Method for treating or preventing proteopathy

Abstract

To provide methods for treating or preventing a proteopathy.SOLUTION: The invention provides methods 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. In the formula, Hal is a halogen atom; R1 is independently a halogen atom, alkyl and the like; R2 is independently a halogen atom, alkyl and the like; and R3 is H, alkyl and the like.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 174,332, filed on June 11, 2015, and U.S. Provisional Application No. 62 / 174,338, filed on June 11, 2015, each of which is hereby incorporated by reference in its entirety.

Background Art

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

[0003] If a protein fails to fold into its correct three-dimensional structure, it can lead to a disease called proteopathy (also sometimes referred to as protein disease or protein conformational disorder). This defect can be caused by one or more mutations in the protein's gene or environmental factors such as oxidative stress, alkalosis, acidosis, pH shift, and osmotic shock. Incorrect folding of the protein can sometimes cause clumping or aggregation into amyloid plaques or fibrils that can exacerbate the disease. Proteopathies include a wide range of diseases, such as neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, polyglutamine diseases, prion diseases); amyloidosis of other non-neuronal proteins such as, inter alia, α1-antitrypsin, immunoglobulin light and heavy chains, lactadherin, apolipoprotein, gelsolin, lysozyme, fibrinogen, atrial natriuretic factor, keratin, lactoferrin, and beta-2 microglobulin; sickle cell disease; cataracts; cystic fibrosis; retinitis pigmentosa; and nephrogenic diabetes insipidus.

[0004] Molecular chaperones are biomolecules that assist in the proper folding, translocation, and / or degradation of proteins. Examples of molecular chaperones include heat shock proteins, which are classified into seven different families in the human genome and include HSPH (Hsp110), HSPC (Hsp90), HSPA (Hsp70), DNAJ (Hsp40), HSPB (low molecular weight Hsp (sHsp)), human chaperonin 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 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] [Chemistry] In the formula, R is fluoro, chloro, iodo, methyl, methoxy, cyano, trifluoromethyl, or -(CO)NH(CH3).

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

[0008] [Chemistry] In the formula, Hal is -Cl, -F, -I, or -Br; x is an integer in the range of 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 in the range of 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] [Chemistry] wherein; 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]

Chemical formula

[0011]

Chemical formula

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

[0013]

Chemical formula

[0014]

Chem.

[0015]

Chem.

[0016]

Chem.

[0017] The present invention further provides a method for treating or preventing 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]

Chem.

[0019] The present invention further provides a method for treating or preventing 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]

Chem.

[0021] The present invention also further provides a method for treating or preventing 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]

Chem.

[0023] The present invention also further provides a method for treating or preventing 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]

Chem.

[0025]

Chem.

[0026]

Chem.

[0027] The present invention further provides a method for treating or preventing 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]

Chemical formula

[0029]

Chemical formula

[0030] The present invention further provides a method for treating or preventing 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]

Chemical formula

[0032]

Chemical formula

[0033] The present invention further provides a method for treating or preventing 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]

Chemical formula

[0035]

Chem.

[0036]

Chem.

[0037] The present invention further provides a method for treating or preventing 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]

Chem.

[0039]

Chem.

[0040] The present invention further provides a method for treating or preventing 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]

Chem.

[0042]

Chem.

[0043] The "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 compound is useful for treating or preventing proteopathy.

Mode for Carrying Out 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 proteopathy, comprising administering an effective amount of a pyrazolopyridazine compound to a subject in need thereof.

[0046] Definition The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. Example alkyl groups include -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -CH2CH2CH2CH3, -CH(CH3)CH2CH3, -CH2CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH2CH3, -CH(CH3)CH2CH2CH3, -CH2CH2CH(CH3)2, -CH2C(CH3)3, -CH2CH2CH2CH2CH3, -CH(CH3)CH2CH2CH3, -CH2CH2CH(CH3)2 and -CH(CH3)C(CH3)3 groups.

[0047] The term "alkylene" refers to an alkyl group bonded to another atom or group. Examples of alkylene groups include -CH2-, -CH2CH2-, -CH2CH2CH2-, -C(CH3)2-, -CH(CH3)-, -CH2CH2CH2CH2-, -CH(CH3)CH2CH2-, -CH2C(CH3)2-, -C(CH3)2CH2-, -CH2CH2CH2CH2CH2-, -CH(CH3)CH2CH2CH2-, -CH2CH2C(CH3)2-, -CH2CH(CH3)CH2CH2-, -CH2CH2CH(CH3)CH2-, -CH2CH2CH2CH2CH2CH2-, -CH(CH3)CH2CH2CH2CH2-, -CH2CH2CH2C(CH3)2-, -CH2CH(CH3)CH2CH2CH2-, -CH2CH2CH2CH(CH3)CH2- and -C(CH3)2C(CH3)2- groups.

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

[0049] The word "about", when preceding a numerical value, means within 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, and so on.

[0050] Abbreviations: APCI Atmospheric Pressure Chemical Ionization DAPI 4’,6-Diamidino-2-phenylindole DCM Dichloromethane DEAD Diethyl Azodicarboxylate DIPEA Diisopropylethylamine DMEM Dulbecco's Modified Eagle 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" means an amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound that is effective to treat or prevent proteopathy in a subject. In some embodiments, when another therapeutic or prophylactic agent is administered before, after, or simultaneously with the administration of the pyrazolopyridazine compound or non-pyrazolopyridazine compound, the "effective amount" is the total amount of (i) the pyrazolopyridazine compound or non-pyrazolopyridazine compound and (ii) another therapeutic or prophylactic agent that is effective to treat or prevent proteopathy in the subject.

[0052] The terms "proteopathy", "proteinopathy", and "protein conformational disease" refer to a disease or disorder resulting from the misfolding of one or more proteins.

[0053] The term "protein aggregate" refers to a biological phenomenon in which misfolded proteins accumulate and aggregate with each other.

[0054] "Subject" is a mammal including a mammalian species-rich eye, for example, a primate such as a human; a rodent species such as a mouse, a rat, or a guinea pig; a carnivorous species such as a dog, a cat, a weasel, a bear, or a seal; a non-human primate such as a monkey, a chimpanzee, a baboon, or a macaque; a bat species such as a bat; a solenodon species such as a solenodon, a mole, or a shrew; and a cetartiodactyl species such as a whale. 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 for treating or preventing 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]

Chemical formula

[0057] In one embodiment, R of formula I is in the para position with respect to the pyrazolopyridazinone ring system. In one embodiment, R of formula I is in the meta position with respect to the pyrazolopyridazinone ring system. In one embodiment, R of formula I is in the ortho position with respect to the pyrazolopyridazinone ring system.

[0058] Pyrazolopyridazine compounds of formula II The present invention also provides a method for treating or preventing 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]

Chemical formula

[0060]

Chemical formula

[0061]

Chemical formula

[0062]

Chemical formula

[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 and: 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 R1 is ortho to the pyrazolopyridazino ring system. In certain embodiments, x is 1 and R1 is para to the pyrazolopyridazino ring system. In further embodiments, x is 1 and R1 is meta to the pyrazolopyridazino ring system.

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

[0068] In certain embodiments, x is 2 and R1 is ortho and meta to the pyrazolopyridazino ring system. In certain embodiments, x is 2 and R1 is ortho and para to the pyrazolopyridazino ring system. In further embodiments, x is 2 and R1 is para and meta to the pyrazolopyridazino ring system.

[0069] In certain embodiments, y is 2 and R2 is ortho and meta to the pyrazolopyridazino ring system. In certain embodiments, y is 2 and R2 is ortho and para to the pyrazolopyridazino ring system. In further embodiments, y is 2 and R2 is para and meta 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 another embodiment, 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 yet another embodiment, R3 is -CH2C(CH3)2OH. In still other embodiments, R3 is -CH2C(O)OCH3. In further embodiments, R3 is -CH2CH(OH)CH3. In yet another embodiment, R3 is -CH2CH2OH. In certain embodiments, R3 is -CH(CH3)CH2OH.

[0077] In further embodiments, R3 is -CH2C(O)R4, where R4 is

[0078]

Chemical formula

[0079]

Chemical formula

[0080] [Chemical formula] is. In still other embodiments, R3 is -CH2C(O)R4 and R4 is

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

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

[0083] [Chemical formula] is. In specific embodiments, R3 is -CH2C(O)R4 and R4 is

[0084] [Chemical formula] is. In still other embodiments, R3 is -CH2C(O)R4 and R4 is

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

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

[0087] [Chemical formula] It is. In certain embodiments, R3 is -CH2C(O)R4, and R4 is

[0088]

Chemical formula

[0089]

Chemical formula

[0090]

Chemical formula

[0091]

Chemical formula

[0092]

Chemical formula

[0093]

Chemical formula

[0094]

Chemical formula

[0095]

Chemical formula

[0096]

Chemical formula

[0097]

Chemical formula

[0098]

Chemical formula

[0099]

Chemical formula

[0100]

Chemical formula

[0101]

Chemical formula

[0102]

Chemical formula

[0103]

Chem.

