Methods for treating or preventing proteopathy

Abstract

Methods for treating or preventing proteopathy are provided. The present invention provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula II or a pharmaceutically acceptable salt thereof. TIFF2023071887000407.tif64158 In the formula, Hal is a halogen atom; R1 is independently a halogen atom, alkyl, etc.; R2 is independently a halogen atom, alkyl, etc.; and R3 is H, alkyl, etc.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 174,332, filed June 11, 2015, and U.S. Provisional Application No. 62 / 174,338, filed June 11, 2015, both of which are incorporated herein by reference in their entireties. [Background technology]

[0002] The proper functioning of organs and cells in living organisms depends on the proper action of proteins. Proteins are biological entities that have a primary amino acid sequence; a secondary structure that forms protein domains and, most importantly, includes alpha helices and beta sheets; and a tertiary structure that is the result of complex folding of the peptide chain in three dimensions, including interactions of the polypeptide chain backbone and amino acid side chains. Some proteins act as complexes of multiple subunits, where the arrangement of multiple proteins into a quaternary structure is crucial for their proper function.

[0003] When proteins do not fold into their correct three-dimensional structure, diseases called proteopathies (sometimes called proteinopathies or protein conformational disorders) can result. This defect can be due to one or more mutations in the protein's gene or environmental factors such as oxidative stress, alkalosis, acidosis, pH shifts, and osmotic shock. Protein misfolding can lead to clumping or aggregation into amyloid plaques or fibrils, which can exacerbate the disease. Proteopathies include a wide range of diseases, including neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, polyglutamine diseases, prion diseases); amyloidoses of other non-neuronal proteins such as alpha 1-antitrypsin, immunoglobulin light and heavy chains, lactadherin, apolipoproteins, gelsolin, lysozyme, fibrinogen, atrial natriuretic factor, keratin, lactoferrin, and beta-2 microglobulin, among others; sickle cell disease; cataracts; cystic fibrosis; retinitis pigmentosa; and nephrogenic diabetes insipidus.

[0004] Molecular chaperones are biomolecules that aid in proper protein folding, protein translocation, and / or protein degradation. Examples of molecular chaperones include heat shock proteins, which are classified into seven different families in the human genome, including HSPH (Hsp110), HSPC (Hsp90), HSPA (Hsp70), DNAJ (Hsp40), HSPB (small Hsp (sHsp)), human chaperonins HSPD / E (HSP60 / HSP10), and CCT (TRiC). Summary of the Invention [Means for solving the problem]

[0005] The present invention provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof:

[0006] [ka] wherein R is fluoro, chloro, iodo, methyl, methoxy, cyano, trifluoromethyl, or —(CO)NH(CH).

[0007] The present invention also provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula II, or a pharmaceutically acceptable salt thereof:

[0008] [ka] wherein Hal is -Cl, -F, -I, or -Br; x is an integer ranging from 0 to 5; each R1 is independently -Cl, -F, -I, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; y is an integer ranging from 0 to 5; each R2 is independently -Cl, -F, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; R3 is -H, -C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-phenyl, -(C1-C6 alkylene)-O-(C1-C6 alkyl), -C2-C6 alkenyl, -(C1-C6 alkylene)-C(O)R4, -(C1-C6 alkylene)-R5,

[0009] [ka] and; R4 is -OH, -O-(C1-C6 alkyl), -NH2, -NH(C1-C6 alkyl), -NH((C1-C6 alkylene)-OH), -NH((C1-C6 alkylene)N(C1-C6 alkyl)2), -N(C1-C6 alkyl)((C1-C6 alkylene)-CN), -N(C1-C6 alkyl)((C1-C6 alkylene)N(C1-C6 alkyl)2), -NH(C1-C6 alkylene)-O-(C1-C6 alkyl),

[0010] [ka] TIFF0007755610000005.tif136161; a is an integer ranging from 0 to 10; b is an integer ranging from 0 to 8; c is an integer ranging from 0 to 6; R5 is

[0011] [ka] is.

[0012] The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula III or a pharmaceutically acceptable salt thereof:

[0013] [ka] wherein Hal is -Cl, -F, -I, or -Br; x is an integer ranging from 0 to 5; each R1 is independently -Cl, -F, -I, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; R3 is -H, -C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-phenyl, -(C1-C6 alkylene)-O-(C1-C6 alkyl), -C2-C6 alkenyl, -(C1-C6 alkylene)-C(O)R4, -(C1-C6 alkylene)-R5,

[0014] [ka] and; R4 is -OH, -O-(C1-C6 alkyl), -NH2, -NH(C1-C6 alkyl), -NH((C1-C6 alkylene)-OH), -NH((C1-C6 alkylene)N(C1-C6 alkyl)2), -N(C1-C6 alkyl)((C1-C6 alkylene)-CN), -N(C1-C6 alkyl)((C1-C6 alkylene)N(C1-C6 alkyl)2), -NH(C1-C6 alkylene)-O-(C1-C6 alkyl),

[0015] [ka] TIFF0007755610000010.tif56163; a is an integer ranging from 0 to 10; b is an integer ranging from 0 to 8; c is an integer ranging from 0 to 6; R5 is

[0016] [ka] and; each R6 and R7 is independently -H or -I, wherein at least one of R6 and R7 is -I; wherein when R3 is -C1-C3 alkyl, R7 is -H.

[0017] The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula IV, or a pharmaceutically effective salt thereof:

[0018] [ka] wherein R8 is —C1-C3 alkyl.

[0019] The present invention further provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound having the structure of the following formula, or a pharmaceutically acceptable salt thereof:

[0020] [ka]

[0021] The present invention still further provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound having the structure of the following formula, or a pharmaceutically acceptable salt thereof:

[0022] [ka] TIFF0007755610000015.tif141163

[0023] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula V, or a pharmaceutically acceptable salt thereof:

[0024] [ka] wherein R1 is:

[0025] [ka] and; R2:

[0026] [ka] and; Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

[0027] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula VI, or a pharmaceutically acceptable salt thereof:

[0028] [ka] wherein R3 is:

[0029] [ka] and; b is 0 or 1; c is 1 or 2.

[0030] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula VII or a pharmaceutically acceptable salt thereof:

[0031] [ka] In the formula, R4 is

[0032] [ka] is.

[0033] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIII or a pharmaceutically acceptable salt thereof:

[0034] [ka] wherein R5 is:

[0035] [ka] and; R6 is:

[0036] [ka] and; Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

[0037] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIV or a pharmaceutically acceptable salt thereof:

[0038] [ka] wherein R7 is:

[0039] [ka] and; b is 0 or 1; c is 1 or 2.

[0040] The present invention still further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XV or a pharmaceutically acceptable salt thereof:

[0041] [ka] wherein R8 is:

[0042] [ka] is.

[0043] A "pyrazolopyridazine compound" is a compound of Formula I, II, III, IV, V, VI, VII, XIII, XIV, or XV; Compounds 1-35, 37-39, 42, 43, 44, 45, 46, 47-97, 98-123, 124a, 124b; or a pharmaceutically acceptable salt of any of the foregoing. The pyrazolopyridazine compounds are useful for treating or preventing proteopathies. DETAILED DESCRIPTION OF THE INVENTION

[0044] In one embodiment, the present invention provides a pyrazolopyridazine compound. In a further embodiment, the present invention provides a pharmaceutical composition comprising an effective amount of a pyrazolopyridazine compound and a pharmaceutically acceptable carrier or vehicle.

[0045] In yet another embodiment, the present invention provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a pyrazolopyridazine compound.

[0046] definition The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. Example alkyl groups include -CH, -CHCH, -CHCHCH, -CH(CH), -CHCHCHCH, -CH(CH)CH, -CH(CH)CHCH, -CHCH(CH), -C(CH), -CHCHCHCHCH, -CH(CH)CHCHCH, -CHCHCH(CH), -CHC(CH), -CHCHCHCHCHCH, -CH(CH)CHCHCH, -CHCHCH(CH), and -CH(CH)C(CH).

[0047] The term "alkylene" refers to an alkyl group that is bonded to another atom or group. Illustrative alkylene groups include -CH-, -CHCH-, -CHCHCH-, -C(CH)-, -CH(CH), -CHCHCHCHCH-, -CH(CH)CHCH-, -CHC(CH)-, -C(CH)CH-, -CHCHCHCHCHCH-, -CH(CH)CHCHCH-, -CHCHC(CH) 2-, -CH2CH(CH3)CH2CH2, -CH2CH2CH(CH3)CH2-, -CH2CH2CH2CH2CH2CH2-, -CH(CH3)CH2CH2CH2CH2-, - There are CH2CH2CH2C(CH3)2-, -CH2CH(CH3)CH2CH2CH2-, -CH2CH2CH2CH(CH3)CH2- and -C(CH3)2C(CH3)2-.

[0048] The term "alkenyl" refers to a straight or branched chain hydrocarbon group having one or more double bonds. Illustrative alkenyl groups include -CH=CH2, -CH2CH=CH2, cis-CH=CHCH3, trans-CH=CHCH3, -C(CH3)=CH2, cis-CH=CHCH2CH3, trans-CH=CHCH2CH3, cis-CH2CH=CHCH3, trans-CH2CH=CHCH3, -CH2CH2CH=CH2, cis-CH=CHCH2CH2CH3, trans-CH=CHCH2CH2CH3, cis-CH 2CH2CH=CHCH3, trans-CH2CH2CH=CHCH3, -CH2CH2CH2CH=CH2, -CH2CH=C(CH3)2, cis-CH=CHCH2CH2CH2CH3, trans-CH=CHCH There are 2CH2CH2CH3, cis-CH2CH2CH2CH=CHCH3, trans-CH2CH2CH2CH=CHCH3, -CH2CH2CH2CH2CH=CH2, and -CH2CH2CH=C(CH3)2 groups.

[0049] The word "about," when used immediately before a numerical value, means a range of plus or minus 10% of that value; for example, "about 100 mg" means 90 mg to 110 mg, "about 300 mg" means 270 mg to 330 mg, etc.

[0050] Abbreviation: APCI Atmospheric Pressure Chemical Ionization DAPI 4',6-diamidino-2-phenylindole DCM dichloromethane DEAD Diethyl azodicarboxylate DIPEA Diisopropylethylamine DMEM Dulbecco's Modified Eagle's Medium DMF Dimethylformamide DMSO dimethyl sulfoxide EDAC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride ESI electrospray ionization ESI-TOF Electrospray Ionization-Time of Flight HATU 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HOPO 2-hydroxypyridine-N-oxide HPLC High Performance Liquid Chromatography LCMS Liquid Chromatography-Mass Spectrometry LDA Lithium diisopropylamide m / z mass-to-charge ratio MALDI-TOF matrix-assisted laser desorption ionization-time of flight MS mass spectrometry PBS Phosphate-buffered saline Rt retention time SDS Sodium dodecyl sulfate TFA trifluoroacetic acid THF tetrahydrofuran

[0051] The term "effective amount" refers to an amount of a pyrazolopyridazine compound or non-pyrazolopyridazine compound effective to treat or prevent a proteopathy in a subject. In some embodiments, when another therapeutic or prophylactic agent is administered before, after, or simultaneously with administration of a pyrazolopyridazine compound or non-pyrazolopyridazine compound, an "effective amount" is the combined amount of (i) the pyrazolopyridazine compound or non-pyrazolopyridazine compound and (ii) the other therapeutic or prophylactic agent effective to treat or prevent a proteopathy in a subject.

[0052] The terms "proteopathy," "proteinopathy," and "protein conformationopathy" refer to diseases or disorders that result from the misfolding of one or more proteins.

[0053] The term "protein aggregate" refers to the biological phenomenon in which misfolded proteins accumulate and clump together.

[0054] A "subject" is a mammal, including species-rich orders, such as primates such as humans; Rodentia species such as mice, rats, or guinea pigs; Carnivora species such as dogs, cats, weasels, bears, or seals; non-human primates such as monkeys, Chinese baboons, or rhesus monkeys; Chiroptera species such as bats; Soricomorpha species such as shrews, moles, or solenodonts; and Cetacean species such as whales. In one embodiment, the subject is a human. In another embodiment, the human is a human fetus.

[0055] Pyrazolopyridazine compounds useful in the present method Pyrazolopyridazine compounds of formula I In one embodiment, the present invention provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof:

[0056] [ka] wherein R is fluoro, chloro, iodo, methyl, methoxy, cyano, trifluoromethyl, or —(CO)NH(CH).

