HK1192249A

HK1192249A - Ire-1a inhibitors

Info

Publication number: HK1192249A
Application number: HK14105694.2A
Authority: HK
Inventors: J．B．帕特森; D．G．洛纳甘; G‧A‧弗林; Q‧曾; P‧V‧帕莱
Original assignee: 复星弘创(苏州)医药科技有限公司
Priority date: 2007-06-08
Filing date: 2014-06-16
Publication date: 2014-08-15

Description

IRE-1 alpha inhibitors

The patent application is a divisional application of an invention patent application with the national application number of PCT/US2008/066310, the international application date of 2008/9.6, and the application number of 200880019035.2 in the Chinese national stage, and the name of IRE-1 alpha inhibitor.

Priority of serial number 60/942,743 filed on 8/6/2007 is claimed and is incorporated herein by reference.

Technical Field

The present invention relates to inhibitors of IRE-1 alpha and their therapeutic uses.

Background

Protein folding stress (protein folding stress) in the endoplasmic reticulum of cells can trigger a signal transduction cascade called the unfolded protein effect or UPR. Phytase 1 (IRE-1. alpha.) is a key enzyme that relieves protein folding stress by enhancing chaperone activity, thus protecting cells from stress-induced apoptosis. Inhibitors of IRE-1 α are useful at least in the treatment of B cell autoimmune diseases, certain cancers and some viral infections.

Disclosure of Invention

The present invention provides IRE-1 alpha inhibitor compounds, and prodrugs and pharmaceutically acceptable salts thereof. The invention also provides pharmaceutical compositions and methods of therapeutically treating diseases associated with the effects of unfolded proteins using IRE-1 alpha inhibitor compounds, prodrugs and pharmaceutically acceptable salts thereof. Treatable patients include those with B-cell autoimmune diseases, certain cancers, and some viral infections.

The present invention includes a number of chemical compounds related by structure and function and methods of their use. Various groupings and applications of these compounds, including any number of them, can be defined and constitute various embodiments of the present invention. Some embodiments expressly include certain compounds, while other embodiments expressly exclude certain compounds. The inclusion or exclusion criteria include a particular structure or structural feature, activity level or range (e.g., IC)₅₀Or EC₅₀) The suitability for administration by a particular route of administration, the disease being treated, and the like.

IRE-1 alpha inhibitor compounds

IRE-1 alpha inhibitor compounds of the present invention are aromatic and heteroaromatic hydroxyaldehydes that directly inhibit the enzyme. It is believed that these compounds act by inhibiting the rnase activity of the enzyme. In a specific embodiment of the invention, the activity is detected as an in vitro cleavage of IRE-1 α to human small-XBP-1 mRNA stem-loop substrate 5'-CAGUCCGCAGGACUG-3' (SEQ ID NO:1) of at least 10, 15, 20, 25, 30, 40, 50, 60, or 75%. Other substrates may also be used to detect cleavage. See US 20070105123.

In some embodiments, the compound inhibits the average IC of IRE-1 α in an in vitro assay₅₀About 20 μ M (20,000nM) or less (e.g., 20000, 15000, 10000, 7500, 7250, 7000, 6750, 6500, 6250, 6000, 5750, 5500, 5250, 5000, 4750, 4500, 4250, 4000, 3750, 3500, 3250, 3000, 2750, 2500, 2250, 2000, 1750, 1500, 1250, 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 15, 10, 5,2 or 1nM or less). In some embodiments, the compounds are used in an in vitro XBP-1 splicing assay (e.g., myeloma cells)Mean EC for inhibition of IRE-1. alpha₅₀Is 80 μ M (80,000nM) or less (e.g., 80000, 75000, 70000, 65000, 60000, 55000, 50000, 45000, 40000, 35000, 30000, 25000, 20000, 15000, 10000, 7500, 7250, 7000, 6750, 6500, 6250, 6000, 5750, 5500, 5250, 5000, 4750, 4500, 4250, 4000, 3750, 3500, 3250, 3000, 2750, 2500, 2250, 2000, 1750, 1500, 1250, 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 15, 10, 5,2, or 1nM or less). IRE-1 α inhibitor compounds may meet any or all of these criteria.

It is well known in the art that the aldehyde group in these compounds can be represented as any of the three equivalent forms shown below:

structural formula (I) includes compounds used in the present invention:

in the formula:

the OH substituent is located ortho to the aldehyde substituent;

q is an aromatic carbocyclic (isocyclic) or heterocyclic ring system selected from: benzene, naphthalene, pyridine N-oxide, thiophene, benzo [ b ]]Thiophene, benzo [ c]Thiophene, furan, pyrrole, pyridazine, pyrimidine (pyrmidine), pyrazine, triazine, isozymeOxazoline,Oxazoline,Thiazoline, pyrazoline, imidazoline, fluorene, biphenyl, quinoline, isoquinoline, cinnoline, 2, 3-naphthyridine, quinazoline, quinoxaline, benzofuran, indole, isoindole, isobenzofuranyl, benzimidazole, 1, 2-benzisoxazolineAzoles and carbazoles;

R^x、R^yand R^zMay or may not be present and is independently selected from: hydrogen, aryl, heteroaryl, -A "R^a、-OH、-OA″R^a、-NO₂、-NH₂、-NHA″R^a、-N(A″R^a)(A′″R^b)、-NHCOA″R^a、-NHCOOA″R^a、-NHCONH₂、-NHCONHA″R^a、-NHCON(A″R^a)(A′″R^b) Halogen, -COOH, -COOA' R^a、-CONH₂、-CONHA″R^a、-CON(A″R^a)(A′″R^b) And

R^aand R^bIndependently are: hydrogen, -COOH, -COOA, -CONH₂、-CONHA、-CONAA′、-NH₂-NHA, -NAA', -NCOA, -NCOOA, -OH or-OA;

y is C₁-C₁₀Alkylene or C₂-C₈Alkenylene radical in which (a)1, 2 or 3 CH₂The group can be O, S, SO₂NH or NR^cAnd/or (b) 1-7H atoms may independently be replaced by F or Cl;

a and A' are:

(a) independently is C₁-C₁₀Alkyl or C₂-C₈Alkenyl wherein (i)1, 2 or 3 CH₂The group can be O, S, SO₂NH or NR^cAnd/or (ii)1 to 7H atoms may independently be replaced by F or Cl, aryl or heteroaryl; or

(b) Or, A and A' together form C₂-C₇Alkylene, wherein 1,2 or 3 CH₂The group can be O, S, SO₂、NH、NR^c、NCOR^cOr NCOOR^cInstead, to form, for example, an alkylenedioxy (alkylenedioxy);

a ', A' are independently (a) absent, (b) C₁-C₁₀Alkylene radical, C₂-C₈Alkenylene or C₃-C₇Cycloalkyl radicals in which 1,2 or 3 CH₂The group can be O, S, SO₂NH or NR^cAnd/or 1 to 7H atoms can be replaced by F and/or Cl; or (C) both may together form C₂-C₇Alkyl radical, in which 1,2 or 3 CH₂The group can be O, S, SO₂、NH、NR^c、NCOR^cOr NCOOR^cInstead of this, the user can either,

R^cis C₁-C₁₀Alkyl radical, C₃-C₇Cycloalkyl radical, C₄-C₈Alkylene cycloalkyl or C₂-C₈An alkenyl group; in which 1,2 or 3 CH₂The group can be O, S, SO₂NH, NMe, NEt and/or-CH ═ CH-groups, 1-7H atoms may be replaced by F and/or Cl, and/or 1H atom may be R^aReplacing;

aryl is phenyl, benzyl, naphthyl, fluorenyl or biphenyl, each of which may be unsubstituted or mono-, di-or trisubstituted by: halogen, -CF₃、-R^f、-OR^d、-N(R^d)₂、-NO₂、-CN、-COOR^d、CON(R^d)₂、-NR^dCOR^e、-NR^dCON(R^e)₂、-NR^dSO₂A、-COR^d、-SO₂N(R^d)₂、-S(O)_mR^fAA' together or-O (aryl),

R^dand R^eIndependently is H or C₁-C₆An alkyl group;

R^fis C₁-C₆An alkyl group;

heteroaryl is a monocyclic or bicyclic saturated, unsaturated or aromatic heterocycle having 1 to 2N, O and/or S atoms, which may be unsubstituted or mono-or disubstituted by: carbonyl oxygen, halogen, R^f、-OR^d、-N(R^d)₂、-NO₂、-CN、-COOR^d、-CON(R^d)₂、-NR^dCOR^e、-NR^dCON(R^e)₂、-NR^fSO₂R^e、-COR^d、-SO₂NR^dand/or-S (O)_mR^f(ii) a And

m is 0,1 or 2.

The group of IRE-1 alpha inhibitor compounds represented by formula (I) includes the following compounds, wherein R^x、R^yAnd R^zAs defined above:

C₁-C₁₀alkyl (i.e., alkyl having 1,2,3,4, 5, 6, 7, 8,9, or 10 carbon atoms) and C₁-C₆Alkyl groups (i.e., alkyl groups having 1,2,3,4, 5, or 6 carbon atoms) can be branched or unbranched, and can be substituted or unsubstituted. Optional substituents include haloElements (e.g., F, Cl, I, Br). Examples include methyl, ethyl, trifluoromethyl, pentafluoroethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl, n-hexyl and n-decyl. In some embodiments, C₁-C₁₀Is methyl, ethyl, trifluoromethyl, propyl, isopropyl, butyl, n-pentyl, n-hexyl or n-decyl.

C₃-C₇Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. In some embodiments, C₃-C₇The cyclopentyl group is cyclopentyl.

In some embodiments, C₂-C₈Alkenyl is vinyl, allyl, 2-butenyl, 3-butenyl, isobutenyl, sec-butenyl, 4-pentenyl, isopentenyl or 5-hexenyl. In some embodiments, C₂-C₈Alkenyl is 4-pentenyl, isopentenyl or 5-hexenyl.

C₁-C₁₀The alkylene group is preferably unbranched, and in some embodiments it is methylene or ethylene, propylene or butylene.

In some embodiments, C₂-C₈Alkenylene is ethenylene or propenylene.

C₂-C₇The alkylene group is preferably unbranched. In some embodiments, C₂-C₇Alkylene is ethylene, propylene or butylene.

In some embodiments, C₄-C₈The alkylidene cycloalkyl is cyclohexylmethyl or cyclopentylethyl.

In some embodiments, R^x、R^yAnd R^zIndependently is-OH, -OA, -NO₂or-NAA'.

In some embodiments, Q is benzene, naphthalene, thiophene, benzo [ b]Thiophene or benzo [ c]Thiophene, R^xAnd R^yIs hydrogen, and R^zIs hydrogen OR-OR^d、-NO₂Pyridyl or pyridyl N-oxide.

In some embodiments, R^xIs hydrogen, OR^d、NO₂、-NH₂or-NHCOOA "R^a。

In some embodiments, R^aIs hydrogen, -COOH, -NHA or-NAA'.

In some embodiments, R^cIs C₁-C₁₀Alkyl or C₁-C₆An alkyl group.

In some embodiments, Y is methylene, ethylene, propylene, or butylene.

In some embodiments, A and A' are independently C₁-C₁₀An alkyl group; c₁-C₁₀Alkyl, wherein 1-7 hydrogen atoms are replaced by F and/or Cl; an aryl group; or a heteroaryl group.

In some embodiments, A "and A'" are independently absent or C₁-C₁₀Alkylene of which 1 CH₂The radical may be NH or NR^cAnd (4) replacing.

In some embodiments, A "and A'" together form C₂-C₇Alkylene chain, in which 1 CH₂The radical may be NH or NR^cAnd (4) replacing.

In some embodiments, aryl is mono-, di-, or tri-substituted with: methyl, ethyl, propyl, butyl, fluorine, chlorine, hydroxyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, nitro, cyanoformyl, acetyl, propionyl, trifluoromethyl, amino, methylamino, ethylamino, dimethylamino, diethylamino, sulfonamido, methylsulfinamido, ethylsulfonamido, propylsulfonamido, butylsulfonamido, dimethylsulfonamido, carboxyl, methoxycarbonyl, ethoxycarbonyl, or aminocarbonyl.

In some embodiments, heteroaryl is selected from: 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 1-pyrrolyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 2-Azolyl, 4-Azolyl, 5-Azolyl, 3-isoAzolyl, 4-isoAzolyl, 5-isoOxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 3-isothiazolyl, 4-isothiazolyl, 5-isothiazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl (pyrmidinyl), 6-pyrimidinyl (pyrimidinyl), 1,2, 3-triazol-1-yl, 1,2, 3-triazol-4-yl, or 1,2, 3-triazol-5-yl, 1,2, 4-triazol-1-yl, 1,2, 4-triazol-3-yl, 1,2, 4-triazol-5-yl, 1-tetrazolyl, 5-tetrazolyl, 1,2,3-Oxadiazol-4-yl, 1,2,3-Oxadiazol-5-yl, 1,2,4-Oxadiazol-3-yl, 1,2,4-Oxadiazol-5-yl, 1,3, 4-thiadiazol-2-yl or 1,3, 4-thiadiazol-5-yl, 1,2, 4-thiadiazol-3-5-yl, 1,2, 3-thiadiazol-4-yl, 1,2, 3-thiadiazol-5-yl, 3-pyridazinyl, 4-pyridazinyl, pyrazinyl, 1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl, 4-isoindolyl, 5-isoindolyl, 1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 1-benzpyrazolyl, 3-benzpyrazolyl, 4-benzpyrazolyl, 5-benzpyrazolyl, 6-benzpyrazolyl, 7-benzpyrazolyl, 2-benzpyrazolylAzolyl, 4-benzoAzolyl, 5-benzoAzolyl, 6-benzoAzolyl, 7-benzoAzolyl, 3-benzisoylAzolyl, 4-benzisoylAzolyl, 5-benzisoylAzolyl, 6-benzisoylAzolyl, 7-benzisoylOxazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl, 2-benzisothiazolyl, 4-benzisothiazolyl, 5-benzisothiazolyl, 6-benzisothiazolyl, 7-benzisothiazolyl, 4-benzo-2, 1,3-Oxadiazolyl, 5-benzo-2, 1,3-Oxadiazolyl, 6-benzo-2, 1,3-Oxadiazolyl, 7-benzo-2, 1,3-Oxadiazolyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl, 1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl, 3-cinnolinyl, 4-cinnolinyl, 5-cinnolinyl, 6-cinnolinyl, 7-cinnolinyl, 8-cinnolinyl, 2-quinazolinyl, 4-quinazolinyl, 5-quinazolinyl, 6-quinazolinyl, 7-quinazolinyl, 8-quinazolinyl, 5-quinoxalinyl, 6-quinoxalinyl, 2-2H-benzo-1, 4-Oxazinyl, 3-2H-benzo-1, 4-Oxazinyl, 5-2H-benzo-1, 4-Oxazinyl, 6-2H-benzo-1, 4-Oxazinyl, 7-2H-benzo-1, 4-Oxazinyl, 8-2H-benzo-1, 4-Azinyl, 1, 3-benzodioxol-5-yl, 1, 4-benzodiAlk-6-yl, 2,1, 3-benzothiadiazol-4-yl, 2,1, 3-benzothiadiazol-5-yl and 2,1, 3-benzothiadiazolOxadiazol-5-yl.

Heterocyclic groups may also be partially or fully hydrogenated. For example, in some embodiments, heteroaryl is 2, 3-dihydro-2-furyl, 2, 3-dihydro-3-furyl, 2, 3-dihydro-4-furyl, 2, 3-dihydro-5-furyl, 2, 5-dihydro-2-furyl, 2, 5-dihydro-3-furyl, 2, 5-dihydro-4-furyl, 2, 5-dihydro-5-furyl, tetrahydro-2-furyl, tetrahydro-3-furyl, 1, 3-dioxolan-4-yl, tetrahydro-2-thienyl, tetrahydro-3-thienyl, 2, 3-dihydro-1-pyrrolyl, 2, 3-dihydro-2-furyl, 2, 5-dihydro-4-furyl, 2, 5-dihydro-2-furyl, 2, 5-dihydro-3-furyl, 1, 3-dioxolan-4-yl, tetrahydro-2-, 2, 3-dihydro-2-pyrrolyl, 2, 3-dihydro-3-pyrrolyl, 2, 3-dihydro-4-pyrrolyl, 2, 3-dihydro-5-pyrrolyl, 2, 5-dihydro-1-pyrrolyl, 2, 5-dihydro-2-pyrrolyl, 2, 5-dihydro-3-pyrrolyl, 2, 5-dihydro-4-pyrrolyl, 2, 5-dihydro-5-pyrrolyl, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, tetrahydro-1-imidazolyl, tetrahydro-2-imidazolyl, tetrahydro-4-imidazolyl, 2, 3-dihydro-1-pyrazolyl, 2, 3-dihydro-2-pyrazolyl, 2, 3-dihydro-3-pyrazolyl, 2, 3-dihydro-4-pyrazolyl, 2, 3-dihydro-5-pyrazolyl, tetrahydro-1-pyrazolyl, tetrahydro-3-pyrazolyl, tetrahydro-4-pyrazolyl, 1, 4-dihydro-1-pyridyl, 1, 4-dihydro-2-pyridyl, 1, 4-dihydro-3-pyridyl, 1, 4-dihydro-4-pyridyl, 1,2,3, 4-tetrahydro-1-, 1,2,3, 4-tetrahydro-2-, 1,2,3, 4-tetrahydro-3-pyridyl, 1,2,3, 4-tetrahydro-4-pyridyl group, 1,2,3, 4-tetrahydro-5-pyridyl group, 1,2,3, 4-tetrahydro-6-pyridyl group, 1-piperidyl group, 2-piperidyl group, 3-piperidyl group, 4-piperidyl group, 2-morpholinyl group, 3-morpholinyl group, 4-morpholinyl group, tetrahydro-2-pyranyl group, tetrahydro-3-pyranyl group, tetrahydro-5-pyridyl group-4-pyranyl, 1, 4-diAlkyl, 1, 3-diAlk-2-yl, 1, 3-diAlk-4-yl, 1, 3-diAlk-5-yl, hexahydro-1-pyridazinyl, hexahydro-3-pyridazinyl, hexahydro-4-pyridazinyl, hexahydro-1-pyrimidinyl, hexahydro-2-pyrimidinyl, hexahydro-4-pyrimidinyl, hexahydro-5-pyrimidinyl, 1-piperazinyl, 2-piperazinyl, 3-piperazinyl, 1,2,3, 4-tetrahydro-1-, 1,2,3, 4-tetrahydro-2-quinolinyl, 1,2,3, 4-tetrahydro-3-quinolinyl, 1,2,3, 4-tetrahydro-4-quinolinyl, 1,2,3, 4-tetrahydro-5-quinolinyl, 1,2,3, 4-tetrahydro-6-quinolinyl, hexahydro-3-pyridazinyl, hexahydro-4-pyridazinyl, hexahydro-1-pyrimidinyl, 1,2,3, 4-tetrahydro-1-quinolinyl, 1,2,3,4-, 1,2,3, 4-tetrahydro-7-quinolinyl, 1,2,3, 4-tetrahydro-8-quinolinyl, 1,2,3, 4-tetrahydro-1-isoquinolinyl, 1,2,3, 4-tetrahydro-2-isoquinolinyl, 1,2,3, 4-tetrahydro-3-isoquinolinyl, 1,2,3, 4-tetrahydro-4-isoquinolinyl, 1,2,3, 4-tetrahydro-5-isoquinolinyl, 1,2,3, 4-tetrahydro-6-isoquinolinyl, 1,2,3, 4-tetrahydro-7-isoquinolinyl, 1,2,3, 4-tetrahydro-8-isoquinolinyl, 2-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 3-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 5-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 6-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 7-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 8-3, 4-dihydro-2H-benzo-1, 4-Oxazinyl, 2, 3-methylenedioxyphenyl, 3, 4-methylenedioxyphenyl, 2, 3-ethylenedioxyphenyl, 3,4- (difluoromethylenedioxy) phenyl, 2, 3-dihydrobenzofuran-5-yl, 2, 3-dihydrobenzofuran-6-yl, 2,3- (2-oxomethylenedioxy) phenyl, 3, 4-dihydro-2H-1, 5-benzodioxepin-6-yl, 3, 4-dihydro-2H-1, 5-benzodioxepin-7-yl, 2, 3-dihydrobenzofuran-yl or 2, 3-dihydro-2-oxofuryl (oxofuryl).

