) in EtOH (3 mL) and Water (1 mL) was dded NH4CI (47.81 mg, 893.84 umol, 31.25 uL, 5 eq) at 25 °C, and then the solution was heated o 90 °C, and then Fe (49.92 mg, 893.84 umol, 5 eq) was added, the solution was hearted to 90 °C or 2 hr. The reaction was monitored by LCMS and HPLC and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (80 mg, crude) was btained as a colorless oil. LCMS: m/z = 418.2 (M+H4").
To a solution of intermediate 4 (0.07 g, 167.65 umol, 1 eq) in MeCN (1 mL) and pyridine (980.00 mg, 12.39 mmol, 1 mL, 73.90 eq) was added SOg-pyridine (80.05 mg, 502.96 umol, 3 eq) at 0 °C, the mixture was stirred at 0 °C for 10 mins. The reaction was monitored by LCMS and HPLC and when complete, the solution was dried with flowing N2. And the crude was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 5%-35%,10min) to yield Compound 28 (19.8 mg, 23%) was obtained as a white solid. LCMS: m/z = 498.1 (Μ+ΙΓ), ‘H-NMR: (400 MHz, DMSO-i&) δ = 8.74 (s, 1 H) 8.48 (d, 7=8.33 Hz, 1 H) 7.66 - 7.91 (m, 1 H) 7.22 - 7.29 (m, 3 H) 7.12 - 7.17 (m, 5 H) 7.07 - 7.09 (m, 1 H) 6.95 - 6.96 (m, 1 H) 6.86 - 6.92 (m, 2 H) 6.77 - 6.83 (m, 2 H) 5.04 - 5.20 (m, 1 H) 4.38 (s, 2 H) 3.38 - 3.45 (m, 2 H) 2.88 - 2.89 (m, 1 H) 2.52 - 2.70 (m, 1 H).
Preparation of intermediate 2 ntermediate 5 (1.01 g, 6.11 mmol, 935.19 uL, 1 eq) in THF (9 mL) and H2O (6 mL) was added LiOH-HzO (513.15 mg, 12.23 mmol, 2 eq) at 0 °C and then the solution was allowed to warm to 5 °C and stirred for 3 h. The reaction was monitored by TLC and when complete, the solution was concentrated in vacuo to yield intermediate 2 (0.8 g, 91%) was obtained as a white solid. H-NMR: (400 MHz, D20) δ = 8.37 (d, 7=1.75 Hz, 2 H) 7.27 (s, 2 H) 3.52 (d, 7=3.07 Hz, 2 H).
To a solution of (S)-l -(2-((lH-imidazol-S-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl) ethanamine ydrochloride salt (0.1 g, 243.73 umol, 1 eq) and intermediate 2 (34.87 mg, 243.73 umol, 245.80 uL, 1 eq) in DMF (3 mL) was added EtsN (147.98 mg, 1.46 mmol, 203.55 uL, 6 eq), and hen the solution was cooled to 0 °C and T3P (23.27 g, 365.60 umol, 21.74 mL, 50% purity, 1.5 q) was added drop-wise, the mixture was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the solution was poured into cold- water (20 mL) and extracted with ethyl acetate (3 x 10 mL), and then combined organic phase was washed with brine (2 x 10 mL), dried with anhydrous Na2S04, filtered and concentrated in acuo. The residue was purified by prep-TLC (dichloromethane: methyl alcohol = 10: 1) to yield ntermediate 3 (20 mg, 18%) was obtained as a white solid. LCMS: m/z = 449.2 (M+H*). ntermediate 3 (0.03 g, 66.89 umol, 1 eq) in EtOH (3 mL) and H20 (1 mL) was added NH4CI 17.89 mg, 334.45 umol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, Fe (3.74 mg, 6.89 umol, 1 eq) was added at this temperature and then the solution was stirred at 90 °C for 2 . The reaction was monitored by LCMS and HPLC and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (40 mg, crude) was btained as a colorless oil. LCMS: m/z = 419.2 (M+FT).
To a solution of intermediate 4 (40 mg, 95.58 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SOs-pyridine (15.21 mg, 95.58 umol, 1 eq) at 0 °C, the mixture was stirred at 0 °C for 0 min. The reaction was monitored by LCMS and HPLC and when complete, the solution was ried by flowing N2. The residue was purified by prep-HPLC (column: Agela Durashell Cl 8 50*305u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 1%-I5%,10min) to yield Compound 29 (9.04 mg, 18 %) was obtained as a white solid. LCMS: m/z = 497.0 (M-H+), 1H- NMR: (400 MHz, D20) δ = 8.35 (d, >5.26 Hz, 2 H) 7.94 (s, 1 H) 7.15 (s, 1 H) 7.04 - 7.10 (m, 3 H) 6.93 - 7.02 (m, 4 H) 5.19 - 5.23 (m, 1 H) 4.30 (s, 2 H) 3.19 (dd, >13.88, 6.05 Hz, 1 H) 2.94 dd, >13.82, 9.54 Hz, 1 H) 2.67 (s, 2 H).
Synthesis of Compound 31
4
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (100 mg, 273.35 umol, 1.2 eq) and intermediate 2 (42.05 mg, 227.79 umol, 1 q) in DMF (2 mL) was added EtsN (1.37 mmol, 190.23 uL, 6 eq), and the mixture was cooled to °C, then T3P (341.68 umol, 203.21 uL, 50% purity, 1.5 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 18 h. The reaction was monitored by LCMS and when complete, the reaction mixture was poured into water (30 mL), and extracted with ethyl acetate (3 x 10 mL). Then combined organic phase was washed with brine (2 x 10 mL), dried with anhydrous Na2S04 and concentrated in vacuo. The residue was purified by prep- TLC (Dichloromethane : Methanol = 10: 1) to yield intermediate 3 (70 mg, 62%) was obtained s a yellow oil. LCMS: m/z = 496.2 (M+H+), 1H-NMR: (400 MHz, CDCb) δ = 8.02 (d, 7=8.681 Hz, 2 H) 7.66 (br s, 1 H) 7.11 - 7.21 (m, 5 H) 7.02 - 7.06 (m, 1 H) 6.96 (s, 1 H) 6.68 (s, 1 H) 5.28 5.33 (m, 1 H) 3.50 (s, 2 H) 3.24 - 3.33 (m, 1 H) 3.14 - 3.23 (m, 1 H) 2.87 - 2.94 (m, 2 H) 2.46 - .52 (m, 2 H).
A solution of intermediate 3 (70 mg, 141.13 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (37.75 mg, 705.67 umol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, Fe (39.41 mg, 705.67 umol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (90 mg, crude) was btained as a yellow solid. LCMS: m/z = 466.2 (M+H+).
To a solution of intermediate 4 (90 mg, 193.13 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SCb-pyridine (92.22 mg, 579.40 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water (0.05% mmonia hydroxide v/v)-ACN];B%: 5%-30%,l lmin) to yield Compound 31 (14 mg, 13%) was btained as a white solid. LCMS: m/z = 546.0 (M+H+), Ή-NMR: (400 MHz, DMSO -de) δ = 4.07 (br s, 1 H) 8.85 (s, 1 H) 8.23 (d, >8.437 Hz, 1 H) 7.54 - 7.98 (m, 1 H) 7.19 - 7.31 (m, 3 H) .08 - 7.17 (m, 3 H) 6.89 (d, >8.436 Hz, 2 H) 6.77 (d, >8.436 Hz, 2 H) 5.08 - 5.17 (m, 1 H) .38 (s, 2 H) 2.93 (dd, >13.510, 6.908 Hz, 1 H) 2.76 - 2.87 (m, 3 H) 2.40 - 2.46 (m, 2 H).
Synthesis of Compound 32
4
To a solution of of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4- itrophenyl)ethanamine hydrochloride salt (100 mg, 273.35 umol, 1.2 eq) and intermediate 2 (41.05 mg, 227.79 umol, 1 eq) in DMF (2 mL) was added EtgN (1.37 mmol, 190.23 uL, 6 eq), and the mixture was cooled to 0 °C, then T3P (341.68 umol, 203.21 uL, 50% purity, 1.5 eq) was dded drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 18 h. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was oured into water (30 mL), and extracted with ethyl acetate (3 x 10 mL). Then combined organic hase was washed with brine (2 x 10 mL), dried with anhydrous Na2SC>4 and concentrated in acuo. The residue was purified by prep-TLC (Dichloromethane: Methanol = 10: 1) to yield ntermediate 3 (70 mg, 63%) was obtained as a yellow oil. LCMS: m/z = 492.2 (M+H4"), 1H- NMR: (400 MHz, Methanol-^) δ = 8.05 (d, >8.803 Hz, 2 H) 7.73 (s, 1 H) 7.31 (d, >8.681 Hz, H) 7.03 - 7.13 (m, 2 H) 6.88 (s, 1 H) 6.66 - 6.73 (m, 3 H) 5.32 - 5.36 (m, 1 H) 4.30 (s, 2 H)
A solution of intermediate 3 (68 mg, 138.33 rnnol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (37.00 mg, 691.67 umol, 5 eq), and then the solution was heated to 90 °C, Fe 38.63 mg, 691.67 umol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was ltered, the filtrate was concentrated in vacuo to yield intermediate 4 (100 mg, crude) was btained as a yellow solid. LCMS: m/z = 462.2 (M+H1"), 1H-NMR: (400 MHz, Methanol-i¾) δ 8.43 (s, 1 H) 7.29 (s, 1 H) 7.13 (s, 1 H) 6.88 - 6.95 (m, 2 H) 6.71 (m, 6 H) 4.41 (s, 2 H) 3.69 - .77 (m, 3 H) 3.57 - 3.64 (m, 1 H) 3.13 (m, 1 H) 2.71 - 2.92 (m, 3 H) 2.46 (m, 2 H).
To a solution of intermediate 4 (100 mg, 216.65 umol, 1 eq) in pyridine (0.5 mL) and MeCN 0.5 mL) was added SCh-pyridine (103.45 mg, 649.94 umol, 3 eq) at 0 °C and stirred for 0.17 h. The reaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide 2 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Agela Durashell C 18 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 5%-40%,10min) to yield Compound 32 (21.4 mg, 18%) was obtained s a white solid. LCMS: m/z = 542.1 (M+FT), Ή-NMR: (400 MHz, DMSO-dfe) δ = 8.62 (s, 1 H) 8.21 (d, >8.436 Hz, 1 H) 7.70 (br s, 1 H) 7.00 - 7.28 (m, 4 H) 6.86 - 6.92 (m, 2 H) 6.70 - .82 (m, 5 H) 5.06 - 5.16 (m, 1 H) 4.35 (s, 2 H) 3.71 (s, 3 H) 2.92 - 2.97 (m, 1 H) 2.79 - 2.84 (m, H) 2.72 - 2.76 (m, 2 H) 2.37 - 2.43 (m, 2 H).
Synthesis of Compound 33
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride sat (100 mg, 273.35 umol, 1.2 eq) and intermediate 2 (37.40 mg, 227.79 umol, 1 q) in DMF (2 mL) was added EtsN (1.37 mmol, 190.23 uL, 6 eq), and the mixture was cooled to °C, then T3P (341.68 umol, 203.21 uL, 50% purity, 1.5 eq) was added drop-wise, and then the olution was allowed to 25 °C at stirred for 13 h. The reaction was monitored by TLC and LCMS nd when complete, the reaction mixture was poured into water (30 mL), and extracted with thyl acetate (3 x 10 mL). Then combined organic phase was washed with brine (2 x 10 mL), ried with anhydrous NazSC^ and concentrated in vacuo. The residue was purified by prep-TLC Dichloromethane : Methanol = 10: 1) to yield intermediate 3 (80 mg, 74%) was obtained as a ellow oil. LCMS: m/z = 476.2 (M+H4), Ή-NMR: (400 MHz, Methanol-^) δ = 8.05 (d, >8.681 Hz, 2 H) 7.75 (s, 1 H) 7.32 (d, >8.559 Hz, 2 H) 7.03 - 7.10 (m, 2 H) 6.92 - 6.97 (m, 2 H) 6.89 (d, >7.458 Hz, 1 H) 6.83 (s, 1 H) 5.31 - 5.35 (m, 1 H) 4.31 (s, 2 H) 3.33 - 3.39 (m, 1 H) .04 - 3.10 (m, 1 H) 2.74 - 2.81 (m, 2 H) 2.41 - 2.49 (m, 2 H) 2.24 (s, 3 H).
A solution of intermediate 3 (80 mg, 168.22 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (44.99 mg, 841.11 umol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (46.97 mg, 841.11 umol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (100 mg, crude) was btained as a yellow solid. LCMS: m/z = 446.3 (M+H+).
To a solution of intermediate 4 (100 mg, 224.43 umol, 1 eq) in pyridine (0.5 mL) and MeCN 0.5 mL) was added SOs-pyridine (107.16 mg, 673.28 umol, 3 eq) at 0°C and stirred for 0.17 h. The reaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide 2 mL) was added drop wise, and then the mixture was dried by flowing Nz, and the residue was urified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 8%-43%,10min) to yield Compound 33 (22.64 mg, 19%) was obtained s a white solid. LCMS: m/z = 526.1 (M+H+), 1H-NMR: (400 MHz, DMSO-i¾) δ = 8.82 (s, 1 H) 8.20 (d, >8.559 Hz, 1 H) 7.40 - 7.96 (m, 1 H) 7.09 - 7.17 (m, 2 H) 7.06 (s, 1 H) 6.93 - 7.01 m, 3 H) 6.89 (d, >8.436 Hz, 2 H) 6.77 (d, >8.437 Hz, 2 H) 5.08 - 5.16 (m, 1 H) 4.38 (s, 2 H) .90 - 2.98 (m, 1 H) 2.79 - 2.86 (m, 1 H) 2.74 (t, >7.642 Hz, 2 H) 2.40 (t, >7.703 Hz, 2 H) 2.25 s, 3 H).
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (100 mg, 273.35 umol, 1.2 eq) and intermediate 2 (37.85 mg, 227.79 umol, 1 eq) in DMF (2 mL) was added EtsN (1.37 mmol, 190.23 uL, 6 eq), and the mixture was cooled to °C, then T3P (341.68 umol, 203.21 uL, 50% purity, 1.5 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 18 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with ethyl acetate (3 x 10 mL). Then combined organic phase was washed with brine 2 x 10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo. The residue was purified y prep-TLC (Dichloromethane : Methanol = 10: 1) to yield intermediate 3 (55 mg, 51%) was btained as a yellow oil. LCMS: m/z = 478.2 (M+FT), 1H-NMR: (400 MHz, Methanol-t¼) δ = .06 (d, >8.681 Hz, 2 H) 7.76 (s, 1 H) 7.32 (d, >8.681 Hz, 2 H) 7.07 (s, 1 H) 6.96 - 7.03 (m, 1 H) 6.87 (s, 1 H) 6.54 - 6.62 (m, 3 H) 5.32 - 5.37 (m, 1 H) 4.32 (s, 2 H) 3.36 - 3.37 (m, 1 H) 3.05 - .11 (m, 1 H) 2.74 (t, >7.275 Hz, 2 H) 2.40 - 2.48 (m, 2 H).
A solution of intermediate 3 (55 mg, 115.17 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (30.80 mg, 575.87 umol, 20.13 uL, 5 eq), and then the solution was heated to 90 C, Fe (32.16 mg, 575.87 umol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C.The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was iltered, the filtrate was concentrated in vacuo to yield intermediate 4 (70 mg, crude) was btained as a yellow solid. LCMS: m/z = 448.2 (M+H+).
To a solution of intermediate 4 (70 mg, 156.41 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SCb-pyridine (74.68 mg, 469.22 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Agela Durashell C 18 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 4%-34%,10min) to yield Compound 34 (14 mg, 17%) was obtained as a white solid. LCMS: m/z = 526.0 (M-H+), 1H-NMR: (400 MHz, DMSO-dk) δ = 9.21 (br s, 1 H) .67 (s, 1 H) 8.21 (d, >8.436 Hz, 1 H) 7.74 (br s, 1 H) 7.14 (s, 1 H) 7.00 - 7.07 (m, 2 H) 6.86 - .92 (m, 2 H) 6.76 - 6.83 (m, 2 H) 6.53 - 6.61 (m, 3 H) 5.06 - 5.13 (m, 1 H) 4.36 (s, 2 H) 2.96 dd, >13.694, 6.847 Hz, 1 H) 2.82 (dd, >13.694, 7.580 Hz, 1 H) 2.67 (t, >7.703 Hz, 2 H) 2.30 2.40 (m, 2 H). Synthesis of Compound 35
To a solution of intermediate 1 (0.1 g, 246.01 umol, 1 eq) and intermediate 2 (41.37 mg, 246.01 mol, 1 eq) in DMF (3 mL) was added EtgN (149.36 mg, 1.48 mmol, 205.45 uL, 6 eq) and then he mixture was cooled to 0 °C, T3P (313.10 mg, 492.02 umol, 292.62 uL, 50% purity, 2 eq) was dded drop-wise, then the reaction was warmed to 25 °C and stirred for 13 h under N2 tmosphere. The reaction was monitored by TLC and LCMS and when complete, the solution was diluted with ice-water (15 mL) and extracted with ethyl acetate (3 x 10 mL). The combined rganic layer was washed with brine (20 mL), dried with anhydrous Na2SC>4, filtered and oncentrated in vacuo. The residue was purified by prep-TLC (Dichloromethane: Methanol = 0:1) to yield intermediate 3 (35 mg, 30%) was obtained as a yellow solid. LCMS: m/z = 480.2 M+FT), ¾-NMR: (400 MHz, Methanol-^) δ = 8.07 (d, >8.68 Hz, 2 H) 7.66 (s, 1 H) 7.34 (d, >8.68 Hz, 2 H) 7.08 - 7.15 (m, 2 H) 7.01 (s, 1 H) 6.86 - 6.93 (m, 3 H) 5.28 - 5.38 (m, 1 H) 4.29 s, 2 H) 3.33 - 3.40 (m, 1 H) 3.06 - 3.15 (m, 1 H) 2.79 - 2.82 (m, 2 H) 2.41 - 2.48 (m, 2 H).
A solution of intermediate 3 (0.04 g, 83.42 umol, 1 eq) and NH4CI (22.31 mg, 417.08 umol, 5 q ) in EtOH (1.5 mL) and H2O (0.5 mL) was heated to 90 °C, then Fe (23.29 mg, 417.08 umol, 5 q) was added, the reaction was stirred at 90 °C for 2 h. The reaction was monitored by TLC and LCMS and when complete, the mixture was filtered and the filtrate was concentrated in vacuo to ield intermediate 4 (0.03 g, crude) was obtained as a yellow solid. LCMS: m/z = 450.3 M+H+).
A solution of intermediate 4 (0.065 g, 144.59 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SOs-pyridine (69.04 mg, 433.77 umol, 3 eq) was added at 0 °C. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when omplete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was ried by flowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell C18 50*25 5u;mobile phase: [water(10mM NH4HCC>3)-ACN];B%: 7%-37%,10min) to yield Compound 35 (7.8 mg, 10%) was obtained as a white solid. LCMS: m/z = 530.1 (M+H4), 1H- NMR: (400 MHz, DMSO-i&) 6 = 8.83 (s, 1 H) 8.24 (d, 7=8.56 Hz, 1 H) 7.74 (br s, 1 H) 7.14 - .22 (m, 3 H) 7.01 - 7.11 (m, 3 H) 6.86 - 6.92 (m, 2 H) 6.77 - 6.79 (m, 2 H) 5.08 - 5.14 (m, 1 H) .38 (s, 2 H) 2.91 - 2.97 (m, 1 H) 2.80 - 2.86 (m, 1 H) 2.73 - 2.77 (m, 2 H) 2.37 - 2.41 (m, 2 H).
Synthesis of Compound 36
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazjol-4-y1)-2-(4-nitrophenyl)ethanamine ydrochloride salt (0.1 g, 273.35 umol, 1 eq) and intermediate 2 (50.46 mg, 273.35 umol, 1 eq) in DMF (2 mL) was added EtsN (165.96 mg, 1.64 mmol, 228.28 uL, 6 eq) and then the mixture was cooled to 0 °C, T3P (34.79 g, 546.69 umol, 32.51 mL, 50% purity, 2 eq) was added drop- wise, the reaction was allowed to warm to 25 °C and stirred for 13 h. The reaction was monitored y LCMS and when complete, the reaction mixture was diluted with water (30 mL) and xtracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with brine (30 mL), dried with anhydrous NazSC^, filtered and concentrated in vacuo to give a residue. The esidue was purified by prep-TLC (Dichloromethane : Methanol= 10: 1) to yield intermediate 3 35 mg, 26%) was obtained as a yellow solid. LCMS: m/z = 496.2 (M+H+), 1H-NMR: (400 MHz, Methanol^) δ = 8.08 (d, >8.80 Hz, 2 H) 7.67 (s, 1 H) 7.34 (d, >8.68 Hz, 2 H) 7.15 - .20 (m, 2 H) 7.07 - 7.13 (m, 2 H) 7.02 (s, 1 H) 6.87 (s, 1 H) 5.30 - 5.33 (m, 1 H) 4.29 (s, 2 H) .33 - 3.39 (m, 1 H) 3.05 - 3.15 (m, 1 H) 2.77 - 2.85 (m, 2 H) 2.42 - 2.49 (m, 2 H).
To a solution of intermediate 3 (0.065 g, 131.05 umol, 1 eg) in EtOH (3 mL) and H2O (1 mL) was added NH4CI (35.05 mg, 655.27 umol, 5 eg) and the mixture was heated to 90 °C, Fe (36.60 mg, 655.27 umol, 5 eg) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate oncentrated in vacuo to yield intermediate 4 (60 mg, 88%) was obtained as a yellow solid. LCMS: m/z = 466.2 (M+H+).
A mixture of intermediate 4 (0.07 g, 150.22 umol, 1 eg) in Py (1 mL) and MeCN (1 mL) was ooled to 0 °C and SCb-pyridine (71.72 mg, 450.65 umol, 3 eg) was added. The mixture was tirred at 0 °C for 10 min. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, then the mixture was dried by flowing N2 to ive a residue. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 u;mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 14%-44%,10min) to yield Compound 6 (19.8 mg, 24%) was obtained as a white solid. LCMS: m/z = 546.1 (M+H+), 1H-NMR: (400 MHz, DMSO-dk) δ = 8.69 (s, 1 H) 8.21 (d, >8.56 Hz, 1 H) 7.71 (br s, 1 H) 7.15 - 7.33 (m, 4 H) .02 - 7.14 (m, 2 H) 6.74 - 6.93 (m, 4 H) 5.07 - 5.13 (m, 1 H) 4.36 (s, 2 H) 2.90 - 3.00 (m, 1 H) .71 - 2.86 (m, 3 H) 2.37 - 2.41 (m, 2 H). Synthesis of Compound 37
To a solution of (S)-l -(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl) ethanamine ydrochloride salt (0.1 g, 273.35 umol, 1 eq) and intermediate 2 (49.26 mg, 273.35 umol, 1 eq) n DMF (2 mL) was added Et¾N (165.96 mg, 1.64 mmol, 228.28 uL, 6 eq) and then the mixture was cooled to 0 °C, T3P (347.89 mg, 546.69 umol, 325.13 uL, 50% purity, 2 eq) was added rop wise, then it was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with water (30 mL) nd extracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with brine 30 mL), dried with NaaStX filtered and concentrated in vacuo to give a residue. The residue was purified by prep-TLC (Dichloromethane: Methanol = 10:1) to yield intermediate 3 (40 mg, 0%) was obtained as a yellow solid. LCMS: m/z = 492.2 (Μ+Ι-Γ), 'H-NMR: (400 MHz, Methanol^) δ = 8.05 - 8.11 (m, 2 H) 7.78 (s, 1 H), 7.36 (d, >8.68 Hz, 2 H) 7.08 (s, 1 H) 7.04 d, >8.56 Hz, 2 H) 6.84 - 687 (m, 1 H) 6.73 - 6.79 (m, 2 H) 5.30 - 5.40 (m, 1 H) 4.33 (s, 2 H) .75 (s, 3 H) 3.35 - 3.40 (m, 1 H) 3.07 - 3.12 (m, 1 H) 2.76 - 2.80 (m, 2 H) 2.43 - 2.47 (m, 2 H).
To a solution of intermediate 3 (0.075 g, 152.58 umol, 1 eq) in EtOH (3 mL) and ¾0 (1 mL) was added NH4CI (40.81 mg, 762.88 umol, 5 eq) and the mixture was heated to 90 °C, and then Fe (42.61 mg, 762.88 umol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The eaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (65 mg, crude) was obtained as a ellow solid. LCMS: m/z = 462.3 (M+H*).
A mixture of intermediate 4 (0.065 g, 140.82 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SOs-pyridine (67.24 mg, 422.46 umol, 3 eq) was added slowly. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, % ammonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by lowing N2 to give a residue. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mMNH4HCO3)-ACN];B%: 5%-35%,10min) to yield Compound 37 ( 11.7 mg, 15%) was obtained as a white solid. LCMS: m/z = 542.1 (M+H÷), *H- NMR: (400 MHz, DMSCMO δ = 8.87 (s, 1 H) 8.17 (s, 1 H) 6.99 - 7.24 (m, 4 H) 6.85 - 6.95 (m, H) 6.74 - 6.84 (m, 4 H) 5.00 - 5.20 (m, 1 H) 4.39 (s, 2 H) 3.71 (s, 3 H) 2.87 - 2.97 (m, 1 H) .77 - 2.86 (m, 1 H) 2.67 - 2.76 (m, 2 H) 2.34 - 2.43 (m, 2 H).
Synthesis of Compound 38
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine hydrochloride salt (0.1 g, 246.01 umol, 1 eq) and intermediate 2 (40.40 mg, 246.01 umol, 1 eq) in DMF (3 mL) was added EtiN (149.36 mg, 1.48 mmol, 205.45 uL, 6 eq) and the mixture was ooled to 0 °C, then T3P (313.10 mg, 492.02 umol, 292.62 uL, 50% purity, 2 eq) was added rop-wise at 0 °C, the reaction was allowed to warm to 25 °C and stirred at 25 °C for 13 h under N2 atmosphere. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 x 10 mL). The ombined organic layer was washed with brine (20 mL), dried with anhydrous NazSO-t, filtered nd concentrated in vacuo to give a residue. The residue was purified by prep-TLC Dichloromethane: Methanol = 10:1) to yield intermediate 3 (40 mg, 34%) was obtained as a ellow solid. LCMS: m/z = 476.2 (M+H+), 1H-NMR: (400 MHz, Methanol-^) δ = 8.02 - 8.07 m, 2 H) 7.65 (d, >0.98 Hz, 1 H) 7.27 - 7.37 (m, 2 H) 7.07 - 7.12 (m, 1 H) 7.02 - 7.06 (m, 1 H) .02 - 7.10 (m, 3 H) 6.78 (s, 1 H) 5.30 - 5.33 (m, 1 H) 4.28 (s, 2 H) 3.35 (dd, >13.75, 5.93 Hz, 1 H) 3.08 (dd, >13.69, 8.80 Hz, 1 H) 2.75 - 2.82 (m, 2 H) 2.42 - 2.48 (m, 2 H) 2.26 (s, 3 H).
To a solution of intermediate 3 (0.04 g, 84.11 umol, 1 eq ) in EtOH (1.5 mL) and H2O (0.5 mL) was added NH4CI (22.50 mg, 420.56 umol, 5 eq) and the mixture was heated to 90 °C, and then e (23.49 mg, 420.56 umol, 5 eq) was added at 90 °C, the reaction was stirred at 90 °C for 2 h. The reaction was monitored by TLC and LCMS and when complete, the mixture was filtered and oncentrated in vacuo to yield intermediate 4 (40 mg, crude) was obtained as a yellow solid. LCMS: m/z = 446.0 (M+H+).
A mixture of intermediate 4 (0.065 g, 145.88 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SCb-pyridine (69.65 mg, 437.63 umol, 3 eq) was added in batches. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when omplete, 7% ammonium hydroxide (1 mL) was added slowly and the mixture was dried by lowing N2 to give a residue. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water( 1 OmM NH4HCO3)- ACN] ;B% : 5%-35%,10min) to yield Compound 38 (28.4 mg, 37%) was obtained as a white solid. LCMS: m/z = 526.1 (M+H4), 1H- NMR: (400 MHz, DMSO-i&) 6 = 8.87 (s, 1 H) 8.19 (d, >8.44 Hz, 1 H) 7.16 (s, 1 H) 7.02 - 7.09 m, 5 H) 6.89 (d, >8.44 Hz, 2 H) 6.77 (d, >8.44 Hz, 2 H) 5.01 - 5.18 (m, 1 H) 4.39 (s, 2 H) .94 (dd, >13.63, 7.15 Hz, 1 H) 2.82 (dd, >13.51, 7.40 Hz, 1 H) 2.71 - 2.74 (m, 2 H) 2.36 - .40 (m, 2 H) 2.25 (s, 3 H). Synthesis of Compound 40
To a solution of (S)-l-(2-((lH-imidazoI-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (400 mg, 874.71 umol, 1 eq) and intermediate 2 (147.09 mg, 874.71 umol, 1 q) in DMF (2 mL) was added EtgN (5.25 mmol, 730.49 uL, 6 eq), and the mixture was cooled to °C, then T3P (1.31 mmol, 780.32 uL, 50% purity, 1.5 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 18 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with dichloromethane (3 x 20 mL). Then combined organic phase was washed with rine (2 x 10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 3 (540 mg, crude) was obtained as a yellow oil. LCMS: m/z = 480.3 (M+FT).
A solution of intermediate 3 (540 mg, 1.13 mmol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (301.19 mg, 5.63 mmol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (314.44 mg, 5.63 mmol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was ltered, the filtrate was concentrated in vacuo to yield intermediate 4 (510 mg, crude) was btained as a yellow oil. LCMS: m/z = 450.3 (M+H÷).
To a solution of intermediate 4 (500 mg, 1.11 mmol, 1 eq) in pyridine ( 1 mL) and MeCN (1 mL) was added SCb-pyridine (531.07 mg, 3.34 mmol, 3 eq) at 0 °C stirred for 0.17 h. The reaction was monitored by TLC and when complete, 7% ammonium hydroxide (3 mL) was added drop- wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC column: YMC-Actus Triart C18 100*30mm *5um ;mobile phase: [water(0.04%NH3H2O+ 1 OmM NHIHC03)-ACN];B%: 5%-35%,10min) to yield Compound 40 (62.1 mg, 10%) was obtained as white solid. LCMS: m/z = 528.1 (M-PL), ‘H-NMR: (400 MHz, DMSO-4.) δ = 8.25 (d, >8.559 Hz, 1 H) 8.05 (s, 1 H) 7.66 (br s, 1 H) 7.14 - 7.29 (m, 3 H) 6.99 - 7.10 (m, 6 H) 6.85 - .90 (m, 2 H) 6.77 - 6.82 (m, 2 H) 5.01 - 5.11 (m, 1 H) 4.25 (s, 2 H) 2.96 (dd, >13.694, 6.236 Hz, 1 H) 2.73 - 2.82 (m, 3 H) 2.35 - 2.41 (m, 2 H).
Synthesis of Compound 41
EtsN (150.92 mg, 1.49 mmol, 207.59 uL, 6 eq) was added into a solution of (S)-l-(2-((lH- midazol-5-yl)methyl)thiazol-4-yJ)-2-(4-nitrophenyl)ethanamine hydrochloride salt (0.1 g, 48.57 umol, 1 eq) and intermediate 2 (45.89 mg, 248.57 umol, 1 eq) in DMF (2 mL), and ooled to 0 °C, then T3P (237.27 mg, 372.86 umol, 221.75 uL, 50% purity, 1.5 eq) was added at °C, then the mixture was allowed to warm to 25 °C and stirred for 16 h. The reaction was monitored by TLC and when complete, the reaction mixture was poured into ice-water (20 mL), nd extracted with ethyl acetate (3 x 10 mL). Then combined organic phase was washed with brine (2 x 10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by prep-TLC (dichloromethane: methyl alcohol = 10:1) to yield intermediate 3 (60 mg, 49%) was obtained as a white solid. LCMS: m/z = 496.0 (M+H4).
A solution of intermediate 3 (0.06 g, 120.97 umol, 1 eq) in EtOH (1.5 mL) and H2O (0.5 mL) was heated to 90 °C, then NH4CI (32.35 mg, 604.86 umol, 21.15 uL, 5 eq ), Fe (33.78 mg, 604.86 mol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by LCMS nd when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to ield intermediate 4 (70 mg, crude) was obtained as a yellow solid. LCMS: m/z = 466.2 M+H4).