[0057] In one embodiment, R of Formula I is in the para position relative to the pyrazolopyridazino ring system. In one embodiment, R of Formula I is in the meta position relative to the pyrazolopyridazino ring system. In one embodiment, R of Formula I is in the ortho position relative to the pyrazolopyridazino ring system.

[0058] Pyrazolopyridazine compounds of formula II The present invention also provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula II, or a pharmaceutically acceptable salt thereof:

[0059] [ka] wherein Hal is -Cl, -F, -I, or -Br; x is an integer ranging from 0 to 5; each R1 is independently -Cl, -F, -I, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; y is an integer ranging from 0 to 5; each R2 is independently -Cl, -F, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; R3 is -H, -C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-phenyl, -(C1-C6 alkylene)-O-(C1-C6 alkyl), -C2-C6 alkenyl, -(C1-C6 alkylene)-C(O)R4, -(C1-C6 alkylene)-R5,

[0060] [ka] and; R4 is -OH, -O-(C1-C6 alkyl), -NH2, -NH(C1-C6 alkyl), -NH((C1-C6 alkylene)-OH), -NH((C1-C6 alkylene)N(C1-C6 alkyl)2), -N(C1-C6 alkyl)((C1-C6 alkylene)-CN), -N(C1-C6 alkyl)((C1-C6 alkylene)N(C1-C6 alkyl)2), -NH(C1-C6 alkylene)-O-(C1-C6 alkyl),

[0061] [ka] TIFF0007755610000034.tif164167; a is an integer ranging from 0 to 10; b is an integer ranging from 0 to 8; c is an integer ranging from 0 to 6; R5 is

[0062] [ka] is.

[0063] In certain embodiments, Hal is -Cl. In yet another embodiment, x and y are 0.

[0064] In certain embodiments, x and y are 0, or x is 0 and y is 1, or x is 1 and y is 2, or x is 1 and y is 0, or x is 1 and y is 1, or x is 1 and y is 2, or x is 2 and y is 0, or x is 2 and y is 1, or x is 2 and y is 2.

[0065] In certain embodiments, Hal is -Cl; x and y are 0, or x is 0 and y is 1, or x is 1 and y is 2, or x is 1 and y is 0, or x is 1 and y is 1, or x is 1 and y is 2, or x is 2 and y is 0, or x is 2 and y is 1, or x is 2 and y is 2.

[0066] In certain embodiments, x is 1 and R is in the ortho position relative to the pyrazolopyridazino ring system. In certain embodiments, x is 1 and R is in the para position relative to the pyrazolopyridazino ring system. In further embodiments, x is 1 and R is in the meta position relative to the pyrazolopyridazino ring system.

[0067] In certain embodiments, y is 1 and R2 is in the ortho position relative to the pyrazolopyridazino ring system. In certain embodiments, y is 1 and R2 is in the para position relative to the pyrazolopyridazino ring system. In further embodiments, y is 1 and R2 is in the meta position relative to the pyrazolopyridazino ring system.

[0068] In certain embodiments, x is 2 and R is in the ortho and meta positions relative to the pyrazolopyridazino ring system. In certain embodiments, x is 2 and R is in the ortho and para positions relative to the pyrazolopyridazino ring system. In further embodiments, x is 2 and R is in the para and meta positions relative to the pyrazolopyridazino ring system.

[0069] In certain embodiments, y is 2 and R2 is in the ortho and meta positions relative to the pyrazolopyridazino ring system. In certain embodiments, y is 2 and R2 is in the ortho and para positions relative to the pyrazolopyridazino ring system. In further embodiments, y is 2 and R2 is in the para and meta positions relative to the pyrazolopyridazino ring system.

[0070] In still other embodiments, R1 is chloro. In certain embodiments, R1 is fluoro. In certain embodiments, R1 is iodo. In other embodiments, R1 is -Br. In further embodiments, R1 is -OCH3. In other embodiments, R1 is -CH3. In still other embodiments, R1 is -C(O)N(H)CH3. In certain embodiments, R1 is -CF3. In further embodiments, R1 is -CN. In additional embodiments, R1 is -C≡CCH2OH.

[0071] In still other embodiments, x is 1 or 2 and R1 is -Cl, -F, -I, -Br, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0072] In still other embodiments, Hal is -Cl, x is 1 or 2, and R1 is -Cl, -F, -I, -Br, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0073] In still other embodiments, R2 is -Cl. In certain embodiments, R2 is -F. In other embodiments, R2 is -Br. In further embodiments, R2 is -OCH3. In other embodiments, R2 is -CH3. In still other embodiments, R2 is -C(O)N(H)CH3. In certain embodiments, R2 is -CF3. In further embodiments, R2 is -CN. In additional embodiments, R2 is -C≡CCH2OH.

[0074] In still other embodiments, y is 1 or 2 and R2 is -Cl, -F, -Br, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0075] In still other embodiments, Hal is -Cl, y is 1 or 2, and R2 is -Cl, -F, -Br, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0076] In certain embodiments, R3 is -H. In certain embodiments, R3 is -CH3. In further embodiments, R3 is -CH2CH3. In yet other embodiments, R3 is -CHCH2. In other embodiments, R3 is -CH2CH2OH. In certain embodiments, R3 is -(CH2)2C6H5. In other embodiments, R3 is -CH2C(O)OH. In still other embodiments, R3 is -CH2C(O)N(H)CH3. In certain embodiments, R3 is -CH2C(O)N(H)((CH2)2N(CH3)2). In still other embodiments, R3 is -CH2C(O)N(H)((CH2)3N(CH3)2). In other embodiments, R3 is -CH2C(O)N(CH3)CH2CN. In certain embodiments, R3 is -CH2C(O)NH2. In certain embodiments, R3 is -CH2C(O)N(H)((CH2)2OH). In other embodiments, R3 is -CH2C(O)N(H)((CH2)2OCH 3 ). In still other embodiments, R3 is -CH2C(CH3)2OH. In still other embodiments, R3 is -CH2C(O)OCH3. In further embodiments, R3 is -CH2CH(OH)CH3. In still other embodiments, R3 is -CH2CH2OH. In certain embodiments, R3 is -CH(CH3)CH2OH.

[0077] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0078] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0079] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0080] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0081] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0082] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0083] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0084] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0085] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0086] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0087] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0088] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0089] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0090] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0091] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0092] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0093] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0094] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0095] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0096] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0097] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0098] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0099] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0100] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0101] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0102] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0103] [ka] In a further embodiment of the invention, R3 is -CH2C(O)R4 and R4 is

[0104] [ka] In certain embodiments of the invention, R3 is -CH2C(O)R4, and R4 is

[0105] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0106] [ka] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0107] [ka] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0108] [ka] is.

[0109] In certain embodiments, R3 is -(CH2)2R5, and R5 is

[0110] [ka] In yet another embodiment, R3 is -(CH2)2R5, and R5 is

[0111] [ka] In certain embodiments, R3 is -(CH2)2R5, and R5 is

[0112] [ka] In another embodiment of the invention, R3 is -(CH2)2R5 and R5 is

[0113] [ka] is.

[0114] In some embodiments, a is an integer ranging from 0 to 5. In some embodiments, b is an integer ranging from 0 to 4. In some embodiments, c is an integer ranging from 0 to 6.

[0115] Illustrative pyrazolopyridazine compounds of formula II In certain embodiments, the pyrazolopyridazine compound of formula II has the following structure:

[0116] [ka] TIFF0007755610000072.tif209159TIFF0007755610000073.tif197163TIFF0007755610000074.tif197164TIFF0007755610000075.tif197155TIFF0007755610000076.tif201164TIFF0007755610000077.tif217160TIFF0007755610000078.tif187164TIFF0007755610000079.tif154160, or a pharmaceutically acceptable salt thereof.

[0117] Pyrazolopyridazine compounds of formula III The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula III or a pharmaceutically acceptable salt thereof:

[0118] [ka] wherein Hal is -Cl, -F, -I, or -Br; x is an integer ranging from 0 to 5; each R1 is independently -Cl, -F, -I, -Br, -C1-C3 alkyl, -O-C1-C3 alkyl, -CN, -CF3, -C(O)NH(CH3), or -C≡CCH2OH; R3 is -H, -C1-C6 alkyl, -(C1-C6 alkylene)-OH, -(C1-C6 alkylene)-phenyl, -(C1-C6 alkylene)-O-(C1-C6 alkyl), -C2-C6 alkenyl, -(C1-C6 alkylene)-C(O)R4, -(C1-C6 alkylene)-R5,

[0119] [ka] and; R4 is -OH, -O-(C1-C6 alkyl), -NH2, -NH(C1-C6 alkyl), -NH((C1-C6 alkylene)-OH), -NH((C1-C6 alkylene)N(C1-C6 alkyl)2), -N(C1-C6 alkyl)((C1-C6 alkylene)-CN), -N(C1-C6 alkyl)((C1-C6 alkylene)N(C1-C6 alkyl)2), -NH(C1-C6 alkylene)-O-(C1-C6 alkyl),

[0120] [ka] TIFF0007755610000083.tif230165; a is an integer ranging from 0 to 10; b is an integer ranging from 0 to 8; c is an integer ranging from 0 to 6; R5 is

[0121] [ka] and; each R6 and R7 is independently -H or -I, wherein at least one of R6 and R7 is -I; When R3 is -C1-C3, R7 is -H.

[0122] In certain embodiments, one R6 in the ortho position relative to the pyrazolopyridazino ring system is iodo, and the remaining R6 and R7 groups are hydrogen. In other embodiments, one R6 in the para position relative to the pyrazolopyridazino ring system is iodo, and the remaining R6 and R7 groups are hydrogen. In further embodiments, one R6 in the ortho position relative to the pyrazolopyridazino ring system and one R6 in the para position relative to the pyrazolopyridazino ring system are iodo, and the remaining R6 and R7 groups are hydrogen. In further embodiments, two R6 groups in the ortho position relative to the pyrazolopyridazino ring system and one R6 in the para position relative to the pyrazolopyridazino ring system are iodo, and the remaining R6 and R7 groups are hydrogen. In still further embodiments, two R6 groups in the para position relative to the pyrazolopyridazino ring system and one R6 in the ortho position relative to the pyrazolopyridazino ring system are iodo, and the remaining R6 and R7 groups are hydrogen. In certain embodiments, all R groups are iodo and R is hydrogen. In still further embodiments, R is iodo and R is hydrogen.

[0123] In certain embodiments, one R6 in the para position relative to the pyrazolopyridazino ring system is iodo and R3 is -CH3.

[0124] In certain embodiments, Hal is -Cl. In yet another embodiment, x is 0. In other embodiments, x is 1. In certain embodiments, x is 2.

[0125] In certain embodiments, x is 1 and R is in the ortho position relative to the pyrazolopyridazino ring system. In certain embodiments, x is 1 and R is in the para position relative to the pyrazolopyridazino ring system. In further embodiments, x is 1 and R is in the meta position relative to the pyrazolopyridazino ring system.

[0126] In certain embodiments, x is 2 and R is in the ortho and meta positions relative to the pyrazolopyridazino ring system. In certain embodiments, x is 2 and R is in the ortho and para positions relative to the pyrazolopyridazino ring system. In further embodiments, x is 2 and R is in the para and meta positions relative to the pyrazolopyridazino ring system.

[0127] In still other embodiments, R1 is -Cl. In certain embodiments, R1 is -F. In certain embodiments, R1 is -I. In further embodiments, R1 is -OCH3. In other embodiments, R1 is -CH3. In still other embodiments, R1 is -C(O)N(H)CH3. In certain embodiments, R1 is -CF3. In further embodiments, R1 is -CN. In additional embodiments, R1 is -C≡CCH2OH.

[0128] In still other embodiments, x is 1 or 2 and R1 is -Cl, -F, -Br, -I, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0129] In still other embodiments, Hal is -Cl, x is 1 or 2, and R1 is -Cl, -F, -Br, -I, -OCH3, -CH3, -C(O)N(H)CH3, -CF3, -CN, or -C≡CCH2OH.

[0130] In certain embodiments, R3 is -H. In certain embodiments, R3 is -CH3. In further embodiments, R3 is -CH2CH3. In yet other embodiments, R3 is -CHCH2. In other embodiments, R3 is -CH2CH2OH. In certain embodiments, R3 is -(CH2)2C6H5. In other embodiments, R3 is -CH2C(O)OH. In still other embodiments, R3 is -CH2C(O)N(H)CH3. In certain embodiments, R3 is -CH2C(O)N(H)((CH2)2N(CH3)2). In still other embodiments, R3 is -CH2C(O)N(H)((CH2)3N(CH3)2). In other embodiments, R3 is -CH2C(O)N(CH3)CH2CN. In certain embodiments, R3 is -CH2C(O)NH2. In certain embodiments, R3 is -CH2C(O)N(H)((CH2)2OH). In other embodiments, R3 is -CH2C(O)N(H)((CH2)2OCH3). In still other embodiments, R3 is -CH2C(CH3)2OH. In still other embodiments, R3 is -CH2C(O)OCH3. In further embodiments, R3 is -CH2CH(OH)CH3. In still other embodiments, R3 is -CH2CH2OH. In particular embodiments, R3 is -CH(CH3)CH2OH.