In some embodiments, heteroaryl is unsubstituted pyridyl, pyridyl N-oxide, thienyl, furyl, pyrrolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, isozylAn azolinyl group,Oxazolinyl, thiazolinyl, pyrazolinyl, imidazolinyl, naphthyl, quinolinyl, isoquinolinyl, cinnolinyl, 2, 3-naphthyridinyl, quinazolinyl or quinoxalinyl. In other embodiments, the heteroaryl is pyridyl.

In some embodiments, heteroaryl is a monocyclic saturated or unsaturated heterocycle having 1-2N and/or O atoms, it may be carbonyloxy, OH or OA mono-or disubstituted, for example 2-oxopiperidin-1-yl, 2-oxopyrrolidin-1-yl, 2-oxo-1H-pyridin-1-yl, 3-oxomorpholin-4-yl, 4-oxo-1H-pyridin-1-yl, 2, 6-dioxopiperidin-1-yl, 2-oxopiperazin-1-yl, 2, 6-dioxopiperazin-1-yl, 2, 5-dioxopyrrolidin-1-yl, 2-oxo-1, 3-.Oxazolidin-3-yl, 3-oxo-2H-pyridazin-2-yl, 2-caprolactam-1-yl (= 2-oxoazepane (o)xoazepan) -1-yl), 2-hydroxy-6-oxopiperazin-1-yl, 2-methoxy-6-oxopiperazin-1-yl, 2-azabicyclo [2.2.2 ]]-oct-3-on-2-yl or 2-oxopiperidin-1-yl. In some embodiments, heteroaryl is 2-oxopiperidin-1-yl.

In other embodiments, heteroaryl is a monocyclic saturated heterocyclic group having 1-2N atoms, which may be C₁-C₆Alkyl is mono-or di-substituted.

The group of IRE-1 alpha inhibitor compounds of formula (I) also includes those of formula (II)

In the formula:

R¹is hydrogen, halogen, -NO₂、-OCH₃or-OCH₂CH₃(ii) a And

orEach of which may be unsubstituted or substituted with 1,2 or 3 substituents independently selected from: halogen, -OH, -COOH, -CH₂OCH₃、C₁-C₃Alkyl radical, C₁-C₃Alkoxy, -CH₂OH, phenoxy and phenyl-C₁-C₃An alkoxy group. The alkoxy group may be straight-chain or branched.

In some embodiments, R¹is-OCH₃。

Representative IRE-1 α inhibitor compounds of formula (II) include those listed in tables 1 and 2.

Table 1.

The group of IRE-1 α inhibitor compounds of formula (I) also includes those of structural formula (III):

in the formula R²、R³And R⁴Independently selected from: hydrogen, halogen, -OH, -COOH, -CH₂OCH₃、C₁-C₃Alkyl radical, C₁-C₃Alkoxy, -CH₂OH, phenoxy and phenyl-C₁-C₃An alkoxy group.

Representative IRE-1 α inhibitor compounds of formula (III) include those listed in Table 2.

TABLE 2

The group of IRE-1 α inhibitor compounds of formula (I) also includes those of structural formula (IV):

in the formula:

R¹selected from hydrogen, -OH, -OCH₃、-OCH₂CH₃-C = O or-NO₂(ii) a And

R⁵and R⁶Independently of one another hydrogen, halogen, C₁-C₃Alkyl or-NO₂。

In some embodiments, the IRE-1 α inhibitor compound is represented by structural formula (IV), but excludes compounds wherein:

R¹、R⁵and R⁶Each is hydrogen;

R¹is-OCH₃，R⁵And R⁶Are all hydrogen;

R¹and R⁵Are each hydrogen, R⁶Is fluorine;

R¹and R⁶Are all-NO₂，R⁵Is hydrogen;

R¹and R⁵Are each hydrogen, R⁶is-CH₃；

R¹is-CH₃，R⁵And R⁶Are all hydrogen;

R¹is-OCH₃，R⁵Is thatR⁶Is hydrogen;

R¹and R⁶Are all Cl, I or F;

R¹is Br, R⁶Is Cl;

R¹is-NO₂，R⁶Is Br;

R¹is a carbonyl group, R⁶Is Cl or methyl;

R¹is methoxy, R⁶is-NO₂Br, methoxy or Cl; and

R¹is methoxy, R⁵Is Br.

Other IRE-1 α inhibitor compounds are represented by the following structural formula:

in the formula R³As defined above. Representative IRE-1 α inhibitor compounds of formula (V) include:

other IRE-1 α inhibitor compounds are represented by structural formula (VI):

in the formula R²As defined above. For example, R²IRE-1 α inhibitor compounds that are phenyl groups are represented by the following structural formula:

in the formula, R⁴And R⁵Independently selected from the above R²And R³A substituent of (1).

Representative IRE-1 α inhibitor compounds of formula (VI) include:

other useful IRE-1 alpha inhibitor compounds are shown in Table 3 below.

In some embodiments, the IRE-1 α inhibitor compound is represented by structural formula (a), which falls within the scope of formula (I):

in the formula:

R¹is hydrogen, halogen or a 5-or 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulfur;

R²is hydrogen,Phenyl or a 5-or 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulphur, wherein the heteroaryl group is optionally benzo-fused and optionally substituted with 1,2 or 3 substituents independently selected from the group consisting of:C₁-C₃a straight or branched alkyl group,C₁-C₃Phenylalkyl, C₁-C₃An alkoxyphenylalkyl group,

R³Is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,OrAnd

q is a 5-or 6-membered heterocyclic ring.

In some compounds of formula (A), R¹Selected from the group consisting of: hydrogen, hydrogen,And Br.

In some compounds of formula (A), R²Selected from the group consisting of: hydrogen, hydrogen,

In some compounds of formula (A), R⁴Selected from the group consisting of: hydrogen, hydrogen,And

in some compounds of formula (A), R⁵Selected from the group consisting of: hydrogen, hydrogen,

In some compounds of formula (A), R⁶Selected from the group consisting of: hydrogen, hydrogen,

In some compounds of formula (A), R⁷Selected from the group consisting of: hydrogen, hydrogen,And

in some compounds of formula (A), R⁸Selected from the group consisting of: hydrogen, hydrogen,Andor R⁸And R⁹And together with the nitrogen atom to which they are attached form

In some compounds of formula (A), R⁹Is hydrogen, or with R⁸And together with the nitrogen atom to which they are attached form

In some compounds of formula (A), R³Selected from the group consisting of: hydrogen, -F, -CF₃、-NO₂、-O、-OCH₃、-CH₂OH,and-OR¹⁰In the formula, R¹⁰Is hydrogen, C₁-C₆Straight or branched alkyl orIn the formula R⁸And R⁹As defined by structure (A) above.

In some embodiments, the compound is represented by structural formula (a1), which falls within the scope of formula (a):

in the formula:

R¹is hydrogen or a 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulfur;

q is an optionally benzo-fused 5-or 6-membered heterocyclic ring;

R⁴、R⁵and R⁶Independently of one another hydrogen, = O, -CH₃、Or

In some compounds of formula (A1), R¹Selected from the group consisting of: hydrogen, hydrogen,

In some compounds of formula (1), Q is selected from the group consisting of:

R⁴、R⁵and R⁶Independently selected from:C₁-C₃a straight or branched alkyl group,C₁-C₃Phenylalkyl, C₁-C₃An alkoxyphenylalkyl group,And

in some compounds of formula (A1), R³Selected from the group consisting of: hydrogen, -F, -CF₃、-NO₂and-OCH₃。

In some embodiments, the compound is represented by structural formula (a2), which falls within the scope of formula (a):

in the formula:

R³is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkyl, C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,OrAnd

in some embodiments, the compound is represented by structural formula (a3), which falls within the scope of formula (a):

in the formula:

q is a 5-or 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulfur;

R¹is hydrogen; and

R³is hydrogen or C₁-C₃An alkoxy group.

In some compounds represented by structural formula (a3), Q is selected from the group consisting of:and

in some compounds of formula (A3), R₃Is that

In some embodiments, the compound is represented by structural formula (a4), which falls within the scope of formula (a):

in the formula:

R¹is hydrogen;

R³is hydrogen, -F, -NO₂Or

R⁸Is thatOr with R⁹And together with the nitrogen atom to which they are attached formAnd

R⁹is hydrogen, or with R⁸And together with the nitrogen atom to which they are attached form

In some embodiments, the compound has one of the following structural formulae:

in some embodiments, the compound is represented by structural formula (B), which falls within the scope of formula (I):

in the formula:

R¹and R²Independently hydrogen, phenyl or an optionally benzo-fused 5-or 6-membered heterocycle, wherein said phenyl or optionally benzo-fused 5-or 6-membered heterocycle is optionally substituted with:-CH₂OH、-CHO、-OCH₃halogen, -OH, -CH₃、

R⁴is hydrogen,

In some embodiments, the compound has one of the following structural formulae:

in some embodiments, the compound is represented by structural formula (C), which falls within the scope of formula (I):

in the formula:

R¹is hydrogen, -CH3 or-OH;

R²and R³Independently hydrogen, phenyl or an optionally benzo-fused 5-or 6-membered heterocycle, wherein said phenyl or optionally benzo-fused 5-or 6-membered heterocycle is optionally substituted with:-CH₂OH、-CHO、-OCH₃halogen, -OH, -CH₃、

The hydroxy substituent in ring a is located ortho to the aldehyde substituent.

In some embodiments, the compound of structural formula (C) has one of the following structures:

in some embodiments, the compound is represented by structural formula (D), which falls within the scope of formula (I):

in the formula R¹Is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkyl, C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,OrIn a compound of formula (D), R¹Is methyl.

Other useful compounds of the invention are shown in Table 11.

A pharmaceutically acceptable salt; a stereoisomer; tautomers

IRE-1 α inhibitor compounds include the compounds in free form or pharmaceutically acceptable salts and stereoisomers thereof. Some particular IRE-1 α inhibitor compounds described herein are protonated salts of amine compounds. The term "free form" refers to the amine compound in a non-salt form. Included pharmaceutically acceptable salts include not only those salts of the particular compounds disclosed herein, but also all typical pharmaceutically acceptable salts of the IRE-1 α inhibitor compounds of formulas I-VII and A-D, in free form, and of the prodrugs of formulas E and F (infra).

The free form of the particular salt compound may be isolated using techniques known in the art. For example, the salt may be treated with a suitable dilute base solution, such as dilute aqueous solutions of NaOH, potassium carbonate, ammonium bicarbonate and sodium bicarbonate, to produce the free form. The free forms may differ from their respective salt forms in certain physical properties, such as solubility in polar solvents, but for the purposes of the present invention, acid and base salts are pharmaceutically equivalent to their respective free forms.

Pharmaceutically acceptable salts of the disclosed IRE-1 α inhibitor compounds can be synthesized from the compounds of the present invention containing a base or acid moiety by conventional chemical methods. Salts of the basic compounds are generally prepared by ion exchange chromatography or by reacting the free base with a stoichiometric amount or an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents. Similarly, salts of acidic compounds can be prepared by reaction with a suitable inorganic or organic base.

Pharmaceutically acceptable salts of IRE-1 α inhibitor compounds include the conventional non-toxic salts of the compounds formed by reacting a basic compound with an inorganic or organic acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, as well as salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic (pamoic), maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, benzenesulfonic, methanesulfonic, ethanedisulfonic (ethane disulphonic), oxalic, isethionic (isethionic), trifluoroacetic and the like.

If the IRE-1. alpha. inhibitor compound is acidic, suitable pharmaceutically acceptable salts include those prepared from pharmaceutically acceptable non-toxic bases, including inorganic and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese salts, manganous, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and salts of basic ion exchange resins, such as arginine, betaine caffeine, choline, N1-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine (tromethamine), and the like. The preparation of the above pharmaceutically acceptable Salts and other typical pharmaceutically acceptable Salts is more fully described by Berg et al ("Pharmaceutical Salts"), J.pharm.Sci., 1977:66: 1-19).

Some IRE-1 α compounds or prodrugs may be internal salts or zwitterions because under physiological conditions, the deprotonated acidic moiety of the compound, e.g., the carboxyl group, may be anionic, and this charge may then be in internal equilibrium with the cationic charge of the protonated or alkylated basic moiety, e.g., the quaternary nitrogen atom.

IRE-1. alpha. inhibitor Compounds or prodrugs thereof may have asymmetric centers, chiral axes and chiral planes (see: E.L. Eliel and S.H. Wilen, "stereochemistry of Carbon Compounds" stereochemistry "Carbon Compounds, John Wiley father (John Wiley & Sons), New York, 1994, p. 1119-1190), racemates, racemic mixtures and individual diastereomers may exist, with all possible isomers including optical isomers and mixtures thereof being within the invention.

Accordingly, an IRE-1 α inhibitor compound or a prodrug thereof may have the following properties; the constituent atoms thereof may be spatially arranged in two or more ways, although having the same bonds. Thus, the compound exists in stereoisomeric forms. Cis/trans isomerism is only one type of stereoisomerism. If these stereoisomers are non-superimposable images and mirror images, they are enantiomers with chiral or chiral nature, due to the presence of one or more asymmetric carbon atoms in the structures that make up them. Enantiomers are optically active and therefore distinguishable because they rotate the plane of polarized light to the same extent but in opposite directions.

If two or more asymmetric carbon atoms are present in the IRE-1 α compound, there are two possible configurations for each of these carbon atoms. For example, if there are two asymmetric carbon atoms, there may be 4 stereoisomers. Furthermore, the 4 possible stereoisomers may be divided into 6 pairs of possible stereoisomers that are different from each other. In order for a pair of molecules with multiple asymmetric carbons to be enantiomers, they must have different configurations at each asymmetric carbon. Those pairs of molecules that do not exhibit enantiomers have different stereochemical relationships, known as diastereorelationships. Stereoisomers that are not enantiomers are referred to as diastereomers, or more commonly, diastereomers.

All of these well known stereochemical aspects of the compounds of the present invention are to be considered as part of the present invention. Accordingly, the present invention encompasses stereoisomers of IRE-1 α inhibitor compounds, if they are enantiomers, individual enantiomers, racemic mixtures of these enantiomers, and artificial (i.e., synthetic) mixtures comprising these enantiomers in various ratios, different from the ratios of these enantiomers observed in the racemic mixture. If the stereoisomers of the IRE-1 α inhibitor compounds are diastereomers, the compounds include each diastereomer as well as mixtures of any two or more of these diastereomers in any desired ratio.

The following is used for illustration: if one asymmetric carbon atom forming the (-) (R) and (+) (S) enantiomers is present in an IRE-1 α inhibitor compound, the IRE-1 α inhibitor compound includes all pharmaceutically acceptable salt forms, prodrugs and metabolites thereof that have therapeutic activity and are useful in the treatment or prevention of the diseases and disorders described further herein. If IRE-1. alpha. inhibitor compounds exist in the form of the (-) (R) and (+) (S) enantiomers, the compounds also include the (+) (S) enantiomer alone or the (-) (R) enantiomer alone if all, substantially all, or the major therapeutic activity is attributed to only one of these enantiomers or if the adverse side effect is attributed to only one of these enantiomers. Such compounds of the invention also include racemic mixtures of both the (+) (S) and (-) (R) enantiomers or non-racemic mixtures of the corresponding parts in optionally desired proportions, provided that there is no substantial difference in the biological properties of the two enantiomers.