To a solution of intermediate 4 (70 mg, 150.22 umol, 1 eq) in pyridine (0.5 mL) and MeCN (0.5 mL) was added SOa-pyridine (71.73 mg, 450.65 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing Nz, and the residue was urified by prep-HPLC (water(10mM NH4HCC>3)-ACN];B%: 5%-38%,12min) to yield Compound 41 (11.5 mg, 14%) was obtained as a white solid. LCMS: m/z = 546.1 (M+H4), 1H- NMR: (400 MHz, DMSO^) 6 = 8.84 (s, 1 H) 8.28 (d, >8.436 Hz, 1 H) 7.72 (br s, 1 H) 7.37 - .43 (m, 1 H) 7.19 - 7.26 (m, 3 H) 7.10 - 7.18 (m, 3 H) 7.07 (s, 1 H) 6.95 (s, 1 H) 6.89 (d, >8.436 Hz, 2 H) 6.78 (d, >8.436 Hz, 2 H) 5.13 (q, >7.254 Hz, 1 H) 4.38 (s, 2 H) 2.81 - 2.97 m, 4 H) 2.41 - 2.46 (m, 2 H). To a solution of (S)-l-(2-((lH-imidazoI-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (200 mg, 546.69 umol, 1.2 eq) and intermediate 2 (82.09 mg, 455.58 umol, 1 q) in DMF (2 mL) was added EtiN (2.73 mmol, 380.47 uL, 6 eq), and the mixture was cooled to °C, then T3P (683.36 umol, 406.42 uL, 50% purity, 1.5 eq) was added dropwise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 18 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with ethyl acetate (3 x 20 mL). Then combined organic phase was washed with brine 2 x 20 mL), dried with anhydrous Na2S(>4 and concentrated in vacuo. The residue was purified y prep-TLC (Dichloromethane: Methanol = 10: 1) to yield intermediate 3 (90 mg, 40% ) was btained as a yellow oil. LCMS: m/z = 492.2 (M+H÷), Έ-ΝΜΚ: (400 MHz, Methanol-^) δ = .07 (d, >8.314 Hz, 2 H) 7.68 (s, 1 H) 7.34 (d, >8.436 Hz, 2 H) 7.14 (t, >8.070 Hz, 1 H) 6.96 7.06 (m, 2 H) 6.85 - 6.93 (m, 2 H) 6.71 - 6.79 (m, 1 H) 5.29 - 5.37 (m, 1 H) 4.29 (s, 2 H) 3.80 s, 3 H) 3.08 (dd, >13.572, 8.803 Hz, 1 H) 2.68 - 2.96 (m, 3 H) 2.38 - 2.48 (m, 2 H).
A solution of intermediate 3 (90 mg, 183.09 rnnol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (48.97 mg, 915.45 umol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (51.12 mg, 915.45 umol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was ltered, the filtrate was concentrated in vacuo to yield intermediate 4 (150 mg, crude) was btained as a yellow solid. LCMS: m/z = 462.3 (M+H*), 1H-NMR: (400 MHz, Methanol-i/») δ 8.02 (br s, 1 H) 7.09 - 7.20 (m, 2 H) 7.02 (d, >5.733 Hz, 1 H) 6.74 - 6.92 (m, 5 H) 6.63 (d, >7.277 Hz, 2 H) 5.14 - 5.24 (m, 1 H) 4.35 (s, 2 H) 3.81 (s, 3 H) 3.07 (br dd, >12.789, 5.513 Hz, 1 H) 2.78 - 2.92 (m, 3 H) 2.42 - 2.45 (m, 2 H).
To a solution of intermediate 4 (150 mg, 324.97 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SOa-pyridine (155.17 mg, 974.92 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NHIHC03)-ACN];B%: 5%-35%,10min) to yield Compound 42 (34.8 mg, 19%) was obtained as white solid. LCMS: m/z = 540.1 (M-FT), 'H-NMR: (400 MHz, DMSO-dk) δ = 8.75 (s, 1 H) 8.20 (d, >8.314 Hz, 1 H) 7.74 (s, 1 H) 7.10 - 7.21 (m, 3 H) 7.07 (d, >7.214 Hz, 1 H) 6.86 - 6.96 m, 3 H) 6.74 - 6.85 (m, 3 H) 5.08 - 5.04 (m, 1 H) 4.37 (s, 2 H) 3.77 (s, 3 H) 2.91 - 2.96 (m, 1 H) .80 - 2.85 (m, 1 H) 2.71 - 2.75 (m, 2 H) 2.33 - 2.37 (m, 2 H).
Synthesis of Compound 43
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)ihiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (0.1 g, 248.57 umol, 1 eq, HC1), intermediate 2 ( 40.82 mg, 248.57 umol, 1 g) in DMF (4 mL) was added EtsN (150.92 mg, 1.49 mmol, 207.59 uL, 6 eq) and the mixture was cooled to 0 °C, and then T3P (237.27 mg, 372.86 umol, 221.75 uL, 50% purity, 1.5 eq) was dded drop-wise. Then the reaction was allowed to warm to 25 °C and stirred at 25 °C for 3 h. The reaction was monitored by TLC and when complete, .The solution was poured into ice-water 20 mL) and extracted with ethyl acetate (3 x 10 mL), washed with brine (10 mL), dried with nhydrous NazSO^ filtered and then the organic phase was concentrated in vacuo to yield ntermediate 3 (0.15 g, crude) was obtained as a yellow oil.
To a solution of intermediate 3 (0.07 g, 147.19 umol, 1 eq) in EtOH (1.5 mL) and H2O (0.5 mL) was added NH4CI (39.37 mg, 735.97 umol, 5 eq) and the mixture was heated to 90 °C, and then e (41.10 mg, 735.97 umol, 5 eq) was added. The reaction was stirred at 90 °C for 2 h. The eaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate concentrated in vacuo to yield intermediate 4 (80 mg, crude) was obtained as a yellow oil. LCMS: m/z = 446.3 (M+H+). A mixture of intermediate 4 (99.56 mg, 223.44 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SOa-pyridine (106.69 mg, 670.31 umol, 3 eq) was added in atches. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, 7% ammonium hydroxide (1 mL) was added drop wise and the mixture was ried by flowing Nz to give a residue. The residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HCOa)-ACN];B%: 4%-34%,10min) o yield Compound 43 (13.5 mg, 11%) was obtained as a white solid. LCMS: m/z = 526.1 Μ+ΕΓ), ¾-NMR: (400 MHz, DMSO-4,) δ = 8.79 (s, 1 H) 8.25 (d, J=8.44 Hz, 1 H) 7.73 (s, 1 H) 7.17 (s, 1 H) 7.04 - 7.14 (m, 5 H) 6.85 - 6.92 (m, 2 H) 6.73 - 6.82 (m, 2 H) 5.09 - 5.14 (m, 1 H) 4.38 (s, 2 H) 2.90 - 2.98 (m, 1 H) 2.79 - 2.87 (m, 1 H) 2.70 - 2.77 (m, 2 H) 2.34 - 2.40 (m, 2 H) 2.25 (s, 3 H).
Synthesis of Compound 44 To a solution of (S)-l-(2-((lH-imidazpl-5-yl)methyl)ihiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (158.02 mg, 902.04 umol, 1.1 q) in DMF (4 mL) was added Et3N (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), then cooled to 0 C, and then T3P (782.76 mg, 1.23 mmol, 731.55 uL, 50% purity, 1.5 eq) was added dropwise at °C under N2, the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS when complete, the reaction mixture was quenched by addition ice water (30 mL) at 25°C, and hen extracted with ethyl acetate (3 x 10 mL). The combined organic layer was washed with brine 10 mL), dried with anhydrous NaaSO^ filtered and concentrated in vacuo, the residue was purified y column chromatography silica gel (S1O2, Petroleum ether: Ethyl acetate = 5: 1 to 1 : 1) to yield ntermediate 3A (193 mg, crude) was obtained as a yellow solid. LCMS: m/z = 644.2 (M+H+).
A solution of intermediate 3A (511 mg, 793.83 umol, 1 eq) in NHs/MeOH (7 M, 2.00 mL, 17.64 q) was stirred at 60 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was dried by flowing N2 to yield intermediate 3 (400 mg, crude) was obtained as a yellow solid. LCMS: m/z = 487.2 (M+tT).
A solution of intermediate 3 (350 mg, 719.36 umol, 1 eq) and NH4CI (192.6 mg, 3.60 mmol, 5 q) in EtOH (8 mL) and H2O (2 mL) was heated to 90 °C, then Fe (200.86 mg, 3.60 mmol, 5 eq) was added, the mixture was stirred at 900 C for 2 h. The reaction was monitored by LCMS when omplete, the solution was filtered and concentrated in vacuo and the residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u; mobile phase: [water(0.04%NH3H20+10mM NH4HC03)-ACN]; B%: 5%-45%,10min) to yield intermediate 4 (85 mg, 25%) was obtained as yellow solid. LCMS: m/z = 457.2 (M+H+), Ή-ΝΜΚ: (400 MHz, DMSO-tifc) δ = 8.22 (d, >8.80 Hz, 1 H) 7.68 (d, >8.19 Hz, 2 H) 7.60 (d, >0.86 Hz, 1 H) 7.30 (d, >8.19 Hz, 2 H) 6.93 7.03 (m, 2 H) 6.79 (d, >8.31 Hz, 2 H) 6.43 (d, >8.31 Hz, 2 H) 4.96 - 5.04 (m, 1 H) 4.82 (br s, H) 4.19 (s, 2 H) 2.96 (br dd, >13.82, 5.38 Hz, 1 H) 2.79 - 2.85 (m, 2 H) 2.71 (br dd, >13.75, .23 Hz, 1 H) 2.39 (br t, >7.46 Hz, 2 H).
To a solution of intermediate 4 (85.00 mg, 186.17 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SCh.pyridine (88.90 mg, 558.52 umol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% ammonium hydroxide (3 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water(0.04%NH3H20+l OmM NH4HC03>ACN]; B%: l%-25%, lOmin) to yield Compound 44 (38 mg, 37%) was obtained as a white solid. LCMS: m/z = 535.1 (M-H+), 1H- NMR: (400 MHz, DMSO-tfc) 5 = 8.56 (s, 1 H) 7.63 (d, 7=8.19 Hz, 2 H) 7.29 (d, 7=8.19 Hz, 2 H) .19 (s, 1 H) 6.95 (s, 1 H) 6.79 - 6.88 (m, 4 H) 5.04 (t, >7.27 Hz, 1 H) 4.33 (s, 2 H) 2.94 (dd, 7=13.76, 6.42 Hz, 1 H) 2.81 (br t, 7=7.40 Hz, 2 H) 2.73 (dd, 7=13.75, 8.25 Hz, 1 H) 2.41 (t, >7.40 Hz, 2 H).
Synthesis of Compound 45
To a solution of (S)-l-(2-((lH-lmidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamlne ydrochloride salt (300 mg, 820.04 umol, 1 eq, HC1) and intermediate 2 (143.66 mg, 820.04 mol, 1 eq) in DMF (8 mL) was added Et3N (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq) and cooled to 0 °C, and then T3P (521.84 mg, 820.04 umol, 487.70 uL, 50% purity, 1 eq) was added t 0 °C, then the mixture was allowed warm to 25 °C and stirred for 14 h. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured into water 30 mL), and extracted with ethyl acetate (3 x 10 mL), the combined organic layer was washed with brine (20 mL), dried with anhydrous NazSCL, filtered and concentrated in vacuo to yield ntermediate 3A (450 mg, crude) was obtained as a yellow oil. LCMS: m/z = 644.3 (M+H+).
A mixture of intermediate 3A (450 mg, 699.07 umol, 1 eg) in NH3/MeOH (7 M, 3 mL) was egassed and purged with N2 for 3 times, and then the mixture was stirred at 60 °C for 1 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to yield intermediate 3 (370 mg, crude) was obtained as a yellow oil. LCMS: m/z = 487.2 (M+H+).
A mixture of intermediate 3 (370 mg, 760.46 umol, 1 eg) and NH4CI (245.54 mg, 4.56 mmol, 6 g) in EtOH (15 mL) and H2O (5 mL) was heated to 90 °C, and then Fe (254.81 mg, 4.56 mmol, eg) was added and stirred at 90 °C for 2 h. The reaction was monitored by LCMS and HPLC when complete, the reaction mixture was concentrated in vacuo. The residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04% NH3H2O+IO mM NH4HC03)-ACN];B%: 5%-45%, lOmin) to yield intermediate 4 (90 mg, 25%) was obtained as yellow oil. LCMS: m/z = 457.1 (M+H+).
To a solution of intermediate 4 (90 mg, 197.13 umol, 1 eg) in pyridine (2 mL) and MeCN (2 mL) was added SCb-pyridine (94.13 mg, 591.38 umol, 3 eg) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mM NH4HC03)-ACN]; B%: l%-30%, lOmin) to yield Compound 45 42.7 mg, 36%) was obtained as a white solid. LCMS: m/z = 535.1 (M-H+), 1H-NMR: (400 MHz, DMSO-ifc) δ = 8.71 (s, 1 H) 8.26 (d, >8.44 Hz, 1 H) 7.55 - 7.67 (m, 3 H) 7.39 - 7.54 (m, H) 7.21 (br s, 1 H) 7.14 (s, 1 H) 7.08 (s, 1 H) 6.93 - 6.99 (m, 1 H) 6.84 - 6.92 (m, 2 H) 6.74 - .79 (m, 2 H) 5.05 - 5.15 (m, 1 H) 4.37 (s, 2 H) 2.92 - 2.97 (m, 1 H) 2.76 - 2.88 (m, 3 H) 2.43 - .47 (m, 2 H).
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (158.02 mg, 902.04 umol, 1.1 q) in DMF (4 mL) was added EUN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), then cooled to 0 C, and then T3P (782.76 mg, 1.23 mmol, 731.55 uL, 50% purity, 1.5 eq) was added dropwise at °C under N2, the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS when complete, the reaction mixture was quenched by addition ice water (30 mL) at 25°C, and xtracted with ethyl acetate (3 x 10 mL). The combined organic layer was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield intermediate 3A 230 mg, crude) was obtained as a yellow solid. LCMS: m/z = 644.2 (M+FT).
A solution of intermediate 3A (406 mg, 630.72 umol, 1 eq) in NHj/MeOH (7 M, 2.00 mL, 22.20 eq) was stirred at 60 °C for 1 h. The reaction was monitored by LCMS when complete, the solution was dried by flowing N2 to yield intermediate 3 (365 mg, crude) was obtained as a yellow solid. LCMS: m/z = 487.2 (M+I Γ).
A solution of intermediate 3 (365 mg, 750.19 umol, 1 eq) and NHtCl (200.62 mg, 3.75 mmol, 5 q) in EtOH (8 mL) and H20 (2 mL) was heated to 90 °C, then Fe (209.47 mg, 3.75 mmol, 5 eq) was added, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when omplete, the solution was filtered and concentrated in vacuo and was purified by prep-HPLC column: Waters Xbridge 150*25 5u; mobile phase: [water(0.04%NH3H20+10mMNH4HC03)- ACN]; B%: 5%-45%, lOmin) to yield intermediate 4 (95 mg, 27%) was obtained as a yellow olid. LCMS: m/z = 455.1 (M-H+), ‘H-NMR: (400 MHz, DMSO-dk) δ = 8.27 (d, >8.68 Hz, 1 H) 7.75 (dd, >7.70, 0.98 Hz, 1 H) 7.60 (s, 1 H) 7.54 (td, >7.64, 1.22 Hz, 1 H) 7.34 - 7.40 (m, 1 H) 7.28 (d, >7.83 Hz, 1 H) 6.93 - 7.05 (m, 2 H) 6.78 (d, >8.31 Hz, 2 H) 6.42 (d, >8.31 Hz, 2 H) 4.95 - 5.05 (m, 1 H) 4.80 (s, 2 H) 4.17 (br s, 2 H) 2.90 - 2.99 (m, 3 H) 2.72 (dd, >13.69, 9.05 Hz, 1 H) 2.43 - 2.48 (m, 2 H).
To a solution of intermediate 4 (95 mg, 208.08 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SO pyridine (99.35 mg, 624.23 umol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water(0.04%NH3H20+10mMNH4HC03>ACN]; B%: l%-25%, lOmin) to yield Compound 46 (43.6 mg, 39%) was obtained as a white solid. LCMS: m/z = 535.1 (M-H÷), Ή- NMR: (400 MHz, DMSO-ifc+D20) δ = 8.56 (s, 1 H) 7.63 (d, >8.19 Hz, 2 H) 7.29 (d, >8.19 Hz, 2 H) 7.19 (s, 1 H) 6.95 (s, 1 H) 6.79 - 6.88 (m, 4 H) 5.04 (t, >7.27 Hz, 1 H) 4.33 (s, 2 H) .94 (dd, >13.76, 6.42 Hz, 1 H) 2.81 (t, >7.40 Hz, 2 H) 2.73 (dd, >13.75, 8.25 Hz, 1 H) 2.41 t, >7.40 Hz, 2 H).
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)ihiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (178.91 mg, 820.04 umol, 1 q)) in DMF (4 mL) was added EtgN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), then cooled to 0 C, and then T3P (782.76 mg, 1.23 mmol, 731.55 uL, 50% purity, 1.5 eq) was added drop-wise at °C under N2, the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition ice-water (30 mL) at 25°C, and xtracted with ethyl acetate (3 x 40 mL). The combined organic layers were washed with brine (2 30 mL), dried with anhydrous NazSCL, filtered and concentrated in vacuo to yield intermediate A (200 mg, crude) was obtained as a yellow solid. LCMS: m/z =730.1(M+H+).
A solution of intermediate 3A (411 mg, 563.25 umol, 1 eq) in NFL/MeOH (7 M, 2.00 mL, 24.86 q) was stirred at 60 °C for 1 h. The reaction was monitored by LCMS when complete, the solution was dried by flowing N2 to yield intermediate 3 (352 mg, crude) was obtained as a yellow solid. LCMS: m/z = 530.1 (M+lT).
A solution of intermediate 3 (352 mg, 664.74 umol, 1 eq) and NH4CI (177.62 mg, 3.75 mmol, 5 q) in EtOH (8 mL) and H2O (2 mL) was heated to 90 °C, then Fe (185.61 mg, 3.32 mmol, 5 eq) was added, the mixture was stirred at 900 C for 2 h. The reaction was monitored by LCMS when omplete, the solution was filtered and concentrated in vacuo and was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile phase: [water (0.04%NH3H20+10mM NH4HCO3)- ACN]; B%: 25%-45%, lOmin) to yield intermediate 4 (94 mg, 25%) was obtained as a yellow olid. LCMS: m/z = 500.1 (M+H+), Ή-ΝΜΚ: (400 MHz, DMSO -ck) δ = 11.92 (br s, 1 H) 8.24 d, J=8.82 Hz, 1 H) 7.52 - 7.63 (m, 3 H) 7.33 (d, J=8.16 Hz, 2 H) 7.02 (s, 1 H) 6.87 (s, 1 H) 6.79 d, J=8.16 Hz, 2 H) 6.42 (d, J=8.38 Hz, 2 H) 4.91 - 5.11 (m, 1 H) 4.16 (s, 2 H) 2.89 - 3.02 (m, 1 H) 2.78 - 2.86 (m, 2 H) 2.68 - 2.74 (m, 1 H) 2.36 - 2.42 (m, 2 H).
To a solution of intermediate 4 (91 mg, 182.16 umol, 1 eq)) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SO3.pyridine (86.98 mg, 546.49 umol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, 7% mmonium hydroxide (1.5 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water(0.04%NH3H20+l OmM NH4HC03>ACN]; B%: 5%-35%, lOmin) to yield Compound 47 (36.04 mg, 33%) was obtained as a white solid. LCMS: m/z = 578.1 (M-lT), 1H- NMR: (400 MHz, DMSO-tfc) δ = 8.73 (s, 1 H) 8.26 (d, J=8.44 Hz, 1 H) 7.72 (br s, 1 H) 7.60 (d, =8.19 Hz, 2 H) 7.39 (d, J=7.95 Hz, 2 H) 7.20 (s, 1 H) 7.13 (s, 1 H) 7.08 (s, 1 H) 6.86 - 6.91 (m, H) 6.79 (d, J=8.44 Hz, 2 H) 5.11 (q, J=7.70 Hz, 1 H) 4.36 (s, 2 H) 2.94 (dd, 1=13.63, 6.91 Hz, H) 2.76 - 2.87 (m, 3 H) 2.38 - 2.46 (m, 2 H).
To a solution of (S)-l-(2-((lH-lmidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamlne ydrochloride salt (300 mg, 820.04 umol, 1 eg, HC1), intermediate 2 (178.91 mg, 820.04 umol, 1 g) and EtsN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eg) in DMF (8 mL) was cooled to 0 °C, and hen T3P (782.76 mg, 1.23 mmol, 731.55 uL, 50% purity, 1.5 eg) was added at 0 °C, then the mixture was allowed warm to 25 °C and stirred for 14 h. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured into water (30 mL), and extracted with ethyl acetate (3 x 10 mL), the combined organic layer was washed with brine 20 mL, dried with anhydrous NazSO-i, filtered and concentrated in vacuo to yield intermediate 3A (420 mg, rude). LCMS: m/z = 730.3 (M+H+).
A mixture of intermediate 3A (420 mg, 575.59 umol, 1 eg) in NHg/MeOH (7 M, 3 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 60 °C for 1 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to yield intermediate 3 (390 mg, crude) was obtained as a yellow oil. LCMS: m/z = 530.2 (M+H").
A mixture of intermediate 3 (390 mg, 736.50 umol, 1 eq) and NH4CI (237.80 mg, 4.42 mmol, 6 q) in EtOH (15 mL) and H2O (5 mL) was heated to 90 °C, and then Fe (246.78 mg, 4.42 mmol, eq) was added and stirred at 90 °C for 2 h. The reaction was monitored by LCMS and HPLC nd when complete, the reaction mixture was concentrated in vacuo. The residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM NHIHC03)-ACN]; B%: 10%-50%, 10 min) to yield intermediate 4 (80 mg, 21%) was obtained s a yellow oil. LCMS: m/z = 500.2 (M+fL).
To a solution of intermediate 4 (80 mg, 160.14 umol, 1 eq) in pyridine (0.5 mL) and MeCN (0.5 mL) was added SOs-pyridine (76.47 mg, 480.43 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (2 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mMNH4HC03)-ACN]; B%: 5%-35%, 10 min) to yield Compound 48 50.1 mg, 52%) was obtained as a white solid. LCMS: m/z = 578.1 (M-H+), 1H-NMR: (400 MHz, DMSO-ifc) 5 = 8.80 (s, 1 H) 8.26 (d, >8.44 Hz, 1 H) 7.75 (s, 1 H) 7.43 - 7.59 (m, 4 H) .21 (s, 1 H) 7.15 (s, 1 H) 7.09 (s, 1 H) 7.06 (s, 1 H) 6.95 (s, 1 H) 6.88 (d, .7=8.31 Hz, 2 H) 6.77 d, >8.31 Hz, 2 H) 5.03 - 5.19 (m, 1 H) 4.20 - 4.45 (s, 2 H) 2.75 - 2.99 (m, 4 H) 2.42 - 2.48 (m, H).
Synthesis of Compound 49
To a solution of (S)-l-(2-((lH-lmidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamine ydrochloride salt (0.3 g, 731.19 umol, 1 eq) and intermediate 2 (159.53 mg, 731.19 umol, 1 eq) n DMF (3 mL) was added EfcN (443.94 mg, 4.39 mmol, 610.64 uL, 6 eq), and then the solution was cooled to 0 °C, then T3P (930.61 mg, 1.46 mmol, 869.73 uL, 50% purity, 2 eq) was added rop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored y LCMS and when complete, the solution was poured into the ice water and extracted with thyl acetate (3 x 10 mL), the combined organic phase was washed with brine (10 mL) dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield intermediate 3 (412 mg, rude) was obtained as yellow solid. LCMS: m/z =730.0 (M+FT). ntermediate 3 (400 mg, 548.18 umol, 1 eq) in NFfc/MeOH (7 M, 2 mL, 25.54 eq) was stirred at 0 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was oncentrated in vacuo to yield intermediate 4 (320 mg, crude) was obtained as yellow solid. LCMS: m/z = 530.2 (Μ+ΗΓ).
To a solution of intermediate 4 (300 mg, 566.54 umol, 1 eq) in EtOH (7 mL) and HzO (5 mL) was added NH4CI (155.26 mg, 2.83 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (158.19 mg, 2.83 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (column: UniSil 120*30* 10um;mobile phase: [water(0.05%HCl)- ACN];B%: 1%-35%,1 lmin) to yield intermediate S (100 mg, 34%) was obtained as yellow olid. LCMS: m/z = 500.2 (M+H+).
To a solution of intermediate 5 (100 mg, 200.15 umol, 1 eq) in pyridine (2 mL) and CH3CN (2 mL) was cooled to 0 °C, then the SOs-pyridine (95.58 mg, 0.6 mmol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing N2, the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 5%- 5%,10min) to yield Compound 49 (55 mg, 15%) was obtained as white solid. LCMS: m/z 578.0 (M-IT). ‘H-NMR: (400 MHz, DMSO-de) δ = 8.66 (s, 1 H) 8.35 (d, >8.33 Hz, 1 H) 7.76 br s, 1 H) 7.65 (d, >7.89 Hz, 1 H) 7.50 - 7.58 (m, 1 H) 7.33 - 7.42 (m, 2 H) 7.21 (s, 1 H) 7.13 - .20 (m, 2 H) 7.09 (s, 1 H) 6.97 (s, 1 H) 6.86 - 6.92 (m, 2 H) 6.80 (d, >8.33 Hz, 2 H) 5.10 - 5.15 m, 1 H) 4.36 (s, 2 H) 2.92 - 2.98 (m, 3 H) 2.80 - 2.87 (m, 1 H) 2.41 - 2.45 (m, 2 H).
Synthesis of Compound SO
To a solution of (S)-l-(2-((lH-!midazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine hydrochloride salt (0.3 g, 731.19 umol, 1 eq) and intermediate 2 (140.57 mg, 731.19 umol, 1 eq) in DMF (3 mL) was added Et3N (443.94 mg, 4.39 mmol, 610.64 uL, 6 eq), and then the solution was cooled to 0 °C, then T3P (930.61 mg, 1.46 mmol, 869.73 uL, 50% purity, 2 eq) was added rop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored y LCMS and when complete, the solution was dried with flowing N2 to yield intermediate 3 456 mg, crude) was obtained as yellow solid, LCMS: m/z = 678.3 (M+H+). ntermediate 3 (450 mg, 663.86 umol, 1 eq) in NH3/MeOH (7 M, 2 mL, 21.09 eq) was stirred at 0 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was oncentrated in vacuo to yield intermediate 4 (341 mg, crude) was obtained as yellow solid. LCMS: m/z = 504.2 (M+I Γ).
To a solution of intermediate 4 (0.33 g, 655.26 umol, 1 eq) in EtOH (7 mL) and H20 (5 mL) was dded NH4CI (179.58 mg, 3.28 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm o 90 °C, and then Fe (182.97 mg, 3.28 mmol, 5 eq ) was added, then the mixture was stirred at 0 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, he reaction mixture was filtered, the filtrate was concentrated in vacuo. The residue was purified y prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: 20%-50%,10min) to yield intermediate 5 80 mg, 26%) was obtained as a light yellow solid. LCMS: m/z = 474.4 (M+H+).
To a solution of intermediate 5 (80 mg, 168.91 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, then the SCh-pyridine (80.65 mg, 506.72 umol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7% ammonium hydroxide (2 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 50*25 5u;mobile phase: [water(0.04%NH3H20+l OmM NH4HC03)-ACN];B%: 5%- 5%,10min) to yield Compound SO (45.6 mg, 49%) was obtained as a white solid. LCMS: m/z 552.4 (M-HT). 1H-NMR: (400 MHz, DMSO-tfc) δ = 8.84 (s, 1 H) 8.21 (d, >8.68 Hz, 1 H) .21 (s, 1 H) 7.15 (s, 1 H) 7.06 - 7.13 (m, 5 H) 7.02 (s, 1 H) 6.95 (s, 1 H) 6.88 (d, >8.31 Hz, 2 H) 6.77 (d, >8.31 Hz, 2 H) 5.06 - 5.18 (m, 1 H) 4.38 (s, 2 H) 2.93 (dd, >13.69, 7.21 Hz, 1 H) .79 - 2.86 (m, 2 H) 2.73 (t, >7.58 Hz, 2 H) 2.39 (t, >7.70 Hz, 2 H) 1.17 (d, >6.85 Hz, 6 H).
To a mixture of (S)-l-(2-((lH-imidazol-S-yl)methyl)thiazol~4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (400 mg, 874.71 umol, 1 eq) and intermediate 2 (199.66 mg, 874.71 umol, 1 q) in DMF (3 mL) was drop-wise added EtgN (531.07 mg, 5.25 mmol, 730.49 uL, 6 eq) and ooled to 0 °C, then T3P (1.11 g, 1.75 mmol, 1.04 mL, 50% purity, 2 eq) was added at 0 °C, and hen the mixture was warmed to 25 °C and stirred for 3 h at 25 °C under N2 atmosphere. The eaction was monitored by LCMS and when complete. The mixture was poured into ice-water (25 mL) then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous NazS04, filtered and concentrated in vacuo o yield intermediate 3 (376 mg, crude) was obtained as a yellow oil . LCMS: m/z = 540.1 (M+H+). A solution of intermediate 3 (345 mg, 639.33 umol, 1 eq) and NH4CI (169.6 mg, 3.20 mmol, 5 eq) n EtOH (3 mL) and H2O (1 mL) was heated to 90 °C and then Fe (178.52 mg, 3.20 mmol, 5 eq) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the solution was filtered and concentrated in vacuo to ield intermediate 4 (433 mg, crude) was obtained as a yellow solid. LCMS: m/z = 510.0 (Μ+ΕΓ). A solution of intermediate 4 (383 mg, 751.51 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SCb-pyridine (358.83 mg, 2.25 mmol, 3 eq) was added in portions and stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water (0.04%NH3H20 + lOmM NILHCO^ACN]; B%: l%-25%, 10 min) to yield Compound 51 (71.4 mg, 15%) was obtained as a white solid. LCMS: m/z = 590.1 (M+H+), 1H- NMR: (400MHz, DMSO-dfc,): 5 = 8.70 (s, 1 H) 8.28 (d, J= 8.6 Hz, 1 H) 7.79 (d, 7=7.9 Hz, 2 H) .65 - 7.76 (m, 1 H) 7.42 (d, J= 7.9 Hz, 2 H) 7.20 (s, 1 H) 7.13 (d, >13.1 Hz, 2 H) 7.08 (s, 1 H) .95 (s, 1 H) 6.84 - 6.92 (m, 2 H) 6.75 - 6.83 (m, 2 H) 5.11 (m, 1 H) 4.36 (s, 2 H) 3.17 (s, 3 H) .77 - 3.00 (m, 4 H) 2.43 - 2.47 (m, 2 H).
To a solution of (S)-l-{2-{(lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamlne ydrochloride salt (400 mg, 874.71 umol, 1 eq) and intermediate 2 (175.14 mg, 874.71 umol, 1 q) in DMF (4 mL) was added Et3N (531.07 mg, 5.25 mmol, 730.49 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (1.11 g, 1.75 mmol, 1.04 mL, 50% purity, 2 eq) was added rop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored y LCMS and when complete, the solution was dried with flowing N2 to yield intermediate 3 776 mg, crude) was obtained as yellow solid. LCMS: m/z = 694.2 (M+H*).
Intermediate 3 (770 mg, 1.11 mmol, 1 eq) in NH3.MeOH (2 mL, 1.00 eq) was stirred at 50 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was dried with lowing N2 to yield intermediate 4 (996 mg, crude) was obtained as yellow solid. LCMS: m/z = 12.3 (Μ+ΙΓ).
To a solution of intermediate 5 (900 mg, 1.76 mmol, 1 eq) in EtOH (20 mL) and ¾0 (5 mL) was added NEtCl (482.11, 8.80 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm o 90 °C, and then Fe (491.21 mg, 8.80 mmol, 5 eq) was added, then the mixture was stirred at 0 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, he reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 5 310 mg, 35%) was obtained as yellow solid. LCMS: m/z = 482.3 (M+FL).
To a solution of intermediate 5 (300 mg, 622.91 umol, 1 eq) in pyridine (2 mL) and CH3CN (2 mL) was cooled to 0 °C, then the SCb-pyridine (297.43 mg, 1.87 mmol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM N¾HC03)-ACN];B%: 1%- 5%,10min) to yield Compound 52 (40 mg, 11%) was obtained as white solid. LCMS: m/z = 60.1 (M-H+). 1H-NMR: (400 MHz, DMSO-tife) 5 = 8.35 - 8.44 (m, 1 H) 7.97 - 8.04 (m, 1 H) .81 - 7.91 (m, 1 H) 7.69 - 7.77 (m, 1 H) 7.42 - 7.59 (m, 2 H) 7.29 - 7.39 (m, 1 H) 7.21 - 7.27 (m, H) 7.10 - 7.17 (m, 1 H) 6.93 - 7.01 (m, 1 H) 6.76 - 6.89 (m, 4 H) 5.01 - 5.10 (m, 1 H) 4.30 (s, 2 H) 3.14 - 3.24 (m, 2 H) 2.88 - 3.00 (m, 1 H) 2.70 - 2.79 (m, 1 H), 2.53 - 2.54 (m, 2 H).
Synthesis of Compound 53
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (400 mg, 874.71 umol, 1 eq) and intermediate 2 (175.14 mg, 874.71 umol, 1 q)) in DMF (5 mL) was added EtgN (531.07 mg, 5.25 mmol, 730.49 uL, 6 eq), then cooled to 0 C, and then T3P (834.94 mg, 1.31 mmol, 780.32 uL, 50% purity, 1.5 eq) was added dropwise at °C under N2, the reaction was stirred at 25 °C for 1 h. The reaction was monitored by LCMS when complete, the reaction mixture was dried by flowing N2 to yield intermediate 3A (220 mg, rude) was obtained as a yellow solid. LCMS: m/z = 694.2(M+H+).
A solution of intermediate 3A (447 mg, 644.27 umol, 1 eq) in NHs/MeOH (7 M, 2.00 mL, 21.73 eq) was stirred at 60 °C for 1 h. The reaction was monitored by LCMS when complete, the solution was dried by flowing N2 to yield intermediate 3 (329 mg, crude) was obtained as a yellow solid. LCMS: m/z = 512.1 (M+I Γ).