[0131] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0132] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0133] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0134] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0135] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0136] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0137] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0138] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0139] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0140] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0141] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0142] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0143] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0144] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0145] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0146] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0147] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0148] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0149] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0150] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0151] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0152] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0153] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0154] [ka] In yet another embodiment, R3 is -CH2C(O)R4, and R4 is

[0155] [ka] In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0156] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0157] [ka] In a further embodiment of the invention, R3 is -CH2C(O)R4 and R4 is

[0158] [ka] In certain embodiments of the invention, R3 is -CH2C(O)R4, and R4 is

[0159] [ka] In other embodiments, R3 is -CH2C(O)R4, and R4 is

[0160] [ka] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0161] [ka] In a further embodiment, R3 is -CH2C(O)R4, and R4 is

[0162] [ka] is.

[0163] In certain embodiments, R3 is -(CH2)2R5, and R5 is

[0164] [ka] In yet another embodiment, R3 is -(CH2)2R5, and R5 is

[0165] [ka] In certain embodiments, R3 is -(CH2)2R5, and R5 is

[0166] [ka] In another embodiment of the invention, R3 is -(CH2)2R5 and R5 is

[0167] [ka] is.

[0168] In some embodiments, a is an integer ranging from 0 to 5. In some embodiments, b is an integer ranging from 0 to 4. In some embodiments, c is an integer ranging from 0 to 6.

[0169] In certain embodiments, the compound of formula III is compound 3, having the following structure, or a pharmaceutically acceptable salt thereof:

[0170] [ka]

[0171] Pyrazolopyridazine compounds of formula IV The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula IV or a pharmaceutically acceptable salt thereof:

[0172] [ka] wherein R8 is —C1-C3 alkyl.

[0173] In certain embodiments of the invention, R8 is -CH3, and in still further embodiments of the invention, R8 is -CH2CH3. In other embodiments of the invention, R8 is -CH2CH2CH3. In other embodiments of the invention, R8 is -CH(CH3)2.

[0174] In certain embodiments, the compound of formula IV is compound 43, having the following structure, or a pharmaceutically acceptable salt thereof:

[0175] [ka]

[0176] Pyrazolopyridazine compounds of formula V The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula V, or a pharmaceutically acceptable salt thereof:

[0177] [ka] In the formula, R1 is

[0178] [ka] and R2 is

[0179] [ka] and Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

[0180] In certain embodiments, R1 is -I. In other embodiments, R1 is -H. In still other embodiments, R1 is -CH3. In certain embodiments, R1 is -CF3.

[0181] In yet other embodiments, R1 is

[0182] [ka] In certain embodiments, R1 is

[0183] [ka] In yet another embodiment, R1 is

[0184] [ka] In certain embodiments, R1 is

[0185] [ka] In another embodiment, R1 is

[0186] [ka] In yet another embodiment, R1 is

[0187] [ka] In certain embodiments, R1 is

[0188] [ka] In certain embodiments, R1 is

[0189] [ka] In certain embodiments, R1 is

[0190] [ka] In yet another embodiment, R1 is

[0191] [ka] In another embodiment, R1 is

[0192] [ka] is.

[0193] In yet other embodiments, R1 is

[0194] [ka] In certain embodiments, R1 is

[0195] [ka] In yet another embodiment, R1 is

[0196] [ka] In another embodiment, R1 is

[0197] [ka] In certain embodiments, R1 is

[0198] [ka] In a further embodiment, R1 is

[0199] [ka] In yet another embodiment, R1 is

[0200] [ka] is.

[0201] In certain embodiments, R2 is -H. In still other embodiments, R2 is

[0202] [ka] In certain embodiments, R2 is

[0203] [ka] In yet another embodiment, R2 is

[0204] [ka] In a further embodiment, R2 is

[0205] [ka] where a=1 and Hal is -F.

[0206] In certain embodiments, R2 is

[0207] [ka] In yet another embodiment, R2 is

[0208] [ka] In certain embodiments, R2 is

[0209] [ka] In another embodiment, R2 is

[0210] [ka] In yet another embodiment, R2 is

[0211] [ka] In certain embodiments, R2 is

[0212] [ka] is.

[0213] In a further embodiment, when a is 2, each Hal is the same or different.

[0214] In certain embodiments, the compound of formula V has the following structure:

[0215] [ka] TIFF0007755610000155.tif198161TIFF0007755610000156.tif203160TIFF0007755610000157.tif207163 or a pharmaceutically acceptable salt thereof.

[0216] Pyrazolopyridazine compounds of formula VI The present invention also provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula VI, or a pharmaceutically acceptable salt thereof:

[0217] [ka] wherein R3 is:

[0218] [ka] and; b is 0 or 1; c is 1 or 2.

[0219] In certain embodiments, b is 0. In other embodiments, b is 1 and -F is in the meta position relative to the pyrazolopyridazino ring system. In yet other embodiments, b is 1 and -F is in the para position relative to the pyrazolopyridazino ring system.

[0220] In certain embodiments, R3 is -CF3. In certain embodiments, R3 is

[0221] [ka] In other embodiments, R3 is

[0222] [ka] In yet another embodiment, R3 is

[0223] [ka] In a further embodiment, R3 is

[0224] [ka] In yet another embodiment, R3 is

[0225] [ka] In certain embodiments, R3 is

[0226] [ka] is.

[0227] In other embodiments, R3 is

[0228] [ka] In yet another embodiment, R3 is

[0229] [ka] In certain embodiments, R3 is

[0230] [ka] In a further embodiment, R3 is

[0231] [ka] In a further embodiment, R3 is

[0232] [ka] is.

[0233] In certain embodiments, R3 is

[0234] [ka] In other embodiments, R3 is

[0235] [ka] In yet another embodiment, R3 is

[0236] [ka] In a further embodiment, R3 is

[0237] [ka] In yet another embodiment, R3 is

[0238] [ka] In certain embodiments, R3 is

[0239] [ka] is.

[0240] In certain embodiments, R3 is

[0241] [ka] In a further embodiment, R3 is

[0242] [ka] and c=1. In yet another embodiment, R3 is

[0243] [ka] In certain embodiments, R3 is

[0244] [ka] In other embodiments, R3 is

[0245] [ka] and c=2. In yet another embodiment, R3 is

[0246] [ka] is.

[0247] In certain embodiments, R3 is

[0248] [ka] In other embodiments, R3 is

[0249] [ka] In yet another embodiment, R3 is

[0250] [ka] is.

[0251] In certain embodiments, the compound of Formula VI has the following structure:

[0252] [ka] TIFF0007755610000187.tif209167TIFF0007755610000188.tif58158 or a pharmaceutically acceptable salt thereof.

[0253] Pyrazolopyridazine compounds of formula VII The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula VII or a pharmaceutically acceptable salt thereof:

[0254] [ka] In the formula, R4 is

[0255] [ka] is.

[0256] In certain embodiments, R4 is

[0257] [ka] In certain embodiments, R4 is

[0258] [ka] In other embodiments, R4 is

[0259] [ka] In yet another embodiment, R4 is

[0260] [ka] is.

[0261] In certain embodiments, the compound of formula VII has the structure:

[0262] [ka] or a pharmaceutically acceptable salt thereof.

[0263] Pyrazolopyridazine compounds of formula XIII The present invention further provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIII or a pharmaceutically acceptable salt thereof:

[0264] [ka] In the formula, R5 is

[0265] [ka] TIFF0007755610000198.tif65159; R6 is:

[0266] [ka] and; Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

[0267] In certain embodiments, R5 is -I. In other embodiments, R5 is -H. In still other embodiments, R5 is -CH3. In certain embodiments, R5 is -CF3.

[0268] In yet other embodiments, R5 is

[0269] [ka] In certain embodiments, R5 is

[0270] [ka] In yet another embodiment, R5 is

[0271] [ka] In certain embodiments, R5 is

[0272] [ka] In other embodiments, R5 is

[0273] [ka] In yet another embodiment, R5 is

[0274] [ka] In certain embodiments, R5 is

[0275] [ka] In certain embodiments, R5 is

[0276] [ka] In certain embodiments, R5 is

[0277] [ka] In yet another embodiment, R5 is

[0278] [ka] In other embodiments, R5 is

[0279] [ka] is.

[0280] In yet other embodiments, R5 is

[0281] [ka] In certain embodiments, R5 is

[0282] [ka] In yet another embodiment, R5 is

[0283] [ka] In other embodiments, R5 is

[0284] [ka] In certain embodiments, R5 is

[0285] [ka] In a further embodiment, R5 is

[0286] [ka] In yet another embodiment, R5 is

[0287] [ka] is.

[0288] In certain embodiments, R5 is

[0289] [ka] In a further embodiment, R5 is

[0290] [ka] In a further embodiment, R5 is

[0291] [ka] In other embodiments, R5 is

[0292] [ka] In yet another embodiment, R5 is

[0293] [ka] In certain embodiments, R5 is

[0294] [ka] is.

[0295] In a further embodiment, R5 is

[0296] [ka] In yet another embodiment, R5 is

[0297] [ka] is.

[0298] In certain embodiments, R5 is

[0299] [ka] In other embodiments, R5 is

[0300] [ka] In yet another embodiment, R5 is

[0301] [ka] In certain embodiments, R5 is

[0302] [ka] is.

[0303] In a further embodiment, R5 is

[0304] [ka] In yet another embodiment, R5 is

[0305] [ka] is.

[0306] In certain embodiments, R5 is

[0307] [ka] In other embodiments, R5 is

[0308] [ka] In yet another embodiment, R5 is

[0309] [ka] In certain embodiments, R5 is

[0310] [ka] is.

[0311] In a further embodiment, R5 is

[0312] [ka] In yet another embodiment, R5 is

[0313] [ka] is.

[0314] In certain embodiments, R5 is

[0315] [ka] is.

[0316] In certain embodiments, R6 is

[0317] [ka] In a further embodiment, R6 is

[0318] [ka] and a=0. In other embodiments, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0319] [ka] In yet another embodiment, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0320] [ka] In certain embodiments, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0321] [ka] In certain embodiments, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0322] [ka] In a further embodiment, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0323] [ka] In other embodiments, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt, and R6 is

[0324] [ka] In certain embodiments, R6 is

[0325] [ka] In a further embodiment, R6 is

[0326] [ka] In certain embodiments, R6 is

[0327] [ka] is.

[0328] In a further embodiment, R6 is

[0329] [ka] In yet another embodiment, R6 is

[0330] [ka] In another embodiment, R6 is

[0331] [ka] In certain embodiments, R6 is

[0332] [ka] In yet another embodiment, R6 is

[0333] [ka] In certain embodiments, R6 is

[0334] [ka] In a further embodiment, R6 is

[0335] [ka] In yet another embodiment, R6 is

[0336] [ka] is.

[0337] In certain embodiments, R6 is

[0338] [ka] In another embodiment, R6 is

[0339] [ka] is.

[0340] In a further embodiment, when a is 2, each Hal is the same or different.

[0341] In certain embodiments, the compound of Formula XIII has the structure:

[0342] [ka] TIFF0007755610000261.tif213162TIFF0007755610000262.tif216164TIFF0007755610000263.tif178157 or a pharmaceutically acceptable salt thereof.

[0343] In other embodiments, the pyrazolopyridazine compound of Formula XIII is a pharmaceutically acceptable salt and has the following structure:

[0344] [ka]

[0345] Pyrazolopyridazine compounds of formula XIV The present invention also provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIV or a pharmaceutically acceptable salt thereof:

[0346] [ka] In the formula, R7 is

[0347] [ka] and; b is 0 or 1; c is 1 or 2.

[0348] In certain embodiments, b is 0. In other embodiments, b is 1 and -F is in the meta position relative to the pyrazolopyridazino ring system. In yet other embodiments, b is 1 and -F is in the para position relative to the pyrazolopyridazino ring system.

[0349] In certain embodiments, R7 is -CF3.

[0350] [ka] In other embodiments, R7 is

[0351] [ka] In yet another embodiment, R7 is

[0352] [ka] In a further embodiment, R7 is

[0353] [ka] In yet another embodiment, R7 is

[0354] [ka] In certain embodiments, R7 is

[0355] [ka] is.