A pair or set of enantiomers (if present) of an IRE-1 alpha inhibitor compound have particular biological effects and/or physical and chemical properties, and it is apparent that these enantiomers can be used in particular ratios, e.g., to form the final therapeutic product. The following is for description only: if a pair of enantiomers is present, these enantiomers may be used, for example, in the following ratios: 90% (R) -10% (S), 80% (R) -20% (S), 70% (R) -30% (S), 60% (R) -40% (S), 50% (R) -50% (S), 40% (R) -60% (S), 30% (R) -70% (S), 20% (R) -80% (S) and 10% (R) -90% (S). Having assessed the properties of the various enantiomers of the IRE-1 alpha inhibitor compound, if present, the respective amounts of one or more of these enantiomers having particular desired properties, which constitute the final therapeutic product, can be determined in a simple manner.

For IRE-1. alpha. inhibitor compounds of the present disclosure in which tautomers may exist, two tautomeric forms are included in the present disclosure, even though only one tautomeric structure is depicted. For example, the keto tautomer compounds depicted below include the enol tautomer (and vice versa) and mixtures thereof.

The invention also includes pharmaceutically useful stereoisomers, E/Z isomers, enantiomers, racemates, diastereomers, hydrates and solvates of the disclosed compounds. A "solvate" is an addition product of an inert solvent molecule and a compound due to the attractive forces between them. Solvates are, for example, monohydrates, dihydrates or alcoholates.

Prodrugs

The invention also provides prodrugs that are metabolized to active IRE-1 α inhibitor compounds after administration. For example, an IRE-1 α inhibitor compound disclosed herein may be modified, e.g., with an alkyl or acyl group, a sugar, or an oligopeptide, to release the active IRE-1 α inhibitor compound upon rapid cleavage in vivo.

Derivatives of the corresponding aromatic alcohols can be used as prodrugs of aromatic aldehydes, since alcohols and aldehydes can be metabolically interchanged according to the following general scheme:

Scheline，1972,Xenobiotica,2,227-36

examples of prodrugs of aldehydes, ketones, alcohols and other functional groups are found in Wermuth et al, 1996,before design Drug and biological precursors I: carrier prodrug "(Designing Prodrugs and Bioprecursors I: Carrier Prodrugs, journal of The medical Chemistry Practice (The Practice of Medicinal Chemistry)Page 672-696; and Wermuth,1996, "Preparation of Water-Soluble Compounds by Covalent Attachment to a solubilizing moiety" (Preparation of Water-solvent Compounds by solvent Attachment of solubilizing Moites), described in Wermuth, edPractice of medicinal chemistry》(The Practice of Medicinal Chemistry) 756, 776. Other versatile aldehyde and alcohol derivatives that may enable prodrug function and methods for their preparation are described in Cheronis et al, 1965, semi-quantitative Organic Analysis, New York: interscience, page 465-518.

Prodrugs of the present invention include compounds having the following structural formula AA, BB, or CC, wherein Q' is the same as Q defined above in all respects, except that the aldehyde substituent of Q is present in the prodrug form shown below, and R is^aAnd R^cAs hereinbefore describedDefining:

in some embodiments, prodrugs of IRE-1 α inhibitor compounds are represented by structural formula (E):

in the formula:

R¹is hydrogen or-OCH₃(ii) a And

in some embodiments, the prodrug of structural formula (E) has one of the following structural formulae:

in some embodiments, the IRE-1 α inhibitor prodrug is represented by structural formula (F):

in the formula:

R¹is hydrogen or Br;

R²is hydrogen, Br orAnd

R³is hydrogen, -OCH₃-COOH or-OCH₂CH₃。

In some embodiments, the IRE-1 α prodrug of structural formula (F) has one of the following structural formulae:

other examples of IRE-1 α inhibitor prodrugs include:

limitations of the Compound claims

To the extent that any of the following compounds are not novel, applicants reserve the right to submit a compound and/or composition claim with the proviso that such compounds and/or their pharmaceutically acceptable salts are excluded from the scope of such claims:

in the formula W²Is halogen; an alkyl group having 1 to 4 carbon atoms; alkoxy having 1 to 4 carbon atoms; an acyloxy group having 2 to 4 carbon atoms; an acyl group having 2 to 4 carbon atoms; a carboxylic acid group; ester group- -COOW⁵Wherein W is⁵Is a straight or branched alkyl group having 1 to 4 carbon atoms; a nitrile group; an OH group; -a CHO group; -NO₂A group; an acetamido group; w¹Is hydrogen or W²One of the defined substituents; w, which may be the same or different³And W⁴Each is a hydrogen atom or W²One of the defined substituents;

in the formula T¹、T²、T³、T⁴And T⁵Independently selected from hydroxy, alkoxy containing 1-6 carbon atoms, alkyl containing 1-6 carbon atoms, phenyl, NO₂COOH, COH, sulfonic acid groups, ketones containing 1 to 6 carbon atoms, F, Cl, Br, I, hydrogen, or salts of any of the above acids or alcohols, wherein at least two or more T groups are hydrogen; or a phenolic mixture thereof;

in the formula of U¹、U²、U³And U⁴Each independently represents a hydrogen or halogen atom or an alkyl, cycloalkyl, aralkyl, aryl, alkaryl, alkoxy, aryloxy, acyl or hydroxyl group;

in the formula V¹、V²、V³And V⁴Represents hydrogen or halogen; or in which V²And V⁴Is hydrogen, V¹And V³Is hydrogen or halogen;

formula (III) Z, Z¹、Z²And Z³May be the same or different and represents a hydrogen atom, an alkyl, aryl, or cycloalkyl group, an alkoxy, hydroxyl or acylamino group, or a halogen;

2-hydroxybenzaldehyde (salicylaldehyde); 2-hydroxy-3-methylbenzaldehyde; 2-hydroxy-3-tert-butylbenzaldehyde; 2-hydroxy-3-tert-butyl-5-methylbenzaldehyde; 2-hydroxy-3, 5-di-tert-butylbenzaldehyde; 2-hydroxy-3-isopropyl-6-methyl-benzaldehyde; 2-hydroxy-3-cyclohexylbenzaldehyde; 2-hydroxy-4-tert-butyl-benzaldehyde; 2-hydroxy-4-chlorobenzaldehyde and 2-hydroxy-6-chlorobenzaldehyde; 2-hydroxy-3-phenylbenzaldehyde; 2-hydroxy-5-methoxybenzaldehyde; 2-hydroxy-3-nonylbenzaldehyde; 2, 5-dihydroxybenzaldehyde; and 2-hydroxy-4-acetylaminobenzaldehyde;

in the formula B¹、B²、B³And B⁴Each is a hydrogen atom, an alkyl group, a cycloalkyl group, an alkoxy group or a hydroxyl group or a halogen atom;

wherein n is 0 or 1, m + n is up to 4 or 3, D is alkyl, alkoxy, hydroxyalkyl, cycloalkyl, aryl, alkoxyalkyl, hydroxy, nitro or halogen;

salicylaldehyde, p-hydroxybenzaldehyde, 2, 3-dihydroxybenzaldehyde, 2, 6-dihydroxybenzaldehyde, 2-hydroxy-3-methoxybenzaldehyde (o-vanillin), 2, 4-diformylphenol, 2, 6-diformylphenol, 1, 2-dihydroxy-3, 5-diformylbenzene, 1, 2-dihydroxy-4, 6-diformylbenzene, 1-hydroxy-2-methoxy-4, 6-diformylbenzene (4, 6-diformylguaiacol), 1-hydroxy-2-ethoxy-4, 6-diformylbenzene, 2, 6-dihydroxy-benzaldehyde and o-hydroxy-p-vanillin;

in the formula E¹Represents a hydroxyl group, a halogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heterocyclic group, an alkoxy group, an aryloxy group, an acylamino group, a sulfonylamino group, a substituted amino group, a monoalkylamino group, a dialkylamino group, an arylamino group or an alkylarylamino group; or E¹May be joined together to represent a 5-or 6-membered ring; e is ortho or para to the formyl group and represents a methylene group substituted with at least one substituent selected from the group consisting of: hydroxyl group, halogen atom, alkoxy group, aryloxy group, alkylthio group (alkylthio), arylthio group (arylthio), acyloxy group, chlorocarbonyloxy group, alkoxycarbonyloxy group and aminocarbonyloxy group; r is an integer from 0 to 3; when r is 2 or more, E¹The same or different;

in the formula E³Represents hydroxy, alkyl, cycloalkyl, aryl, alkoxy, aryloxy, acylamino, sulfonylamino, unsubstituted amino, monoalkylamino, dialkylamino, arylamino or alkylarylamino; or E³May be taken together to represent a 5-or 6-membered ring; -CH₂-in ortho or para position to the formyl group; e²Represents alkylthio, arylthio, chlorocarbonyloxy, alkoxycarbonyloxy or aminocarbonyloxy; s is 0 to 3, and when s is 2 or more, E³The same or different;

2-hydroxybenzaldehyde, 3-methyl-2-hydroxybenzaldehyde, 3-ethyl-2-hydroxybenzaldehyde, 3-n-propyl-2-hydroxybenzaldehyde, 3-isopropyl-2-hydroxybenzaldehyde, 3-n-butyl-2-hydroxybenzaldehyde, 3-sec-butyl-2-hydroxybenzaldehyde, 3-tert-butyl-2-hydroxybenzaldehyde, 3-pentyl-2-hydroxybenzaldehyde, 4-methyl-2-hydroxybenzaldehyde, 4-ethyl-2-hydroxybenzaldehyde, 4-n-propyl-2-hydroxybenzaldehyde, 4-isopropyl-2-hydroxybenzaldehyde, 4-n-butyl-2-hydroxybenzaldehyde, 3-n-propyl-2-hydroxybenzaldehyde, 3-tert-butyl-2-hydroxybenzaldehyde, 3-pentyl-2-hydroxybenzaldehyde, 4-methyl-2-hydroxybenzaldehyde, 4-ethyl-, 4-sec-butyl-2-hydroxybenzaldehyde, 4-tert-butyl-2-hydroxybenzaldehyde, 4-pentyl-2-hydroxybenzaldehyde, 5-methyl-2-hydroxybenzaldehyde, 5-ethyl-2-hydroxybenzaldehyde, 5-n-propyl-2-hydroxybenzaldehyde, 5-isopropyl-2-hydroxybenzaldehyde, 5-n-butyl-2-hydroxybenzaldehyde, 5-sec-butyl-2-hydroxybenzaldehyde, 5-tert-butyl-2-hydroxybenzaldehyde, 5-pentyl-2-hydroxybenzaldehyde, 6-methyl-2-hydroxybenzaldehyde, 6-ethyl-2-hydroxybenzaldehyde, 6-n-propyl-2-hydroxybenzaldehyde, methyl-2-hydroxybenzaldehyde, ethyl-2-hydroxybenzaldehyde, 6-isopropyl-2-hydroxybenzaldehyde, 6-n-butyl-2-hydroxybenzaldehyde, 6-sec-butyl-2-hydroxybenzaldehyde, 6-tert-butyl-2-hydroxybenzaldehyde, 6-pentyl-2-hydroxybenzaldehyde, 3, 5-dinitro-2-hydroxybenzaldehyde, 3, 5-difluoro-2-hydroxybenzaldehyde, 3, 4-diisobutyl-2-hydroxybenzaldehyde, 3, 4-di-tert-butyl-2-hydroxybenzaldehyde, 3, 6-di-tert-butyl-2-hydroxybenzaldehyde, 2-hydroxy-3, 5-dichlorobenzaldehyde, 2, 6-dihydroxybenzaldehyde, 2, 4-dihydroxy-6-methylbenzaldehyde, methyl-2-hydroxy-2-hydroxybenzaldehyde, methyl-2-hydroxy-3, 5-dichlorobenzaldehyde, methyl-2-hydroxy-2-hydroxybenzaldehyde, methyl-2-hydroxy-2-, 2,4, 6-trihydroxybenzaldehyde, 5-chloro-2-hydroxybenzaldehyde, 2-hydroxy-5-bromobenzaldehyde, 2-hydroxy-3, 5-diiodobenzaldehyde, 2, 4-dihydroxy-3-methylbenzaldehyde, 2-hydroxy-3-methoxy-6-bromobenzaldehyde, 2, 4-dihydroxy-5-propylbenzaldehyde, 2, 4-dihydroxy-5-hexylbenzaldehyde, 2-formyl-3, 6-dihydroxy-4, 5-dimethylbenzaldehyde, 2,3, 6-trihydroxybenzaldehyde, 2, 4-dihydroxy-5-acetylbenzaldehyde, 2-formyl-3, 6-dihydroxy-4, 5-dipropylbenzaldehyde, 2-formyl-3-methoxy-4, 5-dimethyl-6-hydroxybenzaldehyde, 2,3, 5-trihydroxybenzaldehyde, 2-hydroxy-6- (oxy-4-methylpentanoic acid) benzaldehyde, 3-formyl-4, 5-dihydroxybenzaldehyde, 2-ethyl-6-hydroxybenzaldehyde, 3-chloro-5- (3, 7-dimethyl-2, 6-octadienyl) -4, 6-dihydroxy-2-methylbenzaldehyde, 2-hydroxy-6- (8-pentadecenyl)) benzaldehyde, 2, 4-dihydroxy-3-ethyl-6- (1-methylpentyl) benzaldehyde, 2-hydroxy-6- (3-hydroxy-4-methylpentenyl) benzaldehyde, 2-hydroxy-6- (2-pentadecenyl) benzaldehyde, 2-hydroxy-3-ethyl-6- (1-methylpentyl) benzaldehyde, 2-hydroxy-5-hydroxybenzaldehyde, 2-hydroxy, 2-pentanoic acid-3-formyl-4, 5-dihydroxybenzaldehyde, 2-propanoic acid-3-formyl-4, 5-dihydroxybenzaldehyde, 2,3, 4-trihydroxy-5-methyl-6-hydroxymethylbenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-carboxybenzaldehyde, 3-carboxy-4-hydroxybenzaldehyde, 2, 3-dihydroxy-4-methoxybenzaldehyde, 2-hydroxy-6-methoxybenzaldehyde, 2, 5-dihydroxybenzaldehyde, 2,3, 4-trihydroxy-6-hydroxymethylbenzaldehyde, 2, 3-dihydroxybenzaldehyde, 2-hydroxy-5-acetylbenzaldehyde, methyl-ethyl-4, 4-trihydroxy-4-methoxybenzaldehyde, 2-hydroxy-5-carboxybenzaldehyde, 2-hydroxy-4-hydroxybenzaldehyde, 2-hydroxy-4-methoxybenzaldehyde, 2-, 2-hydroxy-5-carboxyethylbenzaldehyde, 2-hydroxy-5-carboxypropylbenzaldehyde, 2-hydroxy-5-carboxybutylbenzaldehyde, 2-hydroxy-3-iodo-5-carboxymethylbenzaldehyde and 2-formyl-3, 4, 5-trihydroxybenzaldehyde;

wherein X is halogen;

in the formula G¹、G²、G³And G⁴Independently of one another hydrogen, straight-chain or branched C₁-C₁₀Alkyl radical, C₃-C₈Cycloalkyl, straight or branched C₁-C₁₀Alkoxy, phenyl or halogen, where the alkyl or cycloalkyl radicals may be exclusively para to the hydroxyl group if they do not carry an H atom;

in the formula J¹Is NO₂，J²Is hydrogen; j. the design is a square¹And J²Are all chlorine; or J¹Is hydrogen, J²Is fluorine;

in the formula K¹And K⁴Independently selected from the group consisting essentially of: hydrogen; a hydroxyl group; halogen; a nitro group; a cyano group; a trifluoromethyl group; (C)₁-C₆) An alkyl group; (C)₁-C₆) An alkoxy group; (C)₃-C₆) A cycloalkyl group; (C)₂-C₆) An alkenyl group; - - -C (= O) OK⁷；--OC(=O)K⁷；--S(=O)₂；--S(=O)₂N(K⁷)(K⁹)；--S(=O)₂K⁷；--S(=O)₂OK⁷；--C(=O)NK⁷K⁹；--C(=O)K⁹(ii) a and-N (K)⁷)(K⁹) In which K is⁷Is hydrogen or (C)₁-C₄) Alkyl radical, K⁹Is (C)₁-C₄) An alkyl group; wherein, K is defined¹And K⁴The alkyl, cycloalkyl and alkenyl groups of (a) may be optionally independently substituted with 1 or 2 substituents selected from the group consisting essentially of: halogen; a hydroxyl group; (C)₁-C₂) An alkyl group; (C)₁-C₂) An alkoxy group; (C)₁-C₂) Alkoxy radical- (C₁-C₂) An alkyl group; (C)₁-C₂) An alkoxycarbonyl group; a carboxyl group; (C)₁-C₂) An alkylcarbonyloxy group; a nitro group; a cyano group; (C)₁-C₂) Alkyl disubstituted amino; a sulfonyl group; and (C)₁-C₂) An alkyl disubstituted sulfonamido group; DD and BB independently represent N, CHK²Or CHK³In which K is²And K³Independently selected from the group consisting essentially of: hydrogen; a hydroxyl group; halogen; a nitro group; a cyano group; a trifluoromethyl group; (C)₁-C₆) An alkyl group; (C)₁-C₆) An alkoxy group; (C)₃-C₆) A cycloalkyl group; (C)₂-C₆) An alkenyl group; - - -C (= O) OK¹¹；--OC(=O)K¹¹；--S(=O)₂；--S(=O)₂N(K¹¹)(K¹³) (ii) a and-N (K)¹¹)(K¹³) In which K is¹¹Is hydrogen or (C)₁-C₄) Alkyl radical, K¹³Is (C)₁-C₄) An alkyl group; wherein, K is defined²And K³The alkyl, cycloalkyl and alkenyl groups of (a) may be optionally independently substituted with 1 or 2 substituents selected from the group consisting essentially of: halogen; a hydroxyl group; (C)₁-C₂) An alkyl group; (C)₁-C₂) An alkoxy group; (C)₁-C₂) Alkoxy radical- (C₁-C₂) An alkyl group; (C)₁-C₂) An alkoxycarbonyl group; a carboxyl group; (C)₁-C₂) Alkylcarbonyl-oxy; a nitro group; a cyano group; (C)₁-C₂) Alkyl disubstituted amino; a sulfonyl group; (C)₁-C₂) An alkyl disubstituted sulfonamido group; in the formula K¹And K⁴Independently is hydrogen; a hydroxyl group; a trifluoromethyl group; (C)₁-C₄) An alkyl group; (C)₁-C₄) Alkoxy-; - - -C (= O) OK⁷(ii) a Or- -N (K)⁷)(K⁹) In which K is⁷Is hydrogen or (C)₁-C₂) Alkyl radical, K⁹Is (C)₁-C₂)；K¹And K⁴More preferably independently hydrogen; a hydroxyl group; (C)₁-C₂) An alkyl group; (C)₁-C₂) An alkoxy group; carboxyl or methylamino radical, in this case K⁷Is hydrogen, K⁹Is methyl; in the formula K¹And K⁴Defined as alkyl and substituted with one substituent selected from the group consisting of: a hydroxyl group; (C)₁-C₂) An alkoxy group; a carboxyl group; (C)₁-C₂) Alkyl disubstituted amino; and (C)₁-C₂) Alkyl disubstituted sulfonamido; in the formula K¹And K⁴Defined as alkyl and substituted with one substituent selected from the group consisting of: hydroxy, methoxy, and dimethylamino; wherein one of DD or BB is N and the other is CHK²Or CHK³(ii) a In which DD is CHK²BB is CHK³In which K is²And K³Independently is hydrogen; a hydroxyl group; halogen; a trifluoromethyl group; (C)₁-C₄) An alkyl group; (C)₁-C₄) An alkoxy group; - - -C (= O) OK¹¹；--S(=O)₂N(K¹¹)(K¹³) (ii) a Or- -N (K)¹¹)(K¹³) In which K is¹¹Is hydrogen or (C)₁-C₂) Alkyl radical, K¹³Is (C)₁-C₂) An alkyl group; wherein K²And K³Independently is hydrogen; a hydroxyl group; (C)₁-C₂) An alkyl group; (C)₁-C₂) An alkoxy group; a carbonyl group; or methylamino, K¹¹Is hydrogen, K¹³Is a firstA group; and wherein K²And K³Defined as alkyl and substituted, with one substituent selected from the group consisting of: a hydroxyl group; (C)₁-C₂) An alkoxy group; a carboxyl group; (C)₁-C₂) Alkyl-disubstituted amino; and (C)₁-C₂) Alkyl disubstituted sulfonamido.