A solution of intermediate 3 (329 mg, 643.09 umol, 1 eq) and NH4CI (237.0 mg, 4.43 mmol, 10 q) in EtOH (8 mL) and H20 (2 mL) was heated to 90 °C, then Fe (359.13 mg, 6.43 mmol, 10 q) was added, the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo and was purified by prep- HPLC (column: Phenomenex luna(2) C18250*50 lOu; mobile phase: [water(10mM NH-iHCCh)- ACN]; B%: 40%-50%, 20min) to yield intermediate 4 (153 mg, 47%) was obtained as a blue olid. LCMS: m/z = 480.1 (M-H+), ‘H-NMR: (400 MHz, DMSO-dk) 5 = 8.24 (d, >8.38 Hz, 1 H) 7.83 - 7.89 (m, 1 H) 7.73 - 7.82 (m, 2 H) 7.61 (d, >12.79 Hz, 2 H) 7.41 - 7.49 (m, 2 H) 7.32 dd, >8.38, 1.54 Hz, 1 H) 6.97 (br s, 1 H) 6.87 (s, 1 H) 6.79 (d, >8.16 Hz, 2 H) 6.41 (d, >8.38 Hz, 2 H) 4.97 - 5.04 (m, 1 H) 4.83 (br s, 1 H) 4.17 (s, 2 H) 2.88 - 2.99 (m, 3 H) 2.68 - 2.75 (m, 1 H) 2.42 - 2.47 (m, 2 H).
To a solution of intermediate 4 (153 mg, 317.68 rnnol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, added SCh.pyridine (151.69 mg, 953.05 umol, 3 eq) at 0 °C. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS when complete, % ammonium hydroxide (1.5 mL) was added drop wise, and then the mixture was dried by owing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile phase: [water(0.04%NH3H20+10mM NH4HC03>ACN]; B%: 10%-40%, lOmin) to yield Compound 53 (36.0 mg, 33%) was obtained as a white solid. LCMS: m/z = 560.1 (M-H÷), ¾- NMR: (400 MHz, DMSO-de) 5 = 8.14 (s, 1 H) 7.83 (d, J=7.46 Hz, 1 H) 7.77 (t, J=7.95 Hz, 2 H) .62 (s, 1 H) 7.39 - 7.47 (m, 2 H) 7.32 (d, J=8.56 Hz, 1 H) 7.04 (s, 1 H) 6.85 - 6.92 (m, 2 H) 6.80 6.82 (m, 3 H) 5.04 (t, J=7.27 Hz, 1 H) 4.23 (s, 2 H) 2.93 - 3.00 (m, 1 H) 2.88 - 2.93 (m, 2 H) .73 (dd, 1=13.82, 8.56 Hz, 1 H) 2.39 - 2.48 (m, 2 H).
To a mixture of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (198.67 mg, 820.04 umol, 1 g) in DMF (3 mL) was dropwise added EtsN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq) and cooled o 0 °C, then T3P (521.84 mg, 1.64 mmol, 487.70 uL, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 3 hat 25 °C under N2 atmosphere. The reaction was monitored y LCMS and when complete. The mixture was poured into ice-water (25 mL) and stirred for 10 minutes. Then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic hase was washed with brine (10 mL), dried with anhydrous NazSO^ filtered and concentrated in acuo to yield intermediate 3 (320 mg, crude) was obtained as a yellow oil. LCMS: m/z = 778.3 M+FT).
A solution of intermediate 3 (320 mg, 411.84 umol, 1 eq) in Nth/MeOH (10 mL) was heated to 0 °C, and stirred for 2 h at 60 °C under N2 atmosphere. The reaction was monitored by LCMS nd when complete. The mixture was concentrated in vacuo to yield intermediate 4 (200 mg, rude) was obtained as a yellow oil. LCMS: m/z = 554.2 (M+H+).
To a solution of intermediate 4 (200 mg, 361.66 umol, 1 eq) in EtOAc (5 mL) was added SnCh.2H20 (407.58 mg, 1.81 mmol, 5 eq) and heated to 50 °C and and stirred at 90 °C for 2 h under N2. The reaction was monitored by LCMS and when complete, the solution was added sat. eignette salt (20 mL), then filtered and the filter was extracted with ethyl acetate (2x 10 mL), the ombined organic layer was washed with brine (5 mL), filtered and concentrated in vacuo to yield ntermediate 5 (80 mg, crude) was obtained as a light yellow solid. LCMS: m/z = 546.2 (M+Na+).
A solution of intermediate 5 (80 mg, 152.77 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then S03-pyridine (72.95 mg, 458.32 umol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: 5%-35%,10min) to yield
Compound 54 (7.0 mg, 7%) was obtained as a white solid. LCMS: m/z = 602.0 (M-tL), 1H- NMR: (400 MHz, DMSO-tife). δ = 8.82 (s, 1 H) 8.25 (d, J=8.4 Hz, 1 H) 7.76 (br s, 1 H) 7.26 - 7.44 m, 2 H) 7.14 - 7.25 (m, 3 H) 7.06 - 7.14 (m, 2 H) 6.96 (m, J=8.6, 2 H) 6.84 - 6.93 (m, 4 H) 6.79 m, 2 H) 5.08 - 5.16 (m, 1 H) 4.38 (s, 2 H) 2.90 - 2.97 (m 1 H) 2.70 - 2.87 (m, 3 H) 2.37 - 2.43 (m, H).
To a solution of (S)-l-(2-((lH-lmidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamlne hydrochloride salt (400 mg, 874.71 umol, 1 eq) and intermediate 2 (169.03 mg, 874.71 umol, 1 q) in DMF (4 mL) was added Et3N (531.07 mg, 5.25 mmol, 730.49 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (1.11 g, 1.75 mmol, 1.04 mL, 50% purity, 2 eq) was added rop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored y LCMS and when complete, the solution was dried by flowing N2 to yield intermediate 3 (715 mg, crude) was obtained as yellow solid. LCMS: m/z = 505.2 (M+H+).
To a solution of intermediate 3 (700 mg, 1.39 mmol, 1 eq) in EtOH (16 mL) and H2O (5 mL) was added NH4CI (380.17 mg, 6.94 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (387.35 mg, 6.94 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 4 (628 mg, crude) was obtained as black brown solid. LCMS: m/z = 475.1
To a solution of intermediate 4 (628 mg, 1.32 mmol, 1 eq) in pyridine (3 mL) and MeCN (3 mL) was cooled to 0 °C, then the SCb-pyridine (631.80 mg, 3.97 mmol, 3 eq) was added at 0 °C and he reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 50*25 5u;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 5%- 5%,10min) to yield Compound 55 (10 mg, 1%) was obtained as light yellow solid. LCMS: m/z 553.1 (Μ-ΙΓ-). 1H-NMR: (400 MHz, DMSO-tfc) δ = 8.69 - 8.86 (m, 1 H) 8.10 - 8.27 (m, 1 H) .11 - 7.18 (m, 1 H) 6.97 - 6.99 (m, 3 H) 6.85 - 6.91 (m, 2 H) 6.78 (d, >8.44 Hz, 2 H) 6.63 (d, >8.68 Hz, 2 H) 5.02 - 5.17 (m, 1 H) 4.37 (s, 2 H) 2.90 - 3.00 (m, 1 H) 2.76 - 2.84 (m, 7 H) 2.64 2.66 (m, 2 H) 2.32 - 2.33 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (400 mg, 970.11 umol, 1 eq, HBr), intermediate 2 ( 186.94 mg, 1.07 mmol, .1 eq) in DMF (6 mL) was added EtsN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq), and the eaction mixture was cooled to 0 °C, and then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% urity, 1.5 eq) was added, then it was allowed warm to 25 °C and stirred for 13 h under N2 tmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was uenched by addition ice water (60 mL), and extracted with ethyl acetate (3 x 20 mL). The ombined organic layer was washed with brine (10 mL), dried with anhydrous Na2S(>4, filtered nd concentrated in vacuo to give a residue. The residue was purified by column hromatography silica gel (S1O2, Petroleum ether: Ethyl acetate = 10: 1 to 1 : 1) to yield ntermediate 3 (440 mg, 88%) was obtained as a yellow solid. LCMS: m/z = 489.2 (M+FT), Ή- NMR: (400 MHz, DMSO-de) δ = 8.47 (d, J= 8.68 Hz, 1 H) 8.07 - 8.15 (m, 2 H) 7.72 (dd, >5.07, .04 Hz, 1 H) 7.59 - 7.69 (m, 3 H) 7.45 - 7.50 (m, 2H) 7.24 - 7.34 (m, 3 H) 7.17 (dd, >5.01, .79 Hz, 1 H) 5.21 - 5.30 (m, 1 H) 3.32 - 3.37 (m, 1 H) 3.09 - 3.17 (m, 1 H) 2.80 - 2.84 (m, 2 H) .36 - 2.45 (m, 2 H).
To a solution of intermediate 3 (440 mg, 900.57 umol, 1 eq) in EtOH (7 mL) and H2O (2 mL) was added NHtCl (246.80 mg, 4.50 mmol, 5 eq), then the mixture was heated to 90 °C and Fe 251.46 mg, 4.50 mmol, 5 eq) was added, and the mixture was stirred at 90 °C for 2 h under Nz tmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was ltered and the filtrate was concentrated in vacuo to yield intermediate 4 (453 mg, crude) was btained as a yellow solid. LCMS: m/z = 459.2 (M+H+).
A mixture of intermediate 3 (453 mg, 987.79 umol, 1 eq) in MeCN (3 mL) and pyridine (3 mL) was cooled to 0 °C, then SCh-pyridine (471.66 mg, 2.96 mmol, 3 eq) was added, then the eaction was stirred for at 0 °C for 10 min. The reaction was monitored by TLC and when omplete, 7% ammonium hydroxide (3 mL) was added dropwise, then the mixture was dried by owing N2 to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 50*25 5u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 10%-40%, lOmin) to yield Compound 56 (163 mg, 31%) was obtained as a white solid. LCMS: m/z = 537.0 (M-H4), 1H- NMR: (400 MHz, DMSO-ifc) δ = 8.32 (d, >8.44 Hz, 1 H) 7.63 - 7.73 (m, 4 H) 7.33 (d, >7.82 Hz, 2 H) 7.15 - 7.23 (m, 3 H) 7.08 (s, 1 H) 6.96 (s, 1 H) 6.85 - 6.93 (m, 4 H) 5.04 - 5.12 (m, 1 H) .01 - 3.06 (m, 1 H) 2.79 - 2.88 (m, 3 H) 2.40 - 2.45 (m, 2 H).
Synthesis of Compound 57
To a mixture of of ( S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (400 mg, 970.11 umol, 1 eq, HBr), intermediate 2 (178.45 mg, 1.02 mmol, .05 eq) in DMF (10 mL) was added Et3N (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq) at 25 °C, nd then the solution was cooled to 0 °C, T3P (1.23 g, 1.94 mmol, 1.15 mL, 50% purity, 2 eq) was added drop-wise, then it was allowed warm to 25 °C and stirred for 15 h. The reaction was monitored by TLC and when complete, the reaction mixture was added H2O (50 mL) and xtracted with ethyl acetate (3 x 30 mL), then combined organic phase was washed with brine 50 mL), dried with anhydrous Na2S(>4 and concentrated in vacuo. The residue was purified by olumn chromatography (SiCh, dichloromethane: methyl alcohol = 100:1 to 50:1) to yield ntermediate 3 (420 mg, 74 %) was obtained as a yellow oil. LCMS: m/z = 489.2 (M+H+).
A mixture of intermediate 3 (420 mg, 859.63 umol, 1 eq) and NH4CI (284.06, 5.16 mmol, 6 eq) n EtOH (10 mL) and H2O (2 mL) was heated to 90 °C, and then Fe (288.04 mg, 5.16 mmol, 6 q) was added, and then the mixture was stirred at 90 °C for 2 h under N2 atmosphere. The eaction was monitored by TLC and when complete, the solution was concentrated in vacuo to ield intermediate 4 (450 mg, crude) was obtained as a yellow oil. 1 H-NMR: (400 MHz, DMSO-d>) δ = 8.46 (d, >8.68 Hz, 1 H) 8.09 (d, >8.68 Hz, 2 H) 7.72 (dd, >5.07, 0.92 Hz, 1 H) .65 (dd, >3.67, 0.98 Hz, 1 H) 7.53 - 7.61 (m, 2 H) 7.44 - 7.47 (m, 3 H) 7.36 - 7.42 (m, 1 H) .27 (s, 1 H) 7.17 (dd, >5.01, 3.79 Hz, 1 H) 5.17 - 5.33 (m, 1 H) 3.29 - 3.31 (m, , 1 H) 3.08 - .14 (m, 1 H) 2.74 - 2.85 (m, 2 H) 2.38 - 2.46 (m, 2 H).
To a solution of intermediate 4 (480 mg, 1.05 mmol, 1 eq), pyridine (5 mL) and MeCN (5 mL) was added SCb-pyridine (499.77 mg, 3.14 mmol, 3 eq) at 0 °C, then it was stirred for 10 min at 0 C. The reaction was monitored by LCMS and HPLC and when complete, the solution was dried y flowing N2. And the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 10%-40%,10min) firstly, then it was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: 10%-40%,10min) to yield Compound 57 24.5 mg, 4%). LCMS: m/z = 537.0 (M-H+), ‘H-NMR: (400 MHz, DMSO-4.) δ = 8.30 (d, >8.56 Hz, 1 H) 7.69 - 7.73 (m, 1 H) 7.62 - 7.64 (m, 2 H) 7.60 (d, >7.34 Hz, 1 H) 7.38 - 7.50 m, 2 H) 7.20 - 7.23 (m, 1 H) 7.15 - 7.18 (m, 1 H) 7.12 (s, 1 H) 7.08 (s, 1 H) 6.96 (s, 1 H) 6.83 - .91 (m, 4 H) 4.99 - 5.15 (m, 1 H) 2.98 - 3.07 (m, 1 H) 2.60 - 2.83 (m, 3 H) 2.38 - 2.47 (m, 2 H). Synthesis of Compound 58
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (400 mg, 970.11 umol, 1 eq, HBr), intermediate 2 ( 186.94 mg, 1.07 mmol, .1 eq) inDMF (6 mL) was added EtsN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq), the mixture was cooled to 0 °C and T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 1.5 eq) was added, hen it was allowed warm to 25 °C and stirred for 13 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by ice-water (40 mL), and then extracted with ethyl acetate (3 x 40 mL). The combined organic layer was washed with brine (20 mL), dried with anhydrous NazSO^ filtered and concentrated in vacuo. The esidue was purified by column chromatography silica gel (S1O2, Petroleum ether: Ethyl acetate 10: 1 to 5: 1) to yield intermediate 3 (362 mg, 76%) was obtained as a yellow solid. LCMS: m/z = 489.2 (M+H4), ‘H-NMR: (400 MHz, DMSO-dk) 5 = 8.51 (d, >8.56 Hz, 1 H) 8.09 (d, >8.68 Hz, 2 H) 7.68 - 7.77 (m, 2 H) 7.65 (dd, >3.67, 1.10 Hz, 1 H) 7.44 - 7.53 (m, 3 H) 7.29 - .35 (m, 3 H) 7.17 (dd, >5.01, 3.79 Hz, 1 H) 5.22 - 5.30 (m, 1 H) 3.33 - 3.37 (m, 1 H) 3.09 - .15 (m, 1 H) 2.91 - 2.95 (m, 2 H) 2.44 - 2.49 (m, 2 H).
To a solution of intermediate 3 (362 mg, 740.92 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (204.53 mg, 3.70 mmol, 5 eq) then the mixture was heated to 90 °C, and then e (206.88 mg, 3.70 mmol, 5 eq) was added, and then the mixture was stirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (562 mg, rude) was obtained as a yellow solid. LCMS: m/z = 459.2 (M+H"1"), 1H-NMR: (400 MHz, DMSOntfc) δ = 8.44 (d, >8.56 Hz, 1 H) 7.68 - 7.77 (m, 2 H) 7.62 (d, >3.06 Hz, 1 H) 7.49 - 7.60 m, 2 H) 7.27 - 7.34 (m, 2 H) 7.13 - 7.20 (m, 2 H) 6.80 (d, >7.95 Hz, 2 H) 6.42 (d, >7.95 Hz, 2 H) 4.97 - 5.10 (m, 1 H) 2.91 - 3.12 (m, 4 H) 2.69 - 2.90 (m, 2 H).
A mxiture of intermediate 4 (562 mg, 1.23 mmol, 1 eq) in MeCN (3 mL) and pyridine (3 mL) was cooled to 0 °C, then SCh-pyridine (585.15 mg, 3.68 mmol, 3 eq) was added drop-wise, then was stirred for 10 min. The reaction was monitored by TLC and when complete, 7% mmonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 5%-40%,10min) to yield Compound 58 140 mg, 21%) was obtained as a white solid. LCMS: m/z = 537.0 (M-FT), Ή-ΝΜΚ: (400 MHz, DMSO-dk) δ = 8.37 (d, >8.38 Hz, 1 H) 7.68 - 7.76 (m, 2 H) 7.63 (dd, >3.53, 0.88 Hz, 1 H) 7.52 - 7.58 (m, 1 H) 7.28 - 7.37 (m, 2 H) 7.21 (s, 1 H) 7.14 - 7.18 (m, 2 H) 7.08 (s, 1 H) 6.95 s, 1 H) 6.79 - 6.93 (m, 4 H) 5.01 - 5.11 (m, 1 H) 2.95 - 3.05 (m, 3 H) 2.80 - 2.86 (m, 1 H) 2.51 - .53 (m, 2 H).
Synthesis of Compound 59 To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-y1)lhiazol-4-yl)ethanamine hydrobromide alt (400 mg, 970.11 umol, 1.1 eq) and intermediate 1 (192.41 mg, 881.92 umol, 1 eq) inDMF (5 mL) was drop-wise added EtgN (535.45 mg, 5.29 mmol, 736.52 uL, 6 eq) and cooled to 0 °C, then T3P (420.91 mg, 1.32 mmol, 393.38 uL, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored y LCMS and when complete. The mixture was poured into ice-water (25 mL) and stirred for 10 min. Then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous NaaSO^ filtered and concentrated in vacuo. The residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 10: to 1 : 1) to yield intermediate 2 (430 mg, 87%) was obtained as a yellow solid. LCMS: m/z = 32.1 (M+H1·), IH-NMR: (400 MHz, CDCb) δ = 8.06 (d, >8.82 Hz, 2 H) 7.49 - 7.54 (m, 3 H) .44 (dd, 7=5.07, 1.10Hz, 1 H) 7.31 (d, 7=7.94 Hz, 2 H) 7.17 (d, 7=8.82 Hz, 2 H) 7.11 (dd, 7=4.96, .64 Hz, 1 H) 6.64 (s, 1 H) 6.27 (d, 7=8.38 Hz, 1 H) 5.27 - 5.35 (m, 1 H) 3.28 - 3.35 (m, 1 H) 3.15 3.23 (m, 1 H) 3.04 (t, 7=7.50 Hz, 2 H) 2.55 (td, 7=7.55, 2.09 Hz, 2 H).
A solution of intermediate 2 (430 mg, 808.93 umol, 1 eq) and NH4CI (216.35 mg, 4.04 mmol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (225.87 mg, 4.04 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (405 mg, crude) was obtained as a yellow solid. LCMS: m/z = 502.2 (M+H*), H-NMR: (400 MHz, CDCb) δ = 7.46 - 7.52 (m, 3 H) 7.40 (d, 7=4.85 Hz, 1 H) 7.23 - 7.33 (m, 2 H) 7.09 (d, 7=3.31 Hz, 1 H) 6.77 (d, 7=7.94 Hz, 2 H) 6.58 (s, 1 H) 6.46 - 6.56 (m, 2 H) 6.27 (d, 7=7.50 Hz, 1 H) 5.20 - 5.28 (m, 1 H) 3.07 - 3.15 (m, 1 H) 3.01 (t, 7=7.39 Hz, 2 H) 2.89 - 2.96 (m, H) 2.46 - 2.55 (m, 2 H).
A solution of intermediate 3 (405 mg, 807.44 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (385.54 mg, 2.42 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 5%-35%,10min) to yield Compound 59 (222 mg, 5%) was obtained as a white solid. LCMS: m/z = 580.0 (M-H4), 1H-NMR: (400 MHz, DMSO- ¾>) δ = 8.32 (d, >8.31 Hz, 1 H) 7.71 (d, >5.01 Hz, 1 H) 7.63 (d, >3.30 Hz, 1 H) 7.57 (d, >8.07 Hz, 2H) 7.37 (d, >7.82 Hz, 2 H) 7.21 (s, 1 H) 7.15 - 7.18 (m, l H) 7.12 (s, l H) 7.08 (s, 1 H) 6.95 s, 1 H) 6.85 - 6.92 (m, 4 H) 5.04 - 5.13 (m, 1 H) 3.01 - 3.07 (m, 1 H) 2.79 - 2.89 (m, 3 H) 2.43 (t, >7.46 Hz, 2 H).
To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (350 mg, 848.85 umol, 1 eg, HBr) and intermediate 2 (194.45 mg, 891.29 mol, 1.05 eq) in DMF (10 mL) was added EtsN (515.37 mg, 5.09 mmol, 708.90 uL, 6 eq) at 25 C, and then the solution was cooled to 0 °C, T3P (1.08 g, 1.70 mmol, 1.01 mL, 50% purity, 2 q) was added drop-wise at 0°C, then it was allowed warm to 25 °C and stirred for 15 h. The eaction was monitored by TLC and when complete, the reaction mixture was added H2O (50 mL) and extracted with ethyl acetate (3 x 30 mL), then combined organic phase was washed with rine (50 mL), dried with anhydrous Na2$04 and concentrated in vacuo. The residue was urified by column chromatography (S1O2, dichloromethane: methyl alcohol = 50:1) to yield ntermediate 3 (410 mg, 81%) was obtained as a yellow oil. LCMS: m/z = 532.2 (M+H4), 1H- NMR: (400 MHz, DMSO-de) δ = 8.46 (d, >8.68 Hz, 1 H) 8.08 (d, >8.68 Hz, 2 H) 7.72 (dd, >5.01, 0.98 Hz, 1 H) 7.65 (dd, >3.73, 1.04 Hz, 1 H) 7.38 - 7.53 (m, 6 H) 7.26 (s, 1 H) 7.17 (dd, >5.01, 3.67 Hz, 1 H) 5.22 - 5.28 (m, 1 H) 3.26 - 3.32 (m, 1 H) 3.08 - 3.14 (m, 1 H) 2.78 - 2.87 m, 2 H) 2.40 - 2.48 (m, 2 H).
A mixture of intermediate 3 (410 mg, 771.30 umol, 1 eq) and NH4CI (270.32 mg, 4.63 mmol, 6 q) in EtOH (10 mL) and H2O (2 mL) was heated to 90 °C, and then Fe (258.44 mg, 4.63 mmol, eq) was added, and then the mixture was stirred at 90 °C for 2 h under N2 atmosphere. The eaction was monitored by LCMS and when complete, the solution was concentrated in vacuo to ield intermediate 4 (450 mg, crude) was obtained as a yellow oil. LCMS: m/z = 502.0 (M+H+).
To a solution of intermediate 4 (450 mg, 897.15 umol, 1 eq), pyridine (5 mL) and MeCN (5 mL) was added SOs-pyridine (428.38 mg, 2.69 mmol, 3 eq) at 0 °C, then it was stirred for 10 min at 0 C. The reaction was monitored by LCMS and HPLC and when complete, the solution was dried y flowing N2. And the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 20%-50%,10min) to yield Compound 60 92.1 mg, 17%) was obtained as a white solid. LCMS: m/z = 580.0 (M-H+), 1H-NMR: (400 MHz, DMSO-dk) 5 = 8.31 (d, >8.56 Hz, 1 H) 7.68 - 7.74 (m, 1 H) 7.61 - 7.66 (m, 1 H) 7.55 (s, H) 7.43 - 7.51 (m, 3 H) 7.09 - 7.21(m, 4 H) 6.97 (br s, 1 H) 6.81 - 6.92 (m, 4 H) 5.05 - 5.14 (m, H) 2.98 - 3.10 (m, 1 H) 2.78 - 2.93 (m, 3 H) 2.40 - 2.47 (m, 2 H).
Synthesis of Compound 61 To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-y1)lhiazol-4-yl)ethanamine hydrobromide alt (400 mg, 970.11 umol, 1.1 eq) and intermediate 1 (192.41 mg, 881.92 umol, 1 eq) inDMF (5 mL) was drop wise added EtsN (535.45 mg, 5.29 mmol, 736.52 uL, 6 eq) and cooled to 0 °C, then T3P (420.91 mg, 1.32 mmol, 393.38 uL, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored y LCMS and when complete. The mixture was poured into ice-water (25 mL), and then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous NaaSO^ filtered and concentrated in vacuo. The residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 10: 1 to 1: 1) to ield intermediate 2 (430 mg, 87%) was obtained as a yellow solid. LCMS: m/z = 532.1 (M+H+), H-NMR: (400 MHz, CDCb) δ = 8.07 (d, >8.82 Hz, 2 H) 7.60 (d, >7.94 Hz, 1 H) 7.51 (dd, >3.64, 0.99 Hz, 1 H) 7.39 (br d, >7.28 Hz, 2 H) 7.31 - 7.35 (m, 1 H) 7.24 - 7.28 (m, 1 H) 7.20 d, >8.60 Hz, 2 H) 7.11 (dd, >5.07, 3.75 Hz, 1 H) 6.68 (s, 1 H) 6.25 (d, >8.16 Hz, 1 H) 5.30 - .38 (m, 1 H) 3.30 - 3.36 (m, 1 H) 3.11 - 3.24 (m, 3 H) 2.52 (t, >7.83 Hz, 2 H).
A solution of intermediate 2 (430 mg, 808.93 umol, 1 eq) and NH4CI (216.35 mg, 4.04 mmol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (225.87 mg, 4.04 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (405 mg, crude) was obtained as a yellow solid. LCMS: m/z = 502.2 (M+H*), H-NMR: (400 MHz, CDCb) δ = 7.62 (d, >7.94 Hz, 1 H) 7.52 (d, >2.87 Hz, 1 H) 7.38 - 7.46 m, 2 H) 7.32 - 7.37 (m, 1 H) 7.29 (s, 1 H) 7.11 (br t, >4.19 Hz, 1 H) 6.81 (d, >7.94 Hz, 2 H) .65 (s, 1 H) 6.56 (br d, >8.16 Hz, 2 H) 6.27 (d, >7.94 Hz, 1 H) 5.24 - 5.32 (m, 1 H) 3.09 - 3.21 m, 3 H) 2.95 (m, 1 H) 2.51 (t, >7.61 Hz, 2 H).
A solution of intermediate 3 (405 mg, 807.44 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (385.54 mg, 2.42 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile phase: [water(10mM NH4HCO3)- ACN] ;B% : 5%-35%,10min) to yield Compound 61 (252 mg, 2%) was obtained as a white solid. LCMS: m/z = 580.0 (M-H+), ‘H-NMR: (400MHz, DMSO- ¾>) δ = 8.37 (d, >8.4 Hz, 1 H) 7.71 (d, >4.6 Hz, 1 H) 7.64 (br d, >4.52 Hz, 2 H) 7.56 - 7.49 (m, H) 7.40 - 7.33 (m, 2 H) 7.25 (s, 1 H) 7.21 (s, 1 H) 7.18 - 7.15 (m, 1 H) 7.08 (s, 1 H) 6.95 (s, 1 H) .92 - 6.85 (m, 4 H) 5.15 - 5.06 (m, 1 H) 3.05 (br dd, >13.57, 5.99 Hz, 1 H) 2.97 - 2.83 (m, 3 H) .44 (t, >7.7 Hz, 2 H).
Preparation of intermediate 2
HCOOH (5.00 g, 104.08 mmol, 3 eq) was added drop-wise to DMF (12.5 mL) under stirring and ooling with ice water. EtjN (4.56 g, 45.10 mmol, 6.28 mL, 1.3 eq) was added in the same way ollowed by intermediate 6 (5 g, 34.69 mmol, 1 eq). Then intermediate 5 (5.14 g, 34.69 mmol, .26 mL, 1 eq) was added, the mixture was heated to 80 °C and stirred for 14 h. The reaction was monitored by LCMS and when complete, the solution was cooled down and poured out into ice- water (30 mL) under vigorous stirring. Concentrated HC1 was added till pH = 1 - 2. The recipitate was filtered, washed with water and dried in vacuo to yield intermediate 2 (6.6 g, rude) was obtained as a yellow solid. LCMS: m/z = 191.2 (M-H1"). 1H-NMR: (400 MHz, DMSCMO δ = 7.11 - 7.21 (m, 4 H) 2.84 - 3.03 (m, 3 H) 2.63 - 2.75 (m, 2 H) 1.25 (d, >6.85 Hz, H). S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (0.3 g, 27.58 umol, 1 eq) and intermediate 2 (139.88 mg, 727.58 umol, 1 eq) was added in DMF (3 mL) and then EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) was added at 0 °C, and then T3P 926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added, then the solution was stirred at 5 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was added o ice water (8 mL), then extracted with ethyl acetate (3 x 10 mL), then combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield ntermediate 3 (400 mg, crude) was obtained as yellow solid. LCMS: m/z = 506.0 (M+H+).
To a solution of intermediate 3 (400 mg, 791.06 rnnol, 1 eq ) in EtOH (8.5 mL) and H2O (3 mL) was added NH4CI (216.78 mg, 3.96 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (220.88 mg, 3.96 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 4 (360 mg, crude) was obtained as a yellow solid. LCMS: m/z = 476.3 (M+H+).
To a solution of intermediate 4 (350 mg, 735.81 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then the SCb-pyridine (351.34 mg, 2.21 mmol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7 % ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: YMC- Actus Triart Cl 8 100*30mm*5um;mobile phase: [water(10mM NH4HC03)-ACN];B%: 30%- 0%,12min) to yield Compound 62 (91.2 mg, 21%) was obtained as light yellow solid. LCMS: m/z = 554.0 (M-H+). ‘H-NMR: (400 MHz, DMSO-tik+D20) 5 = 8.29 - 8.39 (m, 1 H) 7.55 - 7.65 m, 2 H) 7.13 (t, >4.22 Hz, 1 H) 6.96 - 7.06 (m, 4 H) 6.82 - 6.94 (m, 5 H) 4.95 - 5.13 (m, 1 H) .95 - 3.09 (m, 1 H) 2.62 - 2.84 (m, 4 H) 2.26 - 2.42 (m, 2 H) 1.08 (br d, >6.85 Hz, 6 H). Synthesis of Compound 63
To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (0.4 g, 970.11 umol, 1 eq) and intermediate 1 (265.73 mg, 1.16 mmol, 1.2 eq) n DMF (5 mL) was added EfcN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq) and the mixture was ooled to 0 °C, and then T3P (1.23 g, 1.94 mmol, 1.15 mL, 50% purity, 2 eq) was added drop- wise. Then the mixture was allowed to warm to 25 °C and stirred for 5 h. The reaction was monitored by TLC and when complete, the reaction mixture was diluted with water (40 mL) and xtracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with brine (40 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to give a residue. The esidue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 2: 1 to : 1) to yield intermediate 2 (0.4 g, 76%) was obtained as a yellow solid. 'H-NMR: (400 MHz, CDCh) δ = 8.04 - 8.11 (m, 2 H) 7.78 - 7.86 (m, 2 H) 7.52 (dd, >3.67, 0.98 Hz, 1 H) 7.38 - 7.44 m, 3 H) 7.17 - 7.23 (m, 2 H) 7.11 (dd, >5.07, 3.73 Hz, 1 H) 6.67 (s, 1 H) 6.38 (d, >8.19 Hz, 1 H) 5.22 - 5.39 (m, 1 H) 3.28 - 3.37 (m, 1 H) 3.17 - 3.22 (m, 1 H) 3.04 - 3.08 (m, 2 H) 3.00 (s, 3 H) 2.54 - 2.59 (m, 2 H).
To a solution of intermediate 2 (0.4 g, 738.47 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (197.50 mg, 3.69 mmol, 5 eq) and the mixture was heated to 90 °C, and then Fe (206.22 mg, 3.69 mmol, 5 eq) was added. And then the mixture was stirred at 90 °C for 2 h. The eaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 3 (0.35 g, crude) was obtained as a ellow solid. LCMS: m/z = 512.0 (M+H+), 1H-NMR: (400 MHz, Methanol-^) δ = 8.00 - 8.20 m, 2 H) 7.60 - 7.85 (m, 4 H) 7.41 (s, 1 H) 7.02 - 7.26 (m, 3 H) 6.80 - 7.00 (m, 2 H) 5.51 (s, 1 H) .11 - 4.44 (m, 4 H) 3.65 (s, 3 H) 2.70 - 2.95 (m, 2 H).
A mixture of intermediate 3 (0.35 g, 684.02 umol, 1 eq) in pyridine (2.5 mL) and MeCN (2.5 mL) was cooled to 0 °C, and then SCfo-pyridine (326.61 mg, 2.05 mmol, 3 eq) was added. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, % ammonium hydroxide (4 mL) was added drop-wise, and then the mixture was dried by flowing N2. The residue was purified by prep-HPLC (column: YMC-Exphere C18 lOum 00*50mm 12nm;mobile phase: [water(10mM NH4HC03)-ACN];B%: 15%-45%,20min) to ield Compound 63 (304.2 mg, 74%) was obtained as a white solid. LCMS: m/z = 590.0 (M- H+), 1H-NMR: (400 MHz, DMSO-tfc) δ = 8.35 (d, >8.77 Hz, 1 H) 7.78 (d, >8.33 Hz, 2 H) .71 (dd, >5.04, 1.10 Hz, 1 H) 7.68 (s, 1 H) 7.64 (dd, >3.73, 1.10 Hz, 1 H) 7.42 (d, >8.33 Hz, H) 7.13 - 7.20 (m, 2 H) 7.08 (s, 4 H) 6.83 - 6.93 (m, 4 H) 5.03 - 5.15 (m, 1 H) 3.15 (s, 3 H) .05 (dd, >13.81, 5.92 Hz, 1 H) 2.79 - 2.91 (m, 3 H) 2.40 - 2.47 (m, 2 H).
