Quinone reductase 2 inhibitor compounds and uses thereof

Abstract

According to some embodiments, provided herein is a compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof. Also provided are compositions comprising the compound, and uses thereof for inhibiting the activity of quinone reductase-2, and uses in methods of treatment.

Description

Background Art

[0001] Aminoquinolines, such as chloroquine (CQ) and hydroxychloroquine (HQ), are quinone reductase 2 (QR2) inhibitors that were originally developed for the treatment of malaria but have subsequently been found to have therapeutic efficacy for other indications, including, inter alia, autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Singer et al., "Update on immunosuppressive therapy," Curr. Opin. Rheumatol. 1998, 10:169-173;Wallace, "The use of chloroquine and hydroxychloroquine for non-infectiousconditions other than rheumatoid arthritis or lupus: a crucial review," Lupus 1996, 5 Suppl 1:S59-64. In SLE and RA, aminoquinolines are the mainstay of first-line therapy and are often used in combination with other drugs. Aminoquinolines not only improve the signs and symptoms of SLE and RA, but also have beneficial effects on lipid metabolism and reduce the occurrence of thrombosis. Similar benefits have been observed in patients with inflammatory or erosive osteoarthritis. Efficacy has also been shown when used as an adjunct therapy for graft-versus-host disease, cancer, and HIV. Savarino et al., " Effects of chloroquine on viral infections: an old drug against today's diseases " Lancet Infect. Dis. 2003, 3(11):722-7; Savarino et al., "Risks and benefits of chloroquine use in anticancer strategies," Lancet Oncol. 2006, 7(10):792-3;Sotelo et al., "Adding chloroquine to conventional treatment for glioblastomamultiforme: a randomized, double-blind, placebo-controlled trial," Ann.Intern. Med. 2006, 144(5):337-43.

[0002] The potential of chloroquine (CQ) in neuroprotection has been previously investigated in preclinical models of stroke, excitotoxicity, and traumatic injury, although the therapeutic mechanisms remain unclear. CQ significantly limits microglia and PMN migration to the injury site in the brain, reduces reactive astrogliosis and neovascularization, and reduces stroke volume by 60% in a permanent MCA occlusion model. Giulian et al., "The role of mononuclearphagocytes in wound healing after traumatic injury to adult mammalian brain," J. Neurosci. 1989, 9:4416-4429; Ivanova et al., "Cerebral ischemia enhancespolyamine oxidation: identification of enzymatically formed 3-aminopropanalas an endogenous mediator of neuronal and glial cell death," J. Exp. Med. 1998, 188:327-340. CQ also reduces cytokine production by microglia in response to various stimuli in vitro. Giulian, "Microglia and the immune pathology of Alzheimer disease," Am. J. Hum. Genet. 1999, 65:13-18.

[0003] Because some malariae are resistant to CQ, derivative compounds have also been explored. For example, US 2006 / 0074105 to Ware et al. describes certain quinoline and quinazoline derivatives that are said to be useful in treating malaria and autoimmune diseases.

[0004] Although CQ and HQ are often used clinically as first-line treatments for autoimmune conditions, their efficacy is limited by severe side effects. The most important and best characterized toxicity is retinal toxicity, where long-term use may lead to "bull's eye maculopathy" and blindness unless dosing is restricted. Although rare, cardiac toxicity may also occur, manifested by conduction disturbances (e.g., bundle branch block) and / or cardiomyopathy associated with congestive heart failure. Electron microscopy of cardiac and retinal biopsies following long-term CQ or HQ treatment reveals characteristic cytoplasmic inclusions, thought to be a direct result of high drug accumulation in lysosomes (and melanosomes in the retina and skin). Notably, during therapeutic dosing, CQ is able to accumulate to mM concentrations in skin, retina, kidney, and hepatocytes, while plasma concentrations remain <1 μM.

[0005] There remains a need to develop additional aminoquinoline quinone reductase 2 (QR2) inhibitors, particularly inhibitors that also have reduced lysosomal accumulation to reduce toxicity. Summary of the invention

[0006] According to some embodiments, provided herein is a compound of formula (I):

[0007]

[0008] in:

[0009] R1 is a nitrogen-containing heterocyclic ring or a nitrogen-containing heteroaryl group,

[0010] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0011] or a pharmaceutically acceptable salt or prodrug thereof.

[0012] In some embodiments, R1 is:

[0013] , or ,

[0014] wherein D1, D2, D3 and D4 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxyl, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl, or

[0015] D1 and D2, D2 and D3, or D3 and D4 together form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0016] wherein the compound (including R1) may be optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0017] or a pharmaceutically acceptable salt or prodrug thereof.

[0018] In some embodiments, D1, D2, D3, and D4 are each hydrogen.

[0019] In some embodiments, the compound is a compound of formula (I)(a)(1) or a compound of formula (I)(a)(2):

[0020]

[0021] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0022] or a pharmaceutically acceptable salt or prodrug thereof.

[0023] In some embodiments, the compound is a compound of formula (I)(a)(2):

[0024]

[0025] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0026] or a pharmaceutically acceptable salt or prodrug thereof.

[0027] In some embodiments, the compound is a compound of formula (I)(a)(2):

[0028]

[0029] or a pharmaceutically acceptable salt or prodrug thereof.

[0030] In some embodiments, R1 is:

[0031] or ,

[0032] wherein X1, X2 and X3 are each independently selected from carbon, nitrogen and oxygen, and

[0033] When present, D5, D6, D7 and D8 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0034] or two of D5, D6, D7 and D8 together form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0035] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0036] or a pharmaceutically acceptable salt or prodrug thereof.

[0037] In some embodiments, the compound is a compound of formula (I)(b)(1):

[0038] ,

[0039] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0040] or a pharmaceutically acceptable salt or prodrug thereof.

[0041] In some embodiments, R1 is:

[0042] , , or

[0043] ,

[0044] wherein X4, X5, X6, X7 and X8 are each independently selected from carbon, nitrogen and oxygen, wherein at least two or at least three of X4, X5, X6, X7 and X8 are carbon, and

[0045] When present, D9, D 10 , D 11 , D 12 , D 13 and D 14 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0046] or D9, D 10 , D 11 , D 12 , D 13 and D 14 Two of the above are taken together to form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0047] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0048] or a pharmaceutically acceptable salt or prodrug thereof.

[0049] In some embodiments, R1 is:

[0050] or ,

[0051] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0052] or a pharmaceutically acceptable salt or prodrug thereof.

[0053] In some embodiments, R1 is:

[0054] , or ,

[0055] X9, X 10 , X 11 and X 12 Each independently selected from carbon, nitrogen and oxygen, wherein X9, X 10 , X 11 and X 12 At least two of are carbon, and

[0056] When present, D 15 , D 16 , D 17 , D 18 and D 19 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0057] or D 15 , D 16 , D 17 , D 18 and D 19 Two of the above are taken together to form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0058] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0059] or a pharmaceutically acceptable salt or prodrug thereof.

[0060] In some embodiments, R1 is:

[0061] ,

[0062] wherein the compound is optionally substituted once, twice or three times with fluoromethyl, difluoromethyl or trifluoromethyl,

[0063] or a pharmaceutically acceptable salt or prodrug thereof.