O-vanillin; salicylaldehyde; 2, 3-dihydroxybenzaldehyde; 2, 6-dihydroxybenzaldehyde; 2-hydroxy-3-ethoxybenzaldehyde; and pyridoxal;

in the formula L¹And L²Represents a halogen atom, in particular a chlorine, bromine or iodine atom, L³Represents a hydrogen or halogen atom, in particular chlorine, L represents a hydroxyl group, an aryl or aralkyl residue substituted by at least one of the following substituents: halogen atom, CF₃、NO₂CN, alkyl, alkoxy, SCN, or tertiary amino;

wherein L is¹And L²Are all Cl, are all Br or are all I;

wherein XX is halogen, n is 2 or 3, YY and ZZ are lower alkyl, which may be the same or different, which may also form a heterocyclic ring with the nitrogen atom and may contain another heteroatom, i.e. N, N or S, and quaternary salts or metal chelates thereof; and

in the formula M¹、M⁴Y 'and X' are defined as follows:

M¹	M⁴	X′	Y′	M²	M³
						H	H	CHM²	CHM³	H	H
H	OH	CHM²	CHM³	H	H
						OH	H	CHM²	CHM³	H	H
CF₃	H	CHM²	CHM³	H	H
						CH₃	H	CHM²	CHM³	H	H
CH₂CH₃	H	CHM²	CHM³	H	H
						OCH₃	H	CHM²	CHM³	H	H
C(=O)OH	H	CHM²	CHM³	H	H
						C(=O)OCH₃	H	CHM²	CHM³	H	H
NHCH₃	H	CHM²	CHM³	H	H
						N(CH₃)₂	H	CHM²	CHM³	H	H
H	OH	CHM²	CHM³	H	H
						H	CH₃	CHM²	CHM³	H	H
H	CF₃	CHM²	CHM³	H	H
						H	CH₂CH₃	CHM²	CHM³	H	H
H	OCH₃	CHM²	CHM³	H	H
						H	C(=O)OH	CHM²	CHM³	H	H

H	C(=O)OCH₃	CHM²	CHM³	H	H
						H	NHCH₃	CHM²	CHM³	H	H
H	N(CH₃)₂	CHM²	CHM³	H	H
						OH	OH	CHM²	CHM³	H	H
CF₃	CF₃	CHM²	CHM³	H	H
						CH₃	CH₃	CHM²	CHM³	H	H
CH₂CH₃	CH₂CH₃	CHM²	CHM³	H	H
						OCH₃	OCH₃	CHM²	CHM³	H	H
C(=O)OH	C(=O)OH	CHM²	CHM³	H	H
						C(=O)OCH₃	C(=O)OCH₃	CHM²	CHM³	H	H
NHCH₃	NHCH₃	CHM²	CHM³	H	H
						N(CH₃)₂	N(CH₃)₂	CHM²	CHM³	H	H
H	H	CHM²	CHM³	OH	H
						H	H	CHM²	CHM³	H	OH
H	H	CHM²	CHM³	OH	OH
						H	H	CHM²	CHM³	CH₃	H
H	H	CHM²	CHM³	H	CH₃
						H	H	CHM²	CHM³	CH₃	CH₃
H	H	CHM²	CHM³	OCH₃	H
						H	H	CHM²	CHM³	H	OCH₃
H	H	CHM²	CHM³	OCH₃	OCH₃
						H	H	CHM²	CHM³	NHCH₃	H
H	H	CHM²	CHM³	H	NHCH₃
						H	H	CHM²	CHM³	NHCH₃	NHCH₃
H	H	CHM²	CHM³	N(CH₃)₂	H
						H	H	CHM²	CHM³	H	N(CH₃)₂
H	H	CHM²	CHM³	N(CH₃)₂	N(CH₃)₂
						CH₃	H	CHM²	CHM³	CH₃	H
H	CH₃	CHM²	CHM³	H	CH₃
						OCH₃	H	CHM²	CHM³	OCH₃	H
OCH₃	H	CHM²	CHM³	H	CH₃
						H	H	CHM²	CHM³	H	OH
H	OH	CHM²	CHM³	CH₃	CH₃
						OCH₃	H	CHM²	CHM³	OCH₃	H
OH	H	CHM²	CHM³	OCH₃	OCH₃
						OCH₃	H	CHM²	CHM³	H	NHCH₃
H	NHCH₃	CHM²	CHM³	NHCH₃	H
						H	OH	CHM²	CHM³	H	NHCH₃
H	OH	CHM²	CHM³	OH	H
						H	OH	CHM²	CHM³	H	OH
N(CH₃)₂	H	CHM²	CHM³	OCH₃	H
						CH₃	H	CHM²	CHM³	H	OCH₃
H	CH₃	CHM²	CHM³	N(CH₃)₂	H
						H	N(CH₃)₂	CHM²	CHM³	CH₃	H

OCH₃	H	CHM²	CHM³	H	OCH₃
						OCH₃	H	CHM²	CHM³	CH₃	CH₃
OCH₃	H	N	CHM³	-	H
						CH₃	H	N	CHM³	-	CH₃
H	N(CH₃)₂	N	CHM³	-	H
						H	CH₃	N	CHM³	-	CH₃
OCH₃	OCH₃	N	CHM³	-	H
						CH₃	H	N	CHM³	-	NHCH₃
CH₃	OCH₃	N	CHM³	-	H
						CH₃	CH₂OH	N	CHM³	-	H
CH₃	CH₂OH	N	CHM³	-	CH₃
						OCH₃	CH₂OH	N	CHM³	-	H

methods of making IRE-alpha inhibitor compounds and prodrugs of the invention

Some IRE-1 α inhibitor compounds that are commercially available for use in the disclosed methods are available, for example, from FC (Fluorochem Ltd.), Aurora Fine Chemicals (Aurora Fine Chemicals), TCI American Organic Chemicals (TCI American Organic Chemicals), AC&S Corp (AKos Consu)longand Solutions) or Maybridge (Maybridge). Other compounds and their starting materials may be prepared by modifications of methods known in the art as described in the literature, e.g. in standard works, e.g. Houben-Weyl (Methoden der organischen Chemie, Georg-Thieme-Verlag, Stuttgart). Can also be retrieved by computerBehcet database of reports (CrossFire Beilstein database) wherein the reaction zones describe the preparation of the substances in detail, see also the following specific examples.

Pharmaceutical preparation

Any of the IRE-1 α inhibitor compounds disclosed herein may be formulated into a medicament using methods well known in the art. Pharmaceutical formulations of the present invention will generally comprise at least one IRE-1 α inhibitor compound or prodrug thereof admixed with a carrier, diluted with a diluent, and/or enclosed or packaged in an ingestible carrier in the form of a capsule, sachet, cachet, paper or other container or in a disposable container such as an ampoule.

The carrier or diluent may be a solid, semi-solid or liquid material. Some examples of diluents or carriers that may be used in the pharmaceutical compositions of the invention are lactose, dextrose, sucrose, sorbitol, mannitol, propylene glycol, liquid paraffin, white soft paraffin, kaolin, microcrystalline cellulose, calcium silicate, silicon dioxide, polyvinylpyrrolidone, cetostearyl alcohol, starch, gum arabic, calcium phosphate, cocoa butter, peanut oil, alginates, tragacanth, gelatin, methyl cellulose, polyoxyethylene sorbitan monolaurate, ethyl lactate, propyl hydroxybenzoate, sorbitan trioleate, sorbitan sesquioleate and oleyl alcohol.

The pharmaceutical compositions of the present invention may be prepared by methods well known in the art, including conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, entrapping or lyophilizing processes.

For injection, the formulations of the present invention may be formulated as aqueous solutions, preferably in physiologically compatible buffers, such as Hanks 'solution, ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art. Any of the IRE-1 α inhibitor compounds disclosed herein, or a prodrug thereof, may be provided, if desired, in a pharmaceutically acceptable pyrogen-free vehicle.

For oral administration, the IRE-1 α inhibitor compound or a prodrug thereof may be mixed with a pharmaceutically acceptable carrier or vehicle, thereby enabling formulation of the IRE-1 α inhibitor compound or prodrug thereof into tablets, pills, lozenges, capsules, liquids, gels, syrups, pastes, suspensions, and the like. Fillers can be utilized, such as gelatin, sugars (e.g., lactose, sucrose, mannitol, or sorbitol); cellulose preparations (e.g., corn starch, wheat starch, rice starch, potato starch, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose); and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate.

The dragee cores may be suitably coated. For this purpose, concentrated sugar solutions may be used, which may optionally contain acacia, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablet or lozenge coating for differentiation.

Pharmaceutical products for oral use include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Push-fit capsules can contain the active ingredient in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the IRE-1 α inhibitor compound or prodrug thereof may be dissolved or suspended in a suitable liquid, such as fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. All formulations for oral administration preferably have dosages suitable for such administration.

For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by inhalation, the pharmaceutical preparations of the present invention may be delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. If desired, a valve may be used to deliver the metered dose. For example, capsules and kits made of gelatin for use in an inhaler or insufflator may be formulated to contain a powder mix of the IRE-1 α inhibitor compound or prodrug thereof and a suitable powder base, such as lactose or starch.

The IRE-1 a inhibitor compound or a prodrug thereof may be formulated to be administered parenterally by injection, for example, by intravenous bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include aqueous solutions of the IRE-1 α inhibitor compounds or prodrugs thereof. Additionally, suspensions of the IRE-1 α inhibitor compounds or prodrugs thereof can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, for example sesame oil, or synthetic fatty acid esters, for example ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. The suspension may optionally further contain suitable stabilizers or agents that increase the solubility of the IRE-1 α inhibitor compound or prodrug thereof, thereby enabling the preparation of highly concentrated solutions.

Alternatively, the IRE-1 α inhibitor compound or prodrug thereof may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

IRE-1 α inhibitor compounds or prodrugs thereof may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described above, IRE-1 α inhibitor compounds or prodrugs thereof may be formulated as long acting articles. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, IRE-1 α inhibitor compounds or prodrugs thereof may be formulated with suitable polymeric or hydrophobic materials (e.g., acceptable oil-formulated emulsions) or ion exchange resins, or as sparingly soluble derivatives, e.g., as a sparingly soluble salt.

The pharmaceutical compositions may also comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to: calcium carbonate, calcium phosphate, various sugars, starch, cellulose derivatives, gelatin, and polymers, such as polyethylene glycol.

In addition to the conventional dosage forms described above, controlled release devices and/or delivery devices, such as those available from azaza CorporationThe IRE-1 alpha inhibitor compound or prodrug thereof is administered by osmotic pump. Suitable delivery devices are described in U.S. patent nos. 3,845,770; 3,916,899; 3,536,809, respectively; 3,598,123, respectively; 3,944,064, and 4,008,719.

Method of treatment

The IRE-1 α inhibitor compounds or prodrugs thereof may be administered to a patient, preferably a human patient, using a pharmaceutical preparation as described above, preferably formulated with a pharmaceutically acceptable pyrogen-free carrier, in an amount effective to treat or alleviate symptoms of a disorder associated with the unfolded protein response.

UPR related diseases

The delicate balance between cell survival and death depends on how the cell controls protein folding stress (protein homeostasis). Imbalances in protein homeostasis lead to a number of metabolic, oncological, neurodegenerative, inflammatory, cardiovascular and infectious diseases (Balch et al, sciences 319,916, 2008). The UPR is specifically associated with protein homeostasis of the endoplasmic reticulum, where all secreted and membrane proteins are translated, folded and processed for delivery to their respective sites of action. Therefore, UPR activation can enhance protein folding in the ER, thereby enabling cell survival. If the protein folding stress in the ER is uncontrolled, the cells begin to apoptosis.

Protein folding stress can be a natural marker for cell types such as insulin-secreting β -islet cells or antibody-secreting plasma cells. In both cases, cells fine-tune the machinery by activating the UPR to cope with stress. Induction or inhibition of UPR may be therapeutically beneficial depending on the type of disease. For example, in type II diabetes or alzheimer's disease, activation of UPR allows β -islet cells to survive the stress of overproducing insulin or neurons to survive the apoptotic action of β -amyloid unfolding aggregates, and thus may be therapeutically beneficial. Modulation of diseases such as cancer, inflammation and viral infections can be treated by inhibition of UPR. In these types of diseases, cell survival due to UPR destruction may be affected. For example, tumor microenvironment conditions such as hypoxia, glucose starvation, amino acid depletion, acidosis and mutant misfolding, and oncogenic proteins negatively affect protein folding in the ER. In addition, chemotherapy, biological therapy and radiotherapy can cause protein folding stress. In these cases, apoptosis may be induced by inhibiting the anti-apoptotic effect of UPR. Myeloma derived from a tumor antibody-secreting plasma cell is an example of a disease to which this method can be applied.

Finally, enveloped viruses must utilize and disrupt the system to ensure the production of progeny of the infected cell. Viruses often produce large amounts of viral membrane glycoproteins that fold and modify in the ER. It is therefore fully conceivable that viruses activate UPRs for this purpose as a survival mechanism. Therefore, it is reasonable that inhibition of UPR during viral infection could beneficially affect disease outcome.

Only specifically differentiated secretory cells and diseased cells activate UPR for their own benefit. Most cells do not experience this protein folding stress and therefore are not affected by UPR inhibitors. Thus, "UPR-associated disease" as used herein refers to a disease in which inhibition of UPR can favorably affect its pathogenesis. In various embodiments of the invention, such inhibition of UPR is achieved by inhibiting IRE-1 α.

In some embodiments, IRE-1 α inhibitor compounds or prodrugs thereof are useful for treating or alleviating the symptoms of B cell autoimmune diseases, certain cancers, and enveloped virus infections that utilize the endoplasmic reticulum as a viral factory to express viral surface and protuberant proteins for budding and infection. IRE-1 alpha inhibitors and prodrugs thereof may be used as single agents or in combination therapy, as described below.

Treatable B-cell autoimmune diseases include, but are not limited to: addison's disease, antiphospholipid syndrome, aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune hypophysitis, autoimmune lymphoproliferative disease, autoimmune myocarditis, churg-Shidi's syndrome, acquired epidermolysis bullosa, giant cell arteritis, Goodpasture's syndrome, Graves ' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, idiopathic thrombocytopenic purpura, IgA nephropathy, myasthenia gravis, pemphigus foliaceus (pemphigus follicaous), pemphigus vulgaris, polyarteritis nodosa, polymyositis/dermatomyositis, rheumatoid arthritis, scleroderma, Yersiglans syndrome, systemic lupus erythematosus, Takayasu's arteritis, and Wegener's granulomatosis.

Cancers that may be treated include, but are not limited to: solid tumors, such as breast tumor, bone tumor, prostate tumor, lung tumor, adrenal tumor (e.g., adrenocortical tumor), bile duct tumor, bladder tumor, bronchial tumor, neural tissue tumor (including neuronal and glioma), gall bladder tumor, stomach tumor, salivary gland tumor, esophageal tumor, small intestine tumor, cervix tumor, colon tumor, rectal tumor, liver tumor, ovary tumor, pancreas tumor, pituitary adenoma, and secretory adenoma. The methods of the invention are particularly useful for treating drug-or radiation-resistant solid tumors.

Hematologic tumors (e.g., lymphomas and leukemias) can also be treated, including but not limited to: multiple myeloma, hodgkin's lymphoma, non-hodgkin's lymphoma (e.g., cutaneous T-cell lymphoma such as sezary syndrome and mycosis fungoides, diffuse large-cell lymphoma, HTLV-1-associated T-cell lymphoma, extranodal peripheral T-cell lymphoma, central nervous system lymphoma, and AIDS-associated lymphoma). Leukemia includes: acute and chronic lymphocytic and myelogenous leukemias (e.g., acute lymphocytic or lymphoblastic leukemia, acute myelogenous leukemia, acute myeloid leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, T-cell prolymphocytic leukemia, adult T-cell leukemia, and hairy cell leukemia). Also, undefined Monoclonal Gammopathy (MGUS), a precursor of myeloma, can be treated.