Synthesis of Compound 64
To a solution of (S)-2-(4-nitrophenyl)-l-(2-lhiophen-2-yl)tkiazol-4-yl)ethanamine hydrobromide salt (0.4 g, 970.11 umol, 1 eq) and intermediate 1 (194.25 mg, 970.11 umol, 1 eq) n DMF (5 mL) was added EtgN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq) and the solution was ooled to 0 °C, and then T3P (1.23 g, 1.94 mmol, 1.15 mL, 50% purity, 2 eq) was added ropwise at 0°C. The mixture was allowed to warm to 25 °C and stirred for 13 h. The reaction was monitored by TLC and when complete, the reaction mixture was diluted with ice water (50 mL) and extracted with ethyl acetate (3 x 25 mL). The combined organic layer was washed with rine (50 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to give a esidue. The residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl cetate = 5: 1 to 0: 1) to yield intermediate 2 (0.5 g, 90%) was obtained as a yellow solid. Ή- NMR: (400 MHz, CDCb) δ = 7.90 - 8.04 (m, 3 H) 7.80 (d, >7.89 Hz, 1 H) 7.60 - 7.69 (m, 1 H) .36 - 7.55 (m, 4 H) 7.24 - 7.35 (m, 2 H) 7.00 - 7.11 (m, 3 H) 6.60 (s, 1 H) 6.30 (d, J=8.33 Hz, 1 H) 5.22 - 5.38 (m, 1 H) 3.31 - 3.51 (m, 2 H) 3.15 - 3.28 (m, 1 H) 3.00 - 3.15 (m, 1 H) 2.62 - 2.66 m, 2 H).
A mixture of intermediate 2 (0.5 g, 876.12 umol, 1 eq) and SnCl2.2H20 (988.48 mg, 4.38 mmol, eq) in ethyl acetate (5 mL) was heated to 50 °C and the reaction was stirred at 50 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with at. seignette salt (50 mL) and extracted with ethyl acetate (50 mL). The combined organic layer was washed with brine (50 mL), dried with anhydrous Na2SC>4, filtered and concentrated in acuo to yield intermediate 3 (0.35 g, 83%) was obtained as a yellow solid. LCMS: m/z = 484.2 Μ+ΡΓ), 'H-NMR: (400 MHz, CDCb) δ = 7.97 (d, >8.31 Hz, 1 H) 7.75 (d, >7.70 Hz, 1 H) .53 - 7.66 (m, 1 H) 7.36 - 7.49 (m, 3 H) 7.23 - 7.34 (m, 3 H) 6.96 - 7.07 (m, 1 H) 6.64 (d,
>8.31 Hz, 2 H) 6.47 (s, 1 H) 6.42 (d, >8.31 Hz, 2 H) 6.04 (d, >8.31 Hz, 1 H) 5.07 - 5.23 (m, 1 H) 3.27 - 3.41 (m, 2 H) 3.01 (dd, >13.33, 5.62 Hz, 1 H) 2.78 (dd, >13.33, 8.56 Hz, 1 H) 2.56 t, >7.70 Hz, 2 H).
A mixture of intermediate 3 (0.33 g, 682.32 umol, 1 eq) in pyridine (3 mL) and MeCN (3 mL) was cooled to 0 °C, and then SCb-pyridine (325.79 mg, 2.05 mmol, 3 eq) was added in batches, he reaction was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (5 mL) was added drop-wise and the mixture was ried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20)-ACN];B%: 15%-45%, lOmin) to yield Compound 64 (16.3 mg, 4%) was obtained as a white solid. LCMS: m/z = 562.1 (M-H4), 'H- NMR: (400 MHz, DMSO-<¾) δ = 8.35 (d, >8.44 Hz, 1 H) 8.07 (d, >8.44 Hz, 1 H) 7.90 (d, >7.95 Hz, 1 H) 7.69 - 7.78 (m, 2 H) 7.61 - 7.67 (m, 2 H) 7.47 - 7.60 (m, 2 H) 7.34 - 7.41 (m, 1 H) 7.26 - 7.32 (m, 1 H) 7.14 - 7.18 (m, 2 H) 7.09 (br s, 3 H) 6.84 - 6.94 (m, 4 H) 5.05 - 5.19 (m, H) 3.20 - 3.26 (m, 2 H) 3.05 (dd, >13.63, 5.81 Hz, 1 H) 2.85 (dd, >13.63, 8.38 Hz, 1 H) 2.53 2.57 (m, 2 H).
Synthesis of Compound 65
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yQethanamine hydrobromide oft (350 mg, 848.85 umol, 1 eq) and intermediate 1 (186.96 mg, 933.73 umol, 1.1 eq) in DMF (3 mL) was drop-wise added EtgN (515.37 mg, 5.09 mmol, 708.90 uL, 6 e<y) and cooled to 0 °C, then T3P (540.17 mg, 1.70 mmol, 504.84 uL, 2 eq) was added at 0 °C, and then the mixture was warmed o 25 °C and stirred for 13 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS nd when complete. The mixture was poured into ice-water (20 mL) and then the mixture was xtracted with ethyl acetate (2 x 10 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield intermediate 2 (200 mg, crude) was obtained as a light yellow solid. LCMS: m/z = 514.2 (M+H+).
To a solution of intermediate 2 (200 mg, 389.39 umol, 1 eq) in EtOAc (10 mL) was added SnCl2.2HzO (439.32 mg, 1.95 mmol, 5 eq) and heated to 50 °C and stirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the solution was added at. seignette salt (40 mL), then filtered and the filter was extracted with ethyl acetate (2x 10 mL), washed with brine (5 mL), filtered and concentrated in vacuo to yield intermediate 5 (100 mg, rude) was obtained as a light yellow solid. LCMS: m/z = 482.3 (M-H+).
A solution of intermediate 3 (100 mg, 207.04 rnnol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SCb-pyridine (177.71 mg, 1.12 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20)-ACN];B%: 5%-40%,10min) to yield Compound 65 (44.1 mg, 0%) was obtained as a white solid. LCMS: m/z = 562.0 (M-H+), ‘H-NMR: (400 MHz, DMSO- d6) δ = 8.33 (d, J= 8.77 Hz, 1 H) 7.76 - 7.85 (m, 3 H) 7.67 - 7.72 (m, 2 H) 7.65 (s, 1 H) 7.62 (dd, >3.51, 1.32 Hz, 1 H) 7.39 - 7.48 (m, 2 H) 7.34 (dd, >8.55, 1.53 Hz, 1 H) 7.16 (dd, >5.04, 3.73 Hz, 1 H) 7.07 (s, 4 H) 7.02 (s, 1 H) 6.85 - 6.92 (m, 4 H) 5.04 - 5.13 (m, 1 H) 3.04 (dd, >13.81, .92 Hz, 1 H) 2.90 - 2.98 (m, 2 H) 2.82 (dd, >13.59, 8.77 Hz, 1 H) 2.52 - 2.56 (m, 2 H). S To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide alt (300 mg, 727.58 umol, 1 eq) and intermediate 1 (176.27 mg, 727.58 umol, 1 eq) in DMF (3 mL) was drop-wise added Et3N (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 3 hat 25 °C under Nz atmosphere. The reaction was monitored y LCMS and when complete, the mixture was poured into ice-water (20 mL) and then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine 5 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield intermediate 2 300 mg, crude) was obtained as a light yellow solid. LCMS: m/z = 556.2 (M+H+).
To a solution of intermediate 2 (300 mg, 539.89 umol, 1 eq) in EtOAc (10 mL) was added nClz.2HzO (609.13 mg, 2.70 mmol, 5 eq) and heated to 50 °C and stirred at 50 °C for 1 h under Nz. The reaction was monitored by LCMS and when complete, the solution was added sat. eignette salt (40 mL), then filtered and the filter was extracted with ethyl acetate (3 x 10 mL), washed with brine (1 x 5 mL), filtered and concentrated in vacuo to yield intermediate 3 (260 mg, rude) was obtained as a light yellow solid. LCMS: m/z = 526.1 (M+H4). ‘H-NMR: (400 MHz, DMSOntfc) δ = 8.28 (d, >8.56 Hz, 1 H) 7.70 (dd, >5.01, 0.98 Hz, 1 H) 7.62 (dd, >3.67, 0.98 Hz, H) 7.30 - 7.43 (m, 2 H) 7.07 - 7.20 (m, 5 H) 6.90 - 6.98 (m, 2 H) 6.85 - 6.89 (m, 2 H) 6.81 - 6.83 m, 2 H) 6.43 (d, >8.31 Hz, 2 H) 5.01 - 5.10 (m, 1 H) 2.98 - 3.03 (m, 1 H) 2.74 - 2.83 (m, 3 H) .35 - 2.42 (m, 2 H).
A solution of intermediate 3 (260 mg, 494.59 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SCb-pyridine (236.16 mg, 1.48 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing Nz, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: 15%-55%,10min) to yield Compound 66 44.1 mg, 20%) was obtained as a white solid. LCMS: m/z = 604.0 (M-H+), 1H-NMR: (400 MHz, DMSO-^e) δ = 8.29 (d, >8.56 Hz, 1 H) 7.69 - 7.70 (m, 1 H) 7.65 (s, 1 H) 7.61 - 7.62 (m, 1 H) .32 - 7.39 (m, 2 H) 7.04 - 7.20 (m, 8 H) 6.82 - 6.98 (m, 8 H) 5.04 - 5.13 (m, 1 H) 3.04 (dd, >13.75, 5.93 Hz, 1 H) 2.71 - 2.89 (m, 3 H) 2.35 - 2.43 (m, 2 H).
To a mixture of of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (400 mg, 970.11 umol, 1.1 eq) and intermediate 1 (170.42 mg, 881.92 umol, 1 q) in DMF (5 mL) was dropwise added EtsN (535.45 mg, 5.29 mmol, 736.52 uL, 6 eq) and cooled o 0 °C, then T3P (420.91 mg, 1.32 mmol, 393.38 uL, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (25 mL) and then he mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 10: 1 to 1: 1) to ield intermediate 2 (350 mg, 78%) was obtained as a yellow solid. LCMS: m/z = 507.2 (M+fL), H-NMR: (400 MHz, CDCb) δ = 8.01 - 8.08 (m, 2 H) 7.49 - 7.52 (m, 1 H) 7.41 - 7.45 (m, 1 H) .09 - 7.16 (m, 3 H) 7.06 (d, >8.60 Hz, 2 H) 6.60 - 6.67 (m, 3 H) 6.16 (d, >8.60 Hz, 1 H) 5.30 - .39 (m, 1 H) 3.27 - 3.34 (m, 1 H) 3.16 - 3.23 (m, 1 H) 2.83 - 2.93 (m, 8 H) 2.50 (t, >7.50 Hz, 2
H).
A solution of intermediate 2 (350 mg, 690.83 umol, 1 eq) and NH4CI (184.76 mg, 3.45 mmol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (192.90 mg, 3.45 mmol, 5 eq) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (329 mg, crude) was obtained as a yellow solid. LCMS: m/z = 477.2 (M+H+).
A solution of intermediate 3 (329 mg, 690.23 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (329.58 mg, 2.07 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water (0.04%NH3H2O)- ACN] ; B%: 15%-45%, lOmin) to yield Compound 67 (182 mg, 5%) was obtained as a white solid. LCMS: m/z = 555.1 (M-H+), 1H-NMR: (400 MHz, Methanol-^) δ = 8.27 (br d, >8.80 Hz, 1 H) 7.57 (dd, >3.67, 1.10 Hz, 1 H) 7.52 - 7.56 (m, 1 H) 7.12 (dd, >5.01, 3.79 Hz, 1 H) 7.00 - 7.09 (m, 6 H) 6.94 (br d, >3.18 Hz, 2 H) 6.86 (br s, 1 H) 5.18 - 5.33 (m, 1 H) 3.17 (dd, >14.00, 5.32 Hz, 1 H) 2.98 (s, 6 H) 2.89 - 2.95 (m, 2 H) 2.73 - .88 (m, 2 H) 2.48 (t, .7=6.91 Hz, 2 H). To a solution of of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol~4-yl)-2-(4- itrophenyl)ethanamine hydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 1 123.96 mg, 820.04 umol, 1 eq) in DMF (4 mL) was added EtsN (4.92 mmol, 684.84 uL, 6 eq), nd the mixture was cooled to 0 °C, then T3P (1.23 mmol, 731.55 uL, 50% purity, 1.5 eq) was dded drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 6 h. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured nto water (30 mL), and extracted with ethyl acetate (3 x 20 mL). Then combined organic phase was washed with brine (2 x 20 mL), dried with anhydrous NazS04 and concentrated in vacuo to ield intermediate 2 (350 mg, crude) was obtained as a yellow oil. LCMS: m/z = 463.2 (M+H4), H-NMR: (400 MHz, CDCh) δ = 8.25 - 8.57 (m, 2 H) 8.03 - 8.05 (m, 1 H) 7.57 - 7.68 (m, 1 H) .46 - 7.54 (m, 1 H) 7.09 - 7.21 (m, 3 H) 6.91 - 6.99 (m, 1 H) 6.65 - 6.70 (m, 1 H) 6.54 - 6.58 (m, H) 5.21 - 5.34 (m, 1 H) 4.30 (s, 2 H) 3.19 - 3.31 (m, 2 H) 3.05 - 3.19 (m, 2 H) 2.42 - 2.60 (m, 2
H).
A solution of intermediate 2 (350 mg, 756.72 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (202.39 mg, 3.78 mmol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (211.29 mg, 3.78 mmol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was ltered, the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: %-35%, 1 Omin) to yield intermediate 3 (100 mg, 29%) was obtained as a yellow oil. LCMS: m/z = 433.3 (M+H4), 1H-NMR: (400 MHz, CDCh) δ = 8.39 (d, >3.791 Hz, 1 H) 8.28 (s, 1 H) .62 (s, 1 H) 7.51 (d, >7.703 Hz, 1 H) 7.19 (dd, >7.641, 4.952 Hz, 1 H) 6.90 (s, 1 H) 6.74 (d, >8.192 Hz, 2 H) 6.68 (s, 1 H) 6.52 (d, >8.314 Hz, 2 H) 6.29 (d, >7.947 Hz, 1 H) 5.15 - 5.22 m, 1 H) 4.30 (s, 2 H) 2.89 - 3.07 (m, 4 H) 2.50 - 2.59 (m, 1 H) 2.38 - 2.46 (m, 1 H).
To a solution of intermediate 3 (100 mg, 231.19 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SOz-pyridine (110.39 mg, 693.58 umol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by LCMS and when complete, 7% ammonium hydroxide (3 mL) was dded drop-wise, and then the mixture was dried by flowing Nz, and the residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM NH4HCC>3)-ACN];B%: l%-20%,10min) to yield Compound 68 (42.3 mg, 36%) was obtained as white solid. LCMS: m/z = 511.1(M-H+), 1H-NMR: (400 MHz, DMSO-tfc) 5 = 8.61 (s, 1 H) .35 - 8.46 (m, 2 H) 8.26 (d, >8.559 Hz, 1 H) 7.76 (br s, 1 H) 7.56 (d, >7.825 Hz, 1 H) 7.26 - .29 (m, 1 H) 7.21 (br s, 1 H) 7.15 (s, 1 H) 7.08 (s, 1 H) 6.96 (br s, 1 H) 6.89 (d, >8.437 Hz, 2 H) 6.75 - 6.84 (m, 2 H) 5.06 - 5.13 (m, 1 H) 4.35 (s, 2 H) 2.94 (dd, >13.633, 6.663 Hz, 1 H) .74 - 2.83 (m, 3 H) 2.43 (t, >7.519 Hz, 2 H).
Synthesis of Compound 69
To a solution of intermediate 1 (300 mg, 565.62 umol, 1 eq) and intermediate 2 (85.50 mg, 65.62 umol, 1 eq) in DMF (4 mL) was added EtsN (3.39 mmol, 472.36 uL, 6 eq), and the mixture was cooled to 0 °C, then T3P (848.43 umol, 504.58 uL, 50% purity, 1.5 eq) was added rop-wise. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured into water (30 mL), and extracted with ethyl acetate (3 x 20 mL). Then ombined organic phase was washed with brine (20 mL), dried with anhydrous NaaSO-t, filtered nd concentrated in vacuo. The residue was purified by silica gel chromatography (Petroleum ther: Ethyl acetate = 5:1 to Dichloromethane: Methanol = 10: 1) to yield intermediate 3 (150 mg, 50 %) was obtained as a yellow oil. LCMS: m/z = 477.2 (Μ+ΕΓ), 1H-NMR: (400 MHz, CDCb) δ = 8.37 - 8.51 (m, 2 H) 8.05 (d, >8.770 Hz, 2 H) 7.43 - 7.54 (m, 2 H) 7.18 (dd, >7.454, 4.823 Hz, 1 H) 7.12 (d, >8.770 Hz, 2 H) 6.99 (s, 1 H) 6.70 (s, 1 H) 6.29 (br d, >8.331 Hz, 1 H) 5.31 - 5.36 (m, 1 H) 4.28 (s, 2 H) 3.55 (s, 3 H) 3.22 - 3.30 (m, 1 H) 3.09 - 3.14 (m, 1 H) .90 - 3.01 (m, 2 H) 2.47 - 2.57 (m, 2 H).
A solution of intermediate 3 (150 mg, 314.76 umol, 1 eq ) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (84.19 mg, 1.57 mmol, 5 eq), and then the solution was heated to 90 °C, Fe (87.89 mg, 1.57 mmol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (200 mg, crude) was obtained as a yellow olid. LCMS: m/z = 447.2 (M+H+), ‘H-NMR: (400 MHz, Methanol-^) δ = 8.22 - 8.59 (m, 3 H) .62 (br d, >7.497 Hz, 1 H) 7.20 - 7.37 (m, 2 H) 7.01 (s, 1 H) 6.85 (d, >7.938 Hz, 2 H) 6.62 (d, >8.379 Hz, 2 H) 5.15 - 5.21 (m, 1 H) 4.48 (s, 2 H) 3.76 (s, 3 H) 3.05 (dd, >13.892, 6.395 Hz, 1 H) 2.77 - 2.94 (m, 3 H) 2.46 - 2.56 (m, 2 H).
To a solution of intermediate 4 (200 mg, 447.86 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SCb-pyridine (213.85 mg, 1.34 mmol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: l%-25%,10min) to yield Compound 69 73.1 mg, 31%) was obtained as a white solid. LCMS: m/z = 525.1 (M-H+), ‘H-NMR: (400 MHz, DMSO-ifc) δ = 8.70 (s, 1 H) 8.34 - 8.44 (m, 2 H) 8.29 (d, >8.437 Hz, 1 H) 7.76 (br s, 1 H) .56 (br d, >7.825 Hz, 1 H) 7.27 (dd, >7.641, 4.829 Hz, 1 H) 7.21 (br s, 1 H) 7.16 (s, 1 H) 7.10 br d, >12.349 Hz, 1 H) 7.00 (s, 1 H) 6.96 (br s, 1 H) 6.87 (d, >8.314 Hz, 2 H) 6.75 (d, >8.436 Hz, 2 H) 5.09 - 5.17 (m, 1 H) 4.44 (s, 2 H) 3.64 (s, 3 H) 2.74 - 2.93 (m, 4 H) 2.42 - 2.47 (m, 2
H).
Synthesis of Compound 70
To a solution of (S)-2-(4-nitrophenyl)-l-(2-lhiophen-2-yl)tkiazol-4-yl)ethanamine ydrobromide salt (0.3 g, 727.58 umol, 1 eq) and intermediate 2 (110.70 mg, 727.58 umol, 1 eq) n DMF (5 mL) was added EtiN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and the mixture was ooled to 0 °C, and then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added rop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water 40 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with brine (40 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield ntermediate 3 (0.3 g, crude) was obtained as a yellow oil. LCMS: m/z = 466.0 (M+H+).
To a solution of intermediate 3 (0.3 g, 644.40 umol, 1 eq) in EtOH (6 mL) and HzO (2 mL) was dded NH4CI (172.35 mg, 3.22 mmol, 5 eq) and then the mixture was heated to 90 °C, then Fe 179.95 mg, 3.22 mmol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (0.3 g, crude) was obtained as a yellow solid. LCMS: m/z = 436.2 (M+FT), ‘H-NMR: (400 MHz, CDCh) δ = 9.02 (s, 1 H) 8.51 - 8.62 (m, 2 H) 8.01 (s, 1 H) 7.49 (d, >3.51 Hz, 1 H) 7.40 (d, >4.82 Hz, 1 H) 7.02 - 7.14 (m, 1 H) 6.77 (d, >7.89 Hz, 2 H) 6.66 (s, 1 H) 6.46 - 6.58 (m, 3 H) 5.14 - 5.31 (m, 1 H) 3.64 - 3.80 (m, 1 H) 3.02 - 3.20 (m, 1 H) 2.46 - 2.56 (m, 2 H) 1.20 - 1.31 (m, 2 H). A mixture of intermediate 4 (0.3 g, 688.76 umol, 1 eg) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, and then SCb-pyridine (328.87 mg, 2.07 mmol, 3 eg) was added. The mixture was stirred at 0 °C for 0.17 hr. The reaction was monitored by TLC and when complete, % ammonium hydroxide (3 mL) was added drop wise and the mixture was dried by flowing N2 o give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 10u;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN];B%: 5%- 5%, 1 lmin) to yield Compound 70 (96.4 mg, 27%) was obtained as a white solid. LCMS: m/z 513.9 (M-FT), Έ-ΝΜΒ: (400 MHz, DMSO-4,) δ = 8.97 (s, 1 H) 8.62 (s, 2 H) 8.32 (d, >8.44 Hz, 1 H) 7.71 (d, >5.01 Hz, 1 H) 7.63 (d, >2.81 Hz, 1 H) 7.21 (s, 1 H) 7.14 - 7.18 (m, 2 H) .08 (s, 1 H) 6.96 (s, 1 H) 6.81 - 6.91 (m, 4 H) 5.04 - 5.12 (m, 1 H) 2.96 - 3.07 (m, 1 H) 2.74 - .87 (m, 3 H) 2.44 - 2.48 (m, 2 H).
Synthesis of Compound 71
To a solution of intermediate 1 (2 g, 7.57 mmol, 1 eqr), intermediate 2 methyl prop-2-enoate (977.85 mg, 11.36 mmol, 1.02 mL, 1.5 eg), phenylurea (206.20 mg, 1.51 mmol, 0.2 eg) and K2CO3 (2.09 g, 15.14 mmol, 2 eg) in anhydrous DMF (20 mL) was added diacetoxypalladium 170.01 mg, 757.24 umol, 0.1 eq), then degassed and purged with N23 times, and stirred at 130 C for 3 h. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was added water 50 mL, extracted with ethyl acetate (2 x 10 mL), the combined organic ayer was washed with brine 5 mL, filtered and concentrated in vacuo to yield intermediate 3 1.73 g, crude) was obtained as a black brown solid. LCMS: m/z = 269.9 (M+H+). 1H-NMR: 400 MHz, CDCI3) δ = 8.39 (d, >2.81 Hz, 1 H) 8.37 (d, >1.59 Hz, 1 H) 7.66 (d, >16.14 Hz, 1 H) 7.34 - 7.47 (m, 6 H) 6.48 (d, >16.14 Hz, 1 H) 5.15 (s, 2 H) 3.83 (s, 3 H)
To a solution of methyl intermediate 3 (1.5 g, 1 eq) in EtOH (5 mL) was added Pd/C (200 mg, 0% purity), then degassed and purged with ¾ (15 Psi) three times, and the reaction mixture was stirred at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the eaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (2 , crude) was obtained as a black brown oil. LCMS: m/z = 182.1 (M+H+).
To a solution of intermediate 4 (500 mg, 2.76 mmol, 1 eq) in THF (1 mL) and ¾0 (1 mL) was dded L1OH.H2O (231.60 mg, 5.52 mmol, 2 eq) , then the reaction was stirred at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was oncentrated in vacuo to yield intermediate 5 (400 mg, crude) was obtained as a light yellow oil. LCMS: m/z = 166.0 (Μ+ΙΓ).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (300 mg, 727.58 umol, 1 eq), intermediate 5 (243.25 mg, 1.46 mmol, 2 eq) nd EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) in DMF (1 mL) was cooled to 0 °C, and then T3P (463.01 mg, 1.46 mmol, 432.72 uL, 50% purity, 1 eq) was added at 0 °C, then the mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC and LCMS and when complete, he reaction mixture was poured into water (30 mL), and extracted with EtOAc (2 x 10 mL). the ombined organic layer was washed with brine 5 mL, dried with anhydrous Na2SC>4, filtered and oncentrated in vacuo to yield intermediate 7 (300 mg, crude) was obtained as a yellow oil. LCMS: m/z = 481.1 (M+H+).
To a solution of intermediate 7 (300 mg, 624.27 umol, 1 eq) in EtOH (5 mL) was added sat. NH4CI (1.53 g, 1 mL), then the mixture heated to 90 °C, then Fe (348.63 mg, 6.24 mmol, 10 eq) was added , the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 8 (200 mg, crude) was obtained as a light yellow solid. LCMS: m/z = 451.0 M+H+). ntermediate 8 (200 mg, 443.88 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added 141.30 mg, 887.75 umol, 2 eq) at 0 °C and stirred for 10 min. The reaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, nd then the mixture was dried by flowing Nz, and the residue was purified by prep-HPLC column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mMNHiHC03)- ACN];B%: l%-30%,10min) to yield Compound 71 (15 mg, 13%) was obtained as a white solid. LCMS: m/z = 529.0 (M-FT), 'H-NMR: (400 MHz, DMSO-ί/β) δ = 9.94 (br s, 1 H) 8.28 (d, >8.56 Hz, 1 H) 7.98 (br d, >1.96 Hz, 1 H) 7.88 (s, 1 H) 7.71 (dd, >5.01, 1.10 Hz, 1 H) 7.64 dd, >3.67, 1.10 Hz, 1 H) 7.21 (br s, 1 H) 7.17 (dd, >5.01, 3.67 Hz, 1 H) 7.14 (s, 1 H) 7.08 (br , 1 H) 7.06 (br s, 1 H) 6.95 (br s, 1 H) 6.88 (q, >8.56 Hz, 4 H) 5.09 (br d, >5.87 Hz, 1 H) 3.05 dd, >13.82, 5.75 Hz, 1 H) 2.82 (dd, >13.88, 8.62 Hz, 1 H) 2.72 (t, >7.52 Hz, 2 H) 2.38 - 2.44 m, 2 H).
Synthesis of Compound 72 HCOOH (1.04 g, 21.19 mmol, 98% purity, 3 eq) was added drop-wise to DMF (3 mL) under tirring and cooling with ice water (0 °C), EtaN (929.30 mg, 9.18 mmol, 1.28 mL, 1.3 eq) was dded in the same way followed by intermediate 2 (1.02 g, 7.06 mmol, 1 eq). After dissolution, ntermediate 1 (1 g, 7.06 mmol, 1 eq) was added, the mixture was heated up to 80 °C for 14 h. The reaction was monitored by LCMS and when complete, the solution was cooled down and oured out into ice-cold (30 mL) water under vigorous stirring. Concentrated HC1 was added till H=l-2. The precipitate was filtered off, washed with water and air-dried to yield intermediate 3 600 mg, 31%) was obtained as a white solid. LCMS: m/z = 270.2 (Μ+ΕΓ).
HC1 (2 M, 1.5 mL, 2.70 eq) was added to a stirred solution of intermediate 3 (0.3 g, 1.11 mmol, eq) in THF (15 mL) and the resultant solution was heated to 70 °C and stirred for 7 h. The eaction was monitored by LCMS and when complete, concentrated in vacuo to yield ntermediate 3A (255.43 mg, crude) was obtained as colorless oil. LCMS: m/z = 227.9 (M-H+). ntermediate 3A (255.43 mg, 1.11 mmol, 1 eq) in DMSO (5 mL) was heated to 140 °C and tirred for 3 h. The reaction was monitored by LCMS and when complete, the solution oncentrated in vacuo to yield intermediate 4 (206.47 mg, crude) was obtained as a light yellow olid. LCMS: m/z = 186.1 (M+H+). S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (458.69 mg, 1.11 mmol, 1 eq) and intermediate 4 (206.48 mg, 1.11 mmol, 1 eq) was added in DCM (10 mL) and then EtgN (675.42 mg, 6.67 mmol, 929.05 uL, 6 eq) was added at 0 °C, and then T3P 1.06 g, 1.67 mmol, 992.42 uL, 50% purity, 1.5 eq) was added, then the solution was stirred at 25 C for 2 h. The reaction was monitored by LCMS and when complete, the solution was oncentrated in vacuo
The residue was purified by prep-TLC (petroleum ether: ethyl acetate = 5:1) to yield ntermediate 6 (180 mg, 32%) was obtained as a white solid. LCMS: m/z = 499.1 (M+H+).
To a solution of intermediate 6 (180 mg, 360.72 umol, 1 eq) in EtOH (4 mL) and H2O (2 mL) was added NH4CI (96.48 mg, 1.80 mmol, 5 eq), then the solution was hearted to 90 °C, Fe 100.72 mg, 1.80 mmol, 5 eq) was added, and then the solution was stirred at 90 °C for 2 h. The eaction was monitored by LCMS and HPLC and when complete, the solution was concentrated n vacuo to yield intermediate 7 (169.18 mg, crude) was obtained as a light yellow solid. LCMS: m/z = 469.2 (M+I f ).
To a solution of intermediate 7 (169.18 mg, 360.71 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added S03-pyridine (172.23 mg, 1.08 mmol, 3 eq) at 0 °C slowly, the reaction was tirred at 0 °C far 0.17 h. The reaction was monitored by LCMS and HPLC and when complete, he solution was dried by flowing N2. And the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%>¾H20+10mMNH4HC03)-ACN];B%: %-40%,10min) to yield Compound 72 (101.9 mg, 51%) as a white solid. LCMS: m/z = 547.4 M-lT), lK NMR: (400 MHz, DMSO-i¾+D20) δ = 8.24 - 8.25 (m, 1 H) 8.25 (d, >2.19 Hz, 1 H) 8.19 - 8.22 (m, 1 H) 7.56 - 7.59 (m, 2 H) 7.10 - 7.14 (m, 1 H) 6.94 (s, 1 H) 6.83 - 6.88 (m, 5 H) 4.98 - 5.06 (m, 1 H) 2.93 - 3.01 (m, 1 H) 2.72 - 2.78 (m, 3 H) 2.39 - 2.44 (m, 2 H).
Synthesis of Compound 73
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (0.3 g, 727.58 umol, 1 eq) and intermediate 2 (101.96 mg, 727.58 umol, 1 eq) n DMF (5 mL) was added Et3N (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and the mixture was ooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added drop- wise. The mixture was allowed to warm to 25 °C and stirred for 13 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water (40 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic layer was washed with rine (20 mL), dried with anhydrous NazSCN, filtered and concentrated in vacuo to yield ntermediate 3 (0.3 g, crude) was obtained as a yellow oil. LCMS: m/z = 454.0 (M+H+).
To a solution of intermediate 3 (0.3 g, 661.47 umol, 1 eq) in EtOH (3 mL) and H2O (1 mL) was dded NH4CI (176.91 mg, 3.31 mmol, 5 eq) and the reaction was heated to 90 °C, then Fe 184.71 mg, 3.31 mmol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (0.3 g, crude) was obtained as a yellow solid. LCMS: m/z = 424.2 (M+H+).
A mixture of intermediate 4 (0.3 g, 708.29 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then SCh-pyridine (338.20 mg, 2.12 mmol, 3 eq) was added. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, 7% mmonium hydroxide (4 mL) was added dropwise and the mixture was dried by flowing N2 to ive a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 50*40 10u;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN];B%: 15%-40%,llmin) o yield Compound 73 (105.9 mg, 28%) was obtained as a white solid. LCMS: m/z = 502.0 (M- H4), 1H-NMR: (400 MHz, DMSO-de) δ = 8.49 (d, >8.93 Hz, 1 H) 8.40 (s, 1 H), 7.90 (br s, 1 H) .72 (d, >5.01 Hz, 1 H) 7.65 (d, >2.81 Hz, 1 H) 7.49 (s, 1 H) 7.40 (s, 1 H) 7.35 (s, 1 H) 7.17 dd, >5.01, 3.79 Hz, 1 H) 6.89 - 6.97 (m, 4 H) 5.10 - 5.21 (m, 1 H) 4.27 - 4.40 (m, 1 H) 4.15 - .26 (m, 1 H) 3.07 (dd, >13.88, 4.22 Hz, 1 H) 2.75 (dd, >13.82, 10.88 Hz, 1 H) 2.64 - 2.70 (m, H).
Synthesis of Compound 74
To a solution of intermediate 5 (1 g, 7.24 mmol, 1 eq) in HzO (20 mL) was added Pd/C (200 mg, 0 % purity, 1.00 eq). The suspension was degassed and purged with H2 several times. Then the mixture was stirred under ¾ (30 psi) at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 2 (964.8 mg, crude) was obtained as white solid. LCMS: m/z = 139.2 (M-H+). 1H- NMR: (400 MHz, DMSO-<¾) δ = 7.50 (s, 1 H) 6.73 (s, 1 H) 2.66 - 2.74 (m, 2 H) 2.44 - 2.50 (m, H). To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt ((400 mg, 970.11 umol, 1 eq)) and intermediate 2 (163.14 mg, 1.16 mmol, .2 eq) in DCM (4 mL) was added EtsN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq\ and then the olution was cooled to 0 °C, then T3P (617.34 mg, 970.11 umol, 576.95 uL, 50% purity, 1 eq) was added drop wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (3 mL) and xtracted with dichloromethane (2 x 10 mL), and then combined organic phase was washed with rine (10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 703.7 mg, crude) was obtained as a yellow oil. LCMS: m/z = 454.0 (Μ+ΙΓ).