[0356] In other embodiments, R7 is

[0357] [ka] In yet another embodiment, R7 is

[0358] [ka] In certain embodiments, R7 is

[0359] [ka] In a further embodiment, R7 is

[0360] [ka] In a further embodiment, R7 is

[0361] [ka] is.

[0362] In certain embodiments, R7 is

[0363] [ka] In other embodiments, R7 is

[0364] [ka] In yet another embodiment, R7 is

[0365] [ka] In a further embodiment, R7 is

[0366] [ka] In yet another embodiment, R7 is

[0367] [ka] In certain embodiments, R7 is

[0368] [ka] is.

[0369] In certain embodiments, R7 is

[0370] [ka] In a further embodiment, R7 is

[0371] [ka] and c=1. In yet another embodiment, R7 is

[0372] [ka] In certain embodiments, R7 is

[0373] [ka] In other embodiments, R7 is

[0374] [ka] and c=2. In yet another embodiment, R7 is

[0375] [ka] is.

[0376] In certain embodiments, R7 is

[0377] [ka] In other embodiments, R7 is

[0378] [ka] In yet another embodiment, R7 is

[0379] [ka] In yet another embodiment, R7 is

[0380] [ka] is.

[0381] In certain embodiments, the compound of formula XIV has the structure:

[0382] [ka] or a pharmaceutically acceptable salt thereof.

[0383] Pyrazolopyridazine compounds of formula XV The present invention also provides a method of treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XV or a pharmaceutically acceptable salt thereof:

[0384] [ka] In the formula, R8 is

[0385] [ka] is.

[0386] In certain embodiments, R8 is

[0387] [ka] In certain embodiments, R8 is

[0388] [ka] In other embodiments, R8 is

[0389] [ka] In yet another embodiment, R8 is

[0390] [ka] In a further embodiment, R8 is

[0391] [ka] In yet another embodiment, R8 is

[0392] [ka] In certain embodiments, R8 is

[0393] [ka] is.

[0394] In certain embodiments, R8 is

[0395] [ka] In certain embodiments, R8 is

[0396] [ka] In other embodiments, R8 is

[0397] [ka] In yet another embodiment, R8 is

[0398] [ka] In a further embodiment, R8 is

[0399] [ka] In yet another embodiment, R8 is

[0400] [ka] is.

[0401] In certain embodiments, the compound of formula XV has the structure:

[0402] [ka] TIFF0007755610000311.tif53157 or a pharmaceutically acceptable salt thereof.

[0403] Other pyrazolopyridazine compounds The present invention further provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a compound having the structure of the following formula, or a pharmaceutically acceptable salt thereof:

[0404] [ka] TIFF0007755610000313.tif124157TIFF0007755610000314.tif190162

[0405] Non-pyrazolopyridazine compounds The compounds in Table 1 below or pharmaceutically acceptable salts of the compounds are non-pyrazolopyridazine compounds.

[0406] In one embodiment, the present invention provides a non-pyrazolopyridazine compound. In a further embodiment, the present invention provides a pharmaceutical composition comprising an effective amount of a non-pyrazolopyridazine compound and a pharmaceutically acceptable carrier or vehicle.

[0407] The present invention further provides a method for treating or preventing a proteopathy, comprising administering to a subject in need thereof an effective amount of a non-pyrazolopyridazine compound.

[0408] [Table 1] TIFF0007755610000316.tif226155TIFF0007755610000317.tif223157TIFF0007755610000318.tif207153TIFF0007755610000319.t if219155TIFF0007755610000320.tif202154TIFF0007755610000321.tif203156TIFF0007755610000322.tif215154TIFF00077556100 00323.tif195155TIFF0007755610000324.tif212157TIFF0007755610000325.tif218157TIFF0007755610000326.tif209155TIFF000 7755610000327.tif212156TIFF0007755610000328.tif221156TIFF0007755610000329.tif210155TIFF0007755610000330.tif139157

[0409] Some of the compounds disclosed herein, for example, compounds 44, 63, 72, 74, 83, 88, 89, 98-101, 111, 124a, 124b, Vp, Vq, Vt, VIj, VIt, VIu, VIx, VIy, XIIIa, XIIIe, XIIIf, XIIIg, XIIIh, XIIIi, XIIIv, and XIIIw, are drawn with a thick wedge or hatched wedge indicating the absolute stereochemistry.

[0410] Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds can be in the form of a salt. In some embodiments, the salt is a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid that forms the acid addition salt can be an organic acid or an inorganic acid. The base that forms the base addition salt can be an organic base or an inorganic base. In some embodiments, the pharmaceutically acceptable salt is a metal salt. In some embodiments, the pharmaceutically acceptable salt is an ammonium salt.

[0411] Acid addition salts can be formed by the addition of an acid to the free base form of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the acid is organic. In some embodiments, the acid is inorganic. Non-limiting examples of suitable acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, nitrous acid, sulfuric acid, sulfurous acid, phosphoric acid, nicotinic acid, isonicotinic acid, lactic acid, salicylic acid, 4-aminosalicylic acid, tartaric acid, ascorbic acid, gentisic acid, gluconic acid, glucuronic acid, saccharinic acid, formic acid, benzoic acid, glutamic acid, pantothenic acid, acetic acid, propionic acid, butyric acid, fumaric acid, succinic acid, citric acid, oxalic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, glycolic acid, malic acid, cinnamic acid, mannitol ... Examples of sulfonic acids include benzoic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, phenylacetic acid, N-cyclohexylsulfamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 2-phosphoglyceric acid, 3-phosphoglyceric acid, glucose-6-phosphate, and amino acids.

[0412] Non-limiting examples of suitable acid addition salts include hydrochloride, hydrobromide, hydroiodide, nitrate, nitrite, sulfate, sulfite, phosphate, hydrogen phosphate, dihydrogen phosphate, carbonate, bicarbonate, nicotinate, isonicotinate, lactate, salicylate, 4-aminosalicylate, tartrate, ascorbate, gentisate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, citrate, oxalate, maleate, hydroxymaleate, methylmaleate, Examples include glycolate, malate, cinnamate, mandelate, 2-phenoxybenzoate, 2-acetoxybenzoate, embonate, phenylacetate, N-cyclohexylsulfamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate, ethane-1,2-disulfonate, 4-methylbenzenesulfonate, naphthalene-2-sulfonate, naphthalene-1,5-disulfonate, 2-phosphoglycerate, 3-phosphoglycerate, glucose-6-phosphate, and amino acid salts.

[0413] Metal salts can be formed by the addition of inorganic bases to pyrazolopyridazine compounds or non-pyrazolopyridazine compounds having a carboxyl group. The inorganic base consists of a metal cation paired with a basic counterion, such as hydroxide, carbonate, bicarbonate, or phosphate. The metal can be an alkali metal, alkaline earth metal, transition metal, or main group metal. Non-limiting examples of suitable metals include lithium, sodium, potassium, cesium, cerium, magnesium, manganese, iron, calcium, strontium, cobalt, titanium, aluminum, copper, cadmium, and zinc.

[0414] Non-limiting examples of suitable metal salts include lithium salts, sodium salts, potassium salts, cesium salts, cerium salts, magnesium salts, manganese salts, iron salts, calcium salts, strontium salts, cobalt salts, titanium salts, aluminum salts, copper salts, cadmium salts, and zinc salts.

[0415] Ammonium salts can be generated by the addition of ammonia or organic amines to pyrazolopyridazine compounds or non-pyrazolopyridazine compounds having a carboxyl group. Non-limiting examples of suitable organic amines include triethylamine, diisopropylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, N-methylmorpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrazole, imidazole, pyrazine, pipyrazine, ethylenediamine, N,N'-dibenzylethylenediamine, procaine, chloroprocaine, choline, dicyclohexylamine, and N-methylglucamine.

[0416] Non-limiting examples of suitable ammonium salts include triethylammonium salts, diisopropylammonium salts, ethanolammonium salts, diethanolammonium salts, triethanolammonium salts, morpholinium salts, N-methylmorpholinium salts, piperidinium salts, N-methylpiperidinium salts, N-ethylpiperidinium salts, dibenzylammonium salts, piperazinium salts, pyridinium salts, pyrazolium salts, imidazolium salts, pyrazinium salts, ethylenediammonium salts, N,N'-dibenzylethylenediammonium salts, procaine salts, chloroprocaine salts, choline salts, dicyclohexylammonium salts, and N-methylglucamine salts.

[0417] Method for making pyrazolopyridazine compounds Methods of making pyrazolopyridazine compounds are disclosed in U.S. Pat. No. 8,765,762, U.S. Patent Application Publication No. 2013 / 0252936, and U.S. Patent Application Publication No. 2014 / 0121197, each of which is incorporated by reference in its entirety.

[0418] Compound 114 can be synthesized according to Vasilevsky, S. F. & Tretyakov, E. V. Cinnolines and pyrazolopyridazines. - Novel synthetic and mechanistic aspects of the Richter reaction. Liebigs Annalen 1995, 775-779 (1995).

[0419] Non-limiting examples of synthetic schemes useful for synthesizing pyrazolopyridazine compounds include the following schemes.

[0420] [ka]

[0421] Scheme 1 outlines the preparation of pyrazolopyridazine compounds bearing a 1-N-methyl group, where R' and R" are independently unsubstituted or substituted phenyl groups. For example, a 2-cyanocarbonyl compound in which R' is unsubstituted or substituted phenyl is condensed with N-methylhydrazine to give a 3-substituted-1-methyl-1H-pyrazol-5-amine. The 5-amino group is acylated, for example, with acetic anhydride in the presence of a base such as pyridine, to give a 5-amide compound. The 5-amide compound is iodinated, for example, with a mixture of iodine and iodic acid in a solvent such as ethanol (EtOH), to give N-(3-substituted-4-iodo-1-methyl-1H-pyrazol-5-yl)acetamide. The 5-amide compound is then iodinated with a mixture of iodine and iodic acid in a solvent such as ethanol (EtOH) to give N-(3-substituted-4-iodo-1-methyl-1H-pyrazol-5-yl)acetamide. The 5-amide compound is then iodinated with dimethylformamide (DMF) containing a base such as triethylamine. Palladium-mediated cross-coupling, such as Sonogashira cross-coupling, of the acetamide with an R"-substituted terminal alkyne, catalyzed by a palladium complex, such as palladium(II) bistriphenylphosphine dichloride, in the presence of copper(I) iodide in a solvent such as Benzyl Alcohol (E), affords a disubstituted alkyne, where R" is unsubstituted or substituted phenyl. Saponification of the alkyne acetamide with a base, such as sodium hydroxide, in a solvent such as ethanol affords a primary amine. Diazotization of the primary amine with sodium nitrite in concentrated hydrochloric acid affords a diazo intermediate, which is cyclized to afford a pyrazolopyridazine compound bearing a 1-N-methyl group, where R' and R" are independently unsubstituted or substituted phenyl groups.

[0422] [ka]

[0423] Scheme 2 outlines the preparation of pyrazolopyridazine compounds having an R group, where R' is an unsubstituted or substituted phenyl group. R' and R3 can be the same or different. For example, 4,6-dichloro-3-phenylpyridazine is deprotonated with a base such as lithium diisopropylamide (LDA) in a solvent such as tetrahydrofuran (THF), and the resulting 5-lithio species is condensed with an unsubstituted or substituted benzaldehyde to give a secondary alcohol. This alcohol is oxidized to a ketone with an oxidizing agent such as manganese dioxide in a solvent such as toluene. This ketone is condensed with an R3-substituted hydrazine in a solvent such as ethanol to give an intermediate hydrazone, which is cyclized to give a pyrazolopyridazine compound having a 1-N-R3 group, where R3 is defined in Formulas II and III, and R' is an unsubstituted or substituted phenyl group.

[0424] [ka]

[0425] Scheme 3 outlines the preparation of pyrazolopyridazine compounds having a 1-N-methyl group, where R' is a cyano group, alkyne, alkene, or aryl group. For example, 1-methyl-3-iodophenyl-4-chloro-5-phenyl-1H-pyrazolo[3,4-c]pyridazine is coupled with a suitable coupling partner, such as a cyanide salt, a terminal alkyne, an alkenyl halide, or an aryl halide, optionally in the presence of a suitable catalyst, such as a palladium complex, optionally in the presence of a non-palladium transition metal salt, such as a zinc or copper salt, and optionally in the presence of an additive, such as triphenylphosphine or an organic amine base, to obtain pyrazolopyridazine compounds having a 1-N-methyl group and where R' is a cyano group, alkyne, alkene, or aryl group. The position of R' in the product, i.e., ortho, meta, or para, is the same as the position of the iodo group in the starting material.