[0064] In some embodiments, the compound has a positive log D value at about pH 4 to pH 5.

[0065] Also provided are compositions comprising a compound described herein and a carrier (eg, a pharmaceutically acceptable carrier).

[0066] Further provided are methods for inhibiting the activity of quinone reductase-2 (QR2), the methods comprising contacting QR2 with a compound or composition taught herein, wherein the contacting is performed in vitro, or the contacting is performed in vivo.

[0067] Also provided is a method of treating malaria in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for use in treating malaria. Still further provided is the use of a compound as taught herein for the preparation of a medicament for treating malaria.

[0068] Also provided is a method of treating an immune disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for treating an immune disorder. Still further provided is the use of a compound as taught herein for preparing a medicament for treating an immune disorder.

[0069] Also provided is a method of treating acute nerve injury in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for use in treating acute nerve injury. Still further provided is the use of a compound as taught herein for the preparation of a medicament for treating acute nerve injury.

[0070] Also provided is a method of treating a chronic neurological disorder (e.g., Parkinson's disease or Alzheimer's disease) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for treating a chronic neurological disorder. Still further provided is the use of a compound as taught herein for preparing a medicament for treating a chronic neurological disorder.

[0071] Also provided is a method of treating lupus in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition taught herein. Further provided is a compound or composition taught herein for use in treating lupus. Still further provided is the use of a compound taught herein for the preparation of a medicament for treating lupus.

[0072] Also provided is a method for treating an infectious disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for treating an infectious disease. Still further provided is the use of a compound as taught herein for preparing a medicament for treating an infectious disease.

[0073] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for use in treating cancer. Still further provided is the use of a compound as taught herein for the preparation of a medicament for treating cancer.

[0074] Also provided is a method of treating CNS lupus in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for use in treating CNS lupus. Still further provided is the use of a compound as taught herein for the preparation of a medicament for treating CNS lupus.

[0075] Also provided is a method of treating a subject at increased risk of cerebrovascular disease, the method comprising administering to the subject a therapeutically effective amount of a compound or composition as taught herein. Further provided is a compound or composition as taught herein for use in treating a subject at increased risk of cerebrovascular disease. Still further provided is the use of a compound as taught herein for the preparation of a medicament for treating a subject at increased risk of cerebrovascular disease.

[0076] In some embodiments of the above methods or uses, the administering comprises chronic administration (eg, over several months or years).

[0077] In some embodiments of the aforementioned methods or uses, the administering is performed once a day. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1A-1B : Figure 1A A graph showing the logD of chloroquine versus pH. Below pH 6, the logD of chloroquine is negative, consistent with a loss of membrane permeability. Figure 1B A graph showing the logD of QR2I-44 versus pH. Above about pH 4, the logD of QR2I-44 is positive, indicating that membrane permeability is retained in the lysosomal pH range. Unlike CQ, QR2I-44 is not trapped in lysosomes.

[0079] Figure 2A-2B : Figure 2A Five-day rotarod results for QR2I-44 versus vehicle are presented. Figure 2BThe 5-day rotarod results of QR2I-44 versus chloroquine (CQ) are presented. The results show that QR2I-44 has a neuroprotective effect after TBI and is therapeutically superior to CQ.

[0080] Figure 3 Comparative testing results of chloroquine (CQ), compound 1, compound 2, and QR2I-44 in a 7-day rotarod test are presented, demonstrating the initial durability of QR2I-44 neuroprotection.

[0081] Figure 4 Presented is a graph of rotarod latency versus time post-traumatic brain injury (TBI) in the 28-day rotarod test for chloroquine (CQ), Compound 1, Compound 2, and QR2I-44, demonstrating the persistence of QR2I-44 therapeutic efficacy almost one month post-TBI.

[0082] Figure 5A-Figure 5B : Figure 5A The results show the rate of QR2I-44 administration versus days after traumatic brain injury (TBI) in the Morris water maze (MWM) test. Figure 5B The effect seen in (i.e., faster learning due to QR2I-44 administration) was not compromised by differences in motor ability. P-value = 0.0165, significance level = 5%.

[0083] Figure 6A-6B : Fig. 6A is a representative high-resolution coronal MRI of the mouse brain with the hippocampal ROI superimposed for volumetric analysis. Figure 6B is a graph of hippocampal volume in normal, treated and untreated groups, showing a statistically significant improvement in volume in mice treated with QR2I-44 relative to vehicle control.

[0084] Figure 7A-7B : Fig. 7A Representative MRI with template overlay ROIs of key subcortical brain structures including the corpus callosum / external capsule (CC / EC). Figure 7B is a plot of the fractional anisotropy (FA) of the CC / EC in normal, treated, and untreated groups after TBI, revealing improvements in FA of these large white matter tracts after QR2I-44 treatment. FA is a general measure of tract integrity using a high-resolution MRI technique called diffusion tensor imaging (DTI).

[0085] Figure 8A-8C : Fig. 8A Shown are the results of inflammatory microparticle (MP) release from RAW 264.7 cells after stimulation with TOLL receptor ligand and treatment with QR2I-44. Figure 8B and Figure 8CResults are shown for TNF α release from RAW264.7 cells after stimulation with TOLL receptor ligands and treatment with QR2I-44. TOLL receptor ligands: PIC = polyinosinic-polycytidylic acid (TLR3); CpG = CpG oligodeoxynucleotide DNA (TLR9).

[0086] Fig. 9 Shown are the results of QR2I-44 treatment on anti-cDNA antibody production in a mouse model of human lupus erythematosus (LPR / mrl) at one and three weeks after the start of treatment.

[0087] Fig.10 Data showing that QR2 inhibitors prevent menadione-induced membrane potential loss are presented. All QR2 inhibitors were able to prevent menadione-induced membrane potential loss, as detected by tetramethylrhodamine ethyl ester (TMRE, a fluorescent dye chelated by live and active mitochondria). Wild-type HEK293 cells were pretreated with QR2 inhibitors (CDL-1, CDL-2, and QR2i-44) for 1 hour, then exposed to menadione for 4 hours and the membrane potential was measured. QR2i-44 significantly prevented menadione-induced membrane potential loss. DETAILED DESCRIPTION

[0088] Provided herein are compounds useful as inhibitors of quinone reductase-2, as well as formulations and methods of use thereof. In some embodiments, the compounds are useful for treating subjects at increased risk for infectious diseases, cancer, immune disorders, acute neurological injuries and chronic neurological disorders, and cerebrovascular disease.

[0089] The disclosure of all patent references cited herein is hereby incorporated by reference to the extent that they are consistent with the disclosure set forth herein. As used in the specification of the present invention and the appended claims, the singular forms "one", "one" and "the" are also intended to include plural forms, unless the context clearly indicates otherwise. As used in the specification of the present invention and the appended claims, the singular forms "one", "one" and "the" are also intended to include plural forms, unless the context clearly indicates otherwise. In addition, when it comes to measurable values ​​(such as the amount of compound, dosage, time, temperature, etc.), the terms "about" and "approximately" as used herein are intended to include 20%, 10%, 5%, 1%, 0.5% or even 0.1% of the change of the specified amount. In addition, as used herein, "and / or" and " / " refer to and cover any and all possible combinations of one or more related listed items, and lack combinations when interpreted in alternative form ("or").