To a solution of intermediate 3 (700 mg, 1.54 mmol, 1 eq) in EtOH (16 mL) and H2O (5 mL) was added NH4CI (422.58 mg, 7.72 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (430.96 mg, 7.72 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 4 (996 mg, crude) was obtained as yellow solid. LCMS: m/z = 424.2 (M+H+).
To a solution of intermediate 4 (990 mg, 2.34 mmol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then the SCb-pyridine (1.12 g, 7.01 mmol, 3 eq) was added at 0 °C and the eaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when omplete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was ried by flowing N2, and the residue was purified by prep-HPLC (column: YMC-Actus Triart C18 100*30mm*5um;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 3%-33%,12min) to yield Compound 74 (90.3 mg, 7%) was obtained as white solid. LCMS: m/z = 502.0 (M-FT). ‘H-NMR: (400 MHz, DMSCM,) δ = 8.77 (d, >1.22 Hz, 1 H) 8.43 (d, >8.80 Hz, 1 H) 7.99 (d, 1 H) 7.70 - 7.74 (m, 1 H) 7.63 - 7.67 (m, 1 H) 7.37 (s, 1 H) 7.17 (dd, >5.01, 3.79 Hz, 1 H) 6.98 - 7.04 (m, 4 H) 6.35 (s, 1 H) 5.06 - 5.30 (m, 1 H) 3.09 - 3.16 (m, 1 H) .74 - 2.86 (m, 2 H) 2.60 - 2.70 (m, 1 H) 2.41 - 2.47 (m, 2 H). o4 •O. o4 Oj
HCVMaOH Mai, NaH pure, Hgpo Ρϋ) UOH
90 °C DMF, 0-29 °C BOH MaOKHgO N ^
U
1 u 1 H i H 4 H 9
Preparation of intermediate 5
A solution of intermediate 1 (5 g, 36.20 mmol, 1 eq) in HCl/MeOH (4 M, 50 mL) was heated to 0 °C and stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete. The eaction mixture was concentrated in vacuo to yield intermediate 2 (2 g, crude) was obtained as a ellow oil. LCMS: m/z = 153.1 (M+H4), ‘H-NMR: (400 MHz, DMSO-4.) δ = 9.27 (s, 1 H) 8.07 s, 1 H) 7.57 (d, >16.26 Hz, 1 H) 6.93 (d, >16.26 Hz, 1 H) 3.72 (s, 3 H).
A solution of intermediate 2 (2 g, 10.60 mmol, 1 eq, HC1) in DMF (15 mL) was cooled to 0 °C, hen NaH (933 mg, 23.33 mmol, 60% purity, 2.2 eq) was added and stirred at 0 °C for 10 min, hen Mel (1.66 g, 11.66 mmol, 726.14 uL, 1.1 eq) was added at 0 °C, then the mixture was warmed o 25 °C and stirred at 25 °C for 1 h under Nz atmosphere. The reaction was monitored by LCMS nd when complete, the mixture was quenched by sat. NHtCl solution (20 mL) and then extracted with ethyl acetate (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield intermediate 3 (1.5 g, crude) was obtained as a black brown solid. LCMS: m/z = 167.1 (M+H4), 1H-NMR: (400 MHz, Methanol-^) δ = 7.67 - 7.86 (m, 1 H) 7.40 - 7.63 (m, 2 H) 6.32 - 6.52 (m, 1 H) 3.72 - 3.82 (m, 6 H). To a solution of intermediate 3 (1.4 g, 8.42 mmol, 1 eq) in EtOH (15 mL) was added Pd/C (1 , 8.42 mmol, 10% purity, 1 eq), then the mixture was stirred under ¾ atmosphere (30 Psi) at 25 C for 2 h. The reaction was monitored by LCMS and when complete, the mixture was filtered nd concentrated in vacuo to yield intermediate 4 (495 mg, crude) was obtained as white solid. LCMS: m/z = 169.1 (M+H4). 1H-NMR: (400 MHz, CDCh) δ = 7.26 (m, 1 H) 6.50 - 6.67 (m, 1 H) 3.43 - 3.59 (m, 6 H) 2.70 - 2.79 (m, 2 H) 2.51 - 2.59 (m, 2 H). To a solution of intermediate 4 495 mg, 2.94 mmol, 1 eq) in MeOH (5 mL) and H2O (5 mL) was added LiOH (140.96 mg, 5.89 mmol, 2 eq), then the mixture was stirred at 25 °C for 1 h. The reaction was monitored by LCMS nd when complete, the mixture was concentrated in vacuo to yield intermediate 5 (480 mg, crude) was obtained as black brown solid. LCMS: m/z = 153.2 (M-H+).
To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide alt (200 mg, 485.06 umol, 1 eq) and intermediate 5 (232.97 mg, 1.46 mmol, 3 eq) in DCM (3 mL) was dropwise added EtsN (294.50 mg, 2.91 mmol, 405.08 uL, 6 eq) and cooled to 0 °C, then T3P (617.34 mg, 970.11 umol, 576.95 uL, 50% purity, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 2 h at 25 °C under Nz atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (20 mL) and tirred for 10 minutes. Then the mixture was extracted with ethyl acetate (3 x 10 mL). The ombined organic phase was washed with brine (5 mL), dried with anhydrous Na2SC>4, filtered nd concentrated in vacuo to yield intermediate 7 (62 mg, 27%) was obtained as a yellow solid. LCMS: m/z = 468.2 (M+H4).
A solution of intermediate 7 (62 mg, 132.60 umol, 1 eq) and NH4CI (35.14 mg, 663.01 umol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (37.03 mg, 663.01 umol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 8 (229 mg, crude) was obtained as a yellow solid. LCMS: m/z = 438.2 (M+H+).
A solution of intermediate 8 (229 mg, 523.33 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SCb-pyridine (249.88 mg, 1.57 mmol, 3 eq) was added in portions and stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [waterC0.04%NH3H20+10mM NH4HC03)-ACN];B%: l%-30%,10min) to yield
Compound 75 (5.9 mg, 2%) was obtained as a white solid. LCMS: m/z = 516.1 (M-H+), 1H- NMR: (400 MHz, DMSO-tife) 5 = 8.71 (s, 1 H) 8.37 (d, >8.68 Hz, 1 H) 7.79 (br s, 1 H) 7.72 (dd, >5.01, 0.98 Hz, 1 H) 7.65 (dd,>3.67, 0.98 Hz, 1 H) 7.33 (s, 1 H) 7.21 (s, 1 H) 7.16 - 7.18 (m, 1 H) 7.09 (s, 1 H) 7.05 (s, 1 H) 6.96 (s, 1 H) 6.88 - 6.94 (m, 4 H) 5.07 - 5.13 (m, 1 H) 3.73 (s, 3 H) .06 (dd, >13.88, 4.71 Hz, 1 H) 2.60 - 2.85 (m, 3 H) 2.38 - 2.45 (m, 2 H).
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide alt (300.00 mg, 727.58 umol, 1 eq) and intermediate 1 (122.35 mg, 727.58 umol, 1 eq) in DMF 5 mL) was drop-wise added Et3N (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, hen T3P (694.51 mg, 1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete, the mixture was poured into ice-water (25 mL) and then he mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 10: 1 to 1: 1) to ield intermediate 2 (320 mg, 91%) was obtained as a yellow solid. LCMS: m/z = 482.1 (M+H+), H-NMR: (400 MHz, CDCI3) δ = 8.06 (d, >8.60 Hz, 2 H) 7.51 (dd, >3.53, 1.10 Hz, 1 H) 7.44 dd, >5.07, 0.88 Hz, 1 H) 7.07 - 7.26 (m, 4 H) 6.96 (d, >7.72 Hz, 1 H) 6.79 - 6.93 (m, 2 H) 6.66 s, 1 H) 6.23 (d, >8.16 Hz, 1 H) 5.29 - 5.36 (m, 1 H) 3.27 - 3.34 (m, 1 H) 3.14 - 3.22 (m, 1 H) .91 - 3.03 (m, 2 H) 2.52 (t, >7.50 Hz, 2 H).
A solution of intermediate 2 (320 mg, 664.50 umol, 1 eq) and NH*C1 (177.73 mg, 3.32 mmol, 5 q) in EtOH (12 mL) and HzO (4 mL) was heated to 90 °C and then Fe (185.55 mg, 3.32 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (300 mg, crude) was obtained as a yellow solid. LCMS: m/z = 452.2 (M+H+).
A solution of intermediate 3 (300 mg, 664.34 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (317.21 mg, 1.99 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (0.8 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H2O 10mM NH4HC03)-ACN];B%: 10%-40%,10min) to yield Compound 76 (169.1 mg, 45%) was obtained as a white solid. LCMS: m/z = 530.0 (M-H+), 1H- NMR: (400 MHz, DMSO-d6) 5 = 8.29 (d, .7=8.44 Hz, 1 H) 7.69 - 7.73 (m, 1 H) 7.62 - 7.65 (m, 1 H) 7.23 - 7.29 (m, 1 H) 7.21 (s, 1 H) 7.15 - 7.18 (m, 1 H) 7.13 (s, 1 H) 7.08 (s, 1 H) 6.94 - 7.03 m, 4 H) 6.84 - 6.92 (m, 4 H) 5.04 - 5.14 (m, 1 H) 3.04 (dd, >13.82, 5.87 Hz, 1 H) 2.74 - 2.88 (m, H) 2.38 - 2.44 (m, 2 H).
Synthesis of Compound 77
To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-y1)ihiazol-4-yl)ethanamine hydrobromide alt (300.00 mg, 727.58 umol, 1 eq) and intermediate 1 (134.33 mg, 727.58 umol, 1 eq) in DMF 5 mL) was drop-wise added EfcN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, hen T3P (694.51 mg, 1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete, the mixture was poured into ice-water (25 mL) and then he mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous NazSO-t, filtered and concentrated in vacuo. The residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 10: 1 to 1: 1) to ield intermediate 2 (360 mg, 99%) was obtained as a yellow solid. LCMS: m/z = 498.1 (M+H+), H-NMR: (400 MHz, CDCb) δ = 8.05 (d, >8.60 Hz, 2 H) 8.01 (s, 1 H) 7.44 - 7.54 (m, 1 H) 7.40 7.46 (m, 1 H) 7.02 - 7.21 (m, 6 H) 6.66 (s, 1 H) 6.34 (d, >8.38 Hz, 1 H) 5.27 - 5.37 (m, 1 H) .25 - 3.34 (m, 1 H) 3.14 - 3.23 (m, 1 H) 2.91 - 3.01 (m, 2 H) 2.51 (t, >7.50 Hz, 2 H).
A solution of intermediate 2 ( 360 mg, 722.87 umol, 1 eq) and NH4CI (193.34 mg, 3.61 mmol, 5 q) in EtOH (12 mL) and HzO (4 mL) was heated to 90 °C and then Fe (201.84 mg, 3.61 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (338 mg, crude) was obtained as a yellow solid. LCMS: m/z = 468.2 (M+H+). A solution of intermediate 3 (338 mg, 722.17 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then S03-pyridine (344.83 mg, 2.17 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (0.8 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+l 0mMNH4HCO3)-ACN]; B%: 15%-45%, lOmin) to yield Compound 77 (158.1 mg, 38%) was obtained as a white solid. LCMS: m/z = 546.0 (M-H+), 1H- NMR: (400 MHz, DMSCMO δ = 8.28 (d, >8.56 Hz, 1 H) 7.71 (dd, >5.07, 0.92 Hz, 1 H) 7.63 dd, >3.67, 0.98 Hz, 1 H) 7.23 - 7.28 (m, 2 H) 7.14 - 7.22 (m, 3 H) 7.07 - 7.13 (m, 3 H) 6.97 (s, H) 6.83 - 6.92 (m, 4 H) 5.01 - 5.14 (m, 1 H) 3.04 (dd, >13.75, 5.93 Hz, 1 H) 2.72 - 2.88 (m, 3 H) 2.41 (td, >7.55, 2.38 Hz, 2 H).
Synthesis of Compound 78
To a solution of of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol~4-yl)ethanamine ydrobromide salt (0.3 g, 727.58 umol, 1 eq, HBr) and intermediate 1 (145.09 mg, 873.10 umol, .2 eq) in DMF (5 mL) was added Et3N (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and the mixture was cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was dded drop-wise. The reaction mixture was allowed to warm to 25 °C and stirred for 13 h. The eaction was monitored by LCMS and TLC and when complete, the reaction mixture was diluted with ice water (40 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic hase was washed with brine (40 mL), dried with anhydrous NazSCU, filtered and concentrated n vacuo to yield intermediate 2 (0.4 g, crude) was obtained as a yellow oil. LCMS: m/z = 480.2 Μ+ΕΓ), ¾-NMR: (400 MHz, CDCh) δ = 7.98 - 8.02 (m, 3 H) 7.49 (d, >2.93 Hz, 1 H) 7.41 (d, >4.52 Hz, 1 H) 7.01 - 7.16 (m, 4 H) 6.60 - 6.78 (m, 4 H) 6.48 (d, >8.44 Hz, 1 H) 5.27 - 5.45 m, 1 H) 3.14 - 3.30 (m, 2 H) 2.81 - 2.88 (m, 2 H) 2.43 - 2.53 (m, 2 H).
To a solution of intermediate 2 (400 mg, 834.08 umol, 1 eqi) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (223.08 mg, 4.17 mmol, 5 eq), then the mixture was heated to 90 °C, and then e (232.90 mg, 4.17 mmol, 5 eq) was added, the reaction was stirred at 90 °C for 2 h. The eaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated to yield intermediate 3 (600 mg, crude) was obtained as a yellow solid. LCMS: m/z = 450.2 (M+H÷), 'H-NMR: (400 MHz, Methanol-^) δ = 7.51 - 7.60 (m, 2 H) 7.12 dd, >5.07, 3.75 Hz, 1 H) 7.00 - 7.07 (m, 1 H) 6.93 - 6.98 (m, 2 H) 6.82 (s, 1 H) 6.67 - 6.74 (m, H) 6.56 - 6.66 (m, 3 H) 5.19 - 5.22 (m, 1 H) 3.57 - 3.65 (m, 1 H) 3.12 - 3.21 (m, 1 H) 2.70 - .81 (m, 2 H) 2.39 - 2.53 (m, 2 H).
A mixture of intermediate 3 (0.35 g, 778.49 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then SO3 -pyridine (371.71 mg, 2.34 mmol, 3 eq) was added in batches. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by TLC and when complete, % ammonium hydroxide (4 mL) was added dropwise and the mixture was dried by flowing N2. The residue was purified by prep-HPLC (column: Phenomenex luna(2) C18250*50 10u;mobile hase: [water(0.04%NH3H20)-ACN];B%: 12%-42%,20min) to yield Compound 78 (78.1 mg, 9%) was obtained as a white solid. LCMS: m/z = 528.0 (M-H+), 1H-NMR: (400 MHz, DMSO- de) δ = 9.21 (s, 1 H) 8.25 (d, >8.56 Hz, 1 H) 7.70 (d, >4.89 Hz, 1 H) 7.63 (d, >3.55 Hz, 1 H) .21 (s, 1 H) 7.14 - 7.18 (m, 1 H) 7.08 (s, 2 H) 6.99 - 7.05 (m, 1 H) 6.95 (s, 1 H) 6.84 - 6.92 (m, H) 6.50 - 6.62 (m, 3 H) 5.01 - 5.12 (m, 1 H) 3.06 (dd, >13.75, 5.69 Hz, 1 H) 2.83 (dd, >13.69, 8.56 Hz, 1 H) 2.61 - 2.72 (m, 2 H) 2.30 - 2.41 (m, 2 H). Synthesis of Compound 79
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (0.3 g, 727.58 umol, 1 eq, HBr) and intermediate 1 (157.33 mg, 873.10 umol, .2 eq) in DMF (5 mL) was added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and the mixture was cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was dded drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 13 h. The eaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water (40 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic phase was washed with brine (40 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to ield intermediate 2 (0.4 g, crude) was obtained as a yellow oil. LCMS: m/z = 494.2 (M+H4), H-NMR: (400 MHz, CDCh) δ = 8.02 - 8.06 (m, 2 H) 7.50 (dd, >3.67, 0.86 Hz, 1 H) 7.42 (dd, >5.01, 0.86 Hz, 1 H) 7.08 - 7.19 (m, 4 H) 6.69 - 6.78 (m, 3 H) 6.64 (s, 1 H) 6.31 (d, >8.44 Hz, H) 5.26 - 5.40 (m, 1 H) 3.75 (s, 3 H) 3.25 - 3.34 (m, 1 H) 3.14 - 3.24 (m, 1 H) 2.89 - 2.94 (m, 2 H) 2.52 (t, >7.52 Hz, 2 H).
To a solution of intermediate 2 (400 mg, 810.38 rnnol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (216.74 mg, 4.05 mmol, 5 eq), then the mixture was heated to 90 °C, then Fe 226.30 mg, 4.05 mmol, 5 eq) was added into the mixture and stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 3 (500 mg, crude) was obtained as a ellow solid. LCMS: m/z = 464.2 (M+H+), ‘H-NMR: (400 MHz, Methanol-^) δ = 7.47 - 7.64 m, 2 H) 7.09 - 7.17 (m, 2 H) 6.93 (br d, >7.95 Hz, 2 H) 6.80 (s, 1 H) 6.63 - 6.76 (m, 5 H) 5.15 - .27 (m, 1 H) 3.71 (s, 3 H) 3.09 - 3.19 (m, 1 H) 2.87 - 2.94 (m, 1 H) 2.74 - 2.83 (m, 2 H) 2.40 - .54 (m, 2 H).
A mixture of intermediate 3 (0.35 g, 754.94 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, and then SCb-pyridine (360.47 mg, 2.26 mmol, 3 eq) was added. The mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by HPLC and LCMS and when complete, 7% ammonium hydroxide (4 mL) was added drop-wise and the mixture was ried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: henomenex Gemini Cl 8250*50 10u;mobile phase: [water (0.05% ammonia hydroxide v/v)- ACN];B%: 15%-45%,20min) to yield Compound 79 (227.6 mg, 55%) was obtained as a white olid. LCMS: m/z = 542.1 (M-H+), ‘H-NMR: (400 MHz, DMSO -φ>) δ = 8.27 (d, >8.44 Hz, 1 H) 7.71 (d, >5.01 Hz, 1 H) 7.63 (d, >2.93 Hz, 1 H) 7.22 (br s, 1 H) 7.12 - 7.18 (m, 2 H) 7.06 - .11 (m, 2 H) 6.96 (br s, 1 H) 6.82 - 6.92 (m, 4 H) 6.67 - 6.77 (m, 3 H) 5.04 - 5.13 (m, 1 H) 3.70 s, 3 H) 3.05 (dd, >13.82, 5.87 Hz, 1 H) 2.84 (dd, >13.75, 8.50 Hz, 1 H) 2.69 - 2.78 (m, 2 H) .35 - 2.44 (m, 2 H).
Synthesis of Compound 80
A mixture of intermediate 1 (5 g, 25.62 mmol, 1 eq) and HCl/MeOH (50 mL) was heated to 60 C, and the mixture was stirred at 60 °C for 1 h. The reaction was monitored by LCMS and when omplete, the reaction mixture was concentrated in vacuo to yield intermediate 2 (5.3 g, crude) was obtained as a white solid. LCMS: m/z = 210.1 (M+H+), ‘H-NMR: (400 MHz, DMSO-de) δ 8.10 (s, 1 H) 8.05 (dd, >8.13, 1.65 Hz, 1 H) 7.71 (d, >7.58 Hz, 1 H) 7.53 - 7.61 (m, 1 H) 3.57 s, 3 H) 2.94 - 3.03 (m, 2 H) 2.66 - 2.75 (m, 2 H).
To a solution of intermediate 2 (4.3 g, 20.55 mmol, 1 eq) in EtOH (40 mL) and H2O (15 mL) was added NH4CI (5.43 g, 102.77 mmol, 5 eq) and the reaction was heated to 90 °C, then Fe (5.74 g, 102.77 mmol, 5 eq) was added, the mixture was stirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and concentrated in vacuo to yield intermediate 3 (3.7 g, crude) was obtained as a yellow olid. LCMS: m/z = 180.2 (M+H+), Ή-ΝΜΚ: (400 MHz, DMSO -ck) δ = 6.91 (t, >7.52 Hz, 1 H) 6.28 - 6.44 (m, 3 H) 4.94 (s, 2 H) 3.58 (s, 3 H) 2.63 - 2.74 (m, 2 H) 2.52 - 2.59 (m, 2 H).
To a solution of intermediate 3 (2.5 g, 13.95 mmol, 1 eq) in THF (15 mL) was added (Boc)20 3.96 g, 18.13 mmol, 4.17 mL, 1.3 eq) and EtsN (1.84 g, 18.13 mmol, 2.52 mL, 1.3 eq), the mixture was stirred at 25 °C for 15 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition water (60 mL) at 5°C, and then extracted with EtOAc (3 x 50 mL). The combined organic layer was washed with rine (100 mL), dried with anhydrous Na2SC>4, filtered and the filtrate concentrated in vacuo, and he residue was purified by column chromatography (S1O2, Petroleum ether: Ethyl acetate = 10: to 5: 1) to yield intermediate 4 (2.9 g, 68%) was obtained as a yellow solid. LCMS: m/z = 02.1 (M+Na+), 'H-NMR: (400 MHz, DMSO^fc) δ = 9.26 (br s, 1 H) 7.33 (s, 1 H) 7.24 (d, = 8.19 Hz, 1 H) 7.10 - 7.17 (m, 1 H) 6.81 (d, >7.46 Hz, 1 H) 3.58 (s, 3 H) 2.73 - 2.83 (m, 2 H) .55 - 2.61 (m, 2 H) 1.47 (s, 9 H).
A solution of intermediate 4 (2.9 g, 10.38 mmol, 1 eq) in DMF (20 mL) was cooled to 0 °C, then NaH (622.86 mg, 15.57 mmol, 60% purity, 1.5 eq) was added, the mixture was stirred for 1 hr at °C and then Mel (11.79 g, 83.06 mmol, 5.17 mL, 8 eq) was added, the reaction was allowed warm to 25 °C and stirred for 3 h under N2 atmosphere. The reaction was monitored by LCMS nd when complete, the reaction mixture was quenched by addition sat. NH4CI (40 mL) at 25 °C, nd then extracted with ethyl acetate (3 x 30 mL). The combined oiganic layer was washed with rine (70 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 5 (3 g, crude) was obtained as a light yellow solid. LCMS: m/z = 316.1 (M+Na+), H-NMR: (400 MHz, DMSO-<¾) δ = 7.12 - 7.24 (m, 1 H) 7.01 - 7.08 (m, 2 H) 6.94 - 6.98 (m, 1 H) 3.63 (s, 3 H) 3.20 (s, 3 H) 2.87 - 2.91 (m, 2 H) 2.54 - 2.64 (m, 2 H) 1.41 (s, 9 H).
To a solution of intermediate 5 (3 g, 10.23 mmol, 1 eq) in MeOH (15 mL) and H2O (15 mL) was dded LiOH (489.81 mg, 20.45 mmol, 2 eq) at 25 °C. The mixture was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the reaction mixture was oncentrated in vacuo to yield intermediate 6(3 g, crude) was obtained as a yellow solid.
LCMS: m/z = 278.1 (M-I Γ"), Ή-ΝΜΚ: (400 MHz, DMSO-i&) δ = 7.13 - 7.22 (m, 1 H) 7.07 (s,
1 H) 6.95 - 7.05 (m, 2 H) 3.15 (s, 3 H) 2.69 - 2.83 (m, 2 H) 2.10 - 2.28 (m, 2 H) 1.38 (s, 9 H). To a solution of intermediate 6 (400 mg, 970.11 umol, 1 eq), (S)-2-(4-nitrophenyl)-l-(2- hiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (415.11 mg, 1.46 mmol, 1.5 eq) in DCM (8 mL) was added EtsN (588.99 mg, 5.82 mmol, 810.17 uL, 6 eq), the reaction was cooled o 0 °C, and then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 1.5 eq) was added ropwise, then it was allowed warm to 25 °C and stirred for 2 h under N2 atmosphere. The eaction was monitored by LCMS and when complete, the reaction mixture was quenched by ddition ice water (40 mL) at 25 °C, and then extracted with Dichloromethane (3 x 50 mL). The ombined organic layer was washed with sat. brine (100 mL), dried with anhydrous Na2S(>4, filtered and concentrated in vacuo to yield intermediate 7 (577 mg, crude) was obtained as a ellow oil. LCMS: m/z = 593.1 (M+H+), ‘H-NMR: (400 MHz, DMSO-dk) δ = 8.46 (d, >8.56 Hz, 1 H) 8.11 (d, >8.19 Hz, 2 H) 7.72 (d, >5.01 Hz, 1 H) 7.65 (d, >3.55 Hz, 1 H) 7.47 (d,
>8.19 FIz, 2 H) 7.26 (s, 1 H), 7.13 - 7.21 (m, 2 H) 7.02 - 7.09 (m, 2 H) 6.92 (d, >7.46 Hz, 1 H) .21 - 5.34 (m, 1 H) 3.34 - 3.39 (m, 1 H) 3.09 - 3.17 (m, 4 FI) 2.68 - 2.75 (m, 2 H) 2.34 - 2.43 (m, H) 1.37 (s, 9 H).
To a solution of intermediate 7 (500 mg, 843.56 umol, 1 eq) in EtOH (15 mL) and H2O (5 mL) was added NH4CI (222.92 mg, 4.22 mmol, 5 eq) and the reaction was heated to 90 °C, then Fe 235.54 mg, 4.22 mmol, 5 eq) was added, the mixture was degassed and purged with N2 for 3 mes and stirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS nd when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo o yield intermediate 8 (450 mg, crude) was obtained as a yellow solid. LCMS: m/z = 563.3 M+FT).
A solution of intermediate 8 (50 mg, 88.85 umol, 1 eq) in MeCN (0.5 mL) and pyridine (0.5 mL) was cooled to 0 °C, then SOi-pyridine (28.28 mg, 177.70 umol, 2 eq) was added, and then he mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when omplete, the compound of intermediate 10 (57 mg, crude) in MeCN and pyridine was obtained s a yellow liquid and was used to the next step directly. LCMS: m/z = 641.1 (M-FT).
A solution of intermediate 10 (57 mg, 88.81 umol, 1 eq) DCM (2 mL) was cooled to 0 °C, then TFA (0.5 mL) was added, and the mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC and when complete, the mixture was dried by flowing N2 to yield intermediate 11 (48 mg, crude) was obtained as a yellow solid. A mixture of intermediate 11 (48 mg, 88.45 umol, 1 eq) and in NH3.H2O (1 mL) was cooled to 0 C and the mixture was stirred at 0 °C for 0.1 h. The reaction was monitored by LCMS and when omplete, the mixture was dried by flowing N2 to give a residue. The residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: 15%-45%,10min) to yield Compound 80 (3.4 mg, 7%) was obtained as a white solid. LCMS: m/z = 541.0 (M-H+), 1H-NMR: (400 MHz, DMSO-i¾) δ = 8.27 (d, >8.31 Hz, 1 H) 7.71 (d, >4.89 Hz, 1 H) 7.63 (d, >3.30 Hz, 1 H) 7.21 (s, 1 H) 7.14 - 7.18 (m, 1 H) .10 (d, >10.51 Hz, 2 H) 7.03 (t, >7.70 Hz, 1 H) 6.96 (s, 1 H) 6.85 - 6.93 (m, 4 H) 6.42 - 6.59 m, 3 H) 5.00 - 5.14 (m, 1 H) 3.06 (dd, >13.76, 5.44 Hz, 1 H) 2.83 (dd, >13.69, 8.93 Hz, 1 H) .63 - 2.71 (m, 5 H) 2.27 - 2.43 (m, 2 H).
Synthesis of Compound 81
HCOOH (1.56 g, 32.57 mmol, 3 eq) was added drop-wise to DMF (5 mL) under stirring and ooling with ice water. EtsN (1.43 g, 14.11 mmol, 1.96 mL, 1.3 eq) was added in the same way, ollowed by intermediate 2 (1.56 g, 10.86 mmol, 1 eq). After dissolution, intermediate 1 (2 g, 0.86 mmol, 1 eq) was added, the mixture was heated to 80 °C and stirred for 14 h. The reaction was monitored by LCMS and when complete, the solution was cooled down and poured out into ce-water (20 mL) under vigorous stirring. Concentrated HC1 was added till pH = 1-2, and then he solution was concentrated in vacuo to yield intermediate 3 (2.4 g, crude) was obtained as a red solid. LCMS: m/z = 227.0 (M-H+). 1H-NMR: (400 MHz, DMSO-ifc) δ = 7.82 (br s, 1 H) .74 (d, J= 6.58 Hz, 1 H) 7.59 - 7.66 (m, 1 H) 7.49 - 7.58 (m, 1 H) 3.36 (s, 2 H) 3.11 - 3.16 (m, 3 H) 2.48 - 2.52 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (300 mg, 727.58 umol, 1 eq) and intermediate 2 (199.30 mg, 873.10 umol, .2 eq) in DCM (1.2 mL) was added Et3N (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50 % purity, 2 eq) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (5 mL) and xtracted with dichloromethane (3 x 10 mL), and then combined organic phase was washed with rine (10 mL), dried with anhydrous NazSQt and concentrated in vacuo to yield intermediate 4 452 mg, crude) was obtained as yellow oil. LCMS: m/z = 542.2 (M+FT).
To a solution of intermediate 4 (500 mg, 923.09 umol, 1 eq) in EtOH (10 mL) and H20 (3 mL) was added NH4CI (240.23 mg, 4.62 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (257.75 mg, 4.62 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 5 (638 mg, crude) was obtained as yellow solid. LCMS: m/z = 510.1 (M-H+).
To a solution of intermediate 5 (638 mg, 1.25 mmol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then the SOs-pyridine (595.37 mg, 3.74 mmol, 3 eq) was added at 0 °C and he reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7 % ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbiidge 50*25 5u;mobile phase: [water(0.04%NH3H20+10mMlS[H4HC()3)-ACN];B%: 1%- 0%,10min) to yield Compound 81 (130 mg, 17%) was obtained as a white solid. LCMS: m/z = 90.0 (M-H+). IH-NMR: (400 MHz, D20) δ = 7.56 - 7.60 (m, 1 H) 7.46 - 7.52 (m, 1 H) 7.39 - .44 (m, 1 H) 7.34 - 7.38 (m, 1 H) 7.22 - 7.33 (m, 2 H) 6.90 - 7.04 (m, 5 H) 6.67 (s, 1 H) 5.00 - .10 (m, 1 H) 3.00 (s, 3 H) 2.89 - 2.97 (m, 1 H) 2.86 - 2.89 (m, 3 H) 2.46 (s, 2 H).
Preparation of intermediate 4
To a solution of intermediate 8 (5 g, 40.01 mmol, 2.84 mL, 1 eq) in DCM (50 mL) was added ntermediate 9 (6.63 g, 43.99 mmol, 5.39 mL, 1.10 eq) followed by EtsN (80.02 mmol, 11.14 mL, 2.00 eq) and finally DMAP (39.07 mg, 319.76 umol, 7.99e-3 eq), then the solution was at 0 °C and stirred for 18 h. The reaction was monitored by LCMS and when complete, the eaction mixture was poured into 1 N HC1 (100 mL), and extracted with ethyl acetate (3 x 50 mL). Then combined organic phase was washed with brine (2 x 50 mL), dried with anhydrous NaiS04 and concentrated in vacuo to yield intermediate 4 (6 g, crude) was obtained as a yellow quid. LCMS: m/z = 239.2 (Μ+1Γ), 'H-NMR: (400 MHz, DMSO-dk) δ = 3.88 (t, >5.685 Hz, H) 3.52 (t, >5.685 Hz, 2 H) 0.88 (s, 9 H) 0.06 (s, 6 H).
EtsN (29.10 mmol, 4.05 mL, 6 eq), T3P (7.28 mmol, 4.33 mL, 50% purity, 1.5 eq) was added into a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (2 g, 4.85 mmol, 1 eq) and intermediate 2 (806.03 mg, 4.85 mmol, 1 eq) in DCM (20 mL) at 0 °C, then the mixture was allowed to warm to 25 °C and stirred for 1 h. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured nto water (50 mL), and extracted with dichloromethane (3 x 20 mL). Then combined organic hase was washed with brine (2 x 20 mL), dried with anhydrous NaaSCU and concentrated in acuo. The residue was purified by silica gel chromatography (Petroleum ether/Ethyl acetate=5:l 3:1) to yield intermediate 3 (1.8 g, 77%) was obtained as a white solid. LCMS: m/z = 480.1 Μ+ΕΓ), *H-NMR: (400 MHz, DMSO-4,) δ = 9.21 (s, 1 H) 8.42 (d, >8.559 Hz, 1 H) 8.10 (d, >8.681 Hz, 2 H) 7.72 (dd, >5.074, 1.039 Hz, 1 H) 7.65 (dd, >3.668, 0.978 Hz, 1 H) 7.46 (d, >8.681 Hz, 2 H) 7.21 (s, 1 H) 7.17 (dd, >5.013, 3.668 Hz, 1 H) 6.96 - 7.03 (m, 1 H) 6.48 - 6.59 m, 3 H) 5.22 - 5.28 (m, 1 H) 3.33 - 3.40 (m, 1 H) 3.13 (dd, >13.694, 9.292 Hz, 1 H) 2.64 (t, >7.642 Hz, 2 H) 2.30 - 2.39 (m, 2 H).