[0426] [ka]

[0427] Scheme 4 generally describes the preparation of pyrazolopyridazine compounds.

[0428] [ka] (i) ethyl hydrazinoacetate hydrochloride, EtOH, reflux, 2 h; (ii) Ac2O, pyridine, 25 °C, 16 h, or AcCl, N-methylmorpholine, CH2Cl2, 25 °C, 3 h; (iii) I2, HIO3, EtOH, 50 °C; (iv) phenylacetylene, Pd(PPh3)2Cl2, CuI, Et3N, DMF, 90 °C.

[0429] Scheme A outlines the preparation of ethyl 2-[5-acetamido-3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-1-yl]acetate and N-[3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-5-yl]acetamide from benzoylacetonitrile.

[0430] Compound 8A of Scheme A: Ethyl 2-(5-amino-3-(phenyl)-1H-pyrazol-1-yl)acetate A mixture of benzoylacetonitrile (7A, 44 g, 304 mmol) and ethyl hydrazinoacetate hydrochloride (47 g, 304 mmol) in ethanol (400 mL) is heated to reflux for 2 h. The reaction mixture is concentrated in vacuo. The crude reaction mixture is partitioned between CHCl (400 mL) and saturated NaHCO(aq). The aqueous phase is extracted with CHCl, and the organic phases are combined, dried over MgSO, filtered, and evaporated to give compound 8A as a solid (70 g, 95% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 7.73(m, 2H), 7.38(m, 2H), 7.26(m, 1H), 5.96(s, 1H), 4.86(s, 2H), 4.25(m, 2H), 3.7(s, 2H), 1.28(s, 3H).

[0431] Compound 10A of Scheme A: Ethyl 2-(5-acetamido-3-phenyl-1H-pyrazol-1-yl)acetate To a solution of ethyl 2-(5-amino-3-(phenyl)-1H-pyrazol-1-yl)acetate (8A, 41.7 g, 0.17 mol) in pyridine (200 mL) under a nitrogen atmosphere at 0 °C, acetic anhydride (17.4 g, 0.17 mol) is added dropwise. The reaction mixture is stirred at room temperature (RT) for 16 h. The reaction mixture is concentrated in vacuo. The residue is diluted with CHCl and water. The layers are separated, and the organic layer is washed with water and brine, dried (MgSO), and concentrated in vacuo. CHCl is added to the residue, and the solid is collected by filtration to give compound 10A as a solid (22 g, 45% yield). The mother liquor is concentrated in vacuo and washed with cold CHCl to give a second batch of compound 10A (15 g, 31% yield). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 10.13(s, 1H), 7.81(d, J=7.6Hz, 2H), 7.45(t, J=7.6Hz, 2H), 7.40-7.32(m, 1H), 6. 80(s, 1H), 5.07(s, 2H), 4.22(q, J=7.1Hz, 2H), 2.14(s, 3H), 1.28(t, J=7.1Hz, 3H).

[0432] Compound 11A of Scheme A: N-(3-phenyl-1H-pyrazol-5-yl)acetamide To a solution of 3-phenyl-1H-pyrazol-5-amine (9, 18.6 g, 0.117 mol) and N-methylmorpholine (30.8 mL, 0.281 mol) in CHCl (250 mL) was added dropwise at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at RT for 3 h. The reaction mixture was diluted with CHCl and water. The layers were separated, and the organic layer was washed with water and brine, dried (phase separator cartridge), and concentrated in vacuo. Diethyl ether was added to the residue, and the solid was collected by filtration to give compound 11A as a solid (25.1 g, 88% yield). 1H NMR (400MHz, DMSO-d6) δ(ppm) 12.79(s, 1H), 10.40(s, 1H), 7.71(d, J=7.5Hz, 2H), 7.44(dd, J=7.6, 7.6Hz, 2H), 7.34(dd, J=7.2, 7.2Hz, 1H), 6.88(s, 1H), 2.02(s, 3H).

[0433] Compound 12A of Scheme A: Ethyl 2-(5-acetamido-4-iodo-3-phenyl-1H-pyrazol-1-yl)acetate A suspension of compound 10A (37 g, 129 mmol), iodic acid (5.6 g, 32 mmol), and iodine (19.7 g, 77 mmol) in ethanol (400 mL) was heated at 50 °C for 2 h and cooled to RT. The reaction mixture was concentrated in vacuo, and the residue was passed through a pad of silica gel and eluted with CHCl / diethyl ether (1:0 to 97:3). The residue was partitioned between CHCl and 2 M NaSO solution (aq). The layers were separated, and the organic layer was washed, dried (MgSO), and concentrated in vacuo to give a residue that was partially purified by chromatography (silica gel, CHCl / isohexane 1:1 to 1:0, then CHCl / diethyl ether 9:1 to 8:2), followed by trituration with diethyl ether to give compound 12A as an off-white solid (43 g, 81% yield). 1 H NMR (400MHz, CDCl3) as a 3:1 mixture of rotamers δ(ppm) 7.81(d, J=7.6Hz, 2H), 7.45-7.35(m, 3H), 7.15(br s, 0.75H), 6.85(br s, 0.25H), 4.97(s, 2H), 4.25(q, J=7.1Hz, 2H), 2.24(s, 2.25H), 2.04(s, 0.75H), 1.30(t, J=7.1Hz, 3H).

[0434] Compound 13A of Scheme A: N-(4-iodo-3-phenyl-1H-pyrazol-5-yl)acetamide A suspension of compound 11A (25.1 g, 0.103 mol), iodic acid (4.5 g, 0.026 mol), and iodine (15.7 g, 0.062 mol) in ethanol (250 mL) was heated at 50 °C for 3 h and cooled to RT. The reaction mixture was concentrated in vacuo and partitioned between CHCl and 2 M NaSO solution (aq). The layers were separated, and the organic layer was washed with brine, dried (phase separator cartridge), and concentrated in vacuo to give a mixture of compound 13A and starting material 11A (2.2:1, 30.3 g). This mixture was reacted again under the same conditions with iodic acid (1.6 g, 9.6 mmol) and iodine (9.7 g, 38 mmol) in ethanol (250 mL) to give compound 13A as a solid (31.9 g, 84% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 11.74-11.74(m, 1H), 7.81(d, J=7.2Hz, 2H), 7.59(s, 1H), 7.49-7.38(m, 3H), 2.31(s, 3H).

[0435] Compound 14A of Scheme A: Ethyl 2-[5-acetamido-3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-1-yl]acetate Nitrogen was bubbled through a mixture of compound 12A (18.6 g, 45 mmol), phenylacetylene (9.2 g, 90 mmol), copper iodide (860 mg, 4.5 mmol), triethylamine (200 mL), and DMF (75 mL) for 15 min. Bis(triphenylphosphine)palladium(II) dichloride (1.6 g, 2.25 mmol) was added, and the reaction mixture was stirred at 90 °C under nitrogen for 4.5 h. The reaction mixture was cooled to RT and diluted with ethyl acetate and water. The organic phase was washed with water and brine, dried (MgSO), filtered, and concentrated in vacuo. The residue was partially purified by column chromatography (silica gel, CHCl, then isohexane / ethyl acetate 1:1, then CHCl / ethyl acetate 9:1 to 8:2) and then triturated with diethyl ether to give compound 14A as a solid (13 g, 75% yield). 1H NMR (400MHz, DMSO-d6) δ(ppm) 10.38(s, 1H), 8.13-8.09(m, 2H), 7.60-7.44(m, 8H), 5.03(s, 2H), 4.23(q, J=7.1Hz, 2H), 2.17(s, 3H), 1.28(t, J=7.1Hz, 3H).

[0436] Compound 15A of Scheme A: N-[3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-5-yl]acetamide Compound 15A (12.5 g, 48% yield) was obtained from compound 13A (31.87 g, 86 mmol) by a procedure similar to that described for the synthesis of compound 14A. 1 H NMR (400MHz, CDCl3) δ(ppm) 11.57-11.57(m, 1H), 8.11(d, J=7.4Hz, 2H), 7.91(s, 1H), 7.55-7.49(m, 2 H), 7.44(dd, J=7.5, 7.5Hz, 2H), 7.37(dd, J=1.9, 5.0Hz, 4H), 2.32(s, 3H).

[0437] [ka] (i) NaOH, EtOH, 80°C; (ii) (a) NaBH4, EtOH, 25°C; (b) NaOH, EtOH, 80°C; (iii) NaNO2, c.HCl, -10°C to 25°C; (iv) MeMgCl, THF, 0°C to 25°C.

[0438] Scheme B outlines the preparation of compounds 3B and 4B.

[0439] Compound 16B of Scheme B: Sodium 2-[5-amino-3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-1-yl]acetate A mixture of compound 14B (13 g, 34 mmol), ethanol (150 mL), and 25% NaOH solution (aq) (150 mL) is stirred and heated to 80 °C for 8 h and cooled to RT. Upon cooling, a precipitate forms. The precipitate is filtered and washed with a cooled mixture of ethyl acetate / water (1:1). The solid is further triturated with diethyl ether, filtered, and dried (MgSO4) to give compound 16B as a solid (9.8 g, 85% yield). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 8.06(d, J=7.8Hz, 2H), 7.56(d, J=7.6Hz, 2H), 7.50-7.39(m, 4H), 7.39-7.31(m, 2H), 4.31(s, 2H).

[0440] Compound 17B of Scheme B: 2-[5-amino-3-phenyl-4-(2-phenylethynyl-1H-)pyrazol-1-yl]ethanol To a suspension of compound 14B (22.4 g, 58 mmol) in ethanol (290 mL) was added sodium borohydride (11 g, 289 mmol), and the reaction mixture was stirred at RT for 16 h. The reaction mixture was partially concentrated to a final volume of 250 mL. 25% NaOH solution (aq) (250 mL) was added, and the reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was cooled to room temperature, and the phases were separated. The aqueous phase was extracted three times with ethyl acetate, and the organic phases were combined, dried (MgSO), filtered, and concentrated in vacuo. The residue was triturated with diethyl ether (20 mL), and the product was filtered and dried in vacuo to give compound 17B as an off-white solid (9.96 g, 57% yield). The mother liquor was concentrated in vacuo and purified by column chromatography (silica gel, gradient 0-100% ethyl acetate / isohexane) to give an additional yield (1.79 g, 10% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 7.78-7.74(m, 4H), 7.55-7.48(m, 6H), 4.98(t, J=4.8Hz, 2H), 4.29(m, 2H), 3.03(t, J=6.4Hz, 1H).

[0441] Compound 18B of Scheme B: 3-phenyl-4-(2-phenylethynyl)-1H-pyrazol-5-amine Compound 18B (5.4 g, 50% yield) was obtained from compound 15B (12.5 g, 41 mmol) by a procedure similar to that described for the synthesis of compound 16B. 1 H NMR (400MHz, CDCl3) δ(ppm) 7.87(d, J=7.2Hz, 2H), 7.51-7.43(m, 4H), 7.42-7.32(m, 4H), 4.09(s, 2H).

[0442] Compound 19B of Scheme B: 2-(4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)acetic acid Sodium nitrite (1.86 g, 26.9 mmol) is added portionwise to cHCl (30 mL) at 0 °C, and after stirring for 15 min, compound 16B (3 g, 8.85 mmol) is added portionwise as a solid to the reaction mixture. The suspension is then stirred at RT for 16 h. The reaction mixture is diluted with CHCl and washed with water and brine. The organic layer is dried (MgSO) and concentrated in vacuo. The residue is purified by column chromatography (silica gel, diethyl ether / CHCl 1:9) to give compound 19B as a solid (1.7 g, 53% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 7.80-7.72(m, 4H), 7.56-7.47(m, 6H), 5.64(s, 2H), 2.10(s, 1H).

[0443] Compound 3B of Scheme B: 2-(4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)ethan-1-ol Sodium nitrite (4.58 g, 66.3 mmol) is added portionwise to CHC1C (220 mL) at -10 °C and stirred for 10 min. Compound 17B (6.7 g, 22.1 mmol) is added as a solid. The reaction mixture is allowed to warm and sonicated for 5 min, then stirred at RT for 2 h. The reaction mixture is diluted with CHCl and water, and the aqueous phase is extracted with CHCl. ​​The organic phases are combined, dried (MgSO), filtered, and concentrated in vacuo. The residue is partially purified by column chromatography (silica gel, gradient 0-100% ethyl acetate / isohexane). The resulting residue is triturated with diethyl ether and then ethyl acetate to give compound 3B as a solid (900 mg, 12% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 7.79-7.75(m, 4H), 7.55-7.46(m, 6H), 4.98(m, 2H), 4.32-4.25(m, 2H), 3.04(t, J=6.4Hz, 1H). 13 C NMR (100 MHz, CDCl3) δ (ppm) 153.09, 151.98, 143.65, 134.48, 130.11, 129.35, 129.33, 129.06, 128.29, 128.17, 127.39, 127.33, 113.70, 60.64, 50.63. LC-MS (Method 1: HPLC (Phenomenex Luna 5 μm C18, 100 × 4.6 mm) with a gradient of 5–95% acetonitrile in water (0.1% formic acid in each mobile phase)). t 4.14 min; m / z 351 [M+H] 99.04% purity.