[0090] I. Definitions

[0091] The following definitions are used in this article.

[0092] As is known in the art, "H" refers to a hydrogen atom. "C" refers to a carbon atom. "N" refers to a nitrogen atom. "O" refers to an oxygen atom.

[0093] "Halogen" refers to F, Cl, Br or I. "Cl" is chlorine, "I" is iodine, "F" is fluorine, and "Br" is bromine.

[0094] "Acyl" is the group -C(O)R, where R is a suitable substituent (eg, acetyl, propionyl, butyryl, benzoyl or alkylbenzoyl).

[0095] As used herein, "alkyl" refers to a straight or branched saturated hydrocarbon containing 1 or 2 to 10 or 20 or more carbon atoms (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, etc.). Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, etc. In some embodiments, the alkyl group is a "lower alkyl" having 1 to 3, 4 or 5 carbon atoms.

[0096] As used herein, "alkenyl" is a straight or branched chain unsaturated hydrocarbon group having one or more double bonds.

[0097] As used herein, "alkynyl" is a straight or branched chain unsaturated hydrocarbon group having one or more triple bonds.

[0098] "Amino" is the group -NH2. "Amide" as used herein refers to an organic functional group having a carbonyl group (C=0) attached to a nitrogen atom (N).

[0099] As used herein, "alkoxy" refers to an alkyl group as defined herein attached to a parent molecule via an oxygen atom (-O-). Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like.

[0100] As used herein, "aryl" refers to a ring system having one or more aromatic rings. Representative examples of aryl include azulenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like.

[0101] As used herein, "haloalkyl" refers to a straight or branched hydrocarbon containing 1 or 2 to 10 or 20 or more carbon atoms (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, etc.) in which at least one hydrogen atom has been replaced by a halogen (F, Cl, Br or I). Representative examples of "haloalkyl" include, but are not limited to, fluoroalkyl (e.g., fluoromethyl (-CH2F), difluoromethyl (-CHF2) or trifluoromethyl (CF3)).

[0102] As used herein, "heterocycle" refers to a monocyclic, bicyclic or tricyclic ring system containing at least one heteroatom selected from O, N and S. Examples of monocyclic heterocyclic ring systems are any 5- or 6-membered rings containing 1, 2, 3 or 4 heteroatoms independently selected from O, N and S. The 5-membered ring has 0-2 double bonds, while the 6-membered ring has 0-3 double bonds. Representative examples of monocyclic ring systems include, but are not limited to, azetidine, aziridine, aziridine, diazepine, 1,3-dioxolane, dioxane, dithiane, furan, imidazole, imidazoline, imidazolidine, isothiazole, isothiazolidine, isoxazole, isoxazoline, isoxazolidine, morpholine, oxadiazole, oxadiazolline, oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine, piperidine, pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridine, pyrimidine, pyridazine, pyrrole, pyrroline, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, tetrazine, tetrazole, thiadiazole, thiadiazoline, thiadiazolidine, thiazole, thiazoline, thiazolidine, thiophene, thiomorpholine, thiomorpholine sulfone, sulfoxide, thiopyran, triazine, triazole, trithiane, and the like. Examples of bicyclic ring systems are any of the above monocyclic ring systems fused to an aryl group as defined herein, a cycloalkyl group as defined herein, or another monocyclic ring system as defined herein. Representative examples of bicyclic ring systems include, but are not limited to, for example, benzimidazole, benzothiazole, benzothiadiazole, benzothiophene, benzoxadiazole, benzoxazole, benzofuran, benzopyran, benzothiopyran, benzodioxine, 1,3-benzodioxole, cinnoline, indazole, indole, indoline, indolizine, naphthyridine, isobenzofuran, isobenzothiophene, isoindole, isoindoline, isoquinoline, phthalazine, pyranopyridine, quinoline, quinolizine, quinoxaline, quinazoline, tetrahydroisoquinoline, tetrahydroquinoline, thiopyranopyridine, and the like. Examples of nitrogen-containing heterocycles include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and the like.

[0103] "Heteroaryl" refers to a cyclic aromatic hydrocarbon in which one or more carbon atoms have been replaced by atoms independently selected from O, N and S. Examples of heteroaryl include pyridyl, pyrimidyl, imidazolyl, thienyl, furyl, pyrazinyl, pyrrolyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, indolyl, isoindolyl, indolizinyl, triazolyl, pyridazinyl, indazolyl, purinyl, quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, isothiazolyl and benzo[b]thienyl. Preferred heteroaryl is a five-membered and six-membered ring and contains 1-3 heteroatoms independently selected from O, N and S. The heteroaryl including each heteroatom can be unsubstituted or substituted by 1-4 suitable substituents, as long as it is chemically feasible. For example, the heteroatom S can be substituted by one or two oxo groups, which can be shown as =O. Examples of nitrogen-containing heteroaryl groups include, but are not limited to, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrrolyl, pyrazolyl, thiazolyl, triazolyl, isothiazolyl, indolyl, benzimidazolyl, benzoxazolyl, quinolyl, isoquinolyl, quinazolinyl, acridinyl, carbazole, azepine, 1,4-diazepine, purinyl, pteridinyl, phthalazinyl, and the like.

[0104] The aryl, heteroaryl and heterocyclyl groups of the present invention may be substituted 1, 2, 3, 4 or 5 times, as far as is chemically feasible, wherein the substituents are independently selected from alkenyl, alkenyloxy, alkoxy, alkoxyalkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, alkylcarbonyloxy, alkylsulfinyl, alkylsulfonyl, alkylthio, alkynyl, aryl, aryloxy, azido, arylalkoxy, arylalkyl, aryloxy, carboxy, cyano, formyl, halogen, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, mercapto, nitro, sulfamoyl, sulfo, sulfonate, NR'R" (wherein R' and R" are independently selected from hydrogen, alkyl, alkylcarbonyl, aryl, arylalkyl and formyl) and -C(O)NR'R" (wherein R' and R" are independently selected from hydrogen, alkyl, alkylcarbonyl, aryl, arylalkyl and formyl).

[0105] "Hydroxy" and "hydroxyl" refer to the group -OH.

[0106] "Nitrile" refers to the radical -CN.

[0107] "Nitro" refers to the radical -NO2.

[0108] "Sulfone" refers to a sulfonyl functional group -SO2R, where R is any covalently linked atom or atoms.

[0109] "Sulfoxide" refers to the group -S(O)R, where R is any covalently linked atom or atoms.

[0110] "Thiol" or "sulfhydryl" refers to the group -SH or its tautomer =S.

[0111] As used herein, "fused ring" refers to a ring system (e.g., "heterocycle," "aryl," or "heteroaryl") that can be formed by two substituents of the formula provided herein. Each of the two substituents can together form a portion of the ring system, as illustrated below as fused ring I or fused ring II, such as substituents R2 and R3, which can be independently selected from C, O, N, or S. The carbon included in fused ring II can also be substituted with heteroatoms independently selected from O, N, and S. When present, the fused ring system including each heteroatom can be unsubstituted or substituted with 1-4 suitable substituents, as long as it is chemically feasible.