A mixture of intermediate 3 (200 mg, 417.04 umol, 1 eq), intermediate 4 (149.65 mg, 625.56 mol, 1.5 eq), K2CO3 (230.55 mg, 1.67 mmol, 4 eq) in DMF (3 mL) was degassed and purged with N2 for 3 times, then the mixture was at 100 °C and stirred for 18 h under N2 atmosphere.
The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was oured into water (30 mL), and extracted with ethyl acetate (3 x 20 mL). Then combined organic hase was washed with brine (2 x 20 mL), dried with anhydrous Na2SC>4 and concentrated in acuo to yield intermediate 5 (320 mg, crude) was obtained as a yellow oil. LCMS: m/z = 638.1 M+H+), ‘H-NMR: (400 MHz, DMSO-4,) δ = 8.43 (d, >8.681 Hz, 1 H) 8.10 (d, >8.803 Hz, 2 H) 7.72 (dd, >5.013, 1.101 Hz, 1 H) 7.64 (dd, >3.729, 1.161 Hz, 1 H) 7.46 (d, >8.803 Hz, 2 H) 7.23 (s, 1 H) 7.17 (dd, >5.074, 3.729 Hz, 1 H) 7.10 (t, >7.947 Hz, 1 H) 6.64 - 6.77 (m, 3 H) .21 - 5.31 (m, 1 H) 3.92 - 4.02 (m, 2 H) 3.80 - 3.91 (m, 2 H) 3.34 - 3.35 (m, 1 H) 3.06 - 3.19 (m, H) 2.65 - 2.72 (m, 2 H) 2.35 - 2.44 (m, 2 H) 0.86 (s, 9 H) 0.04 - 0.08 (m, 6 H).
A solution of intermediate 5 (300 mg, 470.31 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was eated to 90 °C, then NH4CI (125.79 mg, 2.35 mmol, 5 eq), Fe (131.32 mg, 2.35 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by TLC and LCMS nd when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to ield intermediate 6 (400 mg, crude) was obtained as a yellow solid. LCMS: m/z = 608.2 (M+FL). To a solution of intermediate 6 (400 mg, 658.00 umol, 1 eq) in THF (8 mL) was added aq. HC1 2 M, 239.27 uL, 7.27e-1 eq) slowly at 20 °C and stirred for 1 h. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was poured into sat. NaHCCb (30 mL), and extracted with ethyl acetate (3 x 20 mL). Then combined organic phase was washed with brine (2 x 20 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 7 (220 mg, crude) was obtained as a yellow oil. LCMS: m/z = 494.1 (M+H+).
To a solution of intermediate 7 (220 mg, 445.67 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SCb-pyridine (106.40 mg, 668.50 umol, 1.5 eq) at 0 °C and stirred for 0.17 h. The reaction was monitored by TLC and when complete, 7% ammonium hydroxide (5 mL) was dded drop-wise, and then the mixture was dried by flowing Ni, and the residue was purified by rep-HPLC (water(10mM NH4HCO3)- ACN]; B% : 30%-50%,10min) to yield Compound 82 12.0 mg, 5.5%) was obtained as a white solid. LCMS: m/z = 572.0 (M-FT), Ή-ΝΜΚ: (400 MHz, DMSO-ifc) 5 = 8.27 (d, >8.436 Hz, 1 H) 7.71 (dd, >5.074, 1.039 Hz, 1 H) 7.58 - 7.68 m, 2 H) 7.02 - 7.19 (m, 7 H) 6.81 - 6.94 (m, 4 H) 6.67 - 6.78 (m, 3 H) 5.02 - 5.14 (m, 1 H) 4.83 t, >5.563 Hz, 1 H) 3.86 - 4.01 (m, 2 H) 3.68 (q, >5.380 Hz, 2 H) 3.05 (dd, >13.694, 5.869 Hz, 1 H) 2.83 (dd, >13.816, 8.559 Hz, 1 H) 2.69 - 2.78 (m, 2 H) 2.34 - 2.43 (m, 2 H).
Synthesis of Compound 83 To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide sait (250 mg, 606.32 umol, 1 eq), intermediate 2 (126.19 mg, 666.95 umol, 1.1 q) in DCM (6 mL) was added Et3N (368.12 mg, 3.64 mmol, 506.36 uL, 6 eq) and the mixture was cooled to 0 °C, and then T3P (578.76 mg, 909.48 umol, 540.90 uL, 50% purity, 1.5 eq) was dded, then it was allowed warm to 25 °C and stirred for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition ice water (40 mL), and then extracted with Dichloromethane (3 x 40 mL). The combined organic ayer was washed with brine (80 mL), dried with NazSCL, filtered and concentrated in vacuo to ield intermediate 3 (300 mg, crude) was obtained as a yellow solid. LCMS: m/z = 503.1 M+H+), ‘H-NMR: (400 MHz, DMSO-tafc) δ = 10.73 (br s, 1 H) 8.43 (d, >8.56 Hz, 1 H) 8.08 (d, >8.31 Hz, 2 H) 7.72 (d, >5.01 Hz, 1 H) 7.65 (d, >3.55 Hz, 1 H) 7.41 - 7.53 (m, 3 H) 7.32 (d, >8.07 Hz, 1 H) 7.22 (s, 1 H) 7.17 (t, >4.34 Hz, 1 H) 7.02 - 7.08 (m, 2 H) 6.93 - 6.99 (m, 1 H) .24 - 5.33 (m, 1 H) 3.34 - 3.39 (m, 1 H) 3.08 - 3.18 (m, 1 H) 2.81 - 2.85 (m, 2 H) 2.42 - 2.47 (m, H).
A solution of intermediate 3 (360 mg, 716.27 umol, 1 eq) in EtOH (5 mL) and H20 (2 mL) was dded NH4CI (196.35 mg, 3.58 mmol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, e (200.00 mg, 3.58 mmol, 5 eq) was added, and then the mixture was stirred at 90 °C for 2 h nder N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (363 mg, rude) was obtained as a yellow solid. LCMS: m/z = 473.2 (M+H+).
A solution of intermediate 4 (363 mg, 768.05 umol, 1 eq) in MeCN (2 mL) and pyridine (2 mL) was cooled to 0 °C and then S03-pyridine (366.74 mg, 2.30 mmol, 3 e</)was added, the mixture was stirred at 0 °C for 10 min. The reaction was monitored by TLC and when complete, 7% mmonium hydroxide (3 mL) was added dropwise, then the mixture was dried by flowing N2, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 15%-45%,10min) to yield Compound 83 107.2 mg, 25%) was obtained as a light yellow solid. LCMS: m/z = 551.1 (M-If), *H-NMR: 400 MHz, DMSO-dk) δ = 10.66 (br s, 1 H) 8.32 (d, >8.56 Hz, 1 H) 7.79 (br s, 1 H) 7.71 (dd, >5.07, 1.04 Hz, 1 H) 7.64 (dd, >3.67, 0.98 Hz, 1 H) 7.47 (d, >7.70 Hz, 1 H) 7.29 (d, >8.07 Hz, 1 H) 7.14 - 7.22 (m, 3 H) 7.07 - 7.13 (m, 1 H) 7.00 - 7.07 (m, 2 H) 6.95 - 6.99 (m, 5 H) 6.90 6.94 (m, 1 H) 6.67 (d, >1.96 Hz, 1 H) 5.09 - 5.18 (m, 1 H) 3.10 (dd, >13.82, 5.01 Hz, 1 H) .77 - 2.94 (m, 3 H) 2.42 - 2.48 (m, 2 H).
Synthesis of Compound 84
To a solution of (S)-2-(4-nUrophenyQ-l-(2-thiophen-2-yl)thiazol-4-y0ethanamine ydrobromide salt (0.3 g, 7.75 mmol, 1 eq) and intermediate 2 (113.65 mg, 727.58 umol, 1 eq) n DCM (10 mL), EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) was added and cooled to 0 °C, nd then T3P (463.01 mg, 727.58 umol, 432.72 uL, 50% purity, 1 eq) was added dropwise at 0 C under N2, then the reaction was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was dried by flowing N2 to yield intermediate 3 (0.3 g, rude) was obtained as a yellow oil. LCMS: m/z = 470.0 (M+H+). 1H-NMR: (400 MHz, DMSO-<¾) δ = 8.50 (d, >8.56 Hz, 1 H) 8.07 - 8.16 (m, 2 H) 7.71 - 7.75 (m, 1 H) 7.65 (dd, >3.67, 1.10 Hz, 1 H) 7.49 (d, >8.68 Hz, 2 H) 7.31 (s, 1 H) 7.22 (dd, >5.14, 1.22 Hz, 1 H) 7.17 dd, >5.01, 3.67 Hz, 1 H) 6.83 (dd, >5.14, 3.42 Hz, 1 H) 6.72 (dd, >3.42, 0.98 Hz, 1 H) 5.20 - .34 (m, 1 H) 3.34 - 3.38 (m, 1 H) 3.14 (dd, >13.63, 9.35 Hz, 1 H) 2.94 (t, >7.27 Hz, 2 H) 2.37 2.46 (m, 2 H). ntermediates (0.3 g, 638.84 umol, 1 eq) and NH4CI (170.86 mg, 3.19 mmol, 111.68 uL, 5 eq) in EtOH (5 mL) and HzO (2 mL) was heated to 90 °C, and then Fe (178.38 mg, 3.19 mmol, 5 eq) was dded, the mixture was stirred at 90 0 C for 2 h. The reaction was monitored by LCMS when omplete, the solution was filtered and concentrated in vacuo to yield intermediate 4 (400 mg, crude) was obtained as a green solid. LCMS: m/z = 440.2 (M+H4). Intermediate 4 (400 mg, 909.88 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then SO3. pyridine (349.78 mg, 1.36 mmol, 1.5 eq) was added at 0 °C. Then the mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS, HPLC and TLC when complete, 7% ammonium hydroxide (3 mL) was added drop wise, and then the mixture was dried by flowing N2. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(10mM NH4HC03)-ACN];B%: 5%-25%,l lmin) to yield Compound 84 (0.11 g, 22%) was obtained as a white solid. LCMS: m/z = 518.0 (M-H÷), Ή-NMR: (400 MHz, DMSO-dk) δ = 8.36 (d, >8.44 Hz, 1 H) 7.71 (d, >5.01 Hz, 1 H) 7.60 - 7.68 (m, 2 H) 7.26 (d, >5.01 Hz, 1 H) 7.13 - 7.21 (m, 2 H) 7.07 (s, 4 H) 6.83 - 6.93 (m, 5 H) 6.77 (d, >2.81 Hz, 1 H) 5.02 - 5.16 (m, 1 H) 3.06 (dd, >13.69, 5.75 Hz, 1 H) 2.97 (t, >7.34 Hz, 2 H) 2.85 (dd, >13.75, 8.62 Hz, 1 H) 2.40 - 2.46 (m, 2 H).
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yI)thiazol-4-yl)ethanamine hydrobromide alt (300.00 mg, 727.58 umol, 1 eq) and intermediate 1 (159.39 mg, 727.58 umol, 1 eq) in DCM 5 mL) was drop-wise added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, hen T3P (694.51 mg, 1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (25 mL) and then he mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 2 (380 mg, crude) was obtained as a yellow solid. LCMS: m/z = 532.1 (M+H4), Ή- NMR: (400 MHz, CDCI3) δ = 8.06 (d, >8.60 Hz, 2 H) 7.47 - 7.53 (m, 1 H) 7.39 - 7.44 (m, 1 H) .15 - 7.26 (m, 4 H) 7.07 - 7.12 (m, 1 H) 6.99 - 7.05 (m, 1 H) 6.69 (s, 1 H) 6.33 (d, >8.16 Hz, 1 H) 5.28 - 5.37 (m, 1 H) 3.34 - 3.40 (m, 1 H) 3.22 - 3.26 (m, 2 H) 3.02 - 3.12 (m, 1 H) 2.44 - 2.51 m, 2 H).
A solution of intermediate 2 (380 mg, 713.67 umol, 1 eq) and NH4CI (190.88 mg, 3.57 mmol, 5 q) in EtOH (12 mL) and H20 (4 mL) was heated to 90 °C and then Fe (199.27 mg, 3.57 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (300 mg, crude) was obtained as a yellow solid. LCMS: m/z = 502.1 (M+H4).
A solution of intermediate 3 (300 mg, 597.04 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SOs-pyridine (285.08 mg, 1.79 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7 % mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10mM NH4HC03>ACN]; B%: 10%-50%, lOmin) to yield Compound 85 (188.2 mg, 51%) was obtained as a white solid. LCMS: m/z = 580.0 (M-H+), Ή- NMR: (400 MHz, DMSO-ifc) δ = 8.33 - 8.41 (m, 1 H) 7.71 (d, >4.89 Hz, 1 H) 7.60 - 7.66 (m, 1 H) 7.39 - 7.47 (m, 2 H) 7.23 - 7.31 (m, 2 H) 7.21 (s, 1 H) 7.15 - 7.18 (m, 1 H) 7.08 (s, 1 H) 6.96 s, 1 H) 6.83 - 6.92 (m, 3 H) 6.48 (d, >8.31 Hz, 1 H) 5.02 - 5.15 (m, 1 H) 2.97 - 3.10 (m, 3 H) .78 - 2.94 (m, 1 H) 2.26 - 2.38 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazoI-4-yl)ethanamine ydrobromide salt (300 mg, 727.58 umol, 1 eq), intermediate 2 (148.99 mg, 800.34 umol, 1.1 q) in DCM (5 mL) was added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and the mixture was cooled to 0 °C, and then T3P (694.51 mg, 1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was dded drop-wise then the reaction was warmed to 25 °C and stirred for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched y addition ice water (30 mL), and then extracted with ethyl acetate (3 x 40 mL). The combined rganic layer was washed with brine (80 mL), dried with anhydrous Na2SC>4, filtered and oncentrated in vacuo to yield intermediate 3 (350 mg, crude) was obtained as a yellow solid. LCMS: m/z = 500.0 (M+FT), Ή-NMR: (400 MHz, DMSO-tfc) δ = 8.45 (d, .7=8,68 Hz, 1 H) .10 (d, >8.68 Hz, 2 H) 7.68 - 7.75 (m, 1 H) 7.59 - 7.67 (m, 1 H) 7.47 (d, >8.68 Hz, 2 H) 7.30 s, 1 H) 7.11 - 7.25 (m, 3 H) 6.88 - 6.97 (m, 1 H) 5.20 - 5.31 (m, 1 H) 3.32 - 3.36 (m, 1 H) 3.07 - .17 (m, 1 H) 2.73 (t, >7.34 Hz, 2 H) 2.34 - 2.45 (m, 2 H).
A solution of intermediate 3 (350 mg, 700.63 umol, 1 eq ) in EtOH (5 mL) and H2O (2 mL) was dded NH4CI (185.14 mg, 3.50 mmol, 5 eq), and then the solution was heated to 90 °C, Fe (195.63 mg, 3.50 mmol, 5 eq) was added , the reaction was stirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrated was concentrated in vacuo to yield intermediate 4 (350 mg, crude) was btained as a yellow solid. LCMS: m/z = 470.1 (Μ+ΙΓ), 1H-NMR: (400 MHz, DMSO-i&) δ = .36 (d, >8.60 Hz, 1 H) 7.67 - 7.75 (m, 1 H) 7.60 - 7.65 (m, 1 H) 7.13 - 7.24 (m, 3 H) 6.86 - .00 (m, 2 H) 6.80 (d, >8.38 Hz, 2 H) 6.42 (d, >8.16 Hz, 2 H) 4.97 - 5.09 (m, 1 H) 2.88 - 3.05 m, 3 H) 2.71 - 2.82 (m, 1 H) 2.36 - 2.42 (m, 2 H).
A solution of intermediate 4 (350 mg, 745.36 umol, 1 eq) in MeCN (2 mL) and pyridine (1 mL) was cooled to 0 °C and then SCb-pyridine (355.90 mg, 2.24 mmol, 3 eq) was added. The reaction was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (3 mL) was added drop-wise, then the mixture was dried by flowing N2, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 10%-40%,10min) to yield Compound 86 77.2 mg, 19%) was obtained as a white solid. LCMS: m/z = 547.9 (M-H+), 1H-NMR: (400 MHz, DMSO-ifc) 5 = 8.30 (d, >8.31 Hz, 1 H) 7.71 (d, >4.89 Hz, 1 H) 7.64 (d, >3.42 Hz, 1 H) .22 - 7.29 (m, 2 H) 7.21 (br s, 1 H) 7.14 - 7.19 (m, 1 H) 7.08 (br s, 1 H), 6.83 - 6.98 (m, 6 H) .00 - 5.15 (m, 1 H) 3.03 (dd, >13.63, 5.81 Hz, 1 H) 2.83 (dd, >13.57, 8.68 Hz, 1 H) 2.72 - .77 (m, 2 H) 235 - 2.44 (m, 2 H).
Synthesis of Compound 87 To a solution of (S)-2-(4-nitrophenyI)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide sah (300 mg, 727.58 umol, 1 eq) and intermediate 2 (150.79 mg, 727.58 umol, 1 q) in DCM (5 mL) was added EtsN (4.37 mmol, 607.63 uL, 6 eq), and the mixture was cooled o 0 °C, then T3P (1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added drop-wise. Then the mixture was allowed to warm to 25 °C and stirred for 1 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with Dichloromethane (3 x 20 mL). Then combined organic phase was washed with rine (2 x 20 mL), dried with anhydrous Na2S04 and concentrated in vacuo to yield intermediate (320 mg, crude) was obtained as a yellow oil. LCMS: m/z = 521.1 (Μ+ΙΓ), 1H-NMR: (400 MHz, DMSO-dk) δ = 8.64 (d, >8.558 Hz, 1 H) 8.07 (d, >8.681 Hz, 2 H) 7.92 - 7.98 (m, 1 H) .87 (d, >7.703 Hz, 1 H) 7.71 (dd, >5.074, 1.039 Hz, 1 H) 7.64 (dd, >3.668, 1.100 Hz, 1 H) .42 - 7.52 (m, 3 H) 7.33 - 7.40 (m, 2 H) 7.17 (dd, >5.013, 3.790 Hz, 1 H) 5.24 - 5.30 (m, 1 H) .38 (dd, >13.694, 9.414 Hz, 1 H) 3.26 (t, >7.153 Hz, 2 H) 3.16 (dd, >13.694, 9.414 Hz, 1 H) .63 - 2.78 (m, 2 H).
A solution of intermediate 3 (320 mg, 614.62 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (164.39 mg, 3.07 mmol, 5 eq), and then the solution was heated to 90 °C, Fe 171.62 mg, 3.07 mmol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The eaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (370 mg, crude) was btained as a yellow oil. LCMS: m/z = 491.2 (M+H÷).
To a solution of intermediate 4 (370 mg, 754.08 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SCb-pyridine (180.03 mg, 1.13 mmol, 1.5 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and when complete, 7% ammonium hydroxide (5 mL) was dded drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM NH4HC0.3)-ACN];B%: 5%-35%,10min) to yield Compound 87 (93.3 mg, 22%) was obtained as white solid. LCMS: m/z = 568.9 (M-H+), Ή-ΝΜΚ: (400 MHz, DMSO-ifc) δ = 8.47 (d, >8.436 Hz, 1 H) 7.97 - 8.04 (m, 1 H) 7.90 (d, >7.703 Hz, 1 H) 7.70 (dd, >5.074, 1.039 Hz, 1 H) 7.67 (s, 1 H) 7.63 (dd, >3.668, 1.100 Hz, 1 H) 7.46 - 7.48 (m, 1 H) 7.34 - 7.40 (m, 1 H) 7.26 (s, 1 H) 7.07 - 7.17 (m, 1 H) 7.07 (s, 4 H) 6.84 - 6.96 (m, 4 H) 5.01 - 5.15 (m, 1 H) 3.26 - 3.31 m, 2 H) 3.08 (dd, >13.755, 5.808 Hz, 1 H) 2.87 (dd, >13.755, 8.620 Hz, 1 H) 2.65 - 2.80 (m, 2
H).
Synthesis of Compound 88
To a solution of intermediate 1 (0.3 g, 727.58 umol, 1 eq) and intermediate 2 (158.76 mg, 27.58 umol, 1 eq) in DCM (5 mL) was added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) nd the mixture was cooled to 0 °C, then T3P (463.00 mg, 1.46 mmol, 432.71 uL, 2 eq) was dded drop wise at 0 °C. The reaction mixture was allowed to warm to 25 °C and stirred at 25 °C or 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was iluted with ice water (20 mL) and extracted with DCM (3 x 10 mL). The combined organic hase was washed with brine (20 mL), dried with anhydrous Na2SC>4, filtered and concentrated n vacuo to yield intermediate 3 (0.5 g, crude) was obtained as a yellow oil. LCMS: m/z = 532.2 M+FT), !H-NMR: (400 MHz, CDCb) δ = 7.97 - 8.09 (m, 4 H) 7.40 - 7.53 (m, 5 H) 7.20 (d, >8.56 Hz, 2 H) 7.11 (t, >4.34 Hz, 1 H) 6.66 - 6.76 (m, 2 H) 5.32 - 5.39 (m, 1 H) 3.33 - 3.41 m, 1 H) 3.27 - 3.31 (m, 2 H) 3.02 - 3.09 (m, 1 H) 2.80 - 2.89 (m, 2 H).
To a solution of intermediate 3 (0.5 g, 752.44 umol, 1 eq ) in EtOH (9 mL) and H2O (3 mL) was dded NH4CI (201.24 mg, 3.76 mmol, 5 eq) and the solution was heated to 90 °C, then Fe 210.12 mg, 3.76 mmol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 4 (0.5 g, crude) was obtained as a yellow solid. LCMS: m/z = 502.1 (M+H+).
A mixture of intermediate 4 (0.06 g, 119.61 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SOs-pyridine (57.11 mg, 358.84 umol, 3 eq) was added. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when omplete, 7% ammonium hydroxide (1 mL) was added drop-wise and the mixture was dried by flowing N2 to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 50*25 5u;mobile phase: [water(0.04%NH3H20+10mMNH*HC03)-ACN];B%: 10%- 0%,10min) to yield Compound 88 (18.4 mg, 26%) was obtained as a white solid. LCMS: m/z 580.0 (M-H4), !H-NMR: (400 MHz, DMSO-tife+D20) δ = 7.86 - 7.91 (m, 2 H) 7.59 (dd, = 5.07, 0.92 Hz, 1 H) 7.44 - 7.56 (m, 4 H) 7.18 (s, 1 H) 7.12 (dd, .M.95, 3.85 Hz, 1 H) 6.84 - .95 (m, 4 H) 4.99 - 5.10 (m, 1 H) 3.01 - 3.23 (m, 3 H) 2.80 - 2.90 (m, 1 H) 2.60 - 2.79 (m, 2 H).
Synthesis of Compound 89
To a stirred solution of intermediate 1 (0.1 g, 713.57 umol, 1 eq) in anhydrous DMF (2 mL) was dded DIPEA (368.90 mg, 2.85 mmol, 497.17 uL, 4 eq), then (S)-2-(4-nitrophenyl)-l-(2- hiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide salt (294.22 mg, 713.57 umol, 1 eq) and yBOP (427.04 mg, 820.61 umol, 1.15 eq) was added. The reaction was stirred at 20 °C for 1 h nd at 50 °C for another 12 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition water (40 mL) at 25 °C, and then extracted with ethyl acetate (3 x 15 mL). The combined organic phase was washed with brine 40 mL), dried with anhydrous NaaSO-t, filtered and concentrated in vacuo to yield intermediate (0.4 g, crude) was obtained as a yellow solid. LCMS: m/z = 454.0 (M+H+).
To a solution of intermediate 2 (0.38 g, 837.86 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (224.09 mg, 4.19 mmol, 5 eq) and the mixture was heated to 90 °C, then Fe 233.97 mg, 4.19 mmol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (column: Luna C18 100*30 u;mobile phase: [water(0.05%HCl)-ACN];B%: l%-30%,10min) to yield intermediate 3 (0.12 , 34%) was obtained as a yellow solid. LCMS: m/z = 424.0 (M+H+), 1H-NMR: (400 MHz, DMSOntfc) δ = 14.19 (br s, 1 H) 10.38 (br s, 2 H) 8.73 (d, >8.56 Hz, 1 H) 7.72 (dd, >5.01, 1.10 Hz, 1 H) 7.65 (dd, >3.67, 1.10 Hz, 1 H) 7.51 (s, 2 H) 7.41 (s, 1 H) 7.25 - 7.37 (m, 4 H) 7.17 (dd, >5.01, 3.67 Hz, 1 H) 5.00 - 5.30 (m, 1 H) 3.22 (dd, >13.69, 5.50 Hz, 1 H) 2.96 - 3.09 (m, 3 H) .65 - 2.80 (m, 2 H).
A mixture of intermediate 3 (0.12 g, 283.32 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, and then SCb-pyridine (67.64 mg, 424.98 umol, 1.5 eq) was added. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by TLC and when complete, % ammonium hydroxide (2 mL) was added drop-wise and the mixture was dried by flowing N2 o give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 u;mobile phase: [water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: 5%-35%,10min) to ield Compound 89 (50.1 mg, 33%) was obtained as a white solid. LCMS: m/z = 502.0 (M-H+), Ή-ΝΜΚ: (400 MHz, D20) δ = 7.20 - 7.28 (m, 2 H) 6.96 - 7.02 (m, 2 H) 6.88 - 6.95 (m, 4 H) .80 - 6.87 (m, 2 H) 5.05 - 5.20 (m, 1 H) 2.90 - 3.05 (m, 1 H) 2.77 - 2.87 (m, 2 H) 2.65 - 2.77 (m, H) 2.40 - 2.57 (m, 2 H).
Synthesis of Compound 90
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (300 mg, 727.58 umol, 1 eq) and intermediate 2 (138.39 mg, 727.58 umol, 1 q) in DCM (5 mL) was added EtsN (4.37 mmol, 607.63 uL, 6 eq), and the mixture was cooled o 0 °C, then T3P (1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added drop-wise. Then the mixture was allowed to warm to 25 °C and stirred for 1 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with DCM (3 x 20 mL). Then combined organic phase was washed with brine (2 x 20 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 (360 mg, rude) was obtained as a yellow oil. LCMS: m/z = 504.2 (M+H+), 1H-NMR: (400 MHz, CDCb) = 10.01 (s, 1 H) 8.25 (dd, >4.707, 1.284 Hz, 1 H) 7.98 (d, >8.559 Hz, 2 H) 7.92 (dd, = 7.825, 1.100 Hz, 1 H) 7.45 (dd, >3.668, 0.978 Hz, 1 H) 7.37 - 7.42 (m, 1 H) 7.02 - 7.09 (m, 4 H) 6.61 (s, 1 H) 6.45 (d, >8.436 Hz, 1 H) 5.31 - 5.38 (m, 1 H) 3.24 (dd, >13.205, 5.869 Hz, 1 H) 3.06 - 3.17 (m, 3 H) 2.60 (t, >7.214 Hz, 2 H).
A solution of intermediate 3 (360.00 mg, 714.86 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (191.19 mg, 3.57 mmol, 5 eq) at 25 °C, and then the solution was heated to 90 °C, Fe (199.61 mg, 3.57 mmol, 5 eq) was added into the mixture and stirred for 2 h at 90 °C. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (410 mg, crude) was btained as a yellow solid. LCMS: m/z = 474.1 (M+H+).
To a solution of intermediate 4 (410 mg, 865.69 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SCb-pyridine (413.36 mg, 2.60 mmol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and when complete, 7 % ammonium hydroxide (5 mL) was dded dropwise, and then the mixture was dried by flowing N2, and the residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: 10%-40%,10min) to yield Compound 90 (139.6 mg, 29%) was obtained s a white solid. LCMS: m/z = 552.0 (M-H4), Ή-ΝΜΗ: (400 MHz, D20) δ = 7.80 - 7.98 (m, 2 H) 7.50 - 7.62 (m, 2 H) 7.11 - 7.20 (m, 1 H) 7.05 (s, 1 H) 6.91 - 7.00 (m, 3 H) 6.86 (d, >8.379 Hz, 2 H) 6.47 (s, 1 H) 4.91 - 5.05 (m, 1 H) 2.94 - 3.01 (m, 2 H) 2.74 - 2.92 (m, 2 H) 2.48 - 2.60 m, 2 H).
Synthesis of Compound 91
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yQethanamine hydrobromide alt (300.00 mg, 727.58 umol, 1 eq) and intermediate 1 (120.19 mg, 727.58 umol, 1 eq) in DCM 5 mL) was drop-wise added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, hen T3P (694.51 mg, 1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (25 mL) and then he mixture was extracted with DCM (3 x 10 mL). The combined organic phase was washed with rine (10 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 2 (348 mg, crude) was obtained as a yellow solid. LCMS: m/z = 479.2 (M+H+), 1H- NMR (400 MHz, CDCb) δ = 8.26 - 8.34 (m, 1 H) 8.06 (d, >8.60 Hz, 2 H) 7.48 - 7.53 (m, 1 H) .42 - 7.46 (m, 1 H) 7.36 - 7.41 (m, 1 H) 7.18 (d, >8.60 Hz, 2 H) 7.08 - 7.13 (m, 1 H) 6.96 - 7.02 m, 1 H) 6.67 (s, 1 H) 6.36 (d, >8.16 Hz, 1 H) 5.29 - 5.36 (m, 1 H) 3.28 - 3.36 (m, 1 H) 3.15 - .23 (m, 1 H) 2.94 - 3.01 (m, 2 H) 2.55 (s, 3 H) 2.47 - 2.53 (m, 2 H).
A solution of intermediate 2 (348 mg, 727.14 umol, 1 eq) and NH4CI (194.48 mg, 3.64 mmol, 27.11 uL, 5 eq) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (203.04 mg, .64 mmol, 5 eq) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated n vacuo to yield intermediate 3 (326 mg, crude) was obtained as a yellow solid. LCMS: m/z = 49.2 (M+H+), Ή-ΝΜΒ: δ = 8.18 (dd, >4.96, 1.43 Hz, 1 H) 7.58 (dd, >3.75, 1.10 Hz, 1 H) .56 (d, >5.07 Hz, 1 H) 7.48 - 7.53 (m, 1 H) 7.10 - 7.17 (m, 2 H) 7.01 (s, 1 H) 6.93 (d, >8.16 Hz, 2 H) 6.65 (d, >8.38 Hz, 2 H) 5.20 (dd, >8.71, 6.50 Hz, 1 H) 3.13 (dd, >13.78, 6.28 Hz, 1 H) 2.83 - 2.95 (m, 3 H) 2.43 - 2.58 (m, 5 H).
A solution of intermediate 3 (326 mg, 726.70 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (346.99 mg, 2.18 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u; mobile hase: [water(0.04%NH3H20 lOmMNILHCC^ACN]; B%: 10%-40%, lOmin) to yield Compound 91 (54.1 mg, 13%) was obtained as a white solid. LCMS: m/z = 527.0 (M-H÷), 1H- NMR: (400 MHz, DMSO-dfc) δ = 8.33 (d, >8.56 Hz, 1 H) 8.23 - 8.28 (m, 1 H) 7.57 - 7.82 (m, 3 H) 7.46 (d, >7.46 Hz, 1 H) 7.12 - 7.26 (m, 4 H) 7.10 (s, 1 H) 6.96 (s, 1 H) 6.85 - 6.92 (m, 4 H) .03 - 5.13 (m, 1 H) 2.99 - 3.07 (m, 1 H) 2.73 - 2.86 (m, 3 H) 2.44 (s, 3 H) 2.37 - 2.43 (m, 2 H). Synthesis of Compound 92
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (300 mg, 820.04 umol, 1 eg) and intermediate 2 (120.19 mg, 727.58 umol, 1 g) in DCM (3 mL) was added EtgN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eg), and then the olution was cooled to 0 °C, then T3P (521.84 mg, 820.04 umol, 487.70 uL, 50% purity, 1 eg) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (5 mL) and xtracted with DCM (2 x 10 mL), and then combined organic phase was washed with brine (10 mL), dried with anhydrous NazSC^ and concentrated in vacuo to yield intermediate 3 (492.2 mg, rude) was obtained as yellow oil. LCMS: m/z = 479.0 (M+H+).
To a solution of intermediate 3 (490 mg, 1.02 mmol, 1 eg) in EtOH (8 mL) and H2O (3 mL) was dded NH4CI (274.58 mg, 5.12 mmol, 5 eg) at 25 °C, and then the solution was allowed to warm o 90 °C, and then Fe (285.88 mg, 5.12 mmol, 5 eg) was added, then the mixture was stirred at 90 C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and HPLC and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 4 (317 mg, crude) was obtained as yellow soli. LCMS: m/z = 449.2 (M+H+).
To a solution of intermediate 4 (310 mg, 691.03 umol, 1 eg) in pyridine (1 mL) and CH3CN (1 mL) was cooled to 0 °C, then the SCh-pyridine (329.96 mg, 2.07 mmol, 3 eg) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 1%- 0%,10min) to yield Compound 92 (70.6 mg, 19%) was obtained as a white solid. LCMS: m/z 527.0 (Μ-ΙΓ). Ή-NMR: (400 MHz, DMS0-fife+D20) δ = 8.42 (d, >8.44 Hz, 1 H) 8.27 - 8.30 m, 1 H) 7.60 - 7.65 (m, 2 H) 7.30 (s, 1 H) 7.08 - 7.23 (m, 3 H) 6.87 - 6.92 (m, 4 H) 5.01 - 5.07 m, 1 H) 2.99 - 3.04 (m, 1 H) 2.72 - 2.89 (m, 3 H) 2.45 - 2.50 (m, 5 H).