[0444] Compound 21B of Scheme B: N-(1-(2-hydroxy-2-methylpropyl)-3-phenyl-4-(phenylethynyl)-1H-pyrazol-5-yl)acetamide To a solution of compound 14B (1.0 g, 2.58 mmol) in THF (26 mL) is added methylmagnesium chloride (3 M solution in THF, 3 mL, 9 mmol) at 0 °C. The resulting solution is stirred at RT for 3.5 h, then diluted with ethyl acetate and quenched by the addition of 1 M HCl (aq). The aqueous phase is extracted with ethyl acetate, and the combined organic layers are dried (MgSO4) and concentrated in vacuo. The resulting residue is purified using chromatography (silica gel, 0-75% ethyl acetate / isohexane gradient) to give compound 21B as a solid (529 mg, 55% yield). 1 H NMR (400 MHz, CDCl3) for compound 21B δ (ppm) 8.11 (dd, J = 7.5, 12.3 Hz, 2H), 7.51-7.40 (m, 4H), 7.37-7.31 (m, 4H), 4.15-4.07 (m, 2H), 2.24-2.23 (m, 3H), 1.28 (s, 6H) and N-[2-acetonyl-5-phenyl-4-(2-phenylethynyl)pyrazol-3-yl]acetamide δ (ppm) as a 1.5:1 mixture of 8.11 (dd, J = 7.5, 12.3 Hz, 2H), 7.51-7.40 (m, 4H), 7.37-7.31 (m, 4H), 4.95 (s, 2H), 2.24-2.23 (m, 3H), 1.28 (s, 6H).

[0445] Compound 22B of Scheme B: 1-(5-amino-3-phenyl-4-(phenylethynyl)-1H-pyrazol-1-yl)-2-methylpropan-2-ol Compound 22B (310 mg, 59% yield) was synthesized from compound 21B (597 mg, 1.6 mmol) following a similar procedure outlined for the synthesis of compound 16B. 1 H NMR (400MHz, CDCl3) δ(ppm) 8.11-8.08(m, 2H), 7.50-7.47(m, 2H), 7.41(dd, J=7.5, 7.5Hz, 2H), 7.37 -7.29(m, 4H), 4.48(s, 2H), 4.01(s, 2H), 2.70(s, 1H), 1.32-1.31(m, 6H).

[0446] Compound 4B of Scheme B: 1-(4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)-2-methylpropan-2-ol Sodium nitrite (121 mg, 1.75 mmol) was added in one portion to chilled (cooling bath -15 °C) cHCl (9 mL). The suspension was stirred for 10 min, followed by the addition of compound 22B (290 mg, 0.88 mmol). After 5 min, the cooling bath was removed and the reaction mixture was stirred at RT for 3 h. The reaction was cooled again (0 °C) and CHCl was added, followed by water. The aqueous phase was extracted with CHCl, and the organic phases were combined, dried (MgSO), filtered, and concentrated in vacuo. The crude material was purified by column chromatography (silica gel, 0-50% ethyl acetate / isohexane gradient) to give compound 4B as an orange oil (56 mg). The resulting material was further purified by preparative HPLC to give compound 4B as a solid (34 mg, 10% yield). 1 H NMR (400MHz, CDCl3) δ(ppm) 7.81-7.75(m, 4H), 7.55-7.50(m, 6H), 4.85(s, 2H), 3.50(s, 1H), 1.36(s, 6H). 13 C NMR (100 MHz, CDCl3) δ (ppm) 154.36, 152.91, 144.60, 135.42, 130.96, 130.29, 130.28, 129.93, 129.23, 129.07, 128.29, 128.24, 114.16, 71.52, 58.60, 27.26. LCMS (Analytical Method 1: HPLC (Phenomenex Luna 5 μm C18, 100 × 4.6 mm) gradient of 5–95% acetonitrile in water (0.1% formic acid in each mobile phase)) t 4.49 min; m / z 379 [M+H] 99.71% purity.

[0447] Compound 20B of Scheme B: 4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazine Sodium nitrite (2.88 g, 42 mmol) was added portionwise to CHC1C (314 mL) at -15 °C and stirred for 15 min. Compound 18B (5.4 g, 21 mmol) was added as a solid, followed by CHCl (10 mL). The reaction mixture was allowed to warm and stirred at RT for 1 h. The reaction mixture was diluted with CHCl (44 mL) and NaCl (2.7 g) was added. The reaction mixture was heated to 50 °C for 1 day. The layers were separated, and the organic layer was washed with water, dried (phase separator cartridge), and concentrated in vacuo. The residue was purified by column chromatography (silica gel, isohexane / ethyl acetate 4:1, then CHCl / ethyl acetate 1:0 to 4:1) to give compound 20B as a solid (3.0 g, 47% yield). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 15.08(s, 1H), 7.81-7.73(m, 4H), 7.58-7.51(m, 6H).

[0448] [ka] (i) HATU, DIPEA, DMF, 25°C; (ii) TFA, DCM, 25°C; (iii) Compound 19B, HATU, DIPEA, DMF, 25°C.

[0449] Scheme C outlines the preparation of compound 5C.

[0450] Compound 25C of Scheme C: tert-butyl (3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)carbamate To a solution of biotin (23C, 350 mg, 1.43 mmol) in DMF (7.2 mL) was added tert-butyl N-(3-aminopropyl)carbamate (24C, 250 mg, 1.43 mmol), DIPEA (0.375 mL, 2.15 mmol), and HATU (816 mg, 2.15 mmol). The reaction mixture was stirred at RT for 20 h and then diluted with ethyl acetate and 4% aqueous LiCl. The aqueous phase was extracted twice with ethyl acetate, and the combined organic layers were dried (MgSO4) and concentrated in vacuo. The resulting residue was purified using silica gel chromatography (0-12% gradient of 7 M NH3 in MeOH / CHCl2) to give compound 25C as a solid (180 mg, 31% yield). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 7.75(t, J=5.2Hz, 1H), 6.77(s, 1H), 6.43(s, 1H), 6.37(s, 1H), 4.38-4.34(m, 1H), 4. 21-4.16(m, 1H), 3.23(d, J=5.3Hz, 1H), 3.16(dq, J=6.2, 4.3Hz, 1H), 3.07(dd, J=6.8 , 12.9Hz, 2H), 2.96(dd, J=6.6, 13.0Hz, 2H), 2.88(dd, J=5.2, 12.4Hz, 1H), 2.11(t, J =7.5Hz, 2H), 1.73-1.62(m, 1H), 1.61-1.48(m, 5H), 1.43(s, 9H), 1.40-1.29(m, 2H).

[0451] Compound 26C of Scheme C: N-(3-aminopropyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide To a solution of compound 25C (166 mg, 0.415 mmol) in CHCl (1 mL) is added TFA (1 mL). The reaction mixture is stirred at RT for 2 h and then concentrated in vacuo. The resulting residue is dissolved in CHCl (2 mL) and Biotage MP-carbonate resin (550 mg, 1.66 mmol) is added. The reaction mixture is stirred at RT for 30 min. The beads are filtered off and washed with CHCl / MeOH (1:1, 2 mL), and the filtrate is concentrated in vacuo to give compound 26C as a colorless oil (124 mg, 100% yield), which is used directly in the next step.

[0452] Compound 5C of Scheme C: N-(3-(2-(4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)acetamido)propyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide To a solution of compound 26C (124 mg, 0.415 mmol) in DMF (2 mL) was added compound 19B (151 mg, 0.415 mmol), DIPEA (0.11 mL, 0.62 mmol), and HATU (236 mg, 0.62 mmol). The reaction mixture was stirred at RT for 1.5 h and then diluted with CHCl and 4% aqueous LiCl. The aqueous phase was extracted twice with CHCl, and the combined organic layers were dried (phase separation) and concentrated in vacuo. The resulting residue was first purified by preparative HPLC to give 70 mg, which was further purified by silica gel chromatography to give the desired compound 5C as a solid (44 mg, 16% yield). 1H NMR (400MHz, DMSO-d6) δ(ppm) 8.39(dd, J=5.7, 5.7Hz, 1H), 7.87-7.78(m, 5H), 7.64-7.57(m, 6H), 6.45( s, 1H), 6.39(s, 1H), 5.51(s, 2H), 4.33(dd, J=5.3, 7.6Hz, 1H), 4.18-4.13( m, 1H), 3.22-3.08(m, 5H), 2.85(dd, J=5.2, 12.5Hz, 1H), 2.61(d, J=12.4H) z, 1H), 2.10(dd, J=7.5, 7.5Hz, 2H), 1.66-1.48(m, 6H), 1.39-1.28(m, 2H). 13 C NMR(100MHz, CDCl3) δ(ppm) 172.51, 166.25, 163.16, 154.31, 152.75, 144.06, 135.95, 131.35, 130.58, 130.52, 129.58, 129.44, 129.4 2, 128.75, 128.70, 114.18, 61.47, 59.64, 55.85, 50.91, 37.21, 36.66, 35.66, 29.64, 28.66, 28.48, 25.75. LCMS (Analytical Method 1: HPLC (Phenomenex Luna 5 μm C18, 100 × 4.6 mm) gradient of 5-95% acetonitrile in water (0.1% formic acid in each mobile phase)) t 3.52 min; m / z 647 [M+H] 98.38% purity.

[0453] [ka] (i) (a) 1-tert-butoxycarbonyl-4-(2-hydroxyethyl)piperazine, DEAD, PPh3, 1,4-dioxane, 120°C, 1 hour; (b) HCl in 1,4-dioxane, 25°C; (ii) (a) HATU, DIPEA, DMF, 25°C; (b) NaOH, 40°C, 1 hour; (iii) POCl3, 25°C.

[0454] Scheme D outlines the preparation of compound 6D.

[0455] Compound 27D of Scheme D: 4-chloro-3,5-diphenyl-1-(2-(piperazin-1-yl)ethyl)-1H-pyrazolo[3,4-c]pyridazine To a mixture of compound 20B (345 mg, 1.13 mmol), 1-tert-butoxycarbonyl-4-(2-hydroxyethyl)piperazine (520 mg, 2.26 mmol), and triphenylphosphine (888 mg, 2.26 mmol) in 1,4-dioxane (8.4 mL) was slowly added diethyl azodicarboxylate (0.355 mL, 2.26 mmol) at RT. The reaction mixture was then heated to 120 °C using microwave irradiation for 1 h. The reaction mixture was cooled to RT, and 4 M HCl in 1,4-dioxane (4 mL) was added. The reaction mixture was stirred at RT for 4 h, diluted with CHCl (10 mL), and the solution was loaded onto a Biotage SCX-2 cartridge (20 g) and eluted with methanol followed by 7 M NH in methanol. The fractions were concentrated in vacuo to give compound 27D in a 1:1 ratio along with 2-hydroxyethyl)piperazine (950 mg, 100% yield), which was used directly in the next step.