[0104]

Chem.

[0105]

Chem.

[0106]

Chem.

[0107]

Chem.

[0108]

Chem.

[0109] In a particular embodiment, R3 is -(CH2)2R5, and R5 is

[0110]

Chem.

[0111]

Chem.

[0112]

Chemical formula

[0113]

Chemical formula

[0114] In some embodiments, a is an integer in the range of 0 to 5. In some embodiments, b is an integer in the range of 0 to 4. In some embodiments, c is an integer in the range of 0 to 6.

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

[0116]

Chemical formula

[0117] Pyrazolopyridazine compounds of formula III The present invention further provides a method for treating or preventing 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]

Chemical formula

[0119]

Chemical formula

[0120]

Chemical formula

[0121] [Chemical formula] ; each R6 and R7 is independently -H or -I, where at least one of R6 and R7 is -I, when R3 is -C1-C3, R7 is -H.

[0122] In certain embodiments, one R6 that is ortho to the pyrazolopyridazinone ring system is iodine and the remaining R6 and R7 groups are hydrogen. In other embodiments, one R6 that is para to the pyrazolopyridazinone ring system is iodine and the remaining R6 and R7 groups are hydrogen. In further embodiments, one R6 that is ortho to the pyrazolopyridazinone ring system and one R6 that is para to the pyrazolopyridazinone ring system are iodine and the remaining R6 and R7 groups are hydrogen. In further embodiments, two R6 groups that are ortho to the pyrazolopyridazinone ring system and one R6 that is para to the pyrazolopyridazinone ring system are iodine and the remaining R6 and R7 groups are hydrogen. In further embodiments, two R6 groups that are para to the pyrazolopyridazinone ring system and one R6 that is ortho to the pyrazolopyridazinone ring system are iodine and the remaining R6 and R7 are hydrogen. In certain embodiments, all R6 groups are iodine and R7 is hydrogen. In still further embodiments, R7 is iodine and the R6 groups are hydrogen.

[0123] In a particular embodiment, one R6 that is para to the pyrazolopyridazinone ring system is iodine and R3 is -CH3.

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

[0125] In certain embodiments, x is 1 and R1 is ortho to the pyrazolopyridazino ring system. In certain embodiments, x is 1 and R1 is para to the pyrazolopyridazino ring system. In further embodiments, x is 1 and R1 is meta to the pyrazolopyridazino ring system.

[0126] In certain embodiments, x is 2 and R1 is ortho and meta to the pyrazolopyridazino ring system. In certain embodiments, x is 2 and R1 is ortho and para to the pyrazolopyridazino ring system. In further embodiments, x is 2 and R1 is para and meta 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 some embodiments, R3 is -CH3. In further embodiments, R3 is -CH2CH3. In still 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 some 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 some 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 certain embodiments, R3 is -CH(CH3)CH2OH.

[0131] In further embodiments, R3 is -CH2C(O)R4, where R4 is

[0132]

Chemical formula

[0133]

Chemical formula

[0134]

Chem.

[0135]

Chem.

[0136]

Chem.

[0137]

Chem.

[0138]

Chem.

[0139]

Chem.

[0140]

Chem.

[0141]

Chem.

[0142]

Chem.

[0143]

Chem.

[0144]

Chem.

[0145]

Chem.

[0146]

Chem.

[0147]

Chem.

[0148]

Chem.

[0149]

Chem.

[0150] [Chemical formula] In still other embodiments, R3 is -CH2C(O)R4 and R4 is

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

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

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

[0154] [Chemical formula] In still other embodiments, R3 is -CH2C(O)R4 and R4 is

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

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

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

[0158]

Chemical formula

[0159]

Chemical formula

[0160]

Chemical formula

[0161]

Chemical formula

[0162]

Chemical formula

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

[0164]

Chemical formula

[0165]

Chemical formula

[0166]

Chemical formula

[0167]

Chemical formula

[0168] In some embodiments, a is an integer in the range of 0 to 5. In some embodiments, b is an integer in the range of 0 to 4. In some embodiments, c is an integer in the range of 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]

Chemical formula

[0171] Pyrazolopyridazine compounds of formula IV The present invention further provides a method for treating or preventing 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]

Chemical formula

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

[0174] In certain embodiments, the compound of formula IV is compound 43, having the following structure, or a pharma- ceutically 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 pharma- ceutically 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 yet other embodiments, R1 is -CH3. In certain embodiments, R1 is -CF3.

[0181] In yet another embodiment, R1 is

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

[0183]

Chem.

[0184]

Chem.

[0185]

Chem.

[0186]

Chem.

[0187]

Chem.

[0188]

Chem.

[0189]

Chem.

[0190]

Chem.

[0191]

Chem.

[0192]

Chemical formula

[0193] In still other embodiments, R1 is

[0194]

Chemical formula

[0195]

Chemical formula

[0196]

Chemical formula

[0197]

Chemical formula

[0198]

Chemical formula

[0199]

Chemical formula

[0200]

Chemical formula

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

[0202]

Chemical formula

[0203]

Chemical formula

[0204]

Chemical formula

[0205]

Chemical formula

[0206] In certain embodiments, R2 is

[0207]

Chemical formula

[0208]

Chemical formula

[0209]

Chemical formula

[0210]

Chemical formula

[0211] [Chemical formula] is. In certain embodiments, R2 is

[0212] [Chemical formula] 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] [Chemical formula] TIFF2025106385000155.tif198161TIFF2025106385000156.tif203160TIFF2025106385000157.tif207163 or a pharmaceutically acceptable salt thereof.

[0216] Pyrazolopyridazine compounds of formula VI The present invention also provides a method of treating or preventing 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] [Chemical formula] wherein R3 is:

[0218] [Chemical formula] is; 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 still 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]

Chemical Structure

[0222]

Chemical Structure

[0223]

Chemical Structure

[0224]

Chemical Structure

[0225]

Chemical Structure

[0226]

Chemical Structure

[0227] In other embodiments, R3 is

[0228]

Chemical Structure

[0229]

Chemical formula

[0230]

Chemical formula

[0231]

Chemical formula

[0232]

Chemical formula

[0233] In certain embodiments, R3 is

[0234]

Chemical formula

[0235]

Chemical formula

[0236]

Chemical formula

[0237]

Chemical formula

[0238] [Chemical formula] is. In certain embodiments, R3 is

[0239] [Chemical formula] is.

[0240] In certain embodiments, R3 is

[0241] [Chemical formula] is. In a further embodiment, R3 is

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

[0243] [Chemical formula] is. In certain embodiments, R3 is

[0244] [Chemical formula] is. In other embodiments, R3 is

[0245] [Chemical formula] is and c = 2. In still other embodiments, R3 is

[0246] [Chemical formula] is.

[0247] In certain embodiments, R3 is

[0248]

Chem.

[0249]

Chem.

[0250]

Chem.

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

[0252]

Chem.

[0253] Pyrazolopyridazine compounds of formula VII The present invention further provides a method of treating or preventing 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]

Chem.

[0255]

Chem.

[0256] In certain embodiments, R4 is

[0257]

Chem.

[0258]

Chemical formula

[0259]

Chemical formula

[0260]

Chemical formula

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

[0262]

Chemical formula

[0263] Pyrazolopyridazine compounds of formula XIII The present invention further provides a method for treating or preventing 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]

Chemical formula

[0265]

Chemical formula

[0266]

Chemical formula

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

[0268] In still other embodiments, R5 is

[0269] [Chemical formula] is. In certain embodiments, R5 is

[0270] [Chemical formula] is. In yet another embodiment, R5 is

[0271] [Chemical formula] is. In certain embodiments, R5 is

[0272] [Chemical formula] is. In other embodiments, R5 is

[0273] [Chemical formula] is. In still other embodiments, R5 is

[0274] [Chemical formula] is. In certain embodiments, R5 is

[0275] [Chemical formula] is. In certain embodiments, R5 is

[0276] [Chemical formula] is. In certain embodiments, R5 is

[0277] [Chemical formula] is. In yet another embodiment, R5 is

[0278] [Chemical formula] is. In other embodiments, R5 is

[0279] [Chemical formula] is.