Viral infections that may be treated include enveloped virus infections (e.g., measles virus, poxviruses, ebola virus, etc.) that utilize the unfolded protein effector pathway in replicating and forming infectious progeny. Infections also include those caused by epstein-barr virus (EBV), Cytomegalovirus (CMV), flaviviruses (e.g., japanese encephalitis virus and west nile virus), and Hepatitis C Virus (HCV).

Combination therapy

Various physiological stresses induce unfolded protein effects including, but not limited to, hypoxia, nutrient starvation, acidosis, and genetic damage, resulting in mutant or overexpressed misfolded proteins (oncogenic stress). One or more of these events are shown in cancer cells, and may be mediated in part by the tumor microenvironment. It is likely that the cytoprotective effect of the unfolded protein effect (UPR) plays an anti-apoptotic role in tumor survival. In addition, biological and chemotherapeutic drugs and radiation therapies may further influence the protein folding and degradation cycle in the ER, thereby inducing UPR as a protective tolerance mechanism. Patients who die from cancer are in need of new therapies and combination therapies because tumors tolerate conventional treatments or revert to a tolerant form after an initial response to treatment.

Angiogenesis inhibitors block tumor growth by inhibiting neovascularization, a process that enhances the stress effects of the tumor microenvironment. A promising approach to further reduce tumor burden is by administering anti-angiogenic agents in combination with IRE-1 α/XBP-1 inhibitors to obtain effects similar to those shown by RNAi knockdown of GRP78, GRP78 being the major chaperone for ER and the target for XBP-1 (Dong et al, Cancer Res.2007, 7/15/67 (14): 6700-7). Furthermore, IRE-1 α itself regulates angiogenesis by affecting VEGF expression.

It is believed that proteasome inhibitors and Hsp90 inhibitors work, in part, by blocking protein degradation and folding and inducing apoptosis, respectively (Davenport et al, Blood2007, month 10, 1; 110(7): 2641-9). Although it is clear that inhibitors of Hsp90 induce XBP-1 splicing and activate UPR, proteasome inhibitors activate IRE-1 α are not clear. Current scientific literature suggests that IRE-1 α is not or only minimally activated by proteasome inhibitors such as Bozoxami or MG-132 (Davenport et al, Blood2007, month 10, day 1; 110(7): 2641-9). However, the data shown in figure 6 demonstrate activation of this pathway in bauzo-tolerant RPMI8226 cells.

Interference with UPR may sensitize cancer cells to various chemotherapeutic agents that increase cellular stress, and therefore, IRE/XBP-1 inhibitors may become important therapeutic agents in combination with current and future standard cancer care.

Although the level of activated IRE-1 α in solid tumors is currently unknown, induction of GRP78 clearly demonstrates induction of UPR in patient biopsies of drug-resistant tumors (Moenner et al, Cancer Res.2007, 11/15 days; 67(22):10631-4; Lee, Cancer Res.2007, 4/15 days; 67(8): 3496-9).

The effect of inhibiting XBP-1 splicing may be higher than expected because the unspliced form of XBP-1 plays a major negative role in XBP-1 and ATF-6 transcriptional activity. Other inhibitors that block the rnase activity of IRE-1 α but not its kinase activity may have the added benefit of signaling through the JNK pathway, which signaling has a pro-apoptotic consequence.

In some embodiments, an IRE-1 α inhibitor compound or prodrug thereof is administered in combination with a therapeutic agent that induces or upregulates expression of IRE-1 α (e.g., an inhibitor of Η sp90 and Η DAC, both of which induce activation of IRE-1 α and XBP-1 splicing) or that has reduced potency upon expression of IRE-1 α (e.g., 17-AAG: (r) (r))And suberoylanilide hydroxamate (SAHA)).

In some embodiments, the IRE-1 α inhibitor compound or prodrug thereof is administered in combination with a cancer therapeutic, such as radiation therapy or a cancer therapeutic (e.g., a chemotherapeutic or biologic), as described below. The cancer therapeutic agent may be administered separately or together with an IRE-1 alpha inhibitor compound. The cancer therapeutic may be administered at about the same time as the IRE-1 α inhibitor compound, or may be administered before or after the IRE-1 α inhibitor compound.

Cancer therapeutic agents useful in the present invention include, but are not limited to, the following classes of agents (which may overlap):

a. proteasome inhibitors, e.g. porizol ([ (1R) -3-methyl-1- [ [ (2S) -1-oxo-3-phenyl-2- [ (pyrazinylcarbonyl) amino]Propyl radical]Amino group]Butyl radical]Boric acid; MG-341;) MG-132(N- [ (phenylmethoxy) carbonyl)]-L-leucyl-N- [ (1S) -1-formyl-3-methylbutyl]-L-leucinamide);

b. antimetabolites, for example:

i. pyrimidine analogs (e.g., 5-fluorouracil, floxuridine, capecitabine, gemcitabine, and cytarabine);

a purine analog;

folic acid antagonists and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine [ cladribine ]);

folic acid analogs (e.g., methotrexate);

c. an anti-mitotic agent comprising:

i. natural products, such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine);

alkylating agents, such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide and the like, melphalan, toconine), ethyleneimine and methylmelamines (e.g., hexamethylmelamine and thiotepa), alkyl sulfates-busulfan, nitrosoureas (e.g., carmustine (BCNU) and the like, streptozotocin), triazenes (trazenes) -azenamines (DTIC);

d. microtubule disrupting agents, such as taxanes (paclitaxel, docetaxel), vincristine, vinblastine, nocodazole, epothilones (epothilones) and novabine, and etoposide (epipodophyllotoxins) (e.g., teniposide);

dna damaging agents, for example, actinomycin, amsacrine, anthracyclines, bleomycin, busulfan, camptothecin, carboplatin, meclizine, cisplatin, cyclophosphamide (Cytoxan), dactinomycin, daunorubicin, docetaxel, doxorubicin, epirubicin, hexamethylmelamine oxaliplatin, ifosfamide, melphalan, mechlorethamine (merchlorethamine), mitomycin, mitoxantrone, nitrosoureas, paclitaxel, plicamycin, procarbazine, teniposide, triethylenephosphoramide, and etoposide (VP16);

f. antibiotics, such as dactinomycin (actinomycin D), daunorubicin, doxorubicin (adriamycin), idarubicin, anthracyclines, mitoxantrone, bleomycin, mithramycin (plicamycin), and mitomycin;

g. enzymes, such as L-asparaginase;

h. anti-platelet agents;

i. platinum coordination complexes (e.g., cisplatin, carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide;

j. hormones, hormone analogs (e.g., estrogen, tamoxifen, goserelin, bicalutamide, nilutamide);

k. aromatase inhibitors (e.g., letrozole, anastrozole);

anticoagulants (e.g., heparin, synthetic heparin salts, and other inhibitors of thrombin);

fibrinolytic agents (e.g., tissue plasminogen activator, streptokinase, and urokinase), aspirin, COX-2 inhibitors, dipyridamole, ticlopidine, clopidogrel, abciximab;

anti-migratory agents (antimigrating agents);

antisecretory agents (e.g., beliadine (breveldin)); immunosuppressants (e.g., cyclosporin, tacrolimus (FK-506), sirolimus (rapamycin), azathioprine, mycophenolate mofetil);

anti-angiogenic compounds (e.g., TNP-470, genistein) and growth factor inhibitors (e.g., Vascular Endothelial Growth Factor (VEGF) inhibitors, Fibroblast Growth Factor (FGF) inhibitors, Epidermal Growth Factor (EGF) inhibitors);

an angiotensin receptor blocker;

r. a nitric oxide donor;

s. an antisense oligonucleotide;

t. antibodies (e.g., trastuzumab: (a))、、)；

Cell cycle inhibitors and differentiation inducers (e.g., tretinoin);

mtor (mammalian target of rapamycin) inhibitors (e.g., everolimus, sirolimus);

topoisomerase inhibitors (e.g., doxorubicin (adriamycin), amsacrine, camptothecin, daunorubicin, dactinomycin, geniposide, epirubicin, etoposide, idarubicin, irinotecan (CPT-11) and mitoxantrone, topotecan, irinotecan);

corticosteroids (e.g., cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, and prednisolone);

y. growth factor signal transduction kinase inhibitors;

z. mitochondrial dysfunction inducing agent;

a caspase activator;

a chromatin disrupting agent.

In some embodiments, the cancer therapeutic agent is selected from the group consisting of: alemtuzumab, aminoglutethimide, amsacrine, anastrozole, asparaginase, beger (beg), bevacizumab, bicalutamide, bleomycin, porzolamide, buserelin, busulfan, camptothecin, capecitabine, carboplatin, carmustine, CeaVac, cetuximab, oncoclonine, cisplatin, cladribine, clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, daclizumab, dactinomycin, daunorubicin, hexadiene estrol, diethylstilbestrol, docetaxel, doxorubicin, eculizumab, epirubicin, epratuzumab, erlotinib, estradiol, estramustine, etoposide, exemestane, filgrastimethacin, fludarabine, fludrocortisone, fluorouracil, flutolytridazine, gemcitabine, certamide, genistein, gefitinib, morbixin, and gent, huJ591, hydroxyurea, ibritumomab (ibritumomab), idarubicin, ifosfamide, IGN-101, imatinib (imatinib), interferon, irinotecan (ironotecan), letrozole, folinic acid, leuprorelin acetate, levamisole, lintuzumab, lomustine, MDX-210, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, mitolimumab, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, disodium pamidronate, pentostatin, pertuzumab (pertuzumab), mithramycin, porphyrin, procarbazine, ranitidine, rituximab, streptozocin, sunitinib, sultinib, temozoloside, timoloside, timothricin, and a, Thiotepa, cyclopentadienium dichloride, topotecan, tositumomab, trastuzumab, retinoic acid, vatalanib (vatalanib), vinblastine, vincristine, vindesine, and vinorelbine.

Route of administration

The pharmaceutical preparation of the invention may be administered locally or systemically. Suitable routes of administration include oral, pulmonary, rectal, transmucosal, small intestinal, parenteral (including intramuscular, subcutaneous, intramedullary routes), intranodal, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, transdermal, topical and vaginal routes. As detailed above, dosage forms include, but are not limited to, tablets, lozenges, dispersions, suspensions, suppositories, solutions, capsules, creams, patches, micropumps, and the like. Targeted delivery systems (e.g., liposomes coated with target-specific antibodies) can also be utilized.

Dosage form

The pharmaceutical compositions of the present invention comprise a therapeutically effective dose of at least one active ingredient (an IRE-1 α inhibitor compound or a prodrug thereof). "therapeutically effective dose" refers to an amount of an IRE-1 α inhibitor compound or prodrug thereof that, when administered to a patient during treatment, results in a detectable improvement in the characteristics of the disease being treated (e.g., a numerical improvement in the laboratory, a delay in the progression of symptoms, a reduction in the severity of symptoms, or an improvement in the level of an appropriate biological marker).

The therapeutically effective dose is well within the skill of the art. The therapeutically effective dose can first be estimated from in vitro enzyme assays, cell culture assays, and/or animal models. For example, the dose designed in animal models can achieve circulating concentrations in a range of at least the IC50 determined in an in vitro enzyme assay or cell culture (i.e., the concentration of test compound that achieves half-maximal inhibition of IRE-1 α activity). This information can be used to more accurately determine useful human dosages. See FDA Guidance, "Industrial and review guidelines for evaluating Safe Starting doses in Clinical Trials of Therapeutics in adult healthy Volunteers" (guide for Industry and Reviewersystemation) the Safe Starting Dose in Clinical Trials for Therapeutics in humans (HFA-305), which provides a useful formula for calculating Human Equivalent Dose (HED) from in vivo studies in animals.

Suitable animal models for the relevant diseases are known in the art. See, e.g., lupus.1996, month 10; 451-5 (antiphospholipid syndrome); blood.1974 month 7; 44(1) 49-56 (aplastic anemia); autoimmitude.2001; 33(4) 265-74 (autoimmune hypophysitis); methods.2007 month 1; 41(1) 118-22 (autoimmune myocarditis); clin Exp Rheumatol.2003, 11-12 months; 21(6 suppl 32) S55-63 (churg-strauss syndrome, wegener' S granulomatosis); j Clin invest.2005, month 4; 115(4) 870-8 (epidermolysis bullosa acquired); circulation.2005 month 6, 14; 111(23) 3135-40.Epub 6/2005 (giant cell arteritis; takayu' arteritis)); int J Immunopathol pharmacol.2005, 10 months-12 months; 18(4) 701-8(IgA nephropathy); vet Rec.12/5 1984; 114(19) 479 (pemphigus foliaceus); neuropimumunal.98, 130-35,1999 (polymyositis); am.J. Pathol.120,323-25,1985 (dermatomyositis); cell.mol.immunol.2,461-65,2005 (myasthenia gravis); arthritis Rheum.50,3250-59,2004 (lupus erythematosus); clin. exp. Immunol.99,294-302,1995 (Grave); clin. invest.116,961-973,2006 (rheumatoid arthritis); exp Mol Pathol.77,161-67,2004 (Hashimoto thyroiditis); rheumatol.32,1071-75,2005 (Sjogren's syndrome); brain Pathol.12,420-29,2002 (Guillain Barre syndrome); vet. Pathol.32,337-45,1995 (polyarteritis nodosa); immunol. invest.3,47-61,2006 (pemphigus vulgaris); Arch.Dermatol.Res.297,333-44,2006 (scleroderma); exp.med.191,899-906,2000 (goodpasture syndrome); clin. exp. Immunol.99,294-302,1995 (Grave); clin. invest.91,1507-15,1993 (membranous nephropathy); J.Immunol.169,4889-96,2002 (autoimmune hepatitis); surgery128,999-1006,2000 (Addison disease); eur.j.immunol.32,1147-56,2002 (autoimmune hemolytic anemia); and Haematologica88,679-87,2003 (autoimmune thrombocytopenic purpura).

LD can be determined by standard pharmaceutical procedures in cell cultures and/or experimental animals₅₀(dose lethal to 50% of the population) and ED₅₀(a therapeutically effective dose in 50% of the population). Initial human dosages can be determined using data obtained from cell culture assays or animal studies. As is known in the art, the dosage may vary depending on the dosage form and route of administration used.

Useful doses for systemic administration to a human patient are in the range of 1. mu.g/kg to 100mg/kg (e.g., 1-10. mu.g/kg, 20-80. mu.g/kg, 5-50. mu.g/kg, 75-150. mu.g/kg, 100-500. mu.g/kg, 250-750. mu.g/kg, 500-1000. mu.g/kg, 1-10mg/kg, 5-50mg/kg, 25-75mg/kg, 50-100mg/kg, 5mg/kg, 20mg/kg or 50 mg/kg). In some embodiments, a treatment regimen may require that the plasma concentration of an IRE-1 α inhibitor compound be maintained for a certain period (e.g., days or weeks) and then decay after a certain period of rest (e.g., 1,2,3, or 4 weeks). The amount of the composition administered will, of course, depend on the subject being treated, the weight of the subject, the severity of the disease, the mode of administration and the judgment of the prescribing physician.

All patents, patent applications, and references cited herein are expressly incorporated herein by reference. The present invention has been generally described above. It will be more fully understood by reference to the following specific examples, which are provided for purposes of illustration only and are not intended to limit the scope of the invention.

Drawings

FIG. 1 shows the results of cell-based IRE-1. alpha. XBP-1-specific endoribonuclease inhibition using 6-bromo-o-vanillin. 12 μ L DMSO was 1.2%.

FIG. 2 shows the results of the inhibition of cell-based IRE-1. alpha. XBP-1-specific endoribonuclease in human myeloma cells.

FIG. 3 is a photograph of an agarose gel scan showing PCR products from IRE-1. alpha. inhibitor cell assays, showing that the dose-dependence of various IRE-1. alpha. inhibitors inhibits cellular XBP-1 splicing. XBP-1u, unspliced XBP-1; XBP-1s, spliced SBP-1; EC (EC)₅₀IRE-1. alpha. inhibitors inhibit DTT-induced cellular XBP-1 splicing by 50% (μ M). The values on the lanes indicate the concentration of each compound (in. mu.M). Mm.1s myeloma cells were treated with active or inactive compounds for 2 hours and then with DTT for 1 hour. RT-PCR was performed using human XBP-1 specific primers flanked by intron regions. Induction of UPR stress (S) by DTT resulted in the removal of a 26 nucleotide fragment, resulting in a lower band compared to unstressed cells (U) (higher band). EC (EC)₅₀The concentration of XBP-1 induced splicing was determined to inhibit 50% of DTT. EC of Compound 17-1₅₀About 2-3. mu.M.

FIG. 4 shows that IRE-1. alpha. inhibitors reversibly inhibit the activated form of IRE-1. alpha. in cells. Cellular inhibition of XBP-1 splicing was tested in HEK293 cells using 10. mu.M Compound 2. FIG. 4A uses standard RT-PCR to show the relative amounts of spliced XBP-1 after addition of 2mM DTT and maintenance in broth (. tangle-solidup.) or washing out DTT 30 minutes (. diamond-solid.) or 1 hour after induction (■). When cells are stressed with DTT, XBP-1 messenger RNA is rapidly converted to the spliced form. Conversely, when stress is removed, the cells rapidly degrade spliced XBP-1 and replace it in an unspliced form. Figure 4B shows that compound 2 added to DTT stressed cells at 2 hours before induction of DTT (■) or at 1 hour after induction (a-solidup), the unspliced form accumulated rapidly, similar to removal of DTT stress, suggesting that the compound inhibited the activated form of the enzyme. When the compound was washed out to maintain DTT stress, spliced XBP-1 increased within hours after the inhibition was completed, suggesting that the inhibitory effect was reversible (■, X). Percent splicing was determined by scanning the gel for unspliced and spliced XBP-1 bands (as depicted in FIG. 3). The enzyme activity is expressed as the percentage of spliced XBP-1 on the Y axis (calculated as the splice content divided by the total amount of spliced and unspliced XBP-1).