[0456] Compound 28D of Scheme D: (3aS,4S,6aR)-4-(5-(4-(2-(4-hydroxy-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)ethyl)piperazin-1-yl)-5-oxopentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one To a solution of compound 27D (950 mg crude, 1.13 mmol) in DMF (5.7 mL) was added DIPEA (0.59 mL, 3.39 mmol), biotin (678 mg, 2.78 mmol), and HATU (1.29 g, 3.39 mmol). The reaction mixture was stirred at RT for 24 h and then diluted with DMSO (5 mL). NaOH (2 M, 5 mL) was added, and the reaction mixture was heated to 40 °C for 1 h and then stirred at RT for 2 days. The reaction mixture was purified by preparative HPLC to give compound 28D (100 mg, 14% yield). 1 H NMR

[0457] Compound 6D of Scheme D: (3aS,4S,6aR)-4-(5-(4-(2-(4-chloro-3,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-1-yl)ethyl)piperazin-1-yl)-5-oxopentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one A solution of compound 28D (97 mg, 0.155 mmol) in phosphorus oxychloride (6 mL) is stirred at RT for 2 days. The reaction mixture is diluted with CHCl and 2 M NaCO solution. The aqueous phase is extracted twice with CHCl. ​​The combined organic layers are dried (MgSO) and concentrated in vacuo. The resulting residue is purified using chromatography (silica gel, 0-12% gradient of 7 M NH in MeOH / CHCl) to give compound 6D as a solid (42 mg, 42% yield). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 7.86-7.78(m, 4H), 7.66-7.56(m, 6H), 6.47(s, 1H), 6.39(s, 1H), 5.01-4.94(m, 2H), 4.3 8-4.32(m, 1H), 4.20-4.15(m, 1H), 3.17-3.10(m, 1H), 3.08-3.01(m, 2H), 2.87(dd, J=12 .4, 5.1Hz, 1H), 2.63(d, J=12.4Hz, 1H), 2.58(m, 1H), 2.57-2.54(m, 2H), 2.51(m, 2H), 2. 36-2.28(m, 2H), 1.71-1.61(m, 1H), 1.56-1.45(m, 3H), 1.38(m, 2H), 1.27-1.17(m, 1H). 13C NMR(100MHz, CDCl3) δ(ppm) 170.51, 162.70, 153.69, 151.95, 143.18, 135.52, 131.06, 130.15, 130.11, 129.03, 128.94, 128.84, 128. 26, 128.20, 113.42, 61.04, 59.76, 59.19, 56.16, 55.48, 52.71, 52.28, 45.27, 32.05, 28.29, 28.11, 24.85. LCMS (Method 2: HPLC (Hichrom ACE 3 C18-AR mixed-mode column 100 x 4.6 mm) gradient of 2-100% acetonitrile in water (0.1% formic acid in each mobile phase)) t 9.77 min; m / z 645 [M+H] 93.27% purity.

[0458] molecular chaperones Although there are many possible conformations that a particular protein can adopt as it folds, ultimately, a biologically functional protein will fold into a stable state, referred to as its "native state," in which the protein's tendency to aggregate is minimized. The protein folding process is thermodynamically driven by the so-called "hydrophobic effect," which describes the tendency of a protein's hydrophobic amino acid residues to interact with each other and form a hydrophobic core, while its hydrophilic amino acid residues remain on the protein's surface. Nascent, or partially folded, proteins are "sticky" because their hydrophobic amino acids are not completely buried within the protein's core. As a result, sticky proteins can clump together into unmanageable aggregates, especially in the cellular environment where other protein molecules are abundant.

[0459] In the cytoplasm and nucleus of cells, there are quality control systems to ensure that protein folding occurs efficiently. This system includes molecular chaperones and the ubiquitin proteasome system (UPS). The UPS allows proteins to be tagged with ubiquitin to target them for degradation in the proteasome, a complex of protein molecules that degrades ubiquitinated polypeptides and recycles the ubiquitin tag.

[0460] Molecular chaperones are proteins that help other proteins fold efficiently by shielding the sticky hydrophobic surfaces of unfolded or misfolded proteins, thereby minimizing their tendency to aggregate. Molecular chaperones can be found in almost all living organisms and reside in many subcellular compartments. Some molecular chaperones are constitutively expressed and not stress-induced, others are constitutively expressed and stress-induced, and still others are stress-induced. Molecular chaperones, such as heat shock proteins (Hsps), are classified by their molecular weight and include the small Hsp, Hsp40, Hsp60, Hsp70, Hsp90, and Hsp100 families (Table 2).

[0461] [Table 2]

[0462] In some embodiments, the pyrazolopyridazine compound, the non-pyrazolopyridazine compound, or a metabolite thereof is bound to a molecular chaperone. In some embodiments, the pyrazolopyridazine compound, the non-pyrazolopyridazine compound, or a metabolite thereof is covalently bound to a molecular chaperone.

[0463] In some embodiments, binding of a pyrazolopyridazine compound, a non-pyrazolopyridazine compound, or a metabolite thereof with a molecular chaperone results in treating or preventing a proteopathy in a subject in need thereof.

[0464] In another embodiment, the molecular chaperone is a member of the Hsp10 family, Hsp40 family, Hsp60 family, Hsp70 family, Hsp90 family, or Hsp100 family.

[0465] Proteopathy Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds are useful for treating or preventing proteopathies.

[0466] In some embodiments, the proteopathy is a neurodegenerative disease. Illustrative neurodegenerative diseases include, but are not limited to, Alzheimer's disease, progressive supranuclear palsy, dementia pugilistica, frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), Ritiko-Bodding disease, tangle-predominant senile dementia. dementia), ganglioneuroma, gangliocytoma, meningioangiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, lipofuscinosis, Pick's disease, corticobasal degeneration, argyrophilic grain dementia, Huntington's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, neuroaxonal dystrophy, dentatorubral-pallidoluysian atrophy (DRPLA), spinal-bulbar muscular atrophy (SBMA), spinocerebellar ataxia 1 (SCA1), SCA2, SCA3, SCA6, SCA7, SCA17, prion diseases, amyotrophic lateral sclerosis, frontotemporal lobar degeneration (FTLD), and familial encephalopathy with neuroserpin inclusion bodies (FENIB).

[0467] In some embodiments, the proteopathy is amyloidosis. Specific amyloidoses include, but are not limited to, familial British dementia (ABri), familial Danish dementia (ADan), hereditary cerebral hemorrhage with amyloidosis of the Icelandic type (HCHWA-I), familial amyloidotic neuropathy (ATTR), AL (light chain) primary systemic amyloidosis, AH (heavy chain) amyloidosis, AA secondary amyloidosis, Aβ amyloidosis, aortic medial amyloidosis, LECT2 amyloidosis, AIAPP amyloidosis, apolipoprotein AI amyloidosis (AApoAI), apolipoprotein AII amyloidosis ( AApoAII), apolipoprotein AIV amyloidosis (AApoAIV), familial amyloidosis of the Finnish type (FAF), fibrinogen amyloidosis (AFib), lysozyme amyloidosis (ALys), dialysis-associated amyloidosis (Aβ2M), medullary thyroid carcinoma (ACal), atrial amyloidosis (AANF), pituitary prolactinoma (APro), hereditary lattice corneal dystrophy, cutaneous lichen amyloidosis (AKer), Mallory bodies, primary cutaneous amyloidosis, corneal lactoferrin amyloidosis, odontogenic (pinbory) tumor amyloid, or seminal vesicle amyloid.

[0468] In some embodiments, the proteopathy is a lysosomal storage disease, including, but not limited to, activator deficiency / GM2 gangliosidosis, alpha-mannosidosis, aspartylglucosaminuria, cholesteryl ester storage disease, chronic hexosaminidase A deficiency, cystinosis, Danon disease, Fabry disease, Farber disease, fucosidosis, galactosialidosis, Gaucher disease type I, Gaucher disease type II, Gaucher disease type III, and childhood GM1 gangliosidosis. sis, late-onset childhood / juvenile GM1 gangliosidosis, adult / chronic GM1 gangliosidosis, I-cell disease / mucolipidosis II, childhood free sialic acid storage disease / ISSD, juvenile hexosaminidase A deficiency, childhood-onset Krabbe disease, late-onset Krabbe disease, early-onset lysosomal acid lipase deficiency, late-onset lysosomal acid lipase deficiency, metachromatic leukodystrophy, pseudo-Hurler polydystrophy / mucolipidosis IIIA, MPS MPS I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, MPS III A Sanfilippo syndrome, MPS III B Sanfilippo syndrome, MPS III C Sanfilippo syndrome, MPS III D Morquio syndrome, MPS IVA Morquio syndrome, MPS IVB Morquio syndrome, MPS IX Hyaluronidase deficiency, MPS VI Maroto-Lamy, MPS VII Sly syndrome, Mucolipidosis I / sialidosis, Mucolipidosis IIIC, Mucolipidosis IV, Multiple sulfatase deficiency, Niemann-Pick disease type A, Niemann-Pick disease type B, Niemann-Pick disease type C, CLN6 disease-atypical late-onset childhood, CLN6 disease-late-onset variant, CLN6 disease-early juvenile, Batten-Spielmeyer-Vogt / juvenile NCL / CLN3 disease, Finnish variant late-onset childhood CLN5, Jansky-Bielschowski disease / late-onset childhood CLN2 / TPP1 disease, Kufs / adult-onset NCL / CLN4 disease, NorthernEpilepsy / atypical late-onset childhood CLN8, Santavuori-Haltia / childhood CLN1 / PPT disease, beta-mannosidosis, Pompe disease / glycogen storage disease type II, pyknodysostosis, Sandhoff disease / adult-onset / GM2 gangliosidosis, Sandhoff disease / GM2 gangliosidosis-pediatric, Sandhoff disease / GM2 gangliosidosis-juvenile, Schindler disease, Salla disease / sialic acid storage disease, Tay-Sachs / GM2 gangliosidosis, or Wolman disease.

[0469] In some embodiments, the lysosomal storage disease is a mucopolysaccharidoses, including, but not limited to, pseudo-Hurler multiple dystrophy / mucolipidosis IIIA, MPS I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, Sanfilippo syndrome type A / MPS IIIA, Sanfilippo syndrome type B / MPS IIIB, Sanfilippo syndrome type C / MPS IIIC, Sanfilippo syndrome type D / MPS IIID, Morquio syndrome type A / MPS IVA, Morquio syndrome type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Maroto-ramy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC, and mucolipidosis type IV.

[0470] In another embodiment, the lysosomal storage disease is Pompe disease / glycogen storage disease type II.

[0471] In some embodiments, the proteopathy is a retinal degenerative disease. Non-limiting examples of retinal degenerative diseases include: retinitis pigmentosa, Leber's congenital amaurosis, retinal degeneration syndrome, age-related macular degeneration, including wet and dry age-related macular degeneration, and Usher syndrome. In some embodiments, Usher syndrome is a subtype of Usher syndrome. In some embodiments, the subtype is Usher I. In some embodiments, the subtype is Usher II. In some embodiments, the subtype is Usher III.

[0472] In further embodiments of the present invention, the compounds of the present invention can be administered to a subject in need thereof for the treatment of hearing loss associated with Usher syndrome. In some embodiments, the Usher syndrome is a subtype of Usher syndrome. In some embodiments, the subtype is Usher I. In some embodiments, the subtype is Usher II. In some embodiments, the subtype is Usher III.

[0473] Additional specific proteopathies include, but are not limited to, those disclosed in Table 3.

[0474] [Table 3] TIFF0007755610000341.tif252168TIFF0007755610000342.tif250168TIFF00077556100 00343.tif250168TIFF0007755610000344.tif247170TIFF0007755610000345.tif145168

[0475] Cystic fibrosis Cystic fibrosis (CF), also known as mucoviscidosis, is an autosomal recessive disease that primarily affects the lungs but can also affect other organs such as the pancreas, liver, kidneys, and intestines. The name "cystic fibrosis" refers to the characteristic fibrosis and cysts that form within the pancreas. The disease is characterized by frequent lung infections that make breathing difficult and coughing up saliva.

[0476] The most common mutation, found in approximately 70% of CF patients worldwide, is the deletion of a phenylalanine residue at amino acid position 508 of the 1,480-amino acid cystic fibrosis transmembrane conductance regulator (CFTR) protein. This mutation causes the protein to misfold and its degradation by the cell. Other mutations in the CFTR protein can result in truncated forms of the CFTR protein because their production terminates prematurely. Still other mutations produce mutant CFTR proteins that do not use energy normally; do not allow chloride, iodide, or thiocyanate ions to cross the membrane normally; or degrade at a faster-than-normal rate. CFTR protein mutations can also result in fewer copies of the CFTR protein being produced. A list of classes of CFTR protein mutations and example mutations for each class is disclosed in Table 4.

[0477] [Table 4]

[0478] Other CFTR protein mutations include G178R, S549N, S549R, G1244E, S1251N, and S1255P.

[0479] Accordingly, the present invention further provides a method for treating or preventing cystic fibrosis, comprising administering an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound to a subject in need thereof. In some embodiments, the subject has a mutation in the CFTR protein. In one embodiment, the subject has a class I CF mutation. In another embodiment, the subject has a class II CF mutation. In another embodiment, the subject has a class III CF mutation. In another embodiment, the subject has a class IV CF mutation. In another embodiment, the subject has a class V CF mutation. In another embodiment, the subject has a class VI CF mutation.