[0112]

[0113] "Pharmaceutically acceptable salts" are salts that retain the biological effectiveness of the free acids and bases of the specified compound and which are not biologically or otherwise undesirable. Examples of pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, octanoate, acrylate, formate, isobutyrate, hexanoate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate.

[0114] "Prodrugs" known in the art are compounds that can be converted into pharmaceutically active specific compounds under physiological conditions or by solvolysis or metabolism. A comprehensive discussion is provided in T. Higuchi and V. Stella, Prodrugs as Novel delivery Systems, Vol. 14, ACS Symposium Series and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference in their entirety. See also U.S. Patent No. 6,680,299. Examples include prodrugs that are metabolized in vivo by a subject to an active compound described herein, wherein the prodrug is an ester of an alcohol or carboxylic acid group, if such a group is present in the compound; an acetal or ketal of an alcohol group, if such a group is present in the compound; an N-Mannich base or imine of an amine group, if such a group is present in the compound; or a Schiff base, oxime, acetal, enol ester, oxazolidine, or thiazolidine of a carbonyl group, if such a group is present in the compound, as described, for example, in U.S. Pat. No. 6,680,324 and U.S. Pat. No. 6,680,322.

[0115] As understood in the art, the term "optionally substituted" indicates that the specified group is unsubstituted or substituted with one or more suitable substituents. "Substituted" "Substituents" are groups that replace hydrogen atoms on the parent organic molecule.

[0116] II. Active Compounds

[0117] The active compounds provided herein are compounds of formula (I):

[0118]

[0119] in:

[0120] R1 is a nitrogen-containing heterocyclic ring or a nitrogen-containing heteroaryl group,

[0121] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0122] In some embodiments, R1 is:

[0123] , or ,

[0124] wherein D1, D2, D3 and D4 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxyl, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl, or

[0125] D1 and D2, D2 and D3, or D3 and D4 together form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0126] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound of formula (I) may be substituted once, twice or three times by an independently selected suitable group (e.g., fluoromethyl, difluoromethyl or trifluoromethyl). In some embodiments, D1, D2, D3 and D4 are each hydrogen.

[0127] In some embodiments, the compound is a compound of formula (I)(a)(1) or a compound of formula (I)(a)(2):

[0128]

[0129] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0130] In some embodiments, the compound is a compound of formula (I)(a)(1):

[0131]

[0132] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0133] In some embodiments, the compound is a compound of formula (I)(a)(2):

[0134]

[0135] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0136] In some embodiments of Formula (I), R1 is:

[0137] or ,

[0138] wherein X1, X2 and X3 are each independently selected from carbon, nitrogen and oxygen, and

[0139] When present, D5, D6, D7 and D8 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0140] or two of D5, D6, D7 and D8 together form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0141] Or its pharmaceutically acceptable salt or prodrug.In some embodiments, the compound can be substituted once, twice or three times by a suitable group (e.g., fluoromethyl, difluoromethyl or trifluoromethyl).In some embodiments, two of D5, D6, D7 and D8 on adjacent atoms form a condensed ring together.

[0142] In some embodiments, the compound is a compound of formula (I)(b)(1):

[0143] ,

[0144] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0145] In some embodiments of Formula (I), R1 is:

[0146] , , or

[0147] ,

[0148] wherein X4, X5, X6, X7 and X8 are each independently selected from carbon, nitrogen and oxygen, wherein at least two or at least three of X4, X5, X6, X7 and X8 are carbon, and

[0149] When present, D9, D 10 , D 11 , D 12 , D 13 and D 14 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0150] or D9, D 10 , D 11 , D 12 , D13 and D 14 Two of the above are taken together to form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0151] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times by an independently selected suitable group (e.g., fluoromethyl, difluoromethyl or trifluoromethyl). In some embodiments, D9, D 10 , D 11 , D 12 , D 13 and D 14 The two together form a fused ring.

[0152] In some embodiments of Formula (I), R1 is:

[0153] or ,

[0154] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0155] In some embodiments of Formula (I), R1 is:

[0156] , or ,

[0157] X9, X 10 , X 11 and X 12 Each independently selected from carbon, nitrogen and oxygen, wherein X9, X 10 , X 11 and X 12 At least two of are carbon, and

[0158] When present, D 15 , D 16 , D 17 , D 18 and D 19 are each independently selected from hydrogen, halogen, alkyl, acyl, alkoxy, aryl, heteroaryl, amino, amide, nitro, hydroxy, thiol, sulfone, sulfoxide, nitrile, nitro and haloalkyl,

[0159] or D 15 , D 16 , D 17 , D 18 and D 19Two of the above are taken together to form an optionally substituted fused ring (e.g., a cyclohexane or cyclohexene fused ring),

[0160] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times by an independently selected suitable group (e.g., fluoromethyl, difluoromethyl or trifluoromethyl). In some embodiments, the D 15 , D 16 , D 17 , D 18 and D 19 The two together form a fused ring.

[0161] In some embodiments of Formula (I), R1 is:

[0162] ,

[0163] or a pharmaceutically acceptable salt or prodrug thereof. In some embodiments, the compound may be substituted once, twice or three times with independently selected suitable groups such as fluoromethyl, difluoromethyl or trifluoromethyl.

[0164] In some embodiments, the compound has a positive log D value at about pH 4 to pH 5.

[0165] Unless otherwise stated, structures depicted herein are also intended to include all enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure; for example, R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the compounds of the invention are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Tautomeric forms include keto-enol tautomers of the compounds. In addition, unless otherwise stated, all rotational isomeric forms of the compounds of the invention are within the scope of the invention. Unless otherwise stated, structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, a compound having a structure of the invention except that a hydrogen is replaced by a deuterium or tritium, or a carbon is enriched by a 13 C or 14 Such compounds are useful as analytical tools or probes in, for example, biological assays.

[0166] III. How to use

[0167] As described above, the active compounds taught herein can be used as QR2 inhibitors. Such active compounds can also be used to treat subjects with increased risk of infectious diseases, cancer, immune disorders, acute nerve damage and chronic neurological disorders, and cerebrovascular disease. The active compounds can also be used to treat disorders associated with mitochondrial dysfunction.

[0168] Infectious diseases include, but are not limited to, parasitic infections (e.g., malaria and amebiasis), bacterial infections (e.g., Lyme disease), and viral infections (e.g., those of human immunodeficiency virus (HIV), Ebola virus, Chikungunya virus, dengue virus, Zika virus, etc.).

[0169] Cancer treatment includes, but is not limited to, radiosensitization of cancer, chemosensitization of cancer, or a combination thereof. The cancer to be treated may include, but is not limited to, glioblastoma.