[0280] In still other embodiments, R5 is

[0281] [Chemical formula] is. In certain embodiments, R5 is

[0282] [Chemical formula] is. In yet another embodiment, R5 is

[0283] [Chemical formula] is. In other embodiments, R5 is

[0284] [Chemical formula] is. In certain embodiments, R5 is

[0285]

Chemical formula

[0286]

Chemical formula

[0287]

Chemical formula

[0288] In certain embodiments, R5 is

[0289]

Chemical formula

[0290]

Chemical formula

[0291]

Chemical formula

[0292]

Chemical formula

[0293]

Chemical formula

[0294] [Chemical formula] is.

[0295] In a further embodiment, R5 is

[0296] [Chemical formula] is. In yet another embodiment, R5 is

[0297] [Chemical formula] is.

[0298] In certain embodiments, R5 is

[0299] [Chemical formula] is. In other embodiments, R5 is

[0300] [Chemical formula] is. In still other embodiments, R5 is

[0301] [Chemical formula] is. In specific embodiments, R5 is

[0302] [Chemical formula] is.

[0303] In a further embodiment, R5 is

[0304] [Chemical formula] is. In yet another embodiment, R5 is

[0305] [Chemical formula] is as follows.

[0306] In certain embodiments, R5 is

[0307] [Chemical formula] is as follows. In other embodiments, R5 is

[0308] [Chemical formula] is as follows. In still other embodiments, R5 is

[0309] [Chemical formula] is as follows. In certain embodiments, R5 is

[0310] [Chemical formula] is as follows.

[0311] In further embodiments, R5 is

[0312] [Chemical formula] is as follows. In still another embodiment, R5 is

[0313] [Chemical formula] is as follows.

[0314] In certain embodiments, R5 is

[0315] [Chemical formula] It is.

[0316] In certain embodiments, R6 is

[0317]

Chemical formula

[0318]

Chemical formula

[0319]

Chemical formula

[0320]

Chemical formula

[0321]

Chemical formula

[0322]

Chemical formula

[0323]

Chemical formula

[0324]

Chem.

[0325]

Chem.

[0326]

Chem.

[0327]

Chem.

[0328] In further embodiments, R6 is

[0329]

Chem.

[0330]

Chem.

[0331]

Chem.

[0332]

Chem.

[0333] [Chemistry] It is. In certain embodiments, R6 is

[0334] [Chemistry] It is. In a further embodiment, R6 is

[0335] [Chemistry] It is. In yet another embodiment, R6 is

[0336] [Chemistry] It is.

[0337] In certain embodiments, R6 is

[0338] [Chemistry] It is. In other embodiments, R6 is

[0339] [Chemistry] It 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 following structure:

[0342] [Chemistry] TIFF2025106385000261.tif 213162 TIFF2025106385000262.tif 216164 TIFF2025106385000263.tif 178157 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]

Chemical formula

[0345] Pyrazolopyridazine compounds of formula XIV The present invention also provides a method for treating or preventing 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]

Chemical formula

[0347]

Chemical formula

[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. In certain embodiments, R7 is

[0350]

Chemical formula

[0351]

Chemical formula

[0352]

Chemical formula

[0353]

Chemical formula

[0354]

Chemical formula

[0355]

Chemical formula

[0356] In other embodiments, R7 is

[0357]

Chemical formula

[0358]

Chemical formula

[0359]

Chemical formula

[0360] [Chemical formula] is. In a further embodiment, R7 is

[0361] [Chemical formula] is.

[0362] In certain embodiments, R7 is

[0363] [Chemical formula] is. In other embodiments, R7 is

[0364] [Chemical formula] is. In still other embodiments, R7 is

[0365] [Chemical formula] is. In a further embodiment, R7 is

[0366] [Chemical formula] is. In yet another embodiment, R7 is

[0367] [Chemical formula] is. In certain embodiments, R7 is

[0368] [Chemical formula] is.

[0369] In certain embodiments, R7 is

[0370] [Chemical formula] is. In a further embodiment, R7 is

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

[0372] [Chemical formula] is. In a specific embodiment, R7 is

[0373] [Chemical formula] is. In other embodiments, R7 is

[0374] [Chemical formula] is, and c = 2. In still other embodiments, R7 is

[0375] [Chemical formula] is.

[0376] In certain embodiments, R7 is

[0377] [Chemical formula] is. In other embodiments, R7 is

[0378] [Chemical formula] is. In still other embodiments, R7 is

[0379] [Chemical formula] It is. Further, in other embodiments, R7 is

[0380]

Chemical formula

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

[0382]

Chemical formula

[0383] Pyrazolopyridazine compounds of formula XV The present invention also provides a method for treating or preventing 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]

Chemical formula

[0385]

Chemical formula

[0386] In certain embodiments, R8 is

[0387]

Chemical formula

[0388]

Chemical formula

[0389]

Chemical formula

[0390]

Chemical formula

[0391]

Chemical formula

[0392]

Chemical formula

[0393]

Chemical formula

[0394] In certain embodiments, R8 is

[0395]

Chemical formula

[0396]

Chemical formula

[0397]

Chemical formula

[0398]

Chemical formula

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

[0400] [ka] It is.

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

[0402] [ka] TIFF2025106385000311.tif53157 or a pharma- ceutically acceptable salt thereof.

[0403] Other pyrazolopyridazine compounds 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 having the structure of the following formula, or a pharma- ceutically acceptable salt thereof:

[0404] [ka] TIFF2025106385000313.tif124157TIFF2025106385000314.tif190162

[0405] Non-pyrazolopyridazine compounds The compounds in Table 1 below, or pharma- ceutically 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 pharma- ceutically acceptable carrier or vehicle.

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

[0408] [Table 1] TIFF2025106385000316.tif226155TIFF2025106385000317.tif223157TIFF2025106385000318.tif207153TIFF2025106385000319.tif219155TIFF2025106385000320.tif202154TIFF2025106385000321.tif203156TIFF2025106385000322.tif215154TIFF2025106385000323.tif195155TIFF2025106385000324.tif212157TIFF2025106385000325.tif218157TIFF2025106385000326.tif209155TIFF2025106385000327.tif212156TIFF2025106385000328.tif221156TIFF2025106385000329.tif210155TIFF2025106385000330.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 depicted as having thick wedge-shaped or hatched wedge-shaped lines indicating absolute stereochemistry.

[0410] Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds can be in the form of salts. In some embodiments, the salts are pharmaceutically acceptable salts. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid forming the acid addition salt can be an organic acid or an inorganic acid. The base forming 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, saccharic 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, caffeic acid, mandelic 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, gentisinate, gluconate, glucuronate, saccharinate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, citrate, oxalate, maleate, hydroxymaleate, methylmaleate, glycolate, malate, caffeate, 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 an inorganic base to a pyrazolopyridazine compound or a non-pyrazolopyridazine compound having a carboxyl group. The inorganic base consists of, for example, a metal cation paired with a basic counterion such as hydroxide, carbonate ion, bicarbonate ion, or phosphate ion. The metal can be an alkali metal, an alkaline earth metal, a transition metal, or a 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 salt, sodium salt, potassium salt, cesium salt, cerium salt, magnesium salt, manganese salt, iron salt, calcium salt, strontium salt, cobalt salt, titanium salt, aluminum salt, copper salt, cadmium salt, and zinc salt.

[0415] An ammonium salt can be formed by the addition of ammonia or an organic amine to a pyrazolopyridazine compound or a non-pyrazolopyridazine compound 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, chlorprocaine, 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, chlorprocaine salts, choline salts, dicyclohexylammonium salts, and N-methylglucamine salts.

[0417] Methods for preparing pyrazolopyridazine compounds Methods for preparing pyrazolopyridazine compounds are disclosed in U.S. Patent 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 herein 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] The following scheme is a non-limiting example of a synthetic scheme useful for synthesizing pyrazolopyridazine compounds.