FIG. 5 shows the inhibition of multiple myeloma cell proliferation by IRE-1. alpha. inhibitors 11-28 (example 11). RPMI-8226 multiple myeloma cells were seeded at 20,000 cells/well in RPMI medium containing 1% FBS and the desired antibiotic. The plate was heated at 37 deg.C with 95% air and 5% CO₂The mixture was incubated overnight. The following day, compounds 11-28 or medium were added individually to each well to a final volume of 100 microliters/well. The concentration of compounds was 100. mu.M-0. mu.M, with 4-fold dilutions of each compound. After addition of the compound, the plate was incubated at 37 deg.C, 95% air, 5% CO₂The mixture was incubated for 24 hours. Cell proliferation was measured using the Promega CTG assay (CellTiter-Glo assay, Promega) according to the manufacturer's instructions.

FIG. 6 Western blot (FIG. 6A) and agarose gel (FIG. 6B) show the binding of Bortezomib (MG-341;) Treatment of RPMI8226 cells for 24 hours increased the levels of phosphorylated IRE-1 α and XBP 1-splicing. The values indicate the concentration of bauzo in nM.

FIG. 7 shows the enhancement of apoptosis in myeloma cells by proteasome inhibitors MG-132(N- [ (phenylmethoxy) carbonyl ] -L-leucyl-N- [ (1S) -1-formyl-3-methylbutyl ] -L-leucinamide) and IRE-1 α/XBP-1 specific inhibitors as reflected by relative caspase activities (total caspase 3 and caspase 7 activity). FIG. 7A, 100nM MG-132; FIG. 7B, 200nM MG-132.

FIG. 8 results of in vivo experiments of IRE-1. alpha. inhibitors in mouse tissues. FIG. 8A, protocol for tunicamycin and IRE-1. alpha. inhibitor treatment. FIG. 8B, agarose gel showing IRE-1. alpha. specific XBP-1 splicing to most of the inactivated RT-PCR products in the kidney, liver and spleen of NOD-SCID mice. FIG. 8C, treatment with tunicamycin for 6 hours resulted in significant levels of spliced XBP-1(Wu et al, 2007). FIG. 8C, agarose gel of RT-PCR product with reduced levels of spliced XBP-1 in mice treated with IRE-1 α inhibitor 4 hours after intraperitoneal treatment with tunicamycin. FIG. 8D, is a graph showing the average relative percentage of spliced XBP-1 to total XBP-1 for each group of two mice in FIGS. 8B and 8C. The numbers above parentheses in fig. 8B and 8C are the mouse numbers (mouse 3, mouse 4, etc.). FIG. 8D, is a graph showing the average relative percentage of spliced XBP-1 to total XBP-1 for each group of two mice in FIGS. 8B and 8C.

FIG. 9 IgM secretion after LPS stimulation of primary murine B cells was inhibited with selected IRE-1. alpha. inhibitors. Compound 17-1 blocked IgM secretion at all doses tested with a minimum of 100nM when added at the beginning of stimulation and 24 hours post-stimulation. However, when added 40 hours after stimulation, the compound did not work much; there was only slight inhibition at the highest dose. The methods of B cell stimulation, plasma cell differentiation and IgM secretion were performed as previously described by Iwakoshi et al (Nature4,321-29,2003). Primary B cells were isolated from BALB/c spleen using mouse CD43 microbeads (Miltenyi catalog No. 130-049-801) at 1X106 cells per treatment. In a 24-well plate, at a final density of 1 × 10⁶Purified B cells were stimulated with 20. mu.g/ml LPS (Sigma Cat. No. L4391) in a/ml/well B cell culture. Various concentrations (50 μ M, 10 μ M, 2 μ M,0.4 μ M, and 0.08 μ M) were added at the indicated time points (t =0, t =24 hours, t =40 hours, etc.)M) IRE-1. alpha. inhibitor Compound 17-1. Cells were incubated at 37 ℃ for 48 hours. At the end of the incubation, the cells were centrifuged in the plate at 1500rpm for 3 minutes. Supernatants were collected and IgM secretion was quantitatively determined using a mouse IgM ELISA kit (Bethyyl Labs cat # E90-101). B cell culture media included RPMI +10% FBS supplemented with NEAA, HEPES, NaPyr, PSQ, and β -mercaptoethanol.

Detailed Description

Example 1

IRE-1. alpha. assay

A fusion protein comprising Glutathione S Transferase (GST) and human IRE-1 α (GST-IRE-1 α) was obtained from 500ml of baculovirus-infected insect cell culture for in vitro detection of IRE-1 α activity.

Will contain 1 Xreaction buffer (5 Xreaction buffer is 100mM Hepes pH7.5, 250mM KOAc, 2.5mM MgCl₂) Mu.l of reaction mixture, 3mM DTT and 0.4% polyethylene glycol water was added to each well of the 384-well plate. 25 nanoliters of a 1mM test compound solution were added to the test wells. Mu.l of 128ng/ml IRE-1. alpha. preparation was added to each test well and positive control well (final concentration 5.82 ng/well). Negative control wells contained only reaction mixture and test compound.

After centrifugation of the plate at 1200rpm for 30 seconds, 3. mu.l of IRE-1. alpha. human mini (mini) -XBP-1mRNA stem-loop substrate 5'-CAGUCCGCAGCACUG-3' (SEQ ID NO:1) labeled with the fluorescent dye Cy5 at the 5 'end and Black Hole Quencher2(BH2) at the 3' end was added to each well of the control plate. The plate was again centrifuged at 1200rpm for 30 seconds. The final concentrations used for the experiments were: 63nM IRE-1. alpha. substrate, 5.82ng IRE-1. alpha. protein and 2.5. mu.M test compound.

Each plate was covered with a lid and incubated at 30 ℃ for 1 hour. The plates were then transferred to ACQUEST^TMMicroplate reader (microplate reader). The data was analyzed using data analysis software to calculate the percent activity of IRE-1 α.

Example 2

Identification of IRE-1 alpha inhibitor Compounds

Compounds were screened from the fischer bridge laboratory (Maybridge library, Fisher) using the assay described in example 1. Approximately 60 compounds were selected as confirmation hits and re-purified. These compounds are aryl imine or Schiff base adducts of 2-hydroxybenzaldehyde analogs. There was no observable SAR with respect to the R group. However, after re-purification by HPLC, it was noted that these compounds decomposed into their structural components: 2-hydroxybenzaldehyde derivatives and primary amines attached to the R group, which suggests that the aldehyde derivative may be an active component of the compound.

Three purified 2-hydroxybenzaldehydes with halogen (Cl, Br or I) in the 3 and 5 positions were then tested in an IRE-1. alpha. assay. All 3 were active. The most potent is 3,5 iodo 2-hydroxybenzaldehyde (IC)₅₀0.35 μ M), followed by 3,5 bromo 2-hydroxybenzaldehyde (IC)₅₀0.46 μ M), and finally 3,5 chloro 2-hydroxybenzaldehyde (1.05 μ M).

About 20 benzaldehyde derivatives were then purchased and tested in the IRE-1 α test. The results of this test indicate that the compounds require a hydroxyl group in the ortho position to the aldehyde group and also require a hydrophobic electron withdrawing group in the 3,5 or 6 position of the benzene ring. The 3-and 5-positions may be halogen or methoxy or ethoxy. The nitro group at the 3-or 5-position is active, but not both. The most potent compound is o-vanillin containing a bromo substituent at the 5 or 6 position. Without wishing to be limited to the following explanation, the hydrogen of the vicinal hydroxyl group may participate in the binding of the hydrogen of the stable configuration to the aldehyde oxygen.

Example 3

Examples of o-vanillin with SAR and selectivity for IRE-1 alpha in vitro enzyme assays

IRE-1. alpha., T1 RNAse and RNAse A assays were performed in vitro with several o-vanillin derivatives to confirm the selectivity of these derivatives for IRE-1. alpha. The IRE-1. alpha. assay was performed as described in example 1.

The T1 RNase assay was performed as follows. Comprises 1 Xreaction buffer (5 Xreaction buffer is 100mM Hepes pH7.5, 250mM KOAc, 2.5mM MgCl)₂) Mu.l of reaction mixture, 3mM DTT and 0.4% polyethylene glycol water was added to each well of the 384-well plate. 25 nanoliters of a 1mM test compound solution were added to the test wells. Approximately 200,000U/ml of a 3. mu.l 1/48,000 dilution of a preparation of RNase T1 (Wooxington) was added to each test well and positive control well (final concentration 49.5 picograms/well). Negative control wells contained only reaction mixture and test compound.

After centrifuging the plate at 1200rpm for 30 seconds, 3. mu.l of mini-XBP-1 mRNA stem-loop substrate described in example 1 was added to each well of the control plate. Plates were again centrifuged at 1200rpm for 30 seconds. The final concentrations used for this assay were: 63nM substrate, 49.5pg RNase T1 and 2.5. mu.M test compound.

Each plate was covered with a lid and incubated at 30 ℃ for 1 hour. The plates were then transferred to ACQUEST^TMMicroplate reader. The data was analyzed using data analysis software. The percentage of activity of rnase T1 was calculated.

The RNase A assay was performed as described for RNase T1. The final concentrations used for the experiments were: 63nM substrate, 0.4pg RNase A (Qiagen; 100mg/ml or 7000U/ml) and 2.5. mu.M test compound.

Test Compounds are selective for IRE-1, IC₅₀3 μ M (o-vanillin), 1 μ M (3-ethoxyo-vanillin) and 30nm (6-bromoo-vanillin).

Example 4

Cell-based IRE-1 alpha XBP-1-specific endoribonuclease inhibition of 6-bromo-o-vanillin

Preliminary cell-based XBP-1mRNA splicing experiments demonstrated that several potent 5-bromo and 6-bromo ortho-vanillin inhibited IRE-1 α. HEK293 cells were incubated overnight or 2 hours with the compound and IRE-1. alpha. was activated with the UPR inducer thapsigargin. IRE-1 α mediated XBP-1 splicing was detected by RT-PCR using XBP-1 specific primers flanked by 26bp introns cleaved by IRE-1 α. The results are shown in FIG. 1. It can be observed that at higher concentrations, more of unspliced XBP-1 (higher band: substrate) is present compared to the spliced form (lower band: product).

Without wishing to be limited to this explanation, the aldehyde apparently forms a reversible schiff base with the primary amine of lysine at the active site of the enzyme. The ortho-hydroxyl group can accelerate and stabilize the schiff base. Furthermore, the unpaired electron pair may serve as a hydrogen bond acceptor with other amino acids of IRE-1 α. The benzene ring and various R groups may be maintained in the hydrophobic pocket of the enzyme linked by the schiff base of the aldehyde moiety. This ability is greatly facilitated by the electron-withdrawing and hydrophobic nature of the substituents at the 3 and 5 positions. Due to the hydrophobic character of o-vanillin, these compounds may also be suitable for hydrophobic pockets in addition to forming schiff bases.

Example 5

Determination of IC for inhibition of IRE-1. alpha₅₀

Detection of IC of IRE-1. alpha. inhibitory Compounds identified in Table 3 as described in example 1₅₀。

Table 3.

Example 6

Kinase selectivity assays

The ability of compounds to inhibit 86 different kinases was tested at a concentration of 10. mu.M, which is much higher than the IC of each compound₅₀(3.71 and 0.027. mu.M, respectively). The results of the assay demonstrate that these compounds are selective for IRE-1 α.

Example 7

Synthesis of 2' -chloro-4-hydroxy-5-methoxybiphenyl-3-carbaldehyde (carbaldhyde)

In a 5ml microwave vial was added 2-chlorophenylboronic acid (54.73 mg, 0.35 mmol, 1.16 equiv.), tetrakis (triphenylphosphine) palladium (0) (7 mg, 0.006 mmol,2 mol%) as catalyst and a solution of 5-bromo-2-hydroxy-3-methoxy-benzaldehyde (69.3 mg,0.3mmol, 1 equiv.) in 1ml of MeCN. To the resulting solution was added 1M K₂CO₃The solution (0.6 ml, 0.6 mmol,2 equiv.) was then sealed. The reaction mixture was heated at 150 ℃ for 360 seconds using a PCSC Microwave oven (Personal Chemistry Smith Creator Microwave). After completion, the organic layer was transferred to one well of a 96-well plate. The solvent was evaporated and the residue was dissolved in 0.6 ml of 0.5% TFA in DMSO and purified.

Example 8

Synthesis of 2' -chloro-3-hydroxy-4-methoxybiphenyl-2-carbaldehyde

In a 5ml microwave vial was added 2-chlorophenylboronic acid (54.73 mg, 0.35 mmol, 1.16 equiv.), tetrakis (triphenylphosphine) palladium (0) (7 mg, 0.006 mmol,2 mol%) as catalyst and a solution of 6-bromo-2-hydroxy-3-methoxy-benzaldehyde (69.3 mg,0.3mmol, 1 equiv.) in 1ml of MeCN. To the resulting solution was added 1M K₂CO₃The solution (0.6 ml, 0.6 mmol,2 equiv.) was then sealed. The reaction mixture was heated at 150 ℃ for 360 seconds using a PCSC microwave oven. After completion, the organic layer was transferred to one well of a 96-well plate. The solvent was evaporated and the residue was dissolved in 0.6 ml of 0.5% TFA in DMSO and purified.

Example 8

Synthesis of 4-bromo-2- { [ (E) -4-fluoro-phenylimino ] -methyl } -phenol

5-Bromosalicylaldehyde (100mg,0.50mmol), toluene (5ml) and activated molecular sieves (200mg) were added to a 20ml scintillation vial. To the resulting solution was added 4-fluoroaniline (56mg,0.50mmol,2 equiv.). The reaction mixture was heated at 100 ℃ for 16 hours, after which the molecular sieve was filtered off from the solution and washed with dichloromethane. The precipitated product was collected by filtration and washed with hexane. After drying, identification was performed by NMR and TLC.

Example 10

Cell assay

Human melanoma MM.1s cells were incubated with the indicated amounts of compound for 1.25 hours before being stressed with 2mM Dithiothreitol (DTT). And compoundsAfter a further 45 minutes incubation with DTT (2 hours in total), the cells were incubated withCells were harvested (monophasic solution of phenol and guanidinium isothiocyanate) and total RNA was prepared according to the manufacturer's instructions (Invitrogen). Human XBP-1 was amplified by RT-PCR using primers flanking a 26 base non-canonical intron cleaved by IRE-1 α:

CCTGGTTGCTGAAGAGGAGG (SEQ ID NO:2) (forward direction) and

CCATGGGG AGATGTTCTGGAG (SEQ ID NO:3) (reverse).

The results are shown in FIG. 2. In cells that have not been stressed, IRE-1. alpha. is inactive, and thus the 26 base intron remains in the XBP-1 mRNA. RT-PCR of cells not stressed (U) then produced a higher band. When cells are stressed with the Endoplasmic Reticulum (ER) stressor DTT (S), IRE-1. alpha. is activated by accumulation of unfolded protein, resulting in a 26 base pair shorter RT-PCR product (lower band). The transition from the lower band to the higher band demonstrates that increased compound usage blocks IRE-1 α mediated XBP-1 splicing. In vitro enzyme assays, compound potency reflects SAR.

Cellular ED determination of IRE-1 alpha inhibitors₅₀

Examination of cellular EC for Compounds that pass specificity assays in myeloma cells using endogenous XBP-1 splicing₅₀. IRE-1 α highly specific endoribonuclease activity regulates XBP-1 by cleaving a 26 nucleotide intron from XBP-1 mRNA. This splicing event induces a frame shift in the ORF at the C-terminus of XBP-1, resulting in the translation of a larger 54kD active transcription factor, rather than the inactive 33kD form. This splicing event is used to detect IRE-1 α activity on XBP-1mRNA in cells and tissues.

Briefly, compounds are incubated in the presence or absence of an ER stressor (e.g., DTT) and the ratio of XBP-1u (unspliced) to XBP-1s (spliced) is quantitatively determined by RT-PCR. ED (electronic device)₅₀Determined as 50% X relative to the total XPB-1 levelBP-1 (FIG. 3). EC (EC)₅₀Compounds at or below 10. mu.M for standard apoptosis assays including annexin V staining andfig. 5 and 7).

Determination of ED Using a proliferation assay with myeloma cell lines (U266, RPMI8226 and MM.1s)₅₀. The compounds are useful as single agents as well as in combination with other chemotherapeutic agents. As shown in FIG. 5, IRE-1. alpha. inhibitor 11-28 compounds inhibited the proliferation of RPMI8226 myeloma cells that have an endogenous activation pathway and were further induced by the addition of bauzomib (FIG. 6). When IRE-1. alpha. inhibitor compound 2 was used in combination with MG-132, an increase in U266 myeloma apoptosis was observed (FIG. 7).

Example 11

Synthesis of 3' -formyl-4 ' -hydroxy-5 ' -methoxybiphenyl-3-carboxylic acid

5-bromo-2-hydroxy-3-methoxybenzaldehyde (3.00g,13.0mmol), 3-carboxy-phenylboronic acid (2.37g,14.3mmol), sodium carbonate (8.27g,78.0mmol) and tetrakis (triphenylphosphine) palladium (0.728g,0.65mmol) were dissolved in a mixture of 200mL DMF and 200mL water. The reaction was stirred at 105 ℃ for 5 hours under an argon atmosphere. 200mL of 1N sodium hydroxide was added and the solution was extracted with dichloromethane (3X100 mL). The aqueous layer was acidified with 6N hydrochloric acid, the precipitated material was filtered off and washed successively with water and ether to give 11-1(1.70g,6.25mmol, 48%).¹H NMR(400MHz,DMSO-d₆)δppm13.07(br.s,1H),10.34(s,1H),10.44(br.s,1H),8.18(t,J=1.6Hz,1H),7.90-7.97(m,2H),7.59(t,J=7.8Hz,1H),7.55(s,2H),3.97(s,3H).