[0170] Immune disorders include, but are not limited to, autoimmune diseases. Such autoimmune diseases include, but are not limited to, lupus (systemic lupus erythematosus (SLE) and lupus nephritis); autoimmune myopathy; psoriasis; scleroderma; CREST syndrome; inflammatory myositis; Sjogren's syndrome; mixed connective tissue disease; rheumatoid arthritis; psoriatic arthritis; palindromic rheumatism; eosinophilic fasciitis; dermatomyositis; juvenile chronic arthritis, erosive osteoarthritis; calcium pyrophosphate crystal deposition disease; multiple sclerosis; inflammatory bowel disease; colitis; Crohn's disease; acute respiratory distress syndrome; lung inflammation; idiopathic pulmonary fibrosis; osteoporosis; delayed hypersensitivity reaction; autoimmune thyroiditis; Hashimoto's disease; Graves' disease; asthma; primary biliary cirrhosis; idiopathic thrombocytopenic purpura; diabetes; leukopenia; opportunistic infections; thrombosis; atherosclerosis; treatment-induced diseases, such as antibiotic allergy, gene vector hypersensitivity, and chemotherapy-induced human anti-mouse antibody induction. Autoimmune myopathies associated with lupus usually present with proximal weakness and myalgias. There is also involvement of the peripheral nervous system (PNS) in lupus, primarily manifesting as peripheral neuropathy.

[0171] Acute neurological injury includes, but is not limited to, traumatic brain injury and non-traumatic acute brain injury. As known in the art, traumatic brain injury is brain injury and / or dysfunction caused by a single or repeated external mechanical force, such as blunt force or shear force from sudden acceleration or deceleration. Traumatic brain injury includes, but is not limited to, concussion, contusion and hemorrhage, including parenchymal, subdural, epidural and subarachnoid hemorrhage. Other acute neurological injuries include damage from hypoxia or ischemic brain injury, such as from arterial stroke (focal, comprehensive), venous infarction, infection, etc.

[0172] Chronic neurological disorders include, but are not limited to, primary dementias, such as Alzheimer's disease, vascular dementia, dementia with Lewy bodies, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, and secondary dementias associated with chronic inflammatory conditions, such as Behcet's disease, multiple sclerosis, SLE (CNS lupus), celiac disease, and non-celiac gluten sensitivity; movement disorders, such as dystonia, amyotrophic lateral sclerosis (ALS), Parkinson's disease, and Huntington's chorea; and epilepsy. CNS lupus may especially clinically manifest as acute mental confusion, fatigue, headache, subtle cognitive impairment, delirium, coma, dementia, sensory / motor / autonomic defects, and / or epileptic seizures (the latter occurring more frequently in lupus patients than in general populations). CNS lupus may also manifest as psychological disorders, such as depression, mania, and / or psychosis. More focal neurological defects are also possible, and may be secondary to embolism, thrombosis, or vasculitis infarction and cranial neuropathy associated with lupus in the brain and spine. The pathophysiology of CNS lupus may include encephalitis of the brain or spine, transverse myelitis, neuritis, and stroke (embolic, thrombotic, or vasculitic).

[0173] Subjects at increased risk for cerebrovascular disease include, for example, those with prior ischemic stroke or microbleeding; documented carotid or small vessel disease; vascular modifiable risk factors, such as diabetes, hypercoagulable states, hypertension; non-modifiable risk factors, such as age, race, family history, etc.

[0174] There is evidence that the antimalarial hydroxychloroquine can reduce the incidence of cerebrovascular events in patients at high risk for stroke. See, e.g., Sharma et al., "Hydroxychloroquine use is associated with decreased incident cardiovascular events in rheumatoid arthritis patients," J. Amer. Heart Assoc. 2016;5:e002867; Jung et al., "The protective effect of antimalarialdrugs on thrombovascular events in systemic lupus erythrematosus," Arthritis Rheum.2010, 62(3):863-8; Wallace et al., "The relevance of antimalarial therapy with regard to thrombosis, hypercholesterolemia and cytokines in SLE," Lupus1993, Suppl 1:S13-5.

[0175] Disorders known to be associated with mitochondrial dysfunction include, but are not limited to, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Friedreich's ataxia; cardiovascular diseases such as atherosclerosis and other cardiac and vascular conditions; diabetes and metabolic syndrome; autoimmune diseases such as multiple sclerosis, systemic lupus erythematosus, and type 1 diabetes; neurobehavioral and psychiatric disorders such as autism, schizophrenia, bipolar disorder, and mood disorders; gastrointestinal disorders; fatigue disorders such as chronic fatigue syndrome and Gulf War illness; musculoskeletal diseases such as fibromyalgia and skeletal muscle hypertrophy / atrophy; cancer; chronic infection; etc. See, for example, a review by Nicolson, "Mitochondrial Dysfunction and Chronic Disease: Treatment With Natural Supplements," Integrative Medicine, Vol. 13, No. 4, 35-43 (2014).

[0176] As used herein, the term "treatment" refers to any type of treatment that confers a benefit (e.g., improving one or more symptoms such as cognitive dysfunction and / or motor dysfunction or reducing the risk of their development), delays the progression of the injury or symptoms, etc. to a subject having an injury, disease or condition or at risk of an injury, disease or condition.

[0177] In some embodiments, the treatment is used to prevent infectious diseases, cancer, immune disorders, acute nerve damage or chronic neurological disorders, to reduce the risk of their development or to reduce their severity or progression, and for the prophylactic treatment of subjects with increased risk of cerebrovascular disease (i.e., reducing the risk of development). In some embodiments, the treatment (e.g., the compound taught herein) can be administered to the subject daily or otherwise in a chronic manner.

[0178] The present invention is primarily directed to the treatment of human subjects, but the present invention may also be performed on animal subjects, particularly mammalian subjects, such as mice, rats, dogs, cats, livestock and horses, for veterinary purposes, and / or for drug screening and / or drug development purposes.

[0179] IV. Preparation

[0180] In some embodiments, one or more active compounds may be provided in a pharmaceutically acceptable carrier. The carrier should be acceptable in that they are compatible with any other ingredients of the formulation and are not harmful to the recipient thereof. In some embodiments, the pharmaceutically acceptable carrier is sterile (e.g., endotoxin-free or pyrogen-free water, or endotoxin-free or pyrogen-free saline).

[0181] The formulations of the present invention may include short-term, rapid onset, rapid offset, controlled release, sustained release, delayed release, and pulsatile release formulations, so long as the formulation achieves administration of the compounds as described herein. See Remington's Pharmaceutical Sciences (18th ed.; Mack Publishing Company, Eaton, Pa., 1990), which is incorporated herein by reference in its entirety.

[0182] Pharmaceutical formulations according to the present invention may be suitable for various modes of delivery, including oral, parenteral (including intravenous, intramuscular, subcutaneous, intradermal and transdermal), topical (including dermal, buccal and sublingual) and rectal administration.

[0183] The invention is explained in more detail in the following non-limiting examples. Example

[0184] Example 1: Development of non-lysosomal aminoquinoline inhibitors of QR2.

[0185] Chloroquine and hydroxychloroquine are lysosomotropic drugs that preferentially accumulate in cellular lysosomes. For chloroquine, the pKa of the tertiary amine nitrogen is 10.32, while the pKa of the quinoline nitrogen is 7.29. At acidic lysosomal pH between 4-5.5, nearly 100% of chloroquine is doubly protonated, giving the molecule a 2+ charge, making it strongly hydrophilic, membrane-impermeable, and therefore trapped within acidic organelles.

[0186] A quantitative treatment of this trapping phenomenon can be obtained by examining the drug's octanol-water partition coefficient, log D, which describes the relative partitioning properties of all forms of a compound at different pH. For a given pH, compounds with a positive log D are relatively lipophilic and more membrane permeable, while compounds with a negative log D are hydrophilic and less membrane permeable.