[0420] [Chemical formula]

[0421] Scheme 1 describes the outline of the preparation of pyrazolopyridazine compounds having a 1-N-methyl group, where R’ and R” are independently unsubstituted or substituted phenyl groups. For example, a 2-cyanocarbonyl compound where R’ is unsubstituted or substituted phenyl is condensed with N-methylhydrazine to obtain a 3-substituted-1-methyl-1H-pyrazole-5-amine. The 5-amino group is acylated with acetic anhydride in the presence of a base such as pyridine to obtain a 5-amide compound. This 5-amide compound is iodinated with a mixture of iodine and iodic acid in a solvent such as ethanol (EtOH) to obtain N-(3-substituted-4-iodo-1-methyl-1H-pyrazole-5-yl)acetamide. In a solvent such as dimethylformamide (DMF) containing a base such as triethylamine, in the presence of copper(I) iodide, a palladium-mediated cross-coupling such as Sonogashira cross-coupling of the acetamide with a terminal alkyne substituted with R” is catalyzed by a palladium complex such as palladium(II) bistriphenylphosphine dichloride to obtain a disubstituted alkyne where R” is unsubstituted or substituted phenyl. A primary amine is obtained by saponification of the alkyne acetamide with a base such as sodium hydroxide in a solvent such as ethanol. A diazo intermediate is obtained by diazotization of this primary amine with sodium nitrite in concentrated hydrochloric acid, and this is cyclized to obtain a pyrazolopyridazine compound having a 1-N-methyl group. Here, R’ and R” are independently unsubstituted or substituted phenyl groups.

[0422]

Chemical formula

[0423] Scheme 2 describes the outline of the preparation of pyrazolopyridazine compounds having an R3 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 obtain 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 a hydrazine substituted with R3 in a solvent such as ethanol to obtain an intermediate hydrazone, which is cyclized to obtain a pyrazolopyridazine compound having a 1-N-R3 group. Here, R3 is defined by formulas II and III, and R’ is an unsubstituted or substituted phenyl group.

[0424]

Chemical formula

[0425] Scheme 3 describes the outline of 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, terminal alkyne, halogenated alkenyl, or halogenated aryl in the presence of an appropriate catalyst such as a palladium complex, in the presence of a non-palladium transition metal salt such as zinc or copper salt, and in the presence of an additive such as triphenylphosphine or an organic amine base, to obtain a pyrazolopyridazine compound 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 iodine group in the starting material.

[0426]

Chemical formula

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

[0428]

Chemical formula

[0429] Scheme A describes the outline of 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 hours. The reaction mixture is concentrated in vacuo. The crude reaction mixture is partitioned between CH2Cl2 (400 mL) and saturated NaHCO3(aq). The aqueous phase is extracted with CH2Cl2, the organic phases are combined, dried over MgSO4, filtered, and evaporated to give Compound 8A as a solid (70 g, 95% yield). 1 H NMR (400 MHz, 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), acetic anhydride (17.4 g, 0.17 mol) is added dropwise at 0 °C under a nitrogen atmosphere. The reaction mixture is stirred at room temperature (RT) for 16 h. The reaction mixture is concentrated in vacuo. The residue is diluted with CH2Cl2 and water. The layers are separated, and the organic layer is washed with water and brine, dried (MgSO4), and concentrated in vacuo. CH2Cl2 is added to the residue, and the solid is collected by filtration to obtain Compound 10A as a solid (22 g, 45% yield). The mother liquor is concentrated in vacuo and washed with cold CH2Cl2 to obtain a second batch of Compound 10A (15 g, 31% yield). 1 1H NMR (400 MHz, DMSO-d6) δ (ppm) 10.13 (s, 1H), 7.81 (d, J = 7.6 Hz, 2H), 7.45 (t, J = 7.6 Hz, 2H), 7.40 - 7.32 (m, 1H), 6.80 (s, 1H), 5.07 (s, 2H), 4.22 (q, J = 7.1 Hz, 2H), 2.14 (s, 3H), 1.28 (t, J = 7.1 Hz, 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 CH2Cl2 (250 mL), acetyl chloride (20 mL, 0.281 mol) is added dropwise at 0 °C under a nitrogen atmosphere. The reaction mixture is stirred at RT for 3 h. The reaction mixture is diluted with CH2Cl2 and water. The layers are separated, and the organic layer is washed with water and brine, dried (phase separator cartridge), and concentrated in vacuo. Diethyl ether is added to the residue, and the solid is collected by filtration to obtain Compound 11A as a solid (25.1 g, 88% yield). 11H NMR (400 MHz, DMSO-d6) δ (ppm) 12.79 (s, 1H), 10.40 (s, 1H), 7.71 (d, J = 7.5 Hz, 2H), 7.44 (dd, J = 7.6, 7.6 Hz, 2H), 7.34 (dd, J = 7.2, 7.2 Hz, 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) is heated at 50 °C for 2 h and cooled to RT. The reaction mixture is concentrated in vacuo and the residue is eluted through a pad of silica gel with CH2Cl2 / diethyl ether (1:0 to 97:3). The residue is partitioned between CH2Cl2 and 2 M Na2S2O3 solution (aq). The layers are separated, the organic layer is washed, dried (MgSO4) and concentrated in vacuo to give a residue which is partially purified by chromatography (silica gel, CH2Cl2 / isohexane 1:1 to 1:0, then CH2Cl2 / diethyl ether 9:1 to 8:2) and then triturated with diethyl ether to give compound 12A as an off-white solid (43 g, 81% yield). 1 1H NMR (400 MHz, CDCl3) As a 3:1 mixture of rotamers δ (ppm) 7.81 (d, J = 7.6 Hz, 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 = 7.1 Hz, 2H), 2.24 (s, 2.25H), 2.04 (s, 0.75H), 1.30 (t, J = 7.1 Hz, 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) is heated at 50 °C for 3 h and cooled to RT. The reaction mixture is concentrated in vacuo and partitioned between CH2Cl2 and 2 M aqueous Na2S2O3 solution. The layers are separated, the organic layer is washed with brine, dried (phase separator cartridge), and concentrated in vacuo to afford a mixture of compound 13A and starting material 11A (2.2:1, 30.3 g). This mixture is 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 (400 MHz, CDCl3) δ (ppm) 11.74 - 11.74 (m, 1H), 7.81 (d, J = 7.2 Hz, 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 A mixture of compound 12A (18.6 g, 45 mmol), phenylacetylene (9.2 g, 90 mmol), copper(I) iodide (860 mg, 4.5 mmol), triethylamine (200 mL), and DMF (75 mL) is bubbled with nitrogen for 15 min. Bis(triphenylphosphine)palladium(II) dichloride (1.6 g, 2.25 mmol) is added and the reaction mixture is stirred at 90 °C under nitrogen for 4.5 h. The reaction mixture is cooled to RT and diluted with ethyl acetate and water. The organic phase is washed with water and brine, dried (MgSO4), filtered, and concentrated in vacuo. The residue is partially purified by column chromatography (silica gel, CH2Cl2, then isohexane / ethyl acetate 1:1, then CH2Cl2 / ethyl acetate 9:1 - 8:2) and then triturated with diethyl ether to afford compound 14A as a solid (13 g, 75% yield). 11H NMR (400 MHz, 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 = 7.1 Hz, 2H), 2.17 (s, 3H), 1.28 (t, J = 7.1 Hz, 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 the same procedure as described for the synthesis of compound 14A. 1 1H NMR (400 MHz, CDCl3) δ (ppm) 11.57 - 11.57 (m, 1H), 8.11 (d, J = 7.4 Hz, 2H), 7.91 (s, 1H), 7.55 - 7.49 (m, 2H), 7.44 (dd, J = 7.5, 7.5 Hz, 2H), 7.37 (dd, J = 1.9, 5.0 Hz, 4H), 2.32 (s, 3H).

[0437] [Chemical formula] (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 describes the outline of the preparation of compound 3B and compound 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, heated at 80 °C for 8 h, and cooled to RT. A precipitate forms upon cooling. 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 afford compound 16B as a solid (9.8 g, 85% yield). 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 8.06 (d, J = 7.8 Hz, 2H), 7.56 (d, J = 7.6 Hz, 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 Sodium borohydride (11 g, 289 mmol) is added to a suspension of compound 14B (22.4 g, 58 mmol) in ethanol (290 mL), and the reaction mixture is stirred at RT for 16 h. The reaction mixture is partially concentrated to a final volume of 250 mL. 25% NaOH solution (aq) (250 mL) is added, and the reaction mixture is stirred at 80 °C for 4 h. The reaction mixture is cooled to room temperature, and the phases are separated. The aqueous phase is extracted three times with ethyl acetate, the organic phases are combined, dried (MgSO4), filtered, and concentrated in vacuo. The residue is triturated with diethyl ether (20 mL), the product is filtered, and dried in vacuo to afford compound 17B as an off-white solid (9.96 g, 57% yield). The mother liquor is concentrated in vacuo and purified by column chromatography (silica gel, gradient 0 - 100% ethyl acetate / isohexane) to obtain an additional yield (1.79 g, 10% yield). 1 H NMR (400 MHz, CDCl3) δ (ppm) 7.78 - 7.74 (m, 4H), 7.55 - 7.48 (m, 6H), 4.98 (t, J = 4.8 Hz, 2H), 4.29 (m, 2H), 3.03 (t, J = 6.4 Hz, 1H).