The following compounds were prepared by the above method using the corresponding aryl bromide and aryl boronic acid and characterized by LC/MS using Waters UPLC/MS equipped with a UV detector (220nM) and an MS detector (ESI). HPLC column: acquity BEH C181.7 μm (Waters)2.1mm x50 mm. HPLC gradient: 0.6 ml/min, 1.5 min from 95:5 of 20mM ammonium formate buffer (adjusted to pH7.4 with ammonium hydroxide) acetonitrile to 20:80 ammonium formate buffer acetonitrile, held for 1.3 min.

Table 4.

The following compounds were prepared by the above method using the corresponding aryl bromide and aryl boronic acid and characterized by NMR.

TABLE 5

Example 12

Synthesis of N-cyclohexyl-3 ' -formyl-4 ' -hydroxy-5 ' -methoxybiphenyl-3-carboxamide

N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (42mg,0.22mmol), 1-hydroxybenzotriazole (30mg,0.22mmol), triethylamine (140. mu.L, 1mmol) and cyclohexylamine (50. mu.L, 0.44mmol) were added to a solution of 11-1(54mg,0.2mmol) in 2mL THF at room temperature. After 2 hours, the reaction was diluted with 2mL of 2N hydrochloric acid, stirred for 2 hours and then evaporated to dryness. The residue was dissolved in 2mL of chloroform, and extracted with water (1x1.5mL), 1N hydrochloric acid (1x1.5mL), water (1x1.5mL), saturated sodium bicarbonate (1x1.5mL), and water (1 x1.5mL). The organic phase was evaporated and the crude product was purified by preparative HPLC, followed by recrystallization from ether to give 12-1(16mg,0.05mmol, 25%).¹H NMR(400MHz,CDCl₃)δppm11.07(s,1H),10.00(s,1H),8.00(t,J=1.8Hz,1H),7.64-7.69(m,2H),7.50(t,J=7.8Hz,1H),7.42(d,J=2.3Hz,1H),7.35(d,J=2.0Hz,1H),6.01(d,J=7.8Hz,1H),3.97-4.06(m,4H),2.03-2.11(m,2H),1.73-1.82(m,2H),1.63-1.71(m,1H),1.40-1.51(m,2H),1.23-1.32(m,3H)。

The following compounds were prepared by the above method using the corresponding aryl acid and amine and characterized by NMR.

TABLE 6

Example 13

Synthesis of 6-bromo-2-hydroxy-3- (morpholine-4-carbonyl) benzaldehyde

4-bromo-3-formyl-2-hydroxybenzoic acid (122mg,0.5mmol) was dissolved in 5mL dry THF. Phosphorus pentachloride (115mg,0.55mmol) was added at 0 deg.C and the mixture stirred for 20 minutes. The mixture was added dropwise to a solution of morpholine (433 μ L,5mmol) in 20mL dry THF at-10 ℃. The reaction was warmed to room temperature and stirred for 30 minutes. The volatiles were evaporated and the residue was dissolved with 15mL of 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was evaporated and the crude product obtained was purified by column chromatography to give 13-1(25mg,0.08mmol, 16%).¹H NMR(400MHz,CDCl₃)δppm12.33(s,1H),10.34(s,1H),7.41(d,J=8.0Hz,1H),7.25(d,J=8.0Hz,1H),3.78(br.s,4H),3.66(br.s,2H),3.32(br.s,2H)。

The following compounds were prepared by the above method and characterized by LC/MS.

TABLE 7

Example 14

Synthesis of 5- (1, 3-dimethyl-2, 4-dioxo-1, 2,3, 4-tetrahydropyrimidin-5-yl) -2-hydroxy-3-methoxybenzaldehyde

5-bromo-2-hydroxy-3-methoxybenzaldehyde (3.00g;13.0mmol), bis-pinacolato-diboron (3.63g;14.3mmol), potassium acetate (3.80;39.0mmol) and Pd (dppf) Cl2(1.10g;1.50mmol) were dissolved in bisAlkane and argon are added for refluxing for 4 hours. The reaction mixture was cooled, filtered and the filtrate was evaporated to dryness under reduced pressure. The solid residue was purified by column chromatography on silica using dichloromethane as eluent. The collected pale yellow solid was triturated with diisopropyl ether to give 14a (1.45g,5.22mmol, 40%).¹H NMR(400MHz,CDCl₃)δppm11.36(s,1H),9.93(s,1H),7.69(d,J=1.3Hz,1H),7.49(s,1H),3.96(s,3H),1.36(s,12H)。

5-bromo-1, 3-dimethyluracil (88mg,0.4mmol), 14a (117mg,0.4mmol) and anhydrous sodium carbonate (254mg,2.4mmol) were dissolved in a mixture of 6mL DMF and 6mL water. Tetrakis (triphenylphosphine) palladium (22mg,0.02mmol) was added and the reaction heated to 110 ℃ under an argon atmosphere for 1 hour. 40mL of saturated sodium chloride solution was added and the mixture was extracted with chloroform (2X40 mL). The organic layer was dried and evaporated and the residue purified by column chromatography to give 14-1(37mg,0.13mmol,32%).¹H NMR(400MHz,CDCl₃)δppm11.00(s,1H),9.95(s,1H),7.35(d,J=1.8Hz,1H),7.33(dd,J=9.3,2.0Hz,2H),3.96(s,3H),3.51(s,3H),3.44(s,3H)。

The following compounds were prepared by the above method using the corresponding aryl bromide and characterized by LC/MS.

TABLE 8

TABLE 9

Example 15

Synthesis of 2-hydroxy-3-methoxy-5- (pyridin-3-ylethynyl) benzaldehyde

2-hydroxy-5-iodo-3-methoxybenzaldehyde (2.08g;7.5mmol), ethynyl-trimethylsilane (2.65mL,1.8mmol), Pd (PPh)₃)₂Cl₂(158mg;0.23mmol) and copper (I) iodide (43mg;0.23mmol) were dissolved in 40mL of triethylamine and heated at 60 ℃ for 4 hours. The mixture was cooled to room temperature, filtered and the filtrate was evaporated. The solid residue was purified by column chromatography on silica using toluene as eluent to give 15a (0.7g,3.9mmol, 49%).¹H NMR(400MHz,CDCl₃)δppm11.20(s,1H),9.87(s,1H),7.35(d,J=1.8Hz,1H),7.16(d,J=1.8Hz,1H),3.92(s,3H),0.26(s,9H)。

Compound 15a (2.00g;8.06mmol) was dissolved in 150mL of methanol. Sodium carbonate (2.3g,21.7mmol) was added and the mixture was stirred at room temperature overnight. The reaction was evaporated and the residue partitioned between water and dichloromethane. The organic layer was dried and evaporated and the solid residue was purified by silica chromatography using toluene as eluent to give 15b (0.70g,4mmol,50%) as a white powder. 1HNMR (400MHz, CDCl)₃)δppm11.22(s,1H),9.88(s,1H),7.37(d,J=1.8Hz,1H),7.18(d,J=1.8Hz,1H),3.92(s,3H),3.04(s,1H)。

Compound 15b (70mg,0.4mmol), 3-iodopyridine (90mg,0.44mmol), Pd (dppf) Cl₂(15mg,0.02mmol) and copper (I) iodide (5mg,0.02mmol) were dissolved in 5mL triethylamine and 5mL DMF and heated to 80 ℃. After 4 hours, 20mL of 1N hydrochloric acid were added and the mixture was extracted with dichloromethane.The organic layer was evaporated and the residue was purified by column chromatography to give 15-1(9mg,0.04mmol, 9%).¹H NMR(400MHz,CDCl₃)δppm11.24(s,1H),9.93(s,1H),8.77(s,1H),8.57(d,J=3.5Hz,1H),7.81(ddd,J=7.9,1.9,1.8Hz,1H),7.44(d,J=2.0Hz,1H),7.30(dd,J=7.9,4.9Hz,1H),7.24(d,J=1.8Hz,1H),3.97(s,3H)。

Watch 10

The following compounds were prepared by the above method using the corresponding aryl bromide and characterized by NMR.

TABLE 11

Example 16

Synthesis of 6-bromo-2-hydroxy-1-naphthaldehyde

A solution of titanium tetrachloride (231. mu.L, 2.1mmol) and dichloromethyl ether (97. mu.L, 1.1mmol) in 1mL of dichloromethane was stirred at 0 ℃ for 15 minutes. A solution of 6-bromo-2-hydroxy-naphthalene (223mg,1mmol) in 3mL of dichloromethane was added dropwise, the solution was warmed to room temperature and stirred for 12 hours. 10mL of 1N hydrochloric acid was added and the mixture was extracted with dichloromethane. The organic layer was washed with water, dried and evaporated to give 16-1(206mg,0.82mmol, 82%).¹H NMR(400MHz,DMSO-d₆)δppm11.90(s,1H),10.76(s,1H),8.92(d,J=9.3Hz,1H),8.16(d,J=2.0Hz,1H),8.10(d,J=9.3Hz,1H),7.72(dd,J=9.0,2.3Hz,1H),7.30(d,J=9.0Hz,1H)。

The following compounds were prepared by the above method and characterized by NMR.

TABLE 12

Example 17

Synthesis of 4- (5-formyl-6-hydroxynaphthalen-2-yl) -N, N-dimethylbenzamide

Compound 16-1(251mg,1mmol), 4- (N, N-dimethylaminocarbonyl) phenylboronic acid (222mg,1.2mmol) and anhydrous sodium carbonate (424mg,4mmol) were dissolved in a mixture of 20mL DMF and 12mL water. Tetrakis (triphenylphosphine) palladium (56mg,0.05mmol) was added and the reaction was heated at 105 ℃ for 25 minutes under an argon atmosphere. 50mL of a saturated sodium chloride solution and 900. mu.L of acetic acid were added, and the mixture was extracted with chloroform. The organic layer was evaporated and the crude product was purified by column chromatography to give 17-1(186mg,0.58mmol, 58%).¹H NMR(400MHz,CDCl₃)δppm13.15(s,1H),10.85(s,1H),8.44(d,J=9.0Hz,1H),8.05(d,J=9.0Hz,1H),8.01(d,J=2.0Hz,1H),7.88(dd,J=8.8,2.0Hz,1H),7.71-7.75(m,2H),7.56(d,J=8.5Hz,2H),7.19(d,J=9.0Hz,1H),3.15(br.s,3H),3.07(br.s,3H)。

The following compounds were prepared by the above method and characterized by NMR using the corresponding arylboronic acids.

Watch 13

Example 18

Synthesis of 6- (5-formyl-6-hydroxynaphthalen-2-yl) picolinic acid

Compound 16-1(5.00g;19.9 mmol), bis-pinacolatodiboron (5.57g;21.9mmol), potassium acetate (5.86g;59.8mmol) and Pd (dppf) Cl2(1.75g;2.39mmol) were added under an argon atmosphere to a solution of diboronHeated to reflux in an alkane for 4 hours. The reaction mixture was cooled to room temperature, filtered and the filtrate was evaporated to dryness under reduced pressure. The solid residue was purified by column chromatography on silica using dichloromethane as eluent. The collected pale yellow solid was triturated with diisopropyl ether to give 18a (3.56g;11.9mmol, 60%).¹H NMR(400MHz,CDCl₃)δppm13.23(s,1H),10.82(s,1H),8.33(d,J=8.8Hz,1H),8.29(s,1H),8.02(d,J=9.0Hz,1H),7.98(dd,J=8.5,1.3Hz,1H),7.13(d,J=9.0Hz,1H),1.39(s,12H)。

6-Bromopicolinic acid (81mg,0.4mmol), 18a (119mg,0.4mmol) and anhydrous sodium carbonate (339mg,3.2mmol) were dissolved in a mixture of 8mL DMF and 8mL water. Tetrakis (triphenylphosphine) palladium (22mg,0.02mmol) was added, and the reaction was stirred at 110 ℃ under an argon atmosphere for 3 hours. 40mL of 1N sodium hydroxide solution was added and the aqueous layer was extracted with chloroform (2X40 mL). The aqueous layer was acidified to pH5 with 6N hydrochloric acid, the white precipitate was filtered, washed with water, dried in vacuo and recrystallized from ether to give 100mg18-1(0.34mmol, 84%).¹H NMR(400MHz,DMSO-d₆)δppm13.15(br.s,1H),12.08(br.s,1H),10.84(s,1H),9.07(d,J=9.0Hz,1H),8.73(d,J=2.0Hz,1H),8.44(dd,J=9.0,2.0Hz,1H),8.33(dd,J=7.9,0.9Hz,1H),8.28(d,J=9.0Hz,1H),8.11(t,J=7.8Hz,1H),8.02(dd,J=7.8,0.8Hz,1H),7.34(d,J=9.0Hz,1H)。

The following compounds were prepared by the above method and characterized by LC/MS using the corresponding arylboronic acids.

TABLE 14

Watch 15

Example 19

Synthesis of 6- (5-formyl-6-hydroxynaphthalen-2-yl) -N- (2-morpholinoethyl) picolinamide

N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (42mg,0.22mmol), 1-hydroxybenzotriazole (30mg,0.22mmol), triethylamine (140. mu.L, 1mmol) and 1- (2-aminoethyl) morpholine (57. mu.L, 0.44mmol) were added to a solution of 18-1(59mg,0.2mmol) in 2mL THF at room temperature. After 2 hours, 2mL of 2N hydrochloric acid was added, and the reaction was stirred for 2 hours. The mixture was evaporated, the residue was dissolved in 2mL of chloroform, washed with saturated sodium bicarbonate (1X1.5mL) and water (1X1.5 mL). The organic phase was evaporated and the crude product purified by column chromatography to give 7mg of 19-1(0.02mmol, 9%).¹H NMR(400MHz,CDCl₃)δppm13.20(br.s,1H),10.89(s,1H),8.71(br.s,1H),8.49(d,J=8.8Hz,1H),8.46(d,J=2.0Hz,1H),8.38(dd,J=8.8,2.0Hz,1H),8.19(dd,J=7.2,1.6Hz,1H),8.10(d,J=9.0Hz,1H),8.01(dd,J=8.0,1.5Hz,1H),7.97(t,J=7.8Hz,1H),7.23(d,J=9.0Hz,1H),3.78-3.86(m,4H),3.66(q,J=6.0Hz,2H),2.69(t,J=6.1Hz,2H),2.56-2.65(m,4H)。

The following compounds were prepared by the above method and characterized by NMR using the corresponding aryl acid and amine.

TABLE 16

Example 20

Synthesis of 2-hydroxy-6- (5- (morpholine-4-carbonyl) thiophen-2-yl) -1-naphthaldehyde

Compound 18-3(804mg;2.57mmol) was dissolved in 25mL of twoAlkane and 25mL of 1N sodium hydroxide. The mixture was stirred at room temperature for 30 minutes. 75mL of 1N sodium hydroxide was added and the solution was washed with chloroform (2X25 mL). The aqueous layer was acidified with 6N hydrochloric acid and the yellow precipitate was filtered and washed successively with water and ether to give 666mg20-1(2.3mmol, 91%).¹H NMR(400MHz,DMSO-d₆)δppm13.09(br.s,1H),12.08(s,1H),10.78(s,1H),9.04(d,J=8.8Hz,1H),8.28(d,J=2.0Hz,1H),8.20(d,J=9.0Hz,1H),7.98(dd,J=8.8,2.0Hz,1H),7.76(d,J=3.8Hz,1H),7.67(d,J=3.8Hz,1H),7.41(d,J=9.0Hz,1H)。

N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (42mg,0.22mmol), 1-hydroxybenzotriazole (30mg,0.22mmol), triethylamine (140. mu.L, 1mmol) and morpholine (38. mu.L, 0.44mmol) were added to a solution of 20-1(54mg,0.2mmol) in 2mL THF at room temperature. After 2 hours, 2mL of 2N hydrochloric acid was added and the reaction was stirred for 2 hours. The mixture was evaporated to dryness, and the residue was dissolved in 2mL of chloroform and extracted with water (1X1.5mL), 1N hydrochloric acid (1X1.5mL), water (1X1.5mL), saturated sodium bicarbonate (1X1.5mL), and water (1X1.5 mL). The organic phase was evaporated and the crude product was purified by column chromatography to give 20-2(20mg,0.05mmol, 27%).¹H NMR(400MHz,CDCl₃)δppm13.13(s,1H),10.82(s,1H),8.38(d,J=9.0Hz,1H),7.96-8.06(m,2H),7.86(dd,J=8.8,2.0Hz,1H),7.34(d,J=3.8Hz,1H),7.32(d,J=3.8Hz,1H),7.19(d,J=9.3Hz,1H),3.80-3.85(m,4H),3.74-3.79(m,4H)。

The following compounds were synthesized by the methods described above using the corresponding aryl esters and amines (if present) and characterized by NMR.