[0187] exist Figure 1A-1BThe logD of chloroquine and QR2I-44 are shown in Figure 1. The shaded area in the figure represents the likely range of lysosomal pH encountered in vivo. At lysosomal pH, the logD of chloroquine is negative, reflecting the charge accumulated by these molecules at these pHs and the loss of membrane permeability. However, for QR2i-44, the logD remains positive in the lysosomal pH range, thereby retaining some lipophilicity and, therefore, membrane permeability.

[0188] Lysosomal accumulation is believed to be the primary mechanism responsible for the major toxicity (retina, heart) of 4-aminoquinolines such as chloroquine and hydroxychloroquine (Plaquenil). Using cLogP and LogD calculations and LiPE analysis, candidate compounds were identified with the goal of combining QR2 inhibitory properties with a low likelihood of lysosomal accumulation.

[0189] From these analyses, the compound 7-methoxy-4-(pyridin-3-yl)methylaminoquinoline (hereinafter "QR2I-44", "QR2i-44" or "I-44") (structure shown in Example 3 below) was discovered. The log D value of this compound remains positive, indicating that it will remain lipophilic (and therefore membrane permeable) at lysosomal pH between 4-5. As described above, Figure 1A-1B Chloroquine ( Figure 1A ) and QR2I-44 ( Figure 1B ) of log D.

[0190] Although the 4-aminoquinoline skeleton has been studied by several groups since the initial discovery of chloroquine and hydroxychloroquine, this particular compound had not been previously explored. Previous studies may have intentionally avoided extending the 4-aminoquinoline skeleton to include compounds with a methoxy group at the 7-position, as there is extensive literature discussing the need for halogen substitution at the 7-position of 4-aminoquinoline to obtain antimalarial activity (reviewed by Kaschula et al., "Structure-activity relationships in 4-aminoquinoline antiplasmodials," J. Med. Chem. 2002, 45:3531-3539;Shreekant and Bhimanna, "4-aminoquinolines: An Overview of AntimalarialChemotherapy," Med. Chem. 2016, 6:001-011).

[0191] The 7-halogen substituted derivatives were initially shown to be much more active than the unsubstituted analogs (Foley and Tilley, "Quinoline antimalarials: mechanisms of action and resistance and prospects for new agents," Pharmacol. Ther. 1998, 79: 55-87). Several other groups have shown more specifically that 7-chloroquine is actually required for inhibition of β-hematin formation (see, e.g., Vippagunta et al., "Structural specificity of chloroquine-hematin binding related to inhibition of hematin polymerization and parasite growth," J. Med. Chem. 1999, 42: 4630-4639), and replacement of the 7-chloro group with other electron donating groups (e.g., NH2 and OCH3 (methoxy)) substantially weakens or eliminates the inhibition of β-hemoglobin and thereby the antimalarial activity.

[0192] See also Egan TJ (2006) Interactions of quinoline antimalarials withhematin in solution. J Inorg Biochem 100: 916-926; Nsumiwa, S.; Kuter, D.; Wittlin, S.; Chibale, K.; Egan, TJ Bioorg. Med. Chem. 2013, 21, 3738); Egan, TJ; Hunter, R.; Kaschula, CH; Marques, HM; Misplon, A.;Walden, JC, J. Med. Chem. 2000, 43, 283; Kaschula, CH; Egan, TJ;Hunter, R.; Basilico, N.; Parapini, S.; Taramelli, D.; 3531.

[0193] Example 2: Synthesis and characterization of QR2 inhibitors.

[0194]

[0195] A suspension of 4,7-dichloroquinoline (2.0 g, 10.2 mmol) in aqueous methylamine (40% 20 mL 260 mmol, 26 equivalents) was heated in a microwave container at 90 ° C (initial power was set to 150 W) for 2 hours. Analysis of the reaction mixture by TLC (2% MeOH / CH Cl ) indicated that the raw material was completely consumed. The reaction mixture was diluted with H o (100 mL), and insolubles were collected under vacuum. The filter cake was washed with H o and dried in vacuo to obtain the pure product as a white microcrystalline solid (1.8 g, 92%). 1 H NMR (DMSO- d 6, 300 MHz) δ 8.40 (d, J = 5.1 Hz, 1H), 8.16 (d, J = 9.0 Hz, 1H), 7.77(s, 1H), 6.38 (d, J = 5.4 Hz, 1H), 2.86 (d, J = 5.4 Hz, 3H). ESIMS: m / z = 193[(M+H) + ].

[0196]

[0197] General procedure for 7-substituted-4-(pyridin-3-yl)-methylaminoquinolines. A mixture of 7-substituted-4-chloroquinoline (5.1 mmol), 3-aminomethylpyridine (0.70 g, 6.2 mmol, 1.2 equiv.) and 1-butanol (5 mL) was heated in a sealed thick-walled pressure vessel (12 mL) at 130 °C (bath temperature) for 24 h. The vessel was cooled to room temperature and the contents were diluted in Et2O (150 mL). Insoluble material was removed under vacuum. The filter cake was dissolved in a minimum amount of MeOH and the resulting solution was added to silica gel (about 3 g). The mixture was concentrated to dryness under reduced pressure. Flash column chromatography (RediSepR f SiO2 (40 g), 100% CH2Cl2 → 75% (90:10, CH2Cl2: MeOH containing 10% NH3) gave the desired product.

[0198] X = OCH3 (pale yellow solid, 0.60 g, 44%).

[0199] 1 H NMR (DMSO- d6, 400 MHz) δ 8.59 (s, 1H), 8.42 (m, 1H), 8.21 (d, J =5.6 Hz, 1H), 8.14 (d, J = 9.2 Hz, 1H), 7.79-7.72 (m, 2H), 7.32-7.29 (m, 1H), 7.14 (m, 1H), 7.06 (dd, 2.4 Hz, 8.8 Hz, 1H), 6.25 (d, J = 5.4 Hz, 1H), 4.52 (d, J = 5.4 Hz, 2H), 3.84 (s, 3H). ESIMS: m / z 266 [(M+H) + ].

[0200] X = Cl (white solid, 0.92 g, 67%).

[0201] 1 H NMR (DMSO- d 6, 400 MHz) δ 8.61 (s, 1H), 8.42 (s, 1H), 8.27 (m, 2H), 8.00 (s, 1H), 7.75 (m, 2H), 7.46 (d, J = 8.8 Hz, 1H), 7.31 (m, 3H), 6.39 (d,J = 5.4 Hz, 1H), 4.55 (d,J = 5.4 Hz, 2H). ESIMS: m / z = 270 [(M+H) + ].

[0202] Example 3: Comparative testing of QR2 inhibitors.

[0203]

[0204] Table 1: Comparison IC 50 data.