[0441] Compound 18B of Scheme B: 3-Phenyl-4-(2-phenylethynyl)-1H-pyrazol-5-amine Compound 18B (5.4 g, 50% yield) is obtained from compound 15B (12.5 g, 41 mmol) by the same procedure as described for the synthesis of compound 16B. 1 H NMR (400 MHz, CDCl3) δ (ppm) 7.87 (d, J = 7.2 Hz, 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 stirred for 15 minutes. Then, 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 hours. The reaction mixture is diluted with CH2Cl2 and washed with water and brine. The organic layer is dried (MgSO4) and concentrated in vacuo. The residue is purified by column chromatography (silica gel, diethyl ether / CH2Cl2 1:9) to give compound 19B as a solid (1.7 g, 53% yield). 1 H NMR (400 MHz, 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 cHCl (220 mL) at -10 °C and stirred for 10 minutes. Compound 17B (6.7 g, 22.1 mmol) is added as a solid. The reaction mixture is warmed, sonicated for 5 minutes, and then stirred at RT for 2 hours. The reaction mixture is diluted with CH2Cl2 and water, and the aqueous phase is extracted with CH2Cl2. The organic phases are combined, dried (MgSO4), 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 from diethyl ether and then from ethyl acetate to afford Compound 3B as a solid (900 mg, 12% yield). 1 H NMR (400 MHz, 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 = 6.4 Hz, 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 (Analysis method 1: HPLC (Phenomenex Luna 5μm C18, 100×4.6 mm) gradient of 5 - 95% acetonitrile in water (0.1% formic acid in each mobile phase)) R 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) was added methylmagnesium chloride (3 M solution in THF, 3 mL, 9 mmol) at 0 °C. The resulting solution was stirred at RT for 3.5 h, then successively diluted with ethyl acetate and quenched by the addition of 1 M HCl(aq). The aqueous phase was extracted with ethyl acetate, the combined organic layers were dried (MgSO4), and concentrated in vacuo. The resulting residue was purified using chromatography (silica gel, gradient of 0 - 75% ethyl acetate / isohexane) to give compound 21B as a solid (529 mg, 55% yield). 1 H NMR (400 MHz, CDCl3) 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 as a 1.5:1 mixture with N-[2-acetonyl-5-phenyl-4-(2-phenylethynyl)pyrazol-3-yl]acetamide δ (ppm) 8.11 (dd, J = 7.5, 12.3 Hz, 2H), 7.51 - 7.40 (m, 4H), 7.37 - 7.31 (m, 4H), 4.95 (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) according to a similar procedure as outlined for the synthesis of compound 16B. 1 H NMR (400 MHz, CDCl3) δ (ppm) 8.11 - 8.08 (m, 2H), 7.50 - 7.47 (m, 2H), 7.41 (dd, J = 7.5, 7.5 Hz, 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 is added all at once to cooled (cooling bath at -15 °C) cHCl (9 mL) (121 mg, 1.75 mmol), and the suspension is stirred continuously for 10 minutes, after which Compound 22B (290 mg, 0.88 mmol) is added. After 5 minutes, the cooling bath is removed and the reaction mixture is stirred at RT for 3 hours. The reaction is cooled again (0 °C), CH2Cl2 is added, followed by water. The aqueous phase is extracted with CH2Cl2, the organic phases are combined, dried (MgSO4), filtered, and concentrated in vacuo. The crude material is purified by column chromatography (silica gel, 0 - 50% ethyl acetate / isooctane gradient) to give Compound 4B as an orange oil (56 mg). The resulting material is further purified by preparative HPLC to give Compound 4B as a solid (34 mg, 10% yield). 1 H NMR (400 MHz, 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 (Analysis method 1: HPLC (Phenomenex Luna 5μm C18, 100×4.6 mm) gradient of 5 - 95% acetonitrile in water (0.1% formic acid in each mobile phase)) R t 4.49 minutes; 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) is added portionwise to cHCl (314 mL) at -15 °C and stirred for 15 minutes. Compound 18B (5.4 g, 21 mmol) is added as a solid, followed by the addition of CH2Cl2 (10 mL). The reaction mixture is warmed and stirred at RT for 1 hour. The reaction mixture is diluted with CH2Cl2 (44 mL), and NaCl (2.7 g) is added. The reaction mixture is heated to 50 °C for 1 day. The layers are separated, the organic layer is washed with water, dried (phase separator cartridge), and concentrated in vacuo. The residue is purified by column chromatography (silica gel, isohexane / ethyl acetate 4:1, then CH2Cl2 / ethyl acetate 1:0 to 4:1) to afford Compound 20B as a solid (3.0 g, 47% yield). 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 15.08 (s, 1H), 7.81 - 7.73 (m, 4H), 7.58 - 7.51 (m, 6H).

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

[0449] Scheme C depicts the outline of 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)pentaamide)propyl)carbamate To a solution of biotin (23C, 350 mg, 1.43 mmol) in DMF (7.2 mL) are added N-(3-aminopropyl)carbamic acid tert-butyl (24C, 250 mg, 1.43 mmol), DIPEA (0.375 mL, 2.15 mmol), and HATU (816 mg, 2.15 mmol). The reaction mixture is stirred at RT for 20 h and then diluted with ethyl acetate and 4% aqueous LiCl. The aqueous phase is extracted twice with ethyl acetate, and the combined organic layers are dried (MgSO4) and concentrated in vacuo. The resulting residue is purified using chromatography (silica gel, gradient of 0 - 12% 7M NH3 in MeOH / CH2Cl2) to afford compound 25C as a solid (180 mg, 31% yield). 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 7.75 (t, J = 5.2 Hz, 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 = 5.3 Hz, 1H), 3.16 (dq, J = 6.2, 4.3 Hz, 1H), 3.07 (dd, J = 6.8, 12.9 Hz, 2H), 2.96 (dd, J = 6.6, 13.0 Hz, 2H), 2.88 (dd, J = 5.2, 12.4 Hz, 1H), 2.11 (t, J = 7.5 Hz, 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 CH2Cl2 (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 CH2Cl2 (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, washed with CH2Cl2 / 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) are 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 is stirred at RT for 1.5 h and then diluted with CH2Cl2 and 4% aqueous LiCl. The aqueous phase is extracted twice with CH2Cl2, the combined organic layers are dried (phase separation), and concentrated in vacuo. The resulting residue is first purified by preparative HPLC to give 70 mg, which is further purified by silica gel chromatography to give the desired compound 5C as a solid (44 mg, 16% yield). 11H NMR (400 MHz, DMSO-d6) δ (ppm) 8.39 (dd, J = 5.7, 5.7 Hz, 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, 7.6 Hz, 1H), 4.18 - 4.13 (m, 1H), 3.22 - 3.08 (m, 5H), 2.85 (dd, J = 5.2, 12.5 Hz, 1H), 2.61 (d, J = 12.4 Hz, 1H), 2.10 (dd, J = 7.5, 7.5 Hz, 2H), 1.66 - 1.48 (m, 6H), 1.39 - 1.28 (m, 2H). 13 13C NMR (100 MHz, 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, 129.42, 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 (Analysis method 1: HPLC (Phenomenex Luna 5μm C18, 100×4.6 mm) gradient of 5 - 95% acetonitrile in water (0.1% formic acid in each mobile phase)) R t 3.52 minutes; m / z 647 [M + H] 98.38% purity.