Synthesis of Compound 93
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine hydrobromide alt (400 mg, 1.09 mmol, 1 eq) and intermediate 1 (169.77 mg, 1.09 mmol, 1 eq) in DCM (10 mL) was drop-wise added Et3N (330.08 mg, 3.26 mmol, 454.03 uL, 3 eq) and cooled to 0 °C, then T3P 1.04 g, 1.63 mmol, 970.00 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 2 h at 25 °C under N2 atmosphere. The reaction was monitored by TLC and when complete. The mixture was poured into ice- water (15 mL), and then the mixture was extracted with dichloromethane (2 x 15 mL). The combined organic phase was washed with rine (15 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 2 (371 mg, crude) was obtained as a white solid. To a solution of intermediate 2 (341 mg, 726.25 umol, 1 eq) in EtOAc (5 mL) was added nCl2.2H20 (1.31 g, 5.81 mmol, 8 eq) and heated to 50 °C and stirred at 50 °C for 2 h. The reaction was monitored by LCMS and when complete, the solution was added sat. seignette salt (50 mL), hen filtered and the filter was extracted with ethyl acetate (2 x 30 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield intermediate 3 (300 mg, crude) was obtained s a white solid. LCMS: m/z = 440.2 (M+H+).
A solution of intermediate 3 (300 mg, 682.51 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (325.89 mg, 2.05 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: 5%-30%,10min) to yield
Compound 93 (104.3 mg, 28%) was obtained as a white solid. LCMS: m/z = 517.9 (M-H+), *11- NMR (400 MHz, DMSO-dk) δ = 8.47 (d, >8.436 Hz, 1 H) 7.71 (dd, >5.135, 0.978 Hz, 1 H) 7.68 s, 1 H) 7.64 (dd, >3.668, 1.100 Hz, 1 H) 7.29 (s, 1 H) 7.17 (dd, >5.013, 3.790 Hz, 1 H) 7.08 (s, H) 6.85 - 6.92 (m, 4 H) 5.01 - 5.11 (m, 1 H) 2.99 - 3.10 (m, 3 H) 2.86 (dd, >13.755, 8.375 Hz, H) 2.58 - 2.69 (m, 2 H) 2.27 (s, 3 H).
To a mixture of intermediate 1 (4 g, 20.30 mmol, 1 eq) and intermediate 2 (6.99 g, 81.21 mmol, .31 mL, 4 eq) in DMF (20 mL) was added Pd(OAc)2 (1.14 g, 5.08 mmol, 0.25 eq), PPh3 (2.66 g, 0.15 mmol, 0.5 eq) and Et3N (6.16 g, 60.90 mmol, 8.48 mL, 3 eq). Then the mixture was heated o 140 °C and stirred for 16 h at 140 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into water (50 mL) and extracted with ethyl cetate (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried with nhydrous Na2SC>4, filtered and concentrated in vacuo. The residue was purified by column hromatography (Si02, Petroleum ether: Ethyl acetate = 20: 1 to 5: 1) to yield intermediate 3 { 1.3 , 31%) was obtained as a yellow solid. LCMS: m/z = 203.3 (M+H*), 'H-NMR: (400 MHz, DMSCMO δ = 11.89 (s, 1 H) 8.55 (d, >1.96 Hz, 1 H) 8.37 (d, >1.83 Hz, 1 H) 7.80 (d, >16.02 Hz, 1 H) 7.48 - 7.57 (m, 1 H) 6.68 (d, >16.02 Hz, 1 H) 6.45 - 6.53 (m, 1 H) 3.73 (s, 3 H). To a mixture of Pd/C ((1 g, 8.42 mmol, 10% purity, 1.42 eq) in MeOH (10 mL) was added ntermediate 3 (1.2 g, 5.93 mmol, 1 eq) at 25 °C. The suspension was degassed and purged with H2 several times. Then the mixture was stirred under ¾ (15 psi) at 25 °C for 1 h. The reaction was monitored by LCMS and when complete, the solution was filtered and concentrated in vacuo to ield intermediate 4 (805 mg, crude) was obtained as a white solid. LCMS: m/z = 205.3 (M+H4).
To a solution of intermediate 4 (980 mg, 4.80 mmol, 1 eq) in THF (20 mL) and H2O (5 mL) was dded LiOH (459.68 mg, 19.19 mmol, 4 eq) under N2 atmosphere and then stirred at 25 °C for 16 . The reaction was monitored by LCMS and when complete, the solution was filtered and oncentrated in vacuo to yield intermediate 5 (1.9 g, crude) was obtained as a white solid. LCMS: m/z = 191.3 (M+H4).
To a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-y1)thiazol-4-y1)ethanamine hydrobromide alt (300 mg, 727.58 umol, 1 eq) and intermediate 5 (570.80 mg, 2.91 mmol, 4 eq) in DCM (5 mL) was dropwise added EtsN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 3 hat 25 °C under N2 atmosphere. The reaction was monitored y LCMS and when complete. The mixture was poured into ice-water (25 mL) and then the mixture was extracted with DCM (3 x 25 mL). The combined organic phase was washed with rine (25 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 7 (360 mg, crude) was obtained as a yellow solid. LCMS: m/z = 504.2 (M+H*).
A solution of intermediate 2 ( 360 mg, 714.86 umol, 1 eq) and NH4CI (191.19 mg, 3.57 mmol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (199.61 mg, 3.57 mmol, 5 q) was added at 90 °C under N2 atmosphere, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated n vacuo to yield intermediate 8 (330 mg, crude) was obtained as a yellow solid. LCMS: m/z = 74.3 (M+H4).
A solution of intermediate 8 (330 mg, 696.77 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCh-pyridine (332.70 mg, 2.09 mmol, 3 eq) was added in portions and stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10mMNH4HC03)-ACN]; B%: 5%-35%, lOmin]) to yield Compound 94 (52.3 mg, 12%) was obtained as a white solid. LCMS: m/z = 552.0 (Μ-ΙΓ), 1H- NMR: (400 MHz, DMSO-ifc) 5 = 11.61 (s, 1 H) 8.28 (d, >8.56 Hz, 1 H) 7.99 - 8.08 (m, 1 H) .80 (s, 1 H) 7.67 - 7.73 (m, 1 H) 7.58 - 7.65 (m, 1 H) 7.39 - 7.47 (m, 1 H) 7.21 (s, 1 H) 7.13 - .19 (m, 1 H) 7.09 (s, 1 H) 7.00 (s, 1 H) 6.96 (s, 1 H) 6.82 - 6.93 (m, 4 H) 6.31 - 6.43 (m, 1 H) .02 - 5.13 (m, 1 H) 2.99 - 3.07 (m, 1 H) 2.75 - 2.90 (m, 3 H) 2.41 - 2.47 (m, 2 H).
Synthesis of Compound 95
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nltrophenyl)ethanamine ydrochloride salt (500 mg, 1.09 mmol, 1 eq), intermediate 2 (206.88 mg, 1.09 mmol, 1 eq) in DCM (6 mL) was added Et3N (663.84 mg, 6.56 mmol, 913.12 uL, 6 eq) and the mixture was ooled to 0 °C, and then T3P (1.04 g, 1.64 mmol, 975.40 uL, 50% purity, 1.5 eq) was added, then t was warmed to 25 °C and stirred for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was concentrated in vacuo to yield intermediate A (267 mg, crude) was obtained as a yellow solid. LCMS: m/z = 672.2 (M+H*).
A mixture of intermediate 3A (497 mg, 739.84 umol, 1 eq) and NHs/MeOH (7 M, 5 mL) was heated to 60 °C and stirred at 60 °C for 1 h. The reaction was monitored by LCMS and when omplete, the reaction mixture was concentrated in vacuo to yield intermediate 3 (419 mg, rude) was obtained as a yellow solid. LCMS: m/z = 501.0 (M+H+).
A solution of intermediate 3 (460 mg, 918.95 umol, 1 eq ) in EtOH (8 mL) and H2O (3 mL) was dded NH4CI (485.23 mg, 9.19 mmol, 10 eq) at 25 °C, and then the solution was heated to 90 °C, e (513.19 mg, 9.19 mmol, 10 eq) was added, the reaction was stirred at 90 °C for 2 h under N2 tmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC column: Luna C18 100*30 5u;mobile phase: [water(0.05%HCl)-ACN];B%: l%-30%,10min) to ield intermediate 4 (301 mg, 67%) was obtained as a yellow solid. LCMS: m/z = 471.2 M+IF), 'H-NMR: (400 MHz, DMSO-tfc) δ = 10.81 (s, 1 H) 9.11 (s, 1 H) 8.49 (d, >8.8 Hz, H) 7.59 (s, 1 H) 7.50 (d, >7.9 Hz, 1 H) 7.33 (d, >8.2 Hz, 1 H) 7.25 - 7.30 (m, 4 H) 7.15 - 7.19 m, 1 H) 7.05 (t, >7.5 Hz, 1 H) 6.91 - 7.01 (m, 2 H) 5.13 - 5.23 (m, 1 H) 4.49 (s, 2 H) 3.17 - .23 (m, 1 H) 2.89 - 2.98 (m, 1 H) 2.79 - 2.84 (m, 2 H) 2.40 - 2.47 (m, 2 H).
A solution of intermediate 4 (301 mg, 639.63 umol, 1 eq) in MeCN (2 mL) and pyridine (2 mL) was cooled to 0 °C, and then SO3. pyridine (305.41 mg, 1.92 mmol, 3 eq) was added. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H20+10mM NH4HC03>ACN];B%: 5%-35%,10min) to yield Compound 95 (38.1 mg, 11%) was obtained as a white solid. LCMS: m/z = 549.1(M-H+), 1H- NMR: (400 MHz, DMSO-ifc) 5 = 13.90 (br s, 1 H) 10.70 (s, 1 H) 8.64 (s, 1 H) 8.29 (d, >8.60 Hz, 1 H) 7.86 (br s, 1 H) 7.48 (d, >7.72 Hz, 1 H) 7.29 (d, >7.94 Hz, 1 H) 7.22 (s, 1 H) 7.14 (s, H) 7.08 (s,l H) 7.03 (t, >7.50 Hz, 1 H) 6.91 - 6.97 (m, 3 H) 6.86 - 6.91 (m, 2 H) 6.71 (s, 1 H) .08 - 5.21 (m, 1 H) 4.37 (s, 2 H) 3.01 (dd, >13.67, 5.73 Hz, 1 H) 2.74 - 2.95 (m, 3 H) 2.41 - .48 (m, 2 H). Synthesis of Compound 96
To a solution of (S)-l-(2-((lH-lmidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamine ydrochloride salt (0.5 g, 1.37 mmol, 1 eq) and intermediate 2 (213.49 mg, 1.37 mmol, 1 eq) in DCM (5 mL) was added EtsN (8.20 mmol, 1.14 mL, 6 eq), and the mixture was cooled to 0 °C, hen T3P (1.37 mmol, 812.84 uL, 50% purity, 1 eq) was added drop-wise, and then the solution was stirred at 25 °C for 2 h.. The reaction was monitored by LCMS and when complete, then the mixture was dried by flowing N2 to yield intermediate 3 (500 mg, crude) was obtained as a ellow oil. LCMS: m/z = 468.2 (M+H*).
A solution of intermediate 3 (0.5 g, 1.07 mmol, 1 eq) in EtOH (5 mL) and H2O (2 mL) was dded NH4CI (286.00 mg, 5.35 mmol, 5 eq), and then the solution was heated to 90 °C, Fe 298.62 mg, 5.35 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo to yield intermediate 4 (0.8 g, crude) was obtained as a brown solid. LCMS: m/z = 438.2 (M+H*).
To a solution of intermediate 4 (0.4 g, 914.12 umol, 1 eq) in MeCN (2 mL) and pyridine (2 mL) was added SCh-pyridine (351.40 mg, 1.37 mmol, 1.5 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by TLC and when complete, 7 % ammonium hydroxide (2 mL) was added dropwise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Agela Durashell C18 150*25 5u;mobile phase: [water(10mM NH4HCO3)- ACN];B%: 5%-40%,10min) to yield Compound 96 (61.4 mg, 14%) was obtained as a white olid. LCMS: m/z = 516.0 (M-H+), IH-NMR: (400 MHz, DMSO-i&) δ = 8.62 (s, 1 H) 8.31 (d, >8.436 Hz, 1 H) 7.74 (br s, 1 H) 7.28 (dd, >5.135, 1.100 Hz, 1 H) 7.21 (br s, 1 H) 7.11 - 7.17 m, 2 H) 7.08 (br s, 1 H) 6.95 (br s, 1 H) 6.86 - 6.92 (m, 3 H) 6.74 - 6.84 (m, 3 H) 5.12 (q, >7.499 Hz, 1 H) 4.35 (s, 2 H) 3.58 - 3.59 (m, 1 H) 2.94 - 2.99 (m, 3 H) 2.43 - 2.47 (m, 2 H).
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq ) and intermediate 2 (179.64 mg, 820.04 umol, 1 q) in DCM (3 mL) was added EtgN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (521.84 mg, 820.04 umol, 487.70 uL, 50 % purity, 1 eq) was added dropwise at 0 °C and then the solution was stirred at 25 °C far 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (3 mL) and xtracted with dichloromethane (2 x 10 mL), and then combined organic phase was washed with rine (10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 612 mg, crude) was obtained as yellow oil. LCMS: m/z = 530.1 (M+FT). To a solution of intermediate 3 (600 mg, 1.13 mmol, 1 eq) in EtOH (10 mL) and H2O (3 mL) was added NH4CI (295.85 mg, 5.66 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (315.85 mg, 5.66 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and HPLC nd when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo. The esidue was purified by prep-HPLC (column: UniSil 120*30* 10um;mobile phase: water(0.05%HCl)-ACN];B%: 1%-40%,1 lmin) to yield intermediate 4 (167.7 mg, 25%) was btained as a yellow solid. LCMS: m/z = 500.1 (M+FL).
To a solution of intermediate 4 (160 mg, 319.72 rnnol, 1 eq) in pyridine (1 mL) and CH3CN (1 mL) was cooled to 0 °C, then the SOs-pyridine (50.89 mg, 319.72 umol, 1 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd HPLC and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then he mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: UniSil 120*30* 10um;mobile phase: [water(0.05%HCl)-ACN];B%: l%-40%, 11 min) to yield Compound 97 (35.3 mg, 18%) was obtained as a white solid. LCMS: m/z = 578.2 (M-fT). 1H- NMR: (400 MHz, DMSO-ds) δ = 13.19 - 14.46 (m, 1 H) 8.69 (s, 1 H) 8.35 (d, >8.56 Hz, 1 H) .74 (br s, 1 H) 7.42 - 7.46 (m, 2 H) 7.22 - 7.31 (m, 2 H) 7.17 - 7.22 (m, 1 H) 7.13 (s, 1 H) 7.04 - .10 (m, 1 H) 6.92 - 6.98 (m, 1 H) 6.85 - 6.92 (m, 2 H) 6.76 - 6.83 (m, 2 H) 5.10 - 5.18 (m, 1 H) .37 (s, 2 H) 3.00 - 3.09 (m, 2 H) 2.93 - 3.00 (m, 1 H) 2.83 - 2.92 (m, 1 H) 2.30 - 2.36 (m, 2 H).
Synthesis of Compound 98
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nltrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (152.65 mg, 820.04 umol, 1 q) in DCM (3 mL) was added EtaN (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (521.84 mg, 820.04 umol, 487.70 uL, 50% purity, 1 eq) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (10 mL) and xtracted with DCM (2 x 10 mL), and then combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 (406 mg, rude) was obtained as yellow oil. LCMS: m/z = 498.3 (M+FP).
To a solution of intermediate 3 (406 mg, 816.05 umol, 1 eq) in EtOH (8 mL) and H2O (3 mL) was added NH4CI (214.86 mg, 4.08 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (227.86 mg, 4.08 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2 h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (822.0 mg, crude) was obtained as yellow solid. LCMS: m/z = 468.0 (M+FP). To a solution of intermediate 4 (822 mg, 1.76 mmol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then the S03-pyridine (839.50 mg, 5.27 mmol, 3 eq) was added at 0 °C and he reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (2 mL) was added drop-wise, and then the mixture was dried by flowing Nz, and the residue was purified by prep-HPLC (column: Waters Xbridge 50*25 5u;mobile phase: [water(0.04%NH3H2(H10mMNH4HCC>3)-ACN];B%: 1%- 0%,10min) to yield Compound 98 (32.9 mg, 3%) was obtained as white solid. LCMS: m/z = 46.1 (M-H+). ‘H-NMR: (400 MHz, DMSO-4;) δ = 8.65 (s, 1 H) 8.25 (d, >8.56 Hz, 1 H) 7.73 br s, 1 H) 7.21 - 7.34 (m, 2 H) 7.17 - 7.20 (m, 1 H) 7.08 - 7.15 (m, 2 H) 7.07 (br s, 1 H) 6.93 - .03 (m, 2 H) 6.84 - 6.91 (m, 2 H) 6.78 (d, >8.56 Hz, 2 H) 5.02 - 5.19 (m, 1 H) 4.35 (s, 2 H) .91 - 2.96 (m, 1 H) 2.70 - 2.86 (m, 3 H) 2.36 - 2.41 (m, 2 H).
Synthesis of Compound 99
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamine ydrochloride salt (500 mg, 1.37 mmol, 1 eq) and intermediate 2 (283.25 mg, 1.37 mmol, 1 eq) in DCM (5 mL) was added Et3N (8.20 mmol, 1.14 mL, 6 eq), and the mixture was cooled to 0 °C, then T3P (1.30 g, 2.05 mmol, 1.22 mL, 50% purity, 1.5 eq) was added drop-wise. Then the mixture was allowed to warm to 25 °C and stirred for 2 h. The reaction was monitored by LCMS nd when complete, the reaction mixture was poured into water (75 mL), and extracted with DCM (3 x 25 mL). Then combined organic phase was washed with brine (3 x 30 mL), dried with nhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 (705 mg, crude) was btained as a light yellow oil. LCMS: m/z = 519.2 (M+H+), 1H-NMR: (400 MHz, DMSO-<&) δ 8.58 (d, >8.681 Hz, 1 H) 8.02 - 8.09 (m, 3 H) 7.98 (d, >7.825 Hz, 1 H) 7.83 - 7.92 (m, 2 H) .43 - 7.49 (m, 4 H) 7.35 - 7.42 (m, 2 H) 5.17 - 5.28 (m, 1 H) 4.14 - 4.31 (m, 2 H) 3.64 - 3.69 (m, H) 3.50 - 3.55 (m, 1 H) 3.20 - 3.29 (m, 3 H) 3.06 - 3.12 (m, 1 H).
A solution of intermediate 3 (705 mg, 1.36 mmol, 1 eq) in EtOH (12 mL) and H2O (4 mL) was dded NH4CI (363.58 mg, 6.80 mmol, 237.64 uL, 5 eq), and then the solution was heated to 90 C, Fe (379.58 mg, 6.80 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo. The residue was purified by prep-HPLC (column: Luna C18 100*30 u;mobile phase: [water(0.05%HCl)-ACN];B%: l%-30%,10min) to yield intermediate 4 (260 mg, 39% ) was obtained as a light yellow oil. LCMS: m/z = 489.1 (M+H4), 1H-NMR: (400 MHz, DMSCM,) δ = 10.48 (br s, 1 H) 9.10 (s, 1 H) 8.69 (d, >8.600 Hz, 1 H) 8.04 (d, >7.717 Hz, 1 H) 7.90 (d, >7.938 Hz, 1 H) 7.56 (s, 1 H) 7.45 - 7.51 (m, 1 H) 7.37 - 7.42 (m, 1 H) 7.31 s, 1 H) 7.28 (s, 4 H) 5.12 - 5.21 (m, 1 H) 4.47 (s, 2 H) 3.17 - 3.28 (m, 3 H) 2.95 - 3.00 (m, 1 H) .60 - 2.79 (m, 2 H).
To a solution of intermediate 4 (260 mg, 532.10 rnnol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SCb-pyridine (127.04 mg, 798.16 umol, 1.5 eq) at 0 °C and stirred for 0.17 h.
The reaction was monitored by TLC and when complete, 7% ammonium hydroxide (5 mL) was dded dropwise, and then the mixture was dried by flowing N2, and the residue was purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+ 1 OmM NHtHC03)-ACN];B%: l%-30%,10min) to yield Compound 99 (126.4 mg, 41%) was obtained s a white solid. LCMS: m/z = 567.0 (M-H4), Ή-ΝΜΒ: (400 MHz, DMSO-tife) 6 = 8.57 (s, 1 H) .43 (d, >8.436 Hz, 1 H) 7.99 - 8.07 (m, 1 H) 7.90 (d, >7.703 Hz, 1 H) 7.74 - 7.75 (m, 1 H) .45 - 7.49 (m, 1 H) 7.35 - 7.42 (m, 1 H) 7.21 (s, 1 H) 7.12 (s, 1 H) 7.09 (br s, 1 H) 6.96 (br s, 1 H) 6.87 - 6.92 (m, 2 H) 6.78 - 6.85 (m, 2 H) 5.11 (q, >7.580 Hz, 1 H) 4.33 (s, 2 H) 3.29 (t, >7.519 Hz, 2 H) 2.99 (dd, >13.633, 6.541 Hz, 1 H) 2.84 (dd, >13.694, 7.703 Hz, 1 H) 2.65 - 2.79 (m, 2 H). Synthesis of Compound 100
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (0.5 g, 1.09 mmol, 1 eq) and intermediate 2 (238.59 mg, 1.09 mmol, 1 eq) in DCM (7 mL) was added EtgN (663.84 mg, 6.56 mmol, 913.12 uL, 6 eq), and the mixture was ooled to 0 °C, then T3P (1.39 g, 2.19 mmol, 1.30 mL, 50% purity, 2 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with ice water (40 mL) and extracted with DCM (3 x 15 mL). The combined organic phase was washed with brine 40 mL), dried with anhydrous NaiSO^ filtered and concentrated in vacuo to yield intermediate (0.6 g, crude) was obtained as a yellow oil. LCMS: m/z = 530.1 (M+H4).
A solution of intermediate 3 (0.6 g, 1.13 mmol, 1 eq) and SnCl2.2H20 (1.28 g, 5.66 mmol, 5 eq) n EtOAc (10 mL) was heated to 50 °C. The mixture was stirred at 50 °C for 2 h. The reaction was monitored by LCMS and TLC and when complete, the reaction mixture was diluted with at. seignette salt (100 mL) and extracted with ethyl acetate (2 x 50 mL). The combined organic hase was washed with brine (80 mL), dried with anhydrous Na2S04, filtered and concentrated n vacuo. The residue was purified by prep-TLC (Diehl oromethane : Methanol = 10: 1) to yield intermediate 4 (0.08 g, 13%) was obtained as a yellow solid. LCMS: m/z = 500.1 (M+H+), ‘H- NMR: (400 MHz, DMSO-ifc) δ = 8.44 (d, >8.77 Hz, 1 H) 7.92 - 8.01 (m, 2 H) 7.51 - 7.64 (m, 4 H) 7.10 (s, 1 H) 6.98 (s, 1 H) 6.80 (d, >8.33 Hz, 2 H) 6.42 (d, >8.33 Hz, 2 H) 4.92 - 5.06 (m, 1 H) 4.18 (s, 2 H) 3.09 - 3.15 (m, 2 H) 2.99 (dd, >13.59, 5.70 Hz, 1 H) 2.63 - 2.81 (m, 3 H).
A mixture of intermediate 4 (0.08 g, 160.13 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C, then SCb-pyridine (38.23 mg, 240.20 umol, 1.5 eq) was added. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when complete, 7% mmonium (2 mL) was added drop-wise and the mixture was dried by flowing N2 to give a esidue. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile hase: [water(0.04%NH3H2(>H0mM NH4HC03>ACN];B%: l%-30%,10min) to yield Compound 100 (17.6 mg, 19% ) was obtained as a white solid. LCMS: m/z = 578.1 (M-H+), 'H- NMR: (400 MHz, DMSO-de) δ = 8.45 (d, >8.44 Hz, 1 H) 7.93 - 8.02 (m, 2 H) 7.62 - 7.77 (m, 2 H) 7.50 - 7.60 (m, 3 H) 7.09 - 7.15 (m, 1 H) 6.88 - 6.95 (m, 3 H) 6.81 - 6.87 (m, 2 H) 4.97 - 5.12 m, 1 H) 4.10 - 4.30 (m, 2 H) 3.11 - 3.17 (m, 2 H) 2.99 - 3.06 (m, 1 H) 2.79 - 2.86 (m, 1 H) 2.67 - .77 (m, 2 H).
Synthesis of Compound 101
To a stirred solution of intermediate 2 (0.1 g, 713.57 umol, 1 eq) in anhydrous DMF (5 mL) was dded PyBOP (427.04 mg, 820.61 umol, 1.15 eq), then (S)-l-{2-{(lH-lmldazol-5-yl)methyl)thlazol-4- yl)-2-(4-nitrophenyl)ethanamine hydrochloride salt (326.31 mg, 713.57 umol, 1 eq) and DIPEA 368.89 mg, 2.85 mmol, 497.16 uL, 4 eq) was added. The reaction was stirred at 20 °C for 3 h nd stirred at 50 °C for another 10 h under N2 atmosphere. The reaction was monitored by LCMS and when complete, the reaction mixture was quenched by addition water (40 mL), and hen extracted with ethyl acetate (3 x 20 mL). The combined organic phase was washed with rine (40 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 2 (0.4 g, crude) was obtained as a yellow oil. LCMS: m/z = 452.2 (M+H+).
To a solution of intermediate 2 (0.4 g, 885.93 umol, 1 eq ) in EtOH (6 mL) and H2O (2 mL) was dded NH4CI (236.94 mg, 4.43 mmol, 5 eq), then the mixture was heated to 90 °C, and then Fe 247.40 mg, 4.43 mmol, 5 eq) was added. The mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered and the filtrate was concentrated in vacuo to yield intermediate 3 (0.4 g, crude) was obtained as a green oil. LCMS: m/z = 422.1 (M+HT).
A mixture of intermediate 3 (0.4 g, 948.95 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C, then SCb-pyridine (226.55 mg, 1.42 mmol, 1.5 eq) was added in batches.
The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when omplete, 7% ammonium hydroxide (5 mL) was added drop-wise and the mixture was dried by lowing N2 to give a residue. The residue was purified by prep-HPLC (column: Luna C18 00*305u;mobile phase: [water(0.05%HCl)-ACN];B%: 1%-I5%,10min) and re-purified by rep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: l%-25%,10min) to yield Compound 101 (13.6 mg, 3%) was obtained as white solid. LCMS: m/z = 500.0 (M-iF), ‘H-NMR: (400 MHz, D20) 6 = 7.87 (s, 1 H) 7.16 (s, H) 7.07 - 7.10 (m, 1 H) 7.06 (s, 1 H) 7.00 - 7.04 (m, 4 H) 5.12 - 5.22 (m, 1 H) 4.30 (s, 2 H) .14 (dd, >13.88, 6.17 Hz, 1 H) 3.01 - 3.08 (m, 2 H) 2.93 (dd, >13.82, 9.05 Hz, 1 H) 2.62 - .70 (m, 2 H). Synthesis of Compound 102
To a solution of then (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (400 mg, 1.09 mmol, 1 eq) and intermediate 2 (207.96 mg, 1.09 mmol, 1 eq) in DCM (6 mL) was added EtsN (6.56 mmol, 913.12 uL, 6 eq), and the mixture was cooled to 0 °C, hen T3P (1.64 mmol, 487.70 uL, 1.5 eq) was added drop-wise. Then the mixture was allowed to warm to 25 °C and stirred for 1 h. The reaction was monitored by LCMS and when complete, the eaction mixture was poured into water (30 mL), and extracted with ethyl acetate (3 x 20 mL). Then combined organic phase was washed with brine (2 x 20 mL), dried with anhydrous Na2SC>4 nd concentrated in vacuo to yield intermediate 3 (350 mg, crude) was obtained as a yellow oil. LCMS: m/z = 502.1 (M+FT).
A solution of intermediate 3 (350 mg, 697.82 umol, 1 eq) in EtOH (6 mL) and ¾0 (2 mL) was dded NH4CI (186.64 mg, 3.49 mmol, 5 eq), and then the solution was heated to 90 °C, Fe 194.85 mg, 3.49 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by TLC and LCMS and when complete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield intermediate 4 (310 mg, crude) was obtained as a yellow oil. LCMS: m/z = 472.3 (M+H+). To a solution of intermediate 4 (310 mg, 657.37 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was added SCb-pyridine (313.89 mg, 1.97 mmol, 3 eq) at 0 °C and stirred fa- 0.17 h. The eaction was monitored by TLC and LCMS and when complete, 7% ammonium hydroxide (3 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 5%-35%,10min) to yield Compound 102 28.8 mg, 41%) was obtained as a white solid. LCMS: m/z = 550.0 (M-H+), 1H-NMR: (400 MHz, DMSO-dk) δ = 11.25 (s, 1 H) 8.46 (s, 1 H) 8.26 (d, >8.559 Hz, 1 H) 8.15 (dd, >4.646, .589 Hz, 1 H) 7.91 (dd, >7.825, 1.467 Hz, 1 H) 7.78 (s, 1 H) 7.21 (s, 1 H) 7.14 (s, 1 H) 7.09 (s, H) 6.95 - 7.02 (m, 2 H) 6.89 - 6.94 (m, 2 H) 6.81 - 6.88 (m, 2 H) 5.08 - 5.16 (m, 1 H) 4.33 (s, 2 H) 2.95 - 3.00 (m, 1 H) 2.75 - 2.90 (m, 3 H) 2.44 - 2.48 (m, 2 H). To a mixture of (S)-l-(2-((lH-imidazpl-5-yl)methyl)ihiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (500 mg, 1.37 mmol, 1 eq) and intermediate 2 (225.77 mg, 1.37 mmol, 1 eq) n DCM (5 mL) was drop-wise added Et3N (829.80 mg, 8.20 mmol, 1.14 mL, 6 eq) and cooled to °C, then T3P (1.30 g, 2.05 mmol, 1.22 mL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 3 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (75 mL) and then he mixture was extracted with DCM (3 x 25 mL). The combined organic phase was washed with rine (3 x 30 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 3 (500 mg, crude) was obtained as a yellow oil. LCMS: m/z = 624.4 (M+H+).
A solution of intermediate 3 (500 mg, 1.05 mmol, 2.62 eq) in NH3/MeOH (7 M, 2.31 mL, 40.30 q) was heated to 60 °C, and stirred for 1 h at 60 °C under Nz atmosphere. The reaction was monitored by LCMS and when complete. The mixture was concentrated in vacuo to yield ntermediate 4 (690 mg, crude) was obtained as a yellow oil. LCMS: m/z = 477.3 (M+H+).
To a solution of intermediate 4 (690 mg, 1.45 mmol, 1 eq) in EtOH (12 mL) and H2O (3 mL) was dded NH4CI (387.25 mg, 7.24 mmol, 253.11 uL, 5 eq) at 25 °C, and then the solution was heated o 90 °C, Fe (404.29 mg, 7.24 mmol, 5 eq) was added, then the mixture was stirred at 90 °C for 2 . The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, he filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM N¾HC03)-ACN];B%: 20%- 0%,10min) to yield intermediate S (180 mg, 28%) was obtained as a yellow oil. LCMS: m/z = 47.1 (M+iT), Ή-NMR: (400 MHz, DMSO-ifc): δ = 11.92 (br s, 1 H) 8.15 - 8.34 (m, 2 H) 7.53 - .66 (m, 1 H) 7.33 (d, J= 7.580 Hz, 1 H) 6.99 - 7.09 (m, 2 H) 6.96 (s, 1 H) 6.79 (d, >8.070 Hz, 2 H) 6.42 (d, >7.948 Hz, 2 H) 4.93 - 5.07 (m, 1 H) 4.82 (s, 2 H) 2.95 (dd, >13.633, 5.196 Hz, 1 H) 2.72 - 2.76 (m, 1 H) 2.51 - 2.53 (m, 2 H) 2.43 (s, 3 H) 2.30 - 2.35 (m, 2 H).
A solution of intermediate 5 (180 mg, 403.08 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SCb-pyridine (192.46 mg, 1.21 mmol, 3 eq) was added in portions nd stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mM NH4HC03)-ACN];B%: l%-25%,10min) to yield Compound 103 44.2 mg, 20%) was obtained as a white solid. LCMS: m/z = 525.1 (M-H+), ‘H-NMR: (400 MHz, D20): δ = 8.22 (dd, >5.318, 1.528 Hz, 1 H) 7.99 (d, >0.978 Hz, 1 H) 7.57 (dd, >7.825, 1.345 Hz, 1 H) 7.24 (dd, >7.703, 5.380 Hz, 1 H) 7.07 (s, 1 H) 6.90 - 7.02 (m, 5 H) 5.09 (dd, >8.436, .724 Hz, 1 H) 4.29 (s, 2 H) 3.01 (dd, >13.938, 6.480 Hz, 1 H) 2.83 - 2.91 (m, 3 H) 2.49 - 2.57 m, 2 H) 2.43 (s, 3 H).
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nltrophenyl)ethanamine ydrochloride salt (300 mg, 820.04 umol, 1 eq) and intermediate 2 (135.46 mg, 820.04 umol, 1 q) in DCM (3 mL) was added Et3N (497.88 mg, 4.92 mmol, 684.84 uL, 6 eq), and then the olution was cooled to 0 °C, then T3P (521.84 mg, 820.04 umol, 487.70 uL, 50% purity, 1 eq) was added drop-wise at 0 °C and then the solution was stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the solution was poured into water (3 mL) and xtracted with dichloromethane (2 x 10 mL), and then combined organic phase was washed with rine (10 mL), dried with anhydrous Na2SC>4 and concentrated in vacuo to yield intermediate 3 325.3 mg, crude) was obtained as yellow oil. LCMS: m/z = 477.3 (M+H+).