[0205] <![CDATA[ IC 50 (µM) ]]> <![CDATA[ Standard Deviation ]]> QR2 I-44 7.51 0.66 Compound 1 7.23 1.51 Compound 2 15.50 4.84 Chloroquine 70.92 5.27

[0206] We investigated the neuroprotective potential of QR2I-44 in a mouse model of closed head injury (TBI) (Laskowitz et al., “Neuroprotective pentapeptide CN-105 is associated with reduced sterile inflammation and improved functional outcomes in a traumaticbrain injury murine model”, Sci. Rep. 2017 Apr 21;7:46461; Laskowitz et al., “Traumatic brain injury exacerbates neurodegenerative pathology: improvement with an apolipoprotein E-based therapeutic.” Journal of Neurotrauma 2010m 27:1983-1995). The closed head impact model results in injury to selectively vulnerable neurons in the cortex, basal ganglia, and hippocampus, induces diffuse axonal injury, and results in measurable vestibular motor and long-term neurocognitive deficits. The TBI model involves a single impact designed to avoid skull fracture. Injury is primarily produced by acceleration / deceleration forces. For this model, therapeutic compounds were delivered intraperitoneally 4 h after TBI and then once weekly throughout the experiment, which was terminated after 28 days, followed by assessment of spatial learning and memory.

[0207] Figures 2, 3 and 4 summarize the results of vestibular motor function assessment after TBI, comparing the therapeutic efficacy of QR2-44 with three other 4-aminoquinoline compounds that inhibit QR2 (Table 1): (7-chloro-N-methylquinolin-4-amine), which demonstrated similar inhibitory properties, while compound 2 (7-chloro-N-(pyridin-3-yl)quinolin-4-amine) and CQ (chloroquine) are both weaker inhibitors of QR2. Mice were tested on the rotarod for 5 consecutive days after TBI, and then on days 7 and 28. Figure 2A -B shows that mice treated with QR2i-44 had nearly 300% improvement in rotarod latency (time to fall from the rotating rod) compared to vehicle and nearly 200% improvement compared to CQ after 5 days. Figure 3-4 It was shown that QR2I-44 resulted in more sustained improvements in vestibular-motor function than other QR2 inhibitors at 1 and 4 weeks after TBI.

[0208] Figure 5A-B summarizes the results of spatial learning and memory assessments 4 weeks after TBI, using the Morris water maze to test the ability of mice to locate a submerged platform. Mice were tested four times daily for 4 consecutive days (inter-trial interval = 30 minutes). The latency to locate the platform was recorded and the 4 trials per day were averaged. Mice were tested on days 28-31 after injury (n = 11-12 mice / group). A standard control test was also performed to confirm unimpaired vision. Figure 5B Demonstrated that animals treated with QR2-44 had 30-35% improvement in learning and memory compared to vehicle-treated animals, and Figure 5A Improvements in this function were shown not to be confounded by differences in motor function (i.e., swimming speed).

[0209] Figures 6-7 summarize the results of quantitative measurements of volume and axonal tract integrity in brain subregions using high-resolution ex vivo MRI. A total of 15 mice (10-11 week old C57Cl / 6J males). Normal n=3; vehicle n=6; QR2I-44 n=6. TBI and treatment followed the same protocol as for vestibular motor and learning / memory assessments. Mice were sacrificed and perfused 6 weeks after TBI. After fixation, the brain was removed from the skull and stored in 0.5% ProHance-doped formalin to facilitate high-resolution MRI.

[0210] The MRI protocol included: (1) 3D T1-weighted FLASH sequence with FOV = 1.8 cm × 1.8 cm × 1.8 cm; matrix = 256 × 256 × 256; resolution = 70 × 70 × 70 μm / pixel; TE / TR = 6 / 30 ms; average = 16; flip angle = 34; scan time = 6 hours 33 minutes; (2) high-resolution spin-echo-based 3D DTI with FOV = 1.8 cm × 1.8 cm × 1.8 cm; matrix = 128 × 128 × 128; resolution = 141 × 141 × 141 μm / pixel; TE / TR = 25 / 250 ms; diffusion direction = 60; Ao images = 5; B-value for each direction = 1500 S / mm2; scan time = 73 hours 30 minutes. For post-processing, the mouse brain was registered to a reference template from Wake Forest University's Mouse Database and segmented using ITK-SNAP. DTI parameters were calculated using TrackVis software.

[0211] Fig. 6A -B shows that volume loss in the hippocampus after TBI was significantly attenuated in animals treated with QR2I-44 compared to vehicle controls. Fig. 7A-B demonstrates that DTI parameters such as fractional anisotropy (FA), a measure of fiber tract integrity, are significantly improved after QR2I-44 treatment compared to controls in large white matter tracts such as the corpus callosum / external capsule (CC / EC).

[0212] Fig. 8A -C summarizes the results of an investigation into the potential of QR2I-44 to treat lupus. Fig. 8A The efficacy of QR2I-44 in reducing the release of microparticles (MPs) from RAW 264.7 mouse macrophages after stimulation with the Toll receptor ligand CpG oligodeoxynucleotide DNA (CpG). MPs are small membrane-bound vesicles that are produced from activated and dying cells through a blistering process. These particles range in size from 0.1-1.0 microns and contain nuclear, cytoplasmic and membrane components. As shown in in vitro and in vivo experiments, MPs have multiple biological functions and can mediate activities such as inflammation, thrombosis and information exchange between cells. Therefore, MPs play an important role in physiological and pathophysiological environments including lupus erythematosus (Spencer et al., "The Properties of Microparticles from RAW 264.7 Macrophage Cells Undergoing in vitro Activation or Apoptosis" 2014, 20(3): 239-248).

[0213] Figure 8B -C shows the ability of QR2I-44 to inhibit the release of the inflammatory cytokine TNFα from RAW 264.7 murine macrophages following stimulation of the TOLL receptor ligand with CpG and Pl C. In both cases, the inhibition of release was dose-dependent.

[0214] Fig. 9 The MRL / lpr mouse is an established murine model of an autoimmune disease similar to human systemic lupus erythematosus. The MRL / lpr produces several biomarkers also seen in human lupus, including anti-cDNA. Fig. 9 Results are shown for anti-cDNA autoantibody production in MRL / lpr mice 1 and 3 weeks after initiation of QR2I-44 treatment. Comparison of the graphs reveals a gradual therapeutic suppression of autoantibodies in this model that is also dose-dependent.

[0215] We performed a preliminary screening of compound QR2I-44 for drug-like properties, cardiotoxicity, mutagenicity, hepatocyte stability, and cytochrome P450 inhibition and induction. Briefly, we found that I-44 was not a potent inhibitor of the hERG ion channel (IC50 > 10 μM), was not mutagenic in a modified Ames assay, exhibited high solubility in aqueous solution (aqueous solubility > 60 μg / ml), and exhibited high membrane permeability in Caco2 cells (effective permeability > 20 × 10 -6 cm / s), is not a substrate or inhibitor of P-glycoprotein (efflux ratio <0.5; insensitive to 10 μM elacrida), has low to moderate binding to mouse and human plasma proteins (approximately 50-70% binding by ultracentrifugation), exhibits reasonable stability in mouse and human hepatocytes (>90% retention at 1 hour), is not an inducer or inhibitor of CYP3A4 or 2D6, and is likely to be orally bioavailable and CNS penetrant.

[0216] Example 4: Absorption and brain penetration of QR2i-44

[0217] In mice, QR2i-44 was found to be well absorbed and brain penetrant by all dosing methods: intravenous (IV), intraperitoneal (IP), and oral (PO). The results of PO delivery of 25 mg / kg of QR2i-44 in mouse plasma and brain samples are shown in Table 2 below.