[0453]

Chemical formula

[0454] Scheme D describes the outline of 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), diethyl azodicarboxylate (0.355 mL, 2.26 mmol) is slowly added at RT. Next, the reaction mixture is heated to 120 °C for 1 hour using microwave irradiation. The reaction mixture is cooled to RT, and 4M HCl in 1,4-dioxane (4 mL) is added. The reaction mixture is stirred at RT for 4 hours, diluted with CH2Cl2 (10 mL), and the solution is loaded onto a Biotage SCX-2 cartridge (20 g), eluting with methanol and then 7M NH3 in methanol. The fractions are concentrated in vacuo to give Compound 27D in a 1:1 ratio with 2-hydroxyethyl)piperazine (950 mg, 100% yield), which is 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), DIPEA (0.59 mL, 3.39 mmol), biotin (678 mg, 2.78 mmol), and HATU (1.29 g, 3.39 mmol) are added. After the reaction mixture is stirred at RT for 24 hours, it is diluted with DMSO (5 mL). NaOH (2M, 5 mL) is added, and the reaction mixture is heated to 40 °C for 1 hour and then stirred at RT for 2 days. This reaction mixture is 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 CH2Cl2 and 2 M Na2CO3 solution. The aqueous phase is extracted twice with CH2Cl2. The combined organic layers are dried (MgSO4) and concentrated in vacuo. The obtained residue is purified using chromatography (silica gel, gradient of 0 - 12% 7 M NH3 in MeOH / CH2Cl2) to give compound 6D as a solid (42 mg, 42% yield). 1 1H NMR (400 MHz, 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), 4.38 - 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, 5.1 Hz, 1H), 2.63 (d, J = 12.4 Hz, 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). 1313C NMR (100 MHz, 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 (Analysis method 2: HPLC (Hichrom ACE 3 C18-AR mixed-mode column 100×4.6 mm) gradient of 2-100% acetonitrile in water (0.1% formic acid in each mobile phase)) R t 9.77 minutes; m / z 645 [M+H] 93.27% purity.

[0458] Molecular chaperone When a specific protein folds, there are many possible conformations it can adopt, but ultimately the biologically functional protein folds into a stable state called its "native state" where the tendency of the protein to aggregate is minimized. The protein folding process is thermodynamically driven by the so-called "hydrophobic effect" in which the hydrophobic amino acid residues of the protein interact with each other to form a hydrophobic core, while the hydrophilic amino acid residues of the protein tend to remain on the surface of the protein. The initial, i.e., partially folded protein is "sticky" because its hydrophobic amino acids are not fully buried within the core of the protein. As a result, sticky proteins can aggregate together and, especially in the cellular environment where there are many other protein molecules, can form insurmountable aggregates.

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

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

[0461]

Table 2

[0462] In some embodiments, the pyrazolopyridazine compound, non-pyrazolopyridazine compound, or their metabolites bind to a molecular chaperone. In some embodiments, the pyrazolopyridazine compound, non-pyrazolopyridazine compound, or their metabolites covalently bind to a molecular chaperone.

[0463] In some embodiments, as a result of the binding of the pyrazolopyridazine compound, non-pyrazolopyridazine compound, or their metabolites to a molecular chaperone, proteopathy is treated or prevented in a subject in need thereof.

[0464]

[0465] Proteopathy In another embodiment, the molecular chaperone is a member of the Hsp10 family, Hsp40 family, Hsp60 family, Hsp70 family, Hsp90 family, or Hsp100 family.Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds are useful for treating or preventing proteopathy.

[0466] In some embodiments, the proteopathy is a neurodegenerative disease. Exemplary neurodegenerative diseases include, but are not limited to, Alzheimer's disease, progressive supranuclear palsy, boxer dementia, frontotemporal dementia, and Parkinsonism linked to chromosome 17 (FTDP-17), corticobasal degeneration, tangle-predominant dementia, ganglioglioma, gangliocytoma, meningeal angiomatosis, 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, Lewy body dementia, multiple system atrophy, axonal dystrophy, dentatorubral-pallidoluysian atrophy (DRPLA), spinal and bulbar muscular atrophy (SBMA), spinocerebellar ataxia type 1 (SCA1), SCA2, SCA3, SCA6, SCA7, SCA17, prion disease, 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 - Icelandic type (HCHWA - I), familial amyloid 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), Finnish familial amyloidosis (FAF), fibrinogen amyloidosis (AFib), lysozyme amyloidosis (ALys), dialysis amyloidosis (Aβ2M), medullary thyroid carcinoma (ACal), atrial amyloidosis (AANF), pituitary prolactinoma (APro), lattice corneal dystrophy, cutaneous lichen amyloidosis (AKer), Mallory bodies, primary cutaneous amyloidosis, corneal lactoferrin amyloidosis, dental (pindborg) tumor amyloid or seminal vesicle amyloid.

[0468] In some embodiments, the proteopathy is a lysosomal storage disorder. Specific lysosomal storage disorders include, but are 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, infantile GM1 gangliosidosis, late infantile / juvenile GM1 gangliosidosis, adult / chronic GM1 gangliosidosis, I-cell disease / mucolipidosis II, infantile free sialic acid storage disease / ISSD, juvenile hexosaminidase A deficiency, infantile-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 I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, Sanfilippo syndrome type A / MPS III A, Sanfilippo syndrome type B / MPS III B, Sanfilippo syndrome type C / MPS III C, Sanfilippo syndrome type D / MPS III D, Morquio syndrome type A / MPS IVA, Morquio syndrome type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC, mucolipidosis type IV, multiple sulfatase deficiency, Niemann-Pick disease type A, Niemann-Pick disease type B, Niemann-Pick disease type C, CLN6 disease - atypical late infantile, CLN6 disease - late-onset variant, CLN6 disease - early juvenile, Batten-Spielmeyer-Vogt / juvenile NCL / CLN3 disease, Finnish variant late infantile CLN5, Jansky-Bielschowsky disease / late infantile CLN2 / TPP1 disease, Kufs / adult-onset NCL / CLN4 disease, NorthernThere is epilepsy / dysmorphic late-onset infantile CLN8, Santavuori-Haltia / infantile CLN1 / PPT disease, beta-mannosidosis, Pompe disease / glycogen storage disease type II, pycnodysostosis, Sandhoff disease / adult-onset / GM2 gangliosidosis, Sandhoff disease / GM2 gangliosidosis - infantile, 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 mucopolysaccharidosis. Specific mucopolysaccharidoses include, but are not limited to, pseudo-Hurler polydystrophy / mucolipidosis IIIA, MPS I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, Sanfilippo syndrome type A / MPS III A, Sanfilippo syndrome type B / MPS III B, Sanfilippo syndrome type C / MPS III C, Sanfilippo syndrome type D / MPS III D, Morquio syndrome type A / MPS IVA, Morquio syndrome type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC and mucolipidosis IV.

[0470] In other embodiments, 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 congenital amaurosis, retinal degeneration syndromes, age-related macular degeneration including exudative and atrophic age-related macular degeneration, and 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.

[0472] In a further embodiment 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

[0475] Cystic fibrosis Cystic fibrosis (CF), also known as mucoviscidosis, is an autosomal recessive disorder 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 in the pancreas. This disease is characterized by difficulty breathing and coughing up phlegm due to frequent lung infections.

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

[0477]

Table 4

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

[0479] Accordingly, the present invention further provides a method of treating or preventing cystic fibrosis, comprising administering to a subject in need thereof an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. 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 yet another embodiment, the subject has a class III CF mutation. In another embodiment, the subject has a class IV CF mutation. In yet 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 yet another embodiment, the mutation is G551D, G551S, or G1349D. In another embodiment, the mutation is R117H, R334W, or R347P. In yet 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 a genetic, degenerative eye disease that causes severe visual impairment due to the progressive degeneration of rod photoreceptor cells in the retina. After progressive rod degeneration, abnormalities in adjacent retinal pigment epithelium (RPE) and deterioration of cone photoreceptor cells may occur.

[0483] There are multiple genes that can cause RP when mutated. The inheritance patterns of RP have been identified as autosomal dominant, autosomal recessive, X-linked, and maternal (mitochondrial) inheritance. X-linked RP can be recessive and affect mainly males only, or dominant and affect both males and females. Some digenic (controlled by two genes) and mitochondrial forms of RP are also known.

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

[0485] Since the first report of the P23H mutation in the intradiscal domain of the protein, up to 150 RP-related opsin gene mutations have been reported. These mutations are found throughout the opsin gene and are distributed along the three domains of the protein (intradiscal, transmembrane, and cytoplasmic domains). Mutations in the opsin gene are most commonly missense mutations, causing misfolding of the rhodopsin protein. The mutation at amino acid 23 where proline is replaced by histidine in the opsin gene accounts 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 the genes mutated and the types of RP in subjects with RP is given in Table 5.

[0487]

Table 5

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

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

[0490] In one embodiment, retinitis pigmentosa (RP) is 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-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 subject has a mutation in one or more of the following genes: 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 use Pyrazolopyridazine compounds and non-pyrazolopyridazine compounds can be administered to a subject as components of a composition comprising a pharmaceutically acceptable carrier or vehicle. 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, for example, as 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, syrup, methylcellulose, methyl and propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. The composition can further comprise a lubricant, a wetting agent, an emulsifying and suspending agent, a preservative, a sweetening or flavoring agent.