[0480] In one embodiment, the mutation is W1282X, R553X, or G542X. In another embodiment, the mutation is ΔF508 or N1303K. In another embodiment, the mutation is G551D, G551S, or G1349D. In another embodiment, the mutation is R117H, R334W, or R347P. In another embodiment, the mutation is 2789+5G>A or A455E. In another embodiment, the mutation is 120Δ23, N287Y, 4326ΔITC, or 4279insA.

[0481] In one embodiment, the subject has one or more of the following mutations: W1282X, R553X, G542X, ΔF508, N1303K, G551D, G551S, G1349D, R117H, R334W, R347P, 2789+5G>A, A455E, 120Δ23, N287Y, 4326ΔITC, 4279insA, G178R, S549N, S549R, G1244E, S1251N, and S1255P.

[0482] Retinitis pigmentosa Retinitis pigmentosa (RP) is an inherited degenerative eye disease that causes severe vision loss due to the progressive degeneration of rod photoreceptor cells in the retina. Progressive rod degeneration can be followed by abnormalities of the adjacent retinal pigment epithelium (RPE) and deterioration of cone photoreceptor cells.

[0483] There are several genes that, when mutated, can cause RP. Inheritance patterns for RP have been identified as autosomal dominant, autosomal recessive, X-linked, and maternally (mitochondrial) acquired. X-linked RP can be recessive, affecting primarily only males, or dominant, affecting both males and females. Several digenic (controlled by two genes) and mitochondrial forms of RP are also known.

[0484] A mutation has been identified in the gene for rhodopsin, a pigment that plays an essential role in the visual transduction cascade that allows vision in low-light conditions. This mutation results in a substitution of histidine for proline at amino acid position 23. The rhodopsin gene is the major protein of the photoreceptor outer segment of the retina, which consists of opsin, a light-sensitive membrane-bound G protein-coupled receptor, and a reversibly covalently bound cofactor. Mutations in the rhodopsin gene most frequently follow an autosomal dominant inheritance pattern.

[0485] Since the initial description of the P23H mutation in the intradiscal domain of the protein, up to 150 RP-associated opsin gene mutations have been reported. These mutations are found throughout the opsin gene and are distributed along the three domains of the protein (intracerebral, transmembrane, and cytoplasmic). Opsin gene mutations are most commonly missense mutations, causing misfolding of the rhodopsin protein. Mutations at amino acid 23 of the opsin gene, where a proline is replaced by a histidine, account for the largest proportion of rhodopsin mutations in the United States. Other mutations associated with RP include T58R, P347L, P347S, and deletion of Ile255. The rare P23A mutation causes autosomal dominant RP.

[0486] A list of genes mutated in subjects with RP and the type of RP is given in Table 5.

[0487] [Table 5] TIFF0007755610000348.tif247162TIFF0007755610000349.tif176164

[0488] Accordingly, the present invention further provides a method of treating or preventing retinitis pigmentosa, comprising administering an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound to a subject in need thereof. In some embodiments, the subject has a mutation in the RP gene.

[0489] In one embodiment, the retinitis pigmentosa is autosomal dominant, autosomal recessive, X-linked, or mitochondrial acquired.

[0490] In one embodiment, retinitis pigmentosa (RP) is any of RP-1, RP-2, RP-3, RP-4, RP-6, RP-7, RP-9, RP-10, RP-11, RP-12, RP-13, RP-14, RP-15, RP-17, RP-18, RP-19, RP-20, RP-22, RP-23, RP-24, RP-25, RP-26, RP-27, RP-28, RP-30, RP-31, RP-32, RP-33, RP-34, RP-35, RP-36, RP-37, RP-38, RP-39, RP-40, RP-41, RP-42, RP-43, RP-44, RP-45, RP-46, RP-47, RP-48, RP-49, RP-50, RP-51, RP-52, RP-53, RP-54, RP-55, RP-56, RP-57, RP-58, RP-59, RP-60, RP-61, RP-62, RP-63, RP-64, RP-65, RP-66, RP-67, RP-68, RP-69, RP-70, RP-71, RP-72, RP-73, RP-74, RP-75, RP-76, RP-77, RP-78, RP-79, RP-80, RP-81, RP-82, RP-83, RP-84, RP-85, RP-86, RP-87, RP-88, RP-89 -36, RP-37, RP-38, RP-39, RP-40, RP-41, RP-42, RP-43, RP-44, RP-45, RP-46, RP-47, RP-48, RP-49, RP-50, RP-51, RP-53, RP-54, RP-55, RP-56, RP-57, RP-58, RP-59, RP-60, RP-61, RP-62, RP-63, RP-64, RP-66, RP-67, RP-68, RP-69 or RP-70.

[0491] In one embodiment, the following genes are of interest: RP1, RP2, RPGR, RHO, ROM1, RP9, IMPDH1, PRPF31, CRB1, PRPF8, TULP1, CA4, PRPF3, ABCA4, RPE65, OFD1, EYS, CERKL, NRL, FAM161A, FSCN2, TOPORS, SNRNP200, SEMA4A, PRCD, NR2E3, MERTK, USH2A, PDE6B, PROM1, KLHL7, PDE6A , RGR, CNGB1, IDH3B, SAG, GUCA1B, CNGA1, BEST1, TTC8, RDH12, C2orf71, ARL6, IMPG2, PDE6G, ZNF513, DHDDS, PRPF6, CLRN1, MAK, C8ORF37, RBP3, NEK2, SLC7A14, KIZ, PRPF4, PRPH2, LRAT, SPATA7, AIPL1, CRX, MT-TS2, RLBP1, and WDR19.

[0492] Therapeutic or prophylactic uses Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds can be administered to subjects as a composition containing pharmaceutically acceptable carriers or vehicles.Non-limiting examples of suitable pharmaceutical carriers or vehicles include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium carbonate, magnesium stearate, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, buffered water and phosphate buffered saline.These compositions can be administered as, for example, drops, solutions, suspensions, tablets, pills, capsules, powders and sustained-release formulations. In some embodiments, the composition comprises, for example, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methylcellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate, and mineral oil. The composition can further comprise lubricating agents, wetting agents, emulsifying and suspending agents, preservatives, sweetening agents, or flavoring agents.

[0493] The composition can include an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. The composition can be formulated into a unit dosage form containing an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition contains, for example, about 1 ng to about 1,000 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition contains about 100 mg to about 1,000 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition contains about 100 mg to about 500 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition contains about 200 mg to about 300 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound.

[0494] The dosage of the pyrazolopyridazine compound or non-pyrazolopyridazine compound can vary depending on the subject's symptoms, age, and body weight, the type and severity of the proteopathy, the route of administration, and the form of the composition. The compositions described herein can be administered in a single dose or in divided doses. In some embodiments, the dosage of the pyrazolopyridazine compound or non-pyrazolopyridazine compound ranges from about 0.01 ng to about 10 g / kg, about 1 ng to about 0.1 g / kg, or about 100 ng to about 10 mg / kg of the subject's body weight.

[0495] Administration can be, for example, topical, intraauricular, intraocular, parenteral, intravenous, intraarterial, subcutaneous, intramuscular, intracranial, intraorbital, intraventricular, intraarticular, intrathecal, intravesical, intraperitoneal, intranasal, aerosol, suppository, or oral. Oral formulations include tablets containing a pyrazolopyridazine compound or a non-pyrazolopyridazine compound in a mixture with non-toxic pharmaceutically acceptable excipients. These excipients can be, for example, inert diluents or fillers (e.g., sucrose and sorbitol), lubricants, glidants, and anti-adherents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Ophthalmic formulations can be in the form of eye drops.

[0496] The pyrazolopyridazine compounds, non-pyrazolopyridazine compounds, or compositions thereof can be provided in lyophilized form for reconstitution, e.g., in isotonic, aqueous, or saline buffer solutions for parenteral, subcutaneous, intradermal, intramuscular, or intravenous administration. The compositions can also be in the form of liquid preparations useful for oral, otic, nasal, or sublingual administration, such as suspensions, syrups, or elixirs. The compositions can also be in forms suitable for oral administration, such as capsules, tablets, pills, and chewable solid formulations. The compositions can also be formulated as creams for dermal administration, as liquids, viscous liquids, pastes, or powders. The compositions can also be formulated as powders for pulmonary administration, with or without an aerosolizing component.

[0497] The compositions can be in oral, intraaural, intranasal, sublingual, intraduodenal, subcutaneous, buccal, intracolonic, rectal, vaginal, mucosal, pulmonary, transdermal, intradermal, parenteral, intravenous, intramuscular and ocular dosage forms and can cross the blood-brain barrier.

[0498] The compositions can be administered by various means known in the art. For example, the compositions can be administered orally and formulated as tablets, capsules, granules, powders, or syrups. Alternatively, the compositions can be administered parenterally as injections (e.g., intravenous, intramuscular, or subcutaneous), infusion preparations, or suppositories. For ophthalmic use, the compositions can be formulated as eye drops or eye ointments. For otic use, the compositions can be formulated as ear drops, ointments, creams, liquids, gels, or patches for application inside or on the ear. These formulations can be prepared by conventional means, and the compositions can be mixed with any conventional additives, such as excipients, binders, disintegrants, lubricants, solubilizers, suspending aids, emulsifiers, or coating agents.

[0499] The compositions can include wetting agents, emulsifying agents, and lubricants, coloring agents, release agents, coating agents, sweetening agents, flavoring and perfuming agents, preservatives, and antioxidants.

[0500] The compositions may be suitable for, for example, oral, otic, ocular, nasal, topical (including buccal and sublingual), rectal, vaginal, aerosol and / or parenteral administration. The compositions may be presented in unit dosage form and may be prepared by any method known in the art.

[0501] Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges, powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a troche (with an inert base such as gelatin and glycerin, or sucrose and acacia gum). The composition can also be administered as a bolus, electuary, or paste.

[0502] Additional examples of pharmaceutically acceptable carriers or vehicles include: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or gum acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) coloring agents; and (11) buffers. Similar compositions may also be employed as fill materials in soft- or hard-filled gelatin capsules.

[0503] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, gels, solutions, suspensions, syrups, and elixirs. Liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, diethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils such as cottonseed, peanut, corn, germ, olive, castor, and sesame oils, glycerol, tetrahydrofuryl alcohol, polyethylene glycol, fatty acid esters of sorbitan, and mixtures thereof.

[0504] Suspension dosage forms can contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0505] Dosage forms for transdermal administration of the subject compositions include drops, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches. The ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0506] Powders and sprays can contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates, polyamide powder, or mixtures thereof. Sprays can additionally contain conventional propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0507] The composition can be administered by aerosol of solid particles. A non-aqueous (e.g., fluorocarbon propellant) suspension can be used. Sonic nebulizers can be used because they minimize exposure to shear, which can cause degradation.

[0508] Aqueous aerosols can be made by formulating aqueous solutions or suspensions of pyrazolopyridazine compounds or non-pyrazolopyridazine compounds with conventional pharmaceutically acceptable carriers or vehicles such as non-ionic surfactants (Tweens, Pluronics, or polyethylene glycol); proteins such as serum albumin; sorbitan esters; fatty acids; lecithin; amino acids; buffers; salts; sugars; or sugar alcohols.

[0509] Compositions suitable for parenteral administration include pyrazolopyridazine compounds or non-pyrazolopyridazine compounds and one or more pharmaceutically acceptable sterile, isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostats, or solutes to render the formulation isotonic with the subject's blood, and suspending or thickening agents.

[0510] While the invention has been described with reference to certain embodiments, other embodiments will become apparent to those skilled in the art upon consideration of the specification and claims. It will be apparent to those skilled in the art that many modifications, both to materials and methods, can be practiced without departing from the scope of the invention.

[0511] References Each document disclosed in this application is incorporated herein by reference in its entirety.

Claims

1. A compound having the following structure: 【Chemical 1】 or a pharmaceutically acceptable salt thereof, for treating or preventing lipofuscinosis, Parkinson's disease, dementia with Lewy bodies, emphysema, sickle cell disease, Marfan syndrome, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), Alexander disease, Seipinopathy, Santavuori-Haltia / pediatric CLN1 / PPT disease, Jansky-Bielschowski disease / late-onset pediatric CLN2 / TPP1 disease, Batten-Spielmeyer-Vogt / juvenile NCL / CLN3 disease, Finnish variant late-onset pediatric CLN5, CLN6 disease-atypical late-onset pediatric, CLN6 disease-late-onset variant, CLN6 disease-early juvenile, or Northern Epilepsy / variant late-onset pediatric CLN8.

2. The composition of claim 1 , wherein the compound or a pharmaceutically acceptable salt of the compound binds to a molecular chaperone.

3. The composition of claim 2 , wherein the molecular chaperone is a member of the Hsp60 family and the Hsp90 family.

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