[0218] Table 2: Concentrations in mouse plasma and brain samples after oral delivery (PO)

[0219]

[0220] Example 5: Crystal structure of QR2i-44 confirms binding to QR2

[0221] We determined the crystal structure of reduced QR2 in complex with FADH and QR2i-44 to 1.60 Å resolution using molecular replacement. To test the effect of the redox state on the enzyme structure, we also determined the structure of the oxidized complex to 1.40 Å. The structure was readily phased by molecular replacement using the existing crystal structure of QR2 as a search model. The quaternary structure of the enzyme is a constitutive, functional homodimer that has been determined.

[0222] The crystal structure confirmed that QR2i-44 and its related compounds with planar ring structures bind to QR2 and FADH in a manner similar to the natural substrate quinone. In addition, the crystal structure showed that QR2i-44-44 may be superior to other inhibitors of QR2 in its ability to penetrate the substrate binding site and achieve optimal stacking of the planar aromatic functional group with the FAD isoalloxazine.

[0223] Example 6: QR2 inhibition for the treatment of mitochondrial dysfunction

[0224] Parkinson's disease (PD) is the most common motor neurodegenerative disorder. There is an urgent unmet medical need for innovative treatments that can successfully prevent or stop the progression of PD, and there is currently no such treatment. PD is characterized by progressive neurodegeneration, and clinical symptoms include both motor impairment and non-motor features (such as cognitive decline). The exact underlying mechanism of PD pathogenesis is still unclear. However, it is known that mitochondrial dysfunction is one of the key factors contributing to the pathogenesis of both sporadic and familial PD. In subjects with PD, mitochondrial complex I activity is reduced in the brain and systemically. When administered to rodents and non-human primates, inhibitors of complex I (such as rotenone, a mitochondrial toxin) mimic many behaviors, pathologies and clinical features of PD. In addition, many genes that have been reported to cause or increase the risk of developing PD are related to mitochondrial pathways.

[0225] The enzyme quinone reductase 2 (QR2) can contribute to oxidative damage in mitochondria. Inhibition of this enzyme in vivo has been shown to suppress neurotoxic effects in PD models showing mitochondrial dysfunction and oxidative stress, and blocking QR2 activity can protect and restore mitochondrial function in the context of neurodegenerative diseases such as PD.

[0226] Interestingly, associations between polymorphisms in QR2 and the risk of developing human PD have been observed, and some of these polymorphisms have been shown to result in increased QR2 expression, higher enzyme activity, and increased ROS production in the presence of dopamine (Harada et al., "An association between idiopathic Parkinson's disease and polymorphismsof phase II detoxification enzymes: glutathione S-transferase M1 and quinoneoxidoreductase 1 and 2," Biochem Biophys Res Commun. 2001;288(4):887-92; Wang et al., "Association of NRH:quinone oxidoreductase 2 gene promoter polymorphismwith higher gene expression and increased susceptibility to Parkinson'sdisease," J Gerontol A Biol Sci Med Sci. 2008;63(2):127-34).

[0227] Consistent with pathological QR2 activity, previous studies have shown that inhibition of QR2 with drugs such as chloroquine and resveratrol is neuroprotective in different animal models of neural injury, including PD (Boutin et al., "Quinone Reductase 2 Inhibitor: Main Biochemical and Cellular Characterization," Mol Pharmacol. 2019;95(3):269-859-11; Janda et al., "The antidote effect of quinone oxidoreductase 2 inhibitor against paraquat-induced toxicity in vitro and in vivo," Br J Pharmacol. 2013;168(1):46-59; Janda et al., "Parkinsonian toxin-induced oxidative stress inhibits basalautophagy in astrocytes via NQO2 / quinone oxidoreductase 2: Implications for neuroprotection," Autophagy. 2015;11(7):1063-80). For example, QR2 inhibition eliminates PD-associated paraquat-induced toxicity (Janda et al., supra). In addition, QR2 inhibition leads to protection against neurotoxicity induced by the PD-associated toxin MPP+ (Boutin et al., supra). In summary, the role of oxidative stress, mitochondrial dysfunction, the pathophysiology of QR2 in PD, and the report that QR2 inhibition can mediate protection against neurotoxicity by reducing ROS levels support the assumption that QR2 inhibitors are developed as a therapeutic strategy for PD (Cassagnes et al., "Oxidative stress and neurodegeneration: The possible contribution of quinone reductase 2," FreeRadic Biol Med. 2018; 120: 56-61).

[0228] Complex I dysfunction, mitochondrial damage, and oxidative stress are key players in the pathogenesis of PD (Sanders et al., "Oxidative damage to macromolecules in human Parkinson disease and the rotenone model," Free Radic Biol Med. 2013;62:111-20). In fact, environmental toxicants such as rotenone and paraquat, which both cause mitochondrial dysfunction and oxidative stress, have been associated with an increased risk of developing human PD (Tanner et al., "Rotenone, paraquat, and Parkinson's disease," Environ Health Perspect. 2011;119(6):866-72). Therefore, exposure to PD-associated toxicants is a way to mimic mitochondrial dysfunction and oxidative stress in cells and in vivo.

[0229] Wild-type HEK293 cells were pretreated with QR2 inhibitors (CDL-1, CDL-2, or QR2i-44) for 1 h, exposed to menadione or vehicle for 4 h, and mitochondrial membrane potential was measured.

[0230] result :As in Fig.10 As shown in Figure 2, all QR2 inhibitors were able to prevent menadione-induced membrane potential loss, as detected by tetramethylrhodamine ethyl ester (TMRE, a fluorescent dye sequestered by live and active mitochondria). In particular, QR2i-44 significantly prevented menadione-induced membrane potential loss and was able to prevent menadione-induced mitochondrial depolarization.

[0231] The above is illustrative of the present invention and should not be construed as limiting the present invention. The present invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: R1 is: wherein D1, D2, D3 and D4 are each hydrogen.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (I)(a)(1):

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (I)(a)(2):

4. A composition comprising the compound according to any one of claims 1 to 3 and a carrier.

5. The composition of claim 4, wherein the carrier is a pharmaceutically acceptable carrier.

6. An in vitro method for inhibiting the activity of quinone reductase-2 (QR2), the method comprising contacting QR2 with a compound according to any one of claims 1 to 3 or a composition according to claim 4 or 5, wherein the contacting is performed in vitro.

7. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for inhibiting the activity of quinone reductase-2 (QR2).

8. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating malaria.

9. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating an immune disorder.

10. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating acute nerve injury.

11. Use of the compound of any one of claims 1 to 3 or the composition of claim 4 or 5 in the preparation of a medicament for treating chronic neurological disorders.

12. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating lupus.

13. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating an infectious disease.

14. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating cancer.

15. Use of the compound of any one of claims 1 to 3 or the composition of claim 4 or 5 in the preparation of a medicament for treating CNS lupus.

16. Use of the compound according to any one of claims 1 to 3 or the composition according to claim 4 or 5 in the preparation of a medicament for treating cerebrovascular diseases.

17. Use of the compound of any one of claims 1 to 3 or the composition of claim 4 or 5 in the preparation of a medicament for treating a disorder associated with mitochondrial dysfunction.

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