[0493] The composition can comprise an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. The composition can be formulated into unit dosage forms comprising an effective amount of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition comprises, for example, from about 1 ng to about 1,000 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition comprises from about 100 mg to about 1,000 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition comprises from about 100 mg to about 500 mg of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound. In some embodiments, the composition comprises from 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 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 as 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, about 1 ng to about 0.1 g / kg, or about 100 ng to about 10 mg / kg per kg of the subject's body weight.

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

[0496] The pyrazolopyridazine compound, non-pyrazolopyridazine compound, or composition thereof can be provided in a lyophilized form for reconstitution, for example, as an isotonic, aqueous, or saline buffer solution for parenteral, subcutaneous, intradermal, intramuscular, or intravenous administration. The composition can also be in the form of a liquid preparation useful for oral, intratympanic, nasal, or sublingual administration, such as a suspension, syrup, or elixir. The composition can also be in a form suitable for oral administration, such as capsules, tablets, pills, and solid formulations that can be chewed. The composition can also be manufactured as a cream for topical administration as a liquid, viscous liquid, paste, or powder. The composition can also be manufactured as a powder for pulmonary administration with or without an aerosolizing component.

[0497] The composition can be in oral, otic, intranasal, sublingual, duodenal, subcutaneous, buccal, intracolonic, rectal, vaginal, mucosal, pulmonary, transdermal, intradermal, parenteral, intravenous, intramuscular and ophthalmic administration forms and can cross the blood-brain barrier.

[0498] The composition can be administered by various means known in the art. For example, the composition can be administered orally and formulated as tablets, capsules, granules, powders or syrups. Alternatively, the composition can be administered parenterally as an injection solution (e.g., intravenous, intramuscular or subcutaneous), drip preparation or suppository. In the case of ophthalmology, the composition can be formulated as eye drops or ophthalmic ointments. The otic composition can be formulated as ear drops, ointments, creams, liquids, gels, or plasters for the inside or surface of the ear. These formulations can be manufactured by conventional means and the composition can be mixed with any conventional additives such as excipients, binders, disintegrants, lubricants, solubilizers, suspending aids, emulsifiers, or coating agents.

[0499] The composition can include wetting agents, emulsifiers, and lubricants, coloring agents, release agents, coating agents, sweeteners, flavors and fragrances, preservatives and antioxidants.

[0500] The composition can be suitable for, for example, oral, otic, intraocular, nasal, topical (including buccal and sublingual), rectal, vaginal, aerosol and / or parenteral administration. The composition can be provided in unit dosage form and can be manufactured 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 solutions or suspensions in aqueous or non-aqueous liquids, or as water-in-oil or oil-in-water liquid emulsions, or as elixirs or syrups, or as troches (using inert bases such as gelatin and glycerin, or sucrose and acacia gum). The compositions may also be administered as a bolus, a pastille, or a paste.

[0502] Additional examples of pharmaceutically acceptable carriers or vehicles are: (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 acacia gum; (3) humectants such as glycerol; (4) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents such as paraffin; (6) absorption promoters 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) buffering agents. Similar compositions can be used as fillers in soft or hard filled gelatin capsules.

[0503] For oral administration, liquid dosage forms include pharmaceutically acceptable emulsions, microemulsions, gels, solutions, suspensions, syrups, and elixirs. Liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents, 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, tetrahydrofurfuryl 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. Ointments, pastes, creams, and gels can contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, 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 silicate, polyamide powder, or mixtures thereof. Sprays can further contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0507] The composition can be administered by an aerosol of solid particles. A non-aqueous (e.g., fluorocarbon propellant) suspension could be used. A sonic nebulizer can be used as it minimizes exposure to shear that can cause degradation.

[0508] An aqueous aerosol can be prepared by formulating an aqueous solution or suspension of a pyrazolopyridazine compound or a non-pyrazolopyridazine compound with a conventional pharmaceutically acceptable carrier or vehicle such as a nonionic surfactant (Tweens, Pluronics, or polyethylene glycol); a protein such as serum albumin; a sorbitan ester; a fatty acid; lecithin; an amino acid; a buffer; a salt; a sugar; or a sugar alcohol.

[0509] Compositions suitable for parenteral administration include a pyrazolopyridazine compound or a non-pyrazolopyridazine compound, 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 a sterile injectable solution or dispersion immediately before use, which can contain an antioxidant, a buffer, a bacteriostatic agent, or a solute to render the formulation isotonic with the blood of the subject, and a suspending or thickening agent.

[0510] Although the invention has been described with reference to certain embodiments, other embodiments will be 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 can be made to both the materials and methods without departing from the scope of the invention.

[0511] Incorporation by Reference Each document disclosed in this application is hereby incorporated by reference in its entirety into this specification.

Claims

1. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of Formula II or a pharmaceutically acceptable salt thereof: 【Chemical 1】 Wherein, Hal is -Cl, -F, -I, or -Br; x is an integer in the range of 0 to 5; Each R １ is independently -Cl, -F, -I, -Br, -C １ -C ３ alkyl, -O-C １ -C ３ alkyl, -CN, -CF ３ , -C(O)NH(CH ３ ), or -C≡CCH ２ OH; y is an integer in the range of 0 to 5; Each R ２ is independently -Cl, -F, -Br, -C １ -C ３ alkyl, -O-C １ -C ３ alkyl, -CN, -CF ３ , -C(O)NH(CH ３ ), or -C≡CCH ２ OH; R ３ is -H, -C １ -C ６ alkyl, -(C １ -C ６ alkylene)-OH, -(C １ -C ６ alkylene)-phenyl, -(C １ -C ６ alkylene)-O-(C １ -C ６ alkyl), -C ２ -C ６ alkenyl, -(C １ -C ６ alkylene)-C(O)R ４ , -(C １ -C ６ alkylene)-R ５ , [Chemical Formula 2] ; R ４ is —OH, —O—(C １ —C ６ alkyl), —NH ２ , —NH(C １ —C ６ alkyl), —NH((C １ —C ６ alkylene)—OH), —NH((C １ —C ６ alkylene)N(C １ —C ６ alkyl) ２ ), —N(C １ —C ６ alkyl)((C １ —C ６ alkylene)—CN), -N(C １ -C ６ alkyl)((C １ -C ６ alkylene)N(C １ -C ６ alkyl) ２ ), -NH(C １ -C ６ alkylene)-O-(C １ -C ６ alkyl), [Chemical Formula 3] [Chemical] ; a is an integer in the range of 0 to 10; b is an integer in the range of 0 to 8; c is an integer in the range of 0 to 6; R ５ is 【Chemical Formula 4】 .

2. The method according to claim 1, wherein the compound has the structure: [Chemical Formula 5] [Chemical] 【Chem.】 【Chem.】 【Chem.】 [Chemical] 【Chem.】 【Chem.】 【Chem.】 or a pharmaceutically acceptable salt thereof.

3. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of Formula III or a pharmaceutically acceptable salt thereof: ​ Wherein, Hal is -Cl, -F, -I, or -Br; x is an integer in the range of 0 to 5; Each R １ is independently -Cl, -F, -I, -Br, -C １ -C ３ alkyl, -O-C １ -C ３ alkyl, -CN, -CF ３ , -C(O)NH(CH ３ ), or -C≡CCH ２ OH; R ３ is -H, -C １ -C ６ alkyl, -(C １ -C ６ alkylene)-OH, -(C １ -C ６ alkylene)-phenyl, -(C １ -C ６ alkylene)-O-(C １ -C ６ alkyl), -C ２ -C ６ alkenyl, -(C １ -C ６ alkylene)-C(O)R ４ , -(C １ -C ６ alkylene)-R ５ , 【Chemical Formula 7】 ; R ４ is —OH, —O—(C １ —C ６ alkyl), —NH ２ , —NH(C １ —C ６ alkyl), —NH((C １ —C ６ alkylene)—OH), —NH((C １ —C ６ alkylene)N(C １ —C ６ alkyl) ２ ), —N(C １ —C ６ alkyl)((C １ —C ６ alkylene)—CN), —N(C １ —C ６ alkyl)((C １ —C ６ alkylene)N(C １ —C ６ alkyl) ２ ), —NH(C １ —C ６ alkylene)—O—(C １ —C ６ alkyl), [Chemical 8] 【Chem.】 ; a is an integer in the range of 0 to 10; b is an integer in the range of 0 to 8; c is an integer in the range of 0 to 6; R ５ is 【Chemical Formula 9】 ; Each R ６ and R ７ is independently -H or -I, where R ６ and R ７ at least one of which is -I, R ３ is -C １ -C ３ When it is alkyl, R ７ is -H.