To a solution of intermediate 3 (320 mg, 671.49 umol, 1 eq) in EtOH (6 mL) and H2O (2 mL) was added NH4CI (175.20 mg, 3.36 mmol, 5 eq) at 25 °C, and then the solution was allowed to warm to 90 °C, and then Fe (187.50 mg, 3.36 mmol, 5 eq) was added, then the mixture was tirred at 90 °C for 2h under N2 atmosphere. The reaction was monitored by LCMS and when omplete, the reaction mixture was filtered, the filtrate was concentrated in vacuo to yield ntermediate 4 (612.7 mg, crude) was obtained as yellow solid. LCMS: m/z = 447.1 (M+H*).
To a solution of intermediate 4 (610 mg, 1.37 mmol, 1 eq) in pyridine (2 mL) and CH3CN (2 mL) was cooled to 0 °C, then the SCb-pyridine (652.24 mg, 4.10 mmol, 3 eq) was added at 0 °C nd the reaction mixture was stirred at 0 °C for 0.17 h. The reaction was monitored by LCMS nd when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water(0.04%NH3H20+10mM N¾HC03)-ACN];B%: 1%- 0%,10min) to yield Compound 104 (38.7 mg, 5%) was obtained as white solid. LCMS: m/z = 25.1 (M-H+). 1H-NMR: (400 MHz, DMSO-ifc+D20) 5 = 8.16 - 8.41 (m, 2 H) 7.13 (d, >8.56 Hz, 2 H) 7.02 - 7.04 (m, 2 H) 6.85 - 6.91 (m, 2 H) 6.77 - 6.84 (m, 2 H) 4.97 - 5.12 (m, 1 H) 4.30 s, 2 H) 2.90 - 3.01 (m, 1 H) 2.71 - 2.82 (m, 3 H) 2.38 - 2.44 (m, 5 H).
Synthesis of Compound 105
To a solution of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-{4-nitrophenyl)ethanamine ydrochloride salt (0.5 g, 1.37 mmol, 1 eq) and intermediate 2 (213.91 mg, 1.37 mmol, 1 eq) in DCM (5 mL) was added EtgN (831.78 mg, 8.22 mmol, 1.14 mL, 6 eq) and the mixture was ooled to 0 °C, then T3P (1.74 g, 2.74 mmol, 1.63 mL, 50% purity, 2 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 2 h. The reaction was monitored by LCMS and TLC and when complete, the reaction mixture was quenched by ddition ice water (40 mL), and then extracted with DCM (3 x 15 mL). The combined organic hase was washed with brine (40 mL), dried with anhydrous Na2SC>4, filtered and concentrated n vacuo. The residue was purified by column chromatography (S1O2, Dichloromethane : Methanol = 1: 0 to 10: 1) to yield intermediate 3 (0.25 g, 39%) was obtained as a yellow oil. LCMS: m/z = 468.0 (M+ET).
A mixture of intermediate 3 (0.23 g, 491.98 umol, 1 eq) and SnCl2.2H20 (555.07 mg, 2.46 mmol, 5 eq) in EtOAc (5 mL) was heated to 50 °C and stirred at 50 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was diluted with sat. salt (100 mL) and extracted with ethyl acetate (2 x 50 mL). The combined organic phase was washed with rine (100 mL), dried with anhydrous Na2S04, filtered and concentrated in vacuo to yield ntermediate 4 (0.08 g, crude) was obtained as a yellow solid. LCMS: m/z = 438.1 (Μ+ΕΓ). A mixture of intermediate 4 (0.08 g, 182.85 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was cooled to 0 °C, then SCb-pyridine (43.65 mg, 274.28 umol, 1.5 eq) was added. The mixture was stirred at 0 °C for 10 min. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (2 mL) was added drop-wise and the mixture was dried by flowing N2 to ive a residue. The residue was purified by prep-HPLC (column: YMC-Actus Triart C 18 00*30mm*5um;mobile phase: [water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 1%- 5%,10min) to yield Compound 105 (40.2 mg, 41%) was obtained as a white solid. LCMS: m/z 516.1 (M-H4), Έ-ΝΜΒ: (400 MHz, D2O) δ = 7.73 (s, 1 H) 7.08 (s, 1 H) 7.01 - 7.06 (m, 4 H) .01 (s, 1 H) 5.15 - 5.21 (m, 1 H) 4.27 (s, 2 H) 3.06 - 3.18 (m, 3 H) 2.97 - 3.06 (m, 1 H) 2.70 - .79 (m, 2 H) 2.29 (s, 3 H).
Synthesis of Compound 106
To a mixture of (S)-l-(2-((lH-imidazol-5-yl)methyl)thiazol-4-yl)-2-(4-nitrophenyl)ethanamine ydrochloride salt (400 mg, 1.09 mmol, 1 eq) and intermediate 2 (1.07 g, 5.47 mmol, 5 eq) in DCM (10 mL) was added EtsN (663.84 mg, 6.56 mmol, 913.12 uL, 6 eq) and cooled to 0 °C, then T3P (1.04 g, 1.64 mmol, 975.40 uL, 50% purity, 1.5 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 16 h at 25 °C under N2 atmosphere. The reaction was monitored by LCMS and when complete. The mixture was poured into ice-water (75 mL) and then he mixture was extracted with DCM (3 x 25 mL). The combined organic phase was washed with rine (3 x 30 mL), dried with anhydrous Na2SC>4, filtered and concentrated in vacuo to yield ntermediate 3 (900 mg, crude) was obtained as a yellow oil, LCMS: m/z = 502.3 (M+H4).
To a solution of intermediate 3 (900 mg, 1.79 mmol, 1 eq) in ETOH (12 mL) and H2O (3 mL) was dded NH4CI (479.93 mg, 8.97 mmol, 313.68 uL, 5 eq), and then the solution was heated to 90 C, Fe (501.04 mg, 8.97 mmol, 5 eq). Then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo. The residue was purified by prep-HPLC (column: Waters Xbridge 150*25 u;mobile phase: [water(0.04%NH3H20+10Mm NH4HCC>3)-ACN];B%: 5%-35%,10min) to yield ntermediate 4 (30 mg, 4%) was obtained as a light yellow solid. LCMS: m/z = 472.1 (M+H4), H-NMR: (400 MHz, DMSO-de) δ = 11.92 (s, 1 H) 11.47 (s, 1 H) 8.18 (d, >8.558 Hz, 1 H) 8.03 d, >2.078 Hz, 1 H) 7.68 (d, >1.834 Hz, 1 H) 7.59 (s, 1 H) 7.40 (d, >3.179 Hz, 1 H) 7.01 (s, 1 H) 6.75 - 6.82 (m, 3 H) 6.40 (d, >8.314 Hz, 2 H) 6.33 (d, >3.423 Hz, 1 H) 4.93 - 5.03 (m, 1 H) .15 (s, 2 H) 2.95 (dd, >13.755, 5.441 Hz, 1 H) 2.82 (t, >7.519 Hz, 2 H) 2.68 - 2.73 (m, 1 H) .38 - 2.50 (m, 2 H).
A solution of intermediate 4 (30 mg, 63.62 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was ooled to 0 °C and then SCb-pyridine (30.38 mg, 190.85 umol, 3 eq) was added in portions and tirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% mmonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+ 1 OmM NH4HC03)-ACN];B%: l%-25%,10min) to yield Compound 106 6.9 mg, 19% ) was obtained as a white solid. LCMS: m/z = 550.0 (M-H+), 1H-NMR: (400 MHz, D2O) δ = 8.29 (s, 1 H) 7.91 (d, >1.834 Hz, 1 H) 7.76 (d, >1.712 Hz, 1 H) 7.40 (d, >3.546 Hz, H) 7.03 (s, 1 H) 6.87 (d, >8.436 Hz, 2 H) 6.73 (d, >8.436 Hz, 2 H) 6.48 (s, 1 H) 6.43 (d, >3.546 Hz, 1 H) 5.03 (t, >7.214 Hz, 1 H) 4.12 - 4.27 (m, 2 H) 2.86 - 3.05 (m, 2 H) 2.76 - 2.86 m, 2 H) 2.52 - 2.63 (m, 2 H). o a mixture of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol~4-yl)eihanamine hydrobromide alt (300 mg, 727.58 umol, 1 eq) and intermediate 1 (122.35 mg, 727.58 umol, 1 eg) in DMF (3 mL) was ropwise added EtaN (441.74 mg, 4.37 mmol, 607.63 uL, 6 eq) and cooled to 0 °C, then T3P (926.01 mg, .46 mmol, 865.43 uL, 50% purity, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C nd stirred for 3 h at 25 "C under N? atmosphere. The reaction was monitored by LCMS and when omplete, the mixture was poured into ice-water (25 mL) and then the mixture was extracted with ethyl cetate (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous azSO», filtered and concentrated in vacuo to yield intermediate 2 (260 mg, crude) was obtained as a ellow solid. LCMS: m/z = 482.2 (M+H+).
A solution of intermediate 2 (260 mg, 539.91 umol, 1 eq) and NH4CI (143.1 mg, 2.70 mmol, 5 eq) n EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (150.77 mg, 2.70 mmol, 5 eq) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to yield ntermediate 3 (294 mg, crude) was obtained as a yellow solid. LCMS: m/z = 452.2 (M+FT).
A solution of intermediate 3 (294 mg, 651.05 umol, 1 eq) in pyridine (2 mL) and MeCN (2 mL) was cooled to 0 °C and then SOa-pyridine (310.86 mg, 1.95 mmol, 3 eq) was added in portions and stirred for 0.17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% ammonium hydroxide (1 mL) was added dropwise, and then the mixture was dried by flowing N2, nd the residue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water (0.05% ammonia hydroxide v/v)-ACN];B%: 15%-45%,10min) to yield Compound 107 89.5 mg, 24%) was obtained as a white solid. LCMS: m/z = 530.1 (M-H+), ‘H-NMR: (400 MHz, DMSO-de) δ = 8.33 (d, >8.44 Hz, 1 H) 7.71 (d, >4.89 Hz, 1 H) 7.61 - 7.66 (m, 2 H) 7.03 - 7.23 m, 9 H) 6.82 - 6.91 (m, 4 H) 5.05 - 5.10 (m, 1 H) 3.01 - 3.06 (m, 1 H) 2.76 - 2.89 (m, 3 H) 2.38 - .43 (m, 2 H).
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide sah (300 mg, 727.58 umol, 1 eq) and intermediate 2 (134.33 mg, 727.58 umol, 1 q) in DMF (3 mL) was added EtgN (4.37 mmol, 607.62 uL, 6 eq), and the mixture was cooled to °C, then T3P (1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added drop-wise. The mixture was llowed to warm to 25 °C and stirred at 25 °C for 3 h. The reaction was monitored by LCMS and when complete, the reaction mixture was paired into water, and extracted with ethyl acetate (3 x 0 mL). Then combined organic phase was washed with brine, dried with anhydrous Na2S04 and oncentrated in vacuo to yield intermediate 3 (360 mg, crude) was obtained as a yellow oil. LCMS: m/z = 498.1 (M+H+). A solution of intermediate 3 (360 mg, 722.87 umol, 1 eq) in EtOH (3 mL) and H2O (1 mL) was dded NH4CI (201.86 mg, 3.61 mmol, 5 eq), and then the solution was heated to 90 °C, Fe 194.63 mg, 3.61 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo to yield intermediate 4 (419 mg, crude) was obtained as a yellow solid. LCMS: m/z = 468.1 (Μ+ΙΓ).
To a solution of intermediate 4 (419 mg, 895.24 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SOs-pyridine (427.46 mg, 2.69 mmol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by HPLC and LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: [water (0.05% mmonia hydroxide v/v)-ACN];B%: 15%-45%,10min) to yield Compound 108 (79.7 mg, 16%) was obtained as a white solid. LCMS: m/z = 546.0 (M-H4), 1H-NMR: (400 MHz, DMSO-riis) δ 8.35 (d, >8.437 Hz, 1 H) 7.71 (d, >5.013 Hz, 1 H) 7.64 (d, >3.546 Hz, 1 H) 7.34 - 7.41 (m, H) 7.14 - 7.25 (m, 6 H) 7.08 (br s, 1 H) 6.95 (br s, 1 H) 6.83 - 6.92 (m, 4 H) 5.02 - 5.14 (m, 1 H) 3.04 (dd, >13.633, 5.808 Hz, 1 H) 2.81 - 2.91 (m, 3 H) 2.42 (t, >7.458 Hz, 2 H).
Synthesis of Compound 109 To a mixture of (S)-2-(4-nitropheny1)-l-(2-thiophen-2-y1)ihiazol-4-yl)ethanamine hydrobromide alt (300 mg, 727.58 umol, 1 eq) and intermediate 1 (131.11 mg, 727.58 umol, 1 eq) in DCM (3 mL) was drop-wise added Et3N (441.74 mg, 4.37 mmol, 607.62 uL, 6 eq) and cooled to 0 °C, then T3P (926.01 mg, 1.46 mmol, 865.43 uL, 50% purity, 2 eq) was added at 0 °C, and then the mixture was warmed to 25 °C and stirred for 3 hat 25 °C under N2 atmosphere. The reaction was monitored y LCMS and when complete. The mixture was poured into ice-water (25 mL) and stirred for 10 minutes. Then the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic hase was washed with brine (10 mL), dried with anhydrous NaaSO^ filtered and concentrated in acuo to yield intermediate 2 (218 mg, crude) was obtained as a yellow oil. LCMS: m/z = 494.2 M+H+).
A solution of intermediate 2 ( 218 mg, 441.66 umol, 1 eq) andNH+Cl (117.13 mg, 2.21 mmol, 5 q) in EtOH (12 mL) and H2O (4 mL) was heated to 90 °C and then Fe (123.33 mg, 2.21 mmol, 5 q) was added at 90 °C under N2, and then the mixture was stirred at 90 °C for 2 h. The reaction was monitored by LCMS when complete, the solution was filtered and concentrated in vacuo to ield intermediate 3 (353 mg, crude) was obtained as a yellow solid. LCMS: m/z = 464.2 (M+H*).
A solution of intermediate 3 (353 mg, 761.41 umol, 1 eq) in pyridine (1 mL) and MeCN (1 mL) was cooled to 0 °C and then SCh-pyridine (363.56 mg, 2.28 mmol, 3 eq) was added and stirred for .17 h at 0 °C. The reaction was monitored by LCMS and when complete, 7% ammonium ydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the esidue was purified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H20+10mMNH4HC03)-ACN];B%: 10%-50%,10min) to yield Compound 109 80.2 mg, 19%) was obtained as a white solid. LCMS: m/z = 542.1 (M-H+), 1H-NMR: (400 MHz, DMSOntfc) δ = 8.24 (d, >8.44 Hz, 1 H) 7.71 (dd, >5.07, 1.04 Hz, 1 H) 7.63 (dd, >3.67, 1.10 Hz, H) 7.21 (s, 1 H) 7.12 - 7.19 (m, 3 H) 7.04 - 7.09 (m, 2 H) 6.77 - 6.97 (m, 7 H) 5.04 - 5.12 (m, 1 H) 3.77 (s, 3 H) 3.04 (dd, >13.75, 5.93 Hz, 1 H) 2.85 (dd, >13.82, 8.44 Hz, 1 H) 2.68 - 2.76 (m, H) 2.29 - 2.41 (m, 2 H).
Synthesis of Compound 110
To a solution of (S)-2-(4-nitrophenyl)-l-(2-thiophen-2-yl)thiazol-4-yl)ethanamine ydrobromide salt (300 mg, 727.58 umol, 1 eq) and intermediate 2 (119.47 mg, 727.58 umol, 1 q) in DCM (3 mL) was added EtsN (4.37 mmol, 607.62 uL, 6 eq), and the mixture was cooled o 0 °C, then T3P (1.09 mmol, 649.07 uL, 50% purity, 1.5 eq) was added drop-wise. The mixture was allowed to warm to 25 °C and stirred at 25 °C for 3 h. The reaction was monitored by TLC nd LCMS and when complete, the reaction mixture was poured into water (30 mL), and xtracted with ethyl acetate (3 x 10 mL). Then combined organic phase was washed with brine 2 x 10 mL), dried with anhydrous NaaSCfo and concentrated in vacuo. The residue was purified y prep-TLC (Dichloromethane: Methanol = 10: 1) to yield intermediate 3 (70 mg, 63%) was btained as a yellow oil. LCMS: m/z = 478.2 (M+H÷).
A solution of intermediate 3 (245 mg, 512.98 umol, 1 eq) in EtOH (3 mL) and H2O (1 mL) was dded NH4CI (143.25 mg, 2.56 mmol, 5 eq), and then the solution was heated to 90 °C, Fe 138.04 mg, 2.56 mmol, 5 eq) was added into the mixture and stirred for 2 h. The reaction was monitored by LCMS and when complete, the reaction mixture was filtered, the filtrate was oncentrated in vacuo to yield intermediate 4 (269 mg, crude) was obtained as a yellow solid. LCMS: m/z = 448.2 (M+H+).
To a solution of intermediate 4 (269 mg, 600.96 umol, 1 eq) in MeCN (1 mL) and pyridine (1 mL) was added SCb-pyridine (286.95 mg, 1.80 mmol, 3 eq) at 0 °C and stirred for 0.17 h. The eaction was monitored by HPLC and LCMS and when complete, 7% ammonium hydroxide (1 mL) was added drop-wise, and then the mixture was dried by flowing N2, and the residue was urified by prep-HPLC (column: Waters Xbridge 150*25 5u;mobile phase: water(0.04%NH3H2(>H0mMNH4HCO3)-ACN];B%: 10%-50%,10min) to yield Compound 10 (94.6 mg, 28%) was obtained as a white solid. LCMS: m/z = 526.1 (M-H*), 'H-NMR: (400 MHz, DMSO-dk) δ = 8.30 (d, >8.558 Hz, 1 H) 7.71 (dd, >5.074, 1.039 Hz, 1 H) 7.63 (dd, >3.668, 1.101 Hz, 1 H) 7.21 (br s, 1 H) 7.15 - 7.17 (m, 1 H) 7.04 - 7.13 (m, 6 H) 6.96 (br s, 1 H) .82 - 6.92 (m, 4 H) 5.01 - 5.14 (m, 1 H) 3.05 (br dd, >13.694, 5.869 Hz, 1 H) 2.84 (dd, >13.694, 8.559 Hz, 1 H) 2.69 - 2.78 (m, 2 H) 2.33 - 2.40 (m, 2 H).

Claims

CLAIMS . A compound of Formula I wherein
Ri is chosen from wherein
R2 is chosen from substituted or unsubstituted heteroaryl and substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms.
Rs is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6 near alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl; is an integer chosen from 1, 2, 3, 4, 5, and 6;
Rt is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted Ci- Ce branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl;
Rs is chosen from H, substituted or vmsubstituted C1-C6 linear alkyl, substituted or vmsubstituted C1-C6 branched alkyl, and substituted or vmsubstituted Cs-Ce cyclic alkyl; and ach of Xi, X2, and X3 is independently chosen from CH and N, with the proviso that Xi, X2, and X3 are not simultaneously CH. . The compound according to any one of the previous claims, wherein Ri is
. The compound according to any one of the previous claims, wherein Ri is . The compound according to any one of the previous claims, wherein R2 is substituted or unsubstituted heteroaryl. . The compound according to any one of the previous claims, wherein R2 is substituted heteroaryl. . The compound according to any one of the previous claims, wherein R2 is unsubstituted heteroaryl. . The compound according to any one of the previous claims, wherein R2 is 2-thienyl.. The compound according to any one of the previous claims, wherein R2 is substituted Ci- Ce linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. . The compound according to any one of the previous claims, wherein R2 is substituted Ci- Ce linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. 0. The compound according to any one of the previous claims, wherein R2 is a trifluoroalkyl group. 1. The compound according to any one of the previous claims, wherein R2 is trifluoroethyl. 2. The compound according to any one of the previous claims, wherein R3 is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C3-C6 cyclic alkyl. 3. The compound according to any one of the previous claims, wherein R3 is substituted or unsubstituted heteroaryl. 4. The compound according to any one of the previous claims, wherein R3 is substituted heteroaryl.
5. The compound according to any one of the previous claims, wherein Rs is unsubstituted heteroaryl. 6. The compound according to any one of the previous claims, wherein Rs is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl. 7. The compound according to any one of the previous claims, wherein Rs is chosen from substituted or unsubstituted C1-C6 linear alkyl and substituted or unsubstituted Cs-Ce cyclic alkyl. 8. The compound according to any one of the previous claims, wherein Rj is substituted or unsubstituted C1-C6 linear alkyl. 9. The compound according to any one of the previous claims, wherein Rs is substituted Ci- Ce linear alkyl. 0. The compound according to any one of the previous claims, wherein Rs is unsubstituted C1-C6 linear alkyl. 1. The compound according to any one of the previous claims, wherein Rs is substituted Ci- Ce linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. 2. The compound according to any one of the previous claims, wherein Rs is substituted Ci- Ce linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. 3. The compound according to any one of the previous claims, wherein Rs is a trifluoroalkyl group. 4. The compound according to any one of the previous claims, wherein Rs is trifluoroethyl. 5. The compound according to any one of the previous claims, wherein Rs is trifluoropropyl. 6. The compound according to any one of the previous claims, wherein Rj is substituted or unsubstituted Cs-Ce cyclic alkyl.
7. The compound according to any one of the previous claims, wherein R3 is unsubstituted C3-C6 cyclic alkyl. 8. The compound according to any one of the previous claims, wherein R3 is cyclopropyl. 9. The compound according to any one of the previous claims, wherein R3 is substituted Ci- Ce linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a C5-C20 heteroaryl group. 0. The compound according to any one of the previous claims, wherein R3 is chosen from 2- thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl, 1. The compound according to any one of the previous claims, wherein R3 is 2-thienyl. 2. The compound according to any one of the previous claims, wherein 3. The compoimd according to any one of the previous claims, wherein 4. The compound according to any one of the previous claims, wherein R4 is substituted or unsubstituted C1-C6 linear alkyl. 5. The compound according to any one of the previous claims, wherein R* is unsubstituted C1-C6 linear alkyl. 6. The compound according to any one of the previous claims, wherein R4 is methyl. 7. The compound according to any one of the previous claims, wherein R5 is chosen from H and substituted or unsubstituted C1-C6 linear alkyl.
38. The compound according to any one of the previous claims, wherein is R5 is H.
9. The compound according to any one of the previous claims, wherein Rs is substituted Ci- Ce linear alkyl. 0. The compound according to any one of the previous claims, wherein Rs is unsubstituted C1-C6 linear alkyl. 1. The compound according to any one of the previous claims, wherein Rs is methyl. 2. The compound according to any one of the previous claims, wherein Xi and Xz are CH and X3 is N. 3. The compound according to any one of the previous claims, wherein Xi and X3 are CH and Xz is N. 4. The compound according to any one of the previous claims, wherein Xz and X3 are CH and Xi is N. 5. The compound according to any one of the previous claims, wherein Xi is CH and Xz and X3 are N. 6. The compound according to any one of the previous claims, wherein Xz is CH and Xi and X3 are N. 7. The compound according to any one of the previous claims, wherein n is an integer chosen from 1, 2, and 3. 8. The compound according to any one of the previous claims, wherein n is an integer chosen from 1 and 2. 9. The compound according to any one of the previous claims, wherein n is 1. 0. A compound of Formula Π
wherein
Re is substituted or unsubstituted heteroaryl, m is an integer chosen from 1, 2, 3, 4, 5, and 6;
R.7 is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted Ci- Ce branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl;
Rs is chosen from H, substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl; and ach of Rs and Rio is independently selected from H, substituted or unsubstituted C1-C6 linear lkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted C1-C6 yclic alkyl. 1. The compound according to claim 50, wherein Re is substituted heteroaryl. 2. The compound according to any one of claims 50 to the immediately previous claim, wherein Re is unsubstituted heteroaryl. 3. The compound according to any one of claims 50 to the immediately previous claim wherein Re is 2-thienyl. 4. The compound according to any one of claims 50 to the immediately previous claim wherein R? is substituted or unsubstituted C1-C6 branched alkyl. 5. The compound according to any one of claims 50 to the immediately previous claim wherein R? is substituted or unsubstituted C1-C6 cyclic alkyl. 6. The compound according to any one of claims 50 to the immediately previous claim wherein R? is substituted or unsubstituted C1-C6 linear alkyl.
299
7. The compound according to any one of claims 50 through the immediately previous claim, wherein R? is substituted C1-C6 linear alkyl. 8. The compound according to any one of claims 50 to the immediately previous claim wherein R? is unsubstituted C1-C6 linear alkyl. 9. The compound according to any one of claims 50 to the immediately previous claim wherein R? is methyl. 0. The compound according to any one of claims 50 to the immediately previous claim wherein Rs is chosen from H and substituted or unsubstituted C1-C6 linear alkyl. 1. The compound according to any one of claims 50 to the immediately previous claim wherein is Rs is H. 2. The compound according to any one of claims 50 to the immediately previous claim wherein Rs is substituted or unsubstituted C1-C6 linear alkyl. 3. The compound according to any one of claims 50 to the immediately previous claim wherein Rs is substituted C1-C6 linear alkyl. 4. The compound according to any one of claims 50 to the immediately previous claim wherein Rs is unsubstituted C1-C6 linear alkyl. 5. The compound according to any one of claims 50 to the immediately previous claim wherein Rs is methyl. 6. The compound according to any one of claims 50 to the immediately previous claim wherein each of Rs and Rio is independently chosen from H and substituted or unsubstituted C1-C6 linear alkyl. 7. The compound according to any one of claims 50 to the immediately previous claim wherein each of Rs and Rio is H. 8. The compound according to any one of claims 50 to the immediately previous claim wherein each of Rs and Rio is independently substituted C1-C6 linear alkyl. 9. The compound according to any one of claims 50 to the immediately previous claim wherein each of Rs and Rio is independently unsubstituted C1-C6 linear alkyl.
0. The compound according to any one of claims 50 to the immediately previous claim wherein each of Rs and Rio is methyl. 1. The compound according to any one of claims 50 to the immediately previous claim wherein m is an integer chosen from 1, 2, 3, and 4. 2. The compound according to any one of claims 50 to the immediately previous claim wherein m is an integer chosen from 2, 3, and 4. 3. The compound according to any one of claims 50 to the immediately previous claim wherein m is an integer chosen from 3 and 4. 4. The compound according to any one of claims 50 to the immediately previous claim wherein m is 3. 5. The compound according to any one of claims 50 to the immediately previous claim wherein m is 4. 6. A compound of Formula III
Formula ΙΠ wherein
Rn is chosen from substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6 near alkyl, substituted or unsubstituted C1-C6 branched alkyl, substituted or unsubstituted C3-C6 yclic alkyl, and substituted or imsubstituted heterocyclyl; is an integer chosen from 0, 1, 2, 3, 4, 5, and 6;
Y1-Y2 is a group chosen from nothing, -CH2-CH2-, and -CH=CH-; b is an integer chosen from 0, 1, 2, 3, 4, 5, and 6; and
R12 is chosen from substituted or imsubstituted heteroaryl and substituted or imsubstituted aryl.
7. The compound according to claim 76, wherein Rn is substituted or unsubstituted heteroaryl. 8. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted heteroaryl. 9. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is unsubstituted heteroaryl. 0. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted or unsubstituted heterocyclyl. 1. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted heterocyclyl. 2. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is unsubstituted heterocyclyl. 3. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is chosen from substituted or unsubstituted C1-C6 linear alkyl, substituted or unsubstituted C1-C6 branched alkyl, and substituted or unsubstituted Cs-Ce cyclic alkyl. 4. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted or unsubstituted C1-C6 linear alkyl. 5. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted or unsubstituted C1-C6 branched alkyl. 6. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted or unsubstituted Cs-Ce cyclic alkyl. 7. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl. 8. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 branched alkyl. 9. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C3-C6 cyclic alkyl. 0. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is unsubstituted C1-C6 linear alkyl. 1. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is unsubstituted C1-C6 branched alkyl. 2. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is unsubstituted Cs-Ce cyclic alkyl. 3. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more halogen atoms. 4. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with one or more fluorine atoms. 5. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a heteroaryl group. 6. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with a Cs-Ce cyclic alkyl group. 7. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -(CRaRb)zN(RcXRd), wherein each of Ra and Rb is independently chosen from hydrogen and substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl, each of Rc and Rd is independently chosen from hydrogen, substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl, and aryl, or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms, and z is chosen from 0, 1, 2, 3, and 4. 8. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with N(RcXRd), wherein each of Rc and Rd is independently substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl or wherein Rc and Rd together form a ring system comprising 3 to 7 atoms. 9. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with N(RcXRd), wherein each of Rc and Rd is independently substituted or unsubstituted C1-C6 linear, branched, or cyclic alkyl. 00. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with , wherein each of Rc and Rd is independently unsubstituted Ci- Ce linear, branched, or cyclic alkyl. 01. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with N(Rc)(Rd), wherein each of Rc and Rd is independently unsubstituted Ci- Ce linear alkyl. 02. The compoimd according to any one of claims 76 to the immediately previous claim, wherein Rn is substituted C1-C6 linear alkyl, wherein said substituted C1-C6 linear alkyl is substituted with -N(CH3)z 03. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is chosen from 2-thienyl, trifluoroethyl, trifluoropropyl, cyclopropyl,
/
N cyclopropylmethyl, cyclopentylmethyl, +K OMe 04. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 2-thienyl. 05. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is a trifluoroalkyl group. 06. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is trifluoroethyl.
107. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is trifluoropropyl.
304
08. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is cyclopropyl. 09. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is cyclopropylmethyl . 10. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is cyclopentylmethyl. 11. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 12. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 13. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 14. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 15. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is 16. The compound according to any one of claims 76 to the immediately previous claim, wherein Rn is
17. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is substituted or unsubstituted heteroaryl. 18. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is substituted heteroaryl. 19. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is unsubstituted heteroaryl. 20. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is pyridinyl. 21. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is substituted or unsubstituted aryl. 22. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is substituted aryl. 23. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is monosubstituted aryl. 24. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is monosubstituted phenyl. 25. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is unsubstituted aryl. 26. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is phenyl. 27. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is chosen from 1-naphthyl and 2-naphthyl. 28. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is 1-naphthyl. 29. The compound according to any one of claims 76 to the immediately previous claim, wherein R12 is 2-naphthyl.
30. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is nothing. 31. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is -CH2-CH2-. 32. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is -CH=CH-33. The compound according to any one of claims 76 to the immediately previous claim, wherein a is an integer chosen from 0, 1, 2, and 3. 34. The compound according to any one of claims 76 to the immediately previous claim, wherein a is an integer chosen from 0, 1, and 2. 35. The compound according to any one of claims 76 to the immediately previous claim, wherein a is an integer chosen from 0 and 1. 36. The compound according to any one of claims 76 to the immediately previous claim, wherein a is 2. 37. The compound according to any one of claims 76 to the immediately previous claim, wherein a is 1. 38. The compound according to any one of claims 76 to the immediately previous claim, wherein a is 0. 39. The compound according to any one of claims 76 to the immediately previous claim, wherein b is an integer chosen from 0, 1, 2, and 3. 40. The compound according to any one of claims 76 to the immediately previous claim, wherein b is an integer chosen from 0, 1, and 2. 41. The compound according to any one of claims 76 to the immediately previous claim, wherein b is an integer chosen from 0 and 1. 42. The compound according to any one of claims 76 to the immediately previous claim, wherein b is 2. 43. The compound according to any one of claims 76 to the immediately previous claim, wherein b is 1. 44. The compound according to any one of claims 76 to the immediately previous claim, wherein b is 0. 45. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is nothing, a is 1, and b is 1. 46. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is nothing, a is 1, and b is 0. 47. The compound according to any one of claims 76 to the immediately previous claim, wherein Y1-Y2 is -CH=CH-, a is 0, and b is 0. 48. The compound according to any one of claims 76 to the immediately previous claim, with the proviso that, if Y1-Y2 is nothing, then a and b are not simultaneously 0. 49. A compound chosen from: 309 314 315
50. A method for inhibiting VE-PTP in a mammalian subject in need thereof, comprising administering to the subject in need thereof an effective amount of a compound according to any one of claims 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VE- PTP activity is inhibited. 51. The method of claim ISO, wherein the subject suffers from one or more of cancer, glaucoma, occlusive cardiovascular disease, a vascular leaking syndrome, or another vascular-related disease. 52. A method for reducing VE-PTP-mediated signaling in a mammalian subject in need thereof, comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of claims 1 to 149, or a pharmaceutically acceptable salt thereof, whereby VEP TP-mediated signaling is reduced. 53. A method for increasing Tie2-mediated signaling in a mammalian subject in need thereof, comprising administering to the mammalian subject in need thereof an effective amount of a compound according to any one of claims 1 to 149, or a pharmaceutically acceptable salt thereof, whereby Tie2-mediated signaling is increased. 54. A method for treating a Tie2-mediated disorder in a mammalian subject in need thereof, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 149, or pharmaceutically acceptable salt thereof, to the subject in need thereof. 55. The method of claim 154, wherein the subject suffers from one or more of cancer, glaucoma, occlusive cardiovascular disease, a vascular leaking syndrome, or another vascular-related disease.
56. The method according to any one of claims 150 to 155, wherein the “effective amount” is determined in vitro by one or more biochemical and/or biological assays.
EP21825008.2A 2020-06-16 2021-06-16 Small molecule ve-ptp inhibitors Pending EP4164690A1 (en)

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