TABLE 17

Example 21

Synthesis of 3-hydroxyquinoline-4-carbaldehyde

3-Hydroxyquinoline (145mg,1mmol) was added to a well stirred mixture of 5mL chloroform, water (72. mu.L, 4mmol), sodium hydroxide (100mg,2.5mmol) and tetrabutylammonium hydroxide (50. mu.L, 20% water) at room temperature. The resulting suspension was heated to 60 ℃ and stirred for 3 hours. 100mg of sodium hydroxide were added per hour. The reaction mixture was diluted with 5mL chloroform, acidified to pH6 with 10mL1N hydrochloric acid and extracted with chloroform (3 × 10 mL). The combined organic phases were dried and evaporated. The crude product was purified by column chromatography to give 21-1(24mg,0.14mmol, 14%).¹H NMR(400MHz,DMSO-d₆)δppm10.70(s,1H),9.06(s,1H),8.75(d,J=6.3Hz,1H),8.43(d,J=6.0Hz,1H),8.16(d,J=8.8Hz,1H),7.23(d,J=9.3Hz,1H)。

Watch 18

Example 22

Synthesis of 3-hydroxy-2-methylquinoline-4-carbaldehyde

2-methyl-3-hydroxyquinoline-4-carboxylic acid (1.016g,5mmol) was dissolved in 10mL of methanol. Thionyl chloride (730. mu.L, 10mmol) was added at-10 ℃ and the mixture heated to reflux for 20 h, 365. mu.L thionyl chloride (5mmol) was added every 4 h. The reaction mixture was evaporated, dissolved with saturated sodium bicarbonate solution and the mixture was extracted with ethyl acetate. Evaporation of the organic layer and recrystallization of the crude product from hexane gave 22a (258mg,1.1mmol,24%), ESI MS M/e218([ M + H ]]⁺)。

Compound 22a (0.163mg,0.75mmol) was dissolved in 3mL dry THF at-10 deg.C and 1M DIBAL in THF (3.3mL,3.3mmol) was added. After 2H, 5mL of 1M potassium dihydrogen phosphate solution was added and the mixture was extracted with chloroform to give 22b (59mg,0.3mmol,42%), ESI MS M/e191([ M + H ] s)]⁺)。

3-hydroxy-4-hydroxymethylquinoline, 22b, (63mg,0.33mmol) was added to a suspension of manganese dioxide (86mg,1mmol) in 12mL of acetone. The mixture was stirred at room temperature for 48 hours and the other part (86mg,1mmol) of manganese dioxide was added at 12 hour intervals. The suspension was filtered, evaporated and the crude product purified by column chromatography to give 22-1(15mg,0.08mmol, 24%).¹H NMR(400MHz,CDCl₃)δppm12.57(s,1H),10.91(s,1H),8.28-8.34(m,1H),8.00-8.08(m,1H),7.58-7.64(m,2H),2.73(s,3H)。

Example 23

Synthesis of 2- (2-hydroxy-3-methoxy-5- (thien-2-yl) phenyl) thiazolidine-4-carboxylic acid ethyl ester

Compounds 11-28(120mg,0.5mmol), L-cysteine ethyl ester hydrochloride (90mg,0.5mmol) and diisopropylethylamine (85. mu.L, 0.5mmol) were dissolved in 3mL of ethanol and stirred at room temperature for 1 hour. The mixture was filtered to give 23-1(147mg, 0) as a yellow solid.4mmol,80%)。¹HNMR(400MHz,DMSO-d₆Stereoisomers) δ ppm9.43(s,0.4H),9.26(s,0.6H),7.44(dd, J =5.0,1.0Hz,0.4H),7.42(dd, J =5.0,1.0Hz,0.6H),7.39(dd, J =3.5,1.3Hz,0.4H),7.37(dd, J =3.5,1.3Hz,0.6H),7.30(d, J =2.0Hz,0.4H),7.24(d, J =2.0Hz,0.6H),7.15(d, J =2.0Hz,0.4H),7.11(d, J =2.0Hz,0.6H),7.07-7.10(m,1H),5.87(d, J =11.5 = 6H), 7.11(d, J =2.0Hz, 0H), 7.11(d, J =2.0Hz,0.6H),7.07-7.10(m,1H),5.87(d, J = 11.5.5, 6H), 5.5 =0, 3.5H, 3.4H), 3.5H, 3.4H, 3.9H, 3.4H, 7.4H, 3.9H, 3.4H, 3, overlap), 3.26(dd, J =7.0,10.3Hz,0.6H),3.08(dd, J =4.8,10.3Hz,0.6H),3.04(dd, J =8.8,10.0Hz,0.4H),1.24(t, J =7.0Hz,1.2H),1.23(t, J =7.0Hz, 1.8H).

Watch 19

Example 24

Synthesis of 2-methoxy-6- ((4-methoxybenzylimino) methyl) -4- (thien-2-yl) phenol

Compounds 11-28(117mg;0.50mmol) and 4-methoxybenzylamine (65. mu.l; 0.50mmol) were dissolved in 4mL of ethanol andstirred at room temperature for 4 hours. The mixture was filtered to give 24-1(113mg,0.32mmol, 64%).¹H NMR(400MHz,DMSO-d₆)δppm13.82(br.s,1H),8.70(s,1H),7.43(ddd,J=14.3,4.3,1.3Hz,2H),7.25-7.32(m,4H),7.10(dd,J=5.1,3.6Hz,1H),6.92-6.97(m,2H),4.75(s,2H),3.84(s,3H),3.75(s,3H)。

Watch 20

Example 25

Synthesis of 3-hydroxy-4- (morpholinomethyl) -2-naphthaldehyde

3-hydroxy-2-naphthaldehyde (20mg,0.12mmol), morpholine (63 μ L,0.72mmol) and formaldehyde (37 μ L,37% water) were dissolved in 2mL of acetic acid. After evaporation, the solid residue was partitioned between chloroform and saturated sodium bicarbonate solution. The organic layer was washed with water and dried over sodium sulfate. The solvent was removed and the solid residue was recrystallized from diisopropyl ether to give 25-1(18mg,0.07mmol, 55%).¹H NMR(400MHz,CDCl₃)δppm11.79(br.s,1H),10.41(s,1H),8.22(s,1H),7.96(d,J=8.8Hz,1H),7.87(d,J=8.3Hz,1H),7.57(ddd,J=8.5,7.0,1.4Hz,1H),7.36(td,J=7.5,1.0Hz,1H),4.11(s,2H),3.76(t,J=4.5Hz,4H),2.66(t,J=4.5Hz,4H)。

TABLE 21

Example 26

Activity of the Compound

IC₅₀And EC₅₀The results of the tests are shown in tables 26 to 42.

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Table 41

Watch 42

Example 27

Optimization of the test protocol

A series of in vitro ADME assays (absorption-distribution-metabolism-excretion assays, tests such as plasma stability, liver microsome stability, solubility, CaCo) are used₂Permeability, etc.) to optimize the pharmacological properties of IRE-1 α inhibitor compounds. Depending on the activity of the compound analog, the protocol is run in a continuous mode of staged assay. In early optimization, in vitro potency, cell-targeting (cellular-target) XBP-1mRNA splicing, apoptotic caspases 3 and 7, and proteasome inhibitor enhancement assays were employed along with a series of compound property assays: solubility, serum stability andlog P. The activity assay is combined with pharmacological property assays such as serum protein binding, membrane permeability, cell permeability and microsomal stability. Finally, in vitro toxicity and pharmacokinetic assays, such as P450, AMES, hERG and receptor profiling assays, are employed.

Example 28

Animal model/preclinical validation study

Preclinical validation protocols utilize a set of animal models representing normal tissues under chemical stress and multiple myeloma xenografts. The dose-related targeting activity of compounds was demonstrated in tissues sensitive to standard UPR inducers such as tunicamycin using a normal animal model as a surrogate model (Wu et al, Dev cell.2007, 9 months; 13(3): 351-64). As shown in FIG. 8, normal mouse tissues are not ER-stressed, and thus XBP-1mRNA remains in an inactive, unspliced form. Following induction with tunicamycin, tissue induces active XBP-1mRNA splicing, which is inhibited by IRE-1 α inhibitors. The animal model of targeted ER stress is a useful screening and early pharmacokinetic tool.

Antibody production was evaluated in a second surrogate model. However, in cellular models, IRE-1. alpha. inhibitors showed potent inhibition of antibody production.

Final efficacy studies were performed in a myeloma xenograft model, as described below.

Example 29

Model of efficacy of RPMI8226 xenografts

SCID mice were evaluated for their ability to support implantation of the desired tumor cells to support model development and characterization. Mice were injected intravenously or implanted Subcutaneously (SC) or Intraperitoneally (IP). As is well known in the art, to generate relevant animal models that mimic human disease, it is preferable to assess whether all three methods improve engraftment rate and associated disease progression. SC injection is a convenient method to detect tumor growth and efficacy, IV and IP injection represent more physiologically relevant models of human tumor spread. SC injections were given mainly to the flank, while IV injections were given to the tail vein. The mice were restrained by hand for SC and IP injections and Brucem mouse restrictors (Broome mouseterpastainer) for IV injections.

Example 30

Evaluation of IRE-1 alpha inhibitor compounds in a model of xenograft efficacy

According to the results of the xenograft model development study (above), SCID mice were implanted with tumor cells (human RMPI8226 myeloma cells) via IP, IV or SC routes. Mice were treated with compound or mock treated (vehicle) for up to 4-5 weeks. The compounds may be administered by the IV, IP, PO or SC routes. In some cases, tunicamycin is administered via IP injection to stimulate stress in the animal. This stress mimics the stress to which an animal may be subjected during tumor growth. Tunicamycin injection mimics tumor growth during stress and biomarkers indicative of the efficacy of the compound (e.g., XBP-1 splicing) were assessed by RT-PCR, immunohistochemistry or Western blotting.

Mice were monitored for tumor growth, regression, and general health. Tumors were collected and characterized by immunohistochemistry and/or FACS analysis. Tumor growth was detected with calipers, ultrasound or abdominal lavage. Biomarkers (mainly XBP-1 splicing) in blood or tumors can be assessed.

In some experiments, blood samples were collected at various time points during dosing (i.e., day 1 or week 4) to assess pharmacokinetic profiles. The time point of blood collection varies depending on the pharmacokinetic properties of the drug tested. The blood sample volume at each time point was 100 microliters and blood was collected twice via the retro-orbital sinus within 24 hours after dosing. If the same mouse is used, blood is collected once per eye over a 24 hour period.

Tumor cells were cultured and injected into mice in a volume of about 100 microliters using a 21 gauge needle IP, IV (tail vein) or SC (flank). Mice were treated with compound or vehicle alone via IV, IP, SC or PO routes for 5 days a week for up to 4-5 weeks. Blood was collected via retroorbital bleeding (100 microliters) at 2 time points (different eyes). The end point of the study was dependent on the overall health of the mice. Although mice were euthanized at the end of 4-5 weeks in most studies, in a few studies, mice could be maintained up to day 40 if general health conditions allowed. The reason why the study lasted 40 days was to determine whether the tested compounds had a long-term effect of inhibiting tumor growth. Whether or not to euthanasia mice in which tumor regression was observed depends on the experimental design. In the screening mode, the experiment will end when the tumor in the control/untreated group reaches 1.5cm, ulceration or when loss of motility is observed in this group. In subsequent experiments, mice in which tumor regression was observed, could be maintained for longer periods of time until they showed signs of tumor growth or disease.

SCID mice bearing human myeloma RPMI8226 tumor xenografts were administered a therapeutic dose of 0.75mg/kg of Bouzo intravenously twice a week to inhibit tumor growth. However, after cessation of bortezomib treatment, tumors frequently recur and grow into large masses. Thus, mice are treated twice daily with 10-60mg/kg IRE-1 α/XBP-1 inhibitor, such as Compound 17-1, administered orally, IP or IV in combination with Baozomi (shown). Compounds that reduce the rate of tumor recurrence are identified.

Example 31

Combination therapy

Spliced forms of XBP-1, such as homodimers and heterodimers, along with ATF-6, transcriptionally regulate genes involved in adapting to ER stress (Wu et al, Dev cell.2007, 9 months; 13(3): 351-64). Many of these downstream targets are the major chaperones, accessory chaperones and ERAD components of the ER. Chaperones such as GRP78 and GRP94 are stable and have a half-life of up to several days (Wu et al, Dev cell.2007, 9 months; 13(3): 351-64). Thus, each cycle of cancer treatment with an IRE-1 α/XBP-1 inhibitor may require up to 5-6 days of treatment.

In some embodiments, the combination therapy (e.g., a combination proteasome inhibitor) administered in a cycle comprises a2 day pre-treatment with an IRE-1 α/XBP-1 inhibitor administered to the patient, followed by concurrent administration of a chemotherapeutic agent until a pharmacodynamic effect (typically, a potter effect) is achieved24 hours after zolmitm infusion). Botezomib is typically administered on a 3-week cycle, every 1,4, 8 and 11 days (out of 21 days). 1.3mg/m by IV administration². 10-100mg/kg of IRE-1 α/XBP-1 inhibitor can be administered by IV or oral route once, twice or three times daily, 2 days before and 24 hours after the infusion, depending on the PK/PD relationship.

Similar regimens may utilize Hsp90 or HDAC inhibitors. Alternatively, both agents are administered simultaneously in each cycle, depending on the PK/PD relationship of the inhibitor. IRE-1 α/XBP-1 inhibitors can be administered in combination with tamoxifen (Gomez et al, FASEB J.2007, 12 months; 21(14):4013-27) or sorafenib (Sorafini) for a variety of other cancers, including renal and hepatocellular carcinomas (Rahmani et al, Mol Cell biol.2007, 8 months; 27(15): 5499-513).

In summary, since many kinase inhibitors are often not selective for their targeted kinases, and often affect many other kinases; they can lead to nonspecific cellular stress that activates UPR. Thus, a combinatorial approach using IRE-1 α/XBP-1 inhibitors as sensitizers may be useful.

Claims

1. A compound that directly inhibits IRE-1 α activity in vitro as shown in structural formula (B):

in the formula:

R¹is hydrogen, phenyl or an optionally benzo-fused 5-or 6-membered heterocycle, wherein the phenyl or the optionally benzo-fused 5-or 6-membered heterocycleOptionally substituted with:-CH₂OH、-CHO、-OCH₃halogen, -OH, -CH₃、

R²Is phenyl or an optionally benzo-fused 5-or 6-membered heterocycle, wherein the phenyl or the optionally benzo-fused 5-or 6-membered heterocycle is optionally substituted with:-CH₂OH、-CHO、-OCH₃halogen, -OH, -CH₃、

R³Is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkyl, C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,

R⁴Is hydrogen,

2. A compound that directly inhibits IRE-1 α activity in vitro as shown in structural formula (a):

in the formula:

R¹is hydrogen, halogen, or a 5-or 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur;

R²is hydrogen,Phenyl, or a 5-or 6-membered heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulfur, wherein the heteroaryl group is optionally benzo-fused, wherein the heteroaryl group is optionally substituted with 1,2 or 3 substituents independently selected from the group consisting of:C₁-C₃a straight or branched alkyl group,C₁-C₃Phenylalkyl, C₁-C₃An alkoxyphenylalkyl group,

R³Is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,

Q is a 5-or 6-membered heterocyclic ring.

3. A compound that directly inhibits IRE-1 α activity in vitro as shown in structural formula (C):

in the formula:

R¹and R²Independently of one another is hydrogen, -CH₃or-OH; and

the hydroxy substituent in ring a is located ortho to the aldehyde substituent.

4. A compound that directly inhibits IRE-1 α activity in vitro as shown in structural formula (D):

in the formula: r¹Is hydrogen, halogen, -NO₂、C₁-C₃Straight or branched alkyl, C₁-C₃Straight or branched alkoxy, C₁-C₃Straight or branched chain hydroxyalkyl,

5. A prodrug of a compound that directly inhibits IRE-1 α activity in vitro, the prodrug having the formula (E):

in the formula:

R¹is hydrogen or-OCH₃(ii) a And

R²is that

6. A prodrug of a compound that directly inhibits IRE-1 α activity in vitro, the prodrug having the formula (F):

in the formula:

R¹is hydrogen or Br;

R²is hydrogen, Br orAnd

R³is hydrogen, -OCH₃-COOH or-OCH₂CH₃。

7. A pharmaceutical composition comprising a compound of any one of claims 1-6 and a pharmaceutically acceptable carrier.

8. Use of a compound according to any one of claims 1 to 6 or a compound directly inhibiting in vitro IRE-1 α activity as represented by structural formula (I):

in the formula:

the OH substituent is located ortho to the aldehyde substituent;

q is an aromatic carbocyclic or heterocyclic ring system selected from: benzene, naphthalene, pyridine N-oxide, thiophene, benzo [ b ]]Thiophene, benzo [ c]Thiophene, furan, pyrrole, pyridazine, pyrimidine, pyrazine, triazine, iso-isomerOxazoline,Oxazoline, thiazoline, pyrazoline, imidazoline, fluorene, biphenyl, quinoline, isoquinoline, cinnoline, 2, 3-naphthyridine, quinazoline, quinoxaline, benzofuran, indole, isoindole, isobenzofuranyl, benzimidazole, 1, 2-benzisoxazofuranAzoles and carbazoles;

R^x、R^yand R^zMay or may not be present and is independently selected from: hydrogen, aryl, heteroaryl, -A "R^a、-OH、-OA″R^a、-NO₂、-NH₂、-NHA″R^a、-N(A″R^a)(A′″R^b)、-NHCOA″R^a、-NHCOOA″R^a、-NHCONH₂、-NHCONHA″R^a、-NHCON(A″R^a)(A′″R^b) Halogen, -COOH, -COOA' R^a、-CONH₂、-CONHA″R^a、-CON(A″R^a)(A′″R^b)

a and A' are:

(b) Or, A and A' together form C₂-C₇Alkylene, wherein 1,2 or 3 CH₂The group can be O, S, SO₂、NH、NR^c、NCOR^cOr NCOOR^cSubstituted to form, for example, an alkylenedioxy group;

R^cis C₁-C₁₀Alkyl radical, C₃-C₇Cycloalkyl radical, C₄-C₈Alkylene cycloalkyl or C₂-C₈An alkenyl group; in which 1,2 or 3 CH₂The group can be O, S, SO₂NH, NMe, NEt and/or-CH ═ CH-groups, 1 to 7H atoms may be replaced by F and/or Cl, and/or 1H atom may be R^aReplacing;

aryl is phenyl, benzyl, naphthyl, fluorenyl or biphenyl, each of which may be unsubstituted or mono-, di-or trisubstituted by: halogen element、-CF₃、-R^f、-OR^d、-N(R^d)₂、-NO₂、-CN、-COOR^d、CON(R^d)₂、-NR^dCOR^e、-NR^dCON(R^e)₂、-NR^dSO₂A、-COR^d、-SO₂N(R^d)₂、-S(O)_mR^fAA' together or-O (aryl),

R^dand R^eIndependently is H or C₁-C₆An alkyl group;

R^fis C₁-C₆An alkyl group;

m is 0,1 or 2.

9. The use of claim 8, wherein the medicament is formulated for administration with a therapeutic agent that induces or upregulates expression of IRE-1 α.

10. The use of claim 8, wherein the medicament is formulated for administration with a therapeutic agent that decreases potency upon expression of IRE-1 α.

11. The use according to claim 8, wherein the medicament is formulated for administration with a proteasome inhibitor.

12. Use according to claim 8, wherein the compound is a compound according to any one of claims 1 to 4.

13. Use according to claim 8, wherein the compound is a compound according to claim 5 or claim 6.