4. The method according to claim 3, wherein the compound has the structure: 【Chemical 10】 or a pharmaceutically acceptable salt thereof.

5. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of Compound 46 having the structure: 【Chemical 11】 or a pharmaceutically acceptable salt thereof.

6. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of Formula IV or a pharmaceutically acceptable salt thereof: 【Chemical 12】

7. The method according to claim 6, wherein the compound has the structure: 【Chemical 13】 or a pharmaceutically acceptable salt thereof.

8. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of Formula V or a pharmaceutically acceptable salt thereof: 【Chemical 14】 Wherein, R １ is: 【Chemical Formula 15】 ; R ２ is: 【Chemical Formula 16】 ; Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

9. The method according to claim 8, wherein the compound has the structure: 【Chemical 17】 【Chem.】 【Chem.】 【Chem.】 or a pharmaceutically acceptable salt thereof.

10. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of Formula VI or a pharmaceutically acceptable salt thereof: 【Chemical 18】 Wherein, R ３ is: 【Chemical 19】 ; b is 0 or 1; c is 1 or 2.

11. The method according to claim 10, wherein the compound has the structure: 【Chemical 20】 【Chem.】 [Chemical] or a pharmaceutically acceptable salt thereof.

12. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula VII or a pharmaceutically acceptable salt thereof: 【Chemical 21】 wherein, R ４ is 【Chemical 22】 It is.

13. The method according to claim 12, wherein the compound has the structure: 【Chemical 23】 Or a pharmaceutically acceptable salt thereof.

14. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIII or a pharmaceutically acceptable salt thereof: 【Chemical 24】 wherein, R ５ is: 【Chemical 25】 wherein; R ６ is: 【Chemical 26】 wherein; Hal is -Cl, -F, -I, or -Br; a is 0, 1, or 2.

15. The compound is a pharmaceutically acceptable salt, and R ６ is 【Chemical 27】 The method according to claim 14, which is as follows.

16. The method according to claim 14, wherein the compound has the structure: 【Chemical Formula 28】 【Chem.】 【Chem.】 【Chem.】 Or a pharmaceutically acceptable salt thereof.

17. The method according to claim 14, wherein the pharmaceutically acceptable salt has the structure: 【Chemical 29】 having.

18. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XIV or a pharmaceutically acceptable salt thereof: 【Chemical 30】 wherein R ７ is: 【Chemical Formula 31】 wherein; b is 0 or 1; c is 1 or 2.

19. The method according to claim 18, wherein the compound has the structure: 【Chemical Formula 32】 Or a pharmaceutically acceptable salt thereof.

20. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound of formula XV or a pharmaceutically acceptable salt thereof: 【Chemical 33】 where R ８ is: 【Chemical 34】 wherein.

21. The method according to claim 20, wherein the compound has the structure: 【Chemical 35】 【Chem.】 Or a pharmaceutically acceptable salt thereof.

22. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound having the structure: 【Chemical 36】 [Chemical] or a pharmaceutically acceptable salt thereof.

23. A method of treating or preventing proteopathy, comprising administering to a subject in need thereof an effective amount of a compound having the structure: 【Chemical 37】 or a pharmaceutically acceptable salt thereof.

24. The method according to any one of claims 1 to 23, wherein the proteopathy is a neurodegenerative disease.

25. The method according to claim 24, wherein the neurodegenerative disease is Alzheimer's disease, progressive supranuclear palsy, boxer dementia, frontotemporal dementia, Parkinsonism linked to chromosome 17 (FTDP-17), corticobasal degeneration, neuronal tauopathy, ganglioglioma, gangliocytoma, meningeal hemangiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, lipofuscinosis, Pick's disease, corticobasal degeneration, argentophilic grain dementia, Huntington's disease, Parkinson's disease, Lewy body dementia, multiple system atrophy, axonal dystrophy, dentatorubral-pallidoluysian atrophy (DRPLA), spinal and bulbar muscular atrophy (SBMA), spinocerebellar ataxia type 1 (SCA1), SCA2, SCA3, SCA6, SCA7, SCA17, prion disease, amyotrophic lateral sclerosis, frontotemporal lobar degeneration (FTLD) or familial encephalopathy with neuroserpin inclusions (FENIB).

26. The method according to any one of claims 1 to 23, wherein the proteopathy is amyloidosis.

27. The method according to claim 26, wherein the amyloidosis is familial British dementia (ABri), familial Danish dementia (ADan), hereditary cerebral hemorrhage with amyloidosis - Icelandic type (HCHWA-I), familial amyloid polyneuropathy (ATTR), AL (light chain) primary systemic amyloidosis, AH (heavy chain) amyloidosis, AA secondary amyloidosis, Aβ amyloidosis, medial aortic amyloidosis, LECT2 amyloidosis, AIAPP amyloidosis, apolipoprotein AI amyloidosis (AapoAI), apolipoprotein AII amyloidosis (AapoAII), apolipoprotein AIV amyloidosis (AapoAIV), Finnish type familial amyloidosis (FAF), fibrinogen amyloidosis (AFib), lysozyme amyloidosis (ALys), dialysis amyloidosis (Aβ2M), medullary thyroid carcinoma (ACal), atrial amyloidosis (AANF), pituitary prolactinoma (APro), lattice corneal dystrophy, cutaneous lichen amyloidosis (AKer), Mallory body, primary cutaneous amyloidosis, corneal lactoferrin amyloidosis, dental (pindborg) tumor amyloid or seminal vesicle amyloid.

28. The method according to any one of claims 1 to 23, wherein the proteopathy is a lysosomal storage disorder.

29. The method according to claim 28, wherein the lysosomal storage disorder is a mucopolysaccharidosis.

30. The method according to claim 29, wherein the mucopolysaccharidosis is pseudo-Hurler polydystrophy / mucolipidosis IIIA, MPS I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, type A Sanfilippo syndrome / MPS IIIA, type B Sanfilippo syndrome / MPS IIIB, type C Sanfilippo syndrome / MPS IIIC, type D Sanfilippo syndrome / MPS IIID, Morquio syndrome type A / MPS IVA, Morquio syndrome type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC or mucolipidosis IV.

31. The method according to claim 28, wherein the lysosomal storage disorder is Pompe disease / glycogen storage disease type II.

32. Lysosomal storage diseases include activator deficiency / GM2 gangliosidosis, alpha-mannosidosis, aspartylglucosaminuria, cholesteryl ester storage disease,Chronic hexosaminidase A deficiency, cystinosis, Danon disease, Fabry disease, Farber disease, fucosidosis, galactosialidosis, type I Gaucher disease, type II Gaucher disease, type III Gaucher disease, infantile GM1 gangliosidosis, late infantile / juvenile GM1 gangliosidosis, adult / chronic GM1 gangliosidosis, I-cell disease / mucolipidosis II, infantile free sialic acid storage disease / ISSD, juvenile hexosaminidase A deficiency, infantile-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 I Hurler syndrome, MPS I Scheie syndrome, MPS I Hurler-Scheie syndrome, MPS II Hunter syndrome, type A Sanfilippo syndrome / MPS IIIA, type B Sanfilippo syndrome / MPS IIIB, type C Sanfilippo syndrome / MPS IIIC, type D Sanfilippo syndrome / MPS IIID, Morquio syndrome type A / MPS IVA, Morquio syndrome type B / MPS IVB, MPS IX hyaluronidase deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly syndrome, mucolipidosis I / sialidosis, mucolipidosis IIIC, mucolipidosis IV type, multiple sulfatase deficiency, Niemann-Pick disease type A, Niemann-Pick disease type B, Niemann-Pick disease type C, CLN6 disease - atypical late infantile, CLN6 disease - late-onset variant, CLN6 disease - early juvenile, Batten-Spielmeyer-Vogt / juvenile NCL / CLN3 disease, Finnish variant late infantile CLN5, Jansky-Bielschowsky disease / late infantile CLN2 / TPP1 disease, Kufs / adult-onset NCL / CLN4 disease, Northern Epilepsy / atypical late infantile CLN8, Santavuori-Haltia / infantile CLN1 / PPT disease, beta-mannosidosis, Pompe disease / glycogen storage disease type II, pycnodysostosis, Sandhoff disease / adult-onset / GM2 gangliosidosis, Sandhoff disease / GM2 gangliosidosis - infantile, Sandhoff disease / GM2 gangliosidosis - juvenile, Schindler disease, Salla disease / sialic acid storage disease, Tay-Sachs / GM2 gangliosidosis or Wolman disease, the method according to claim 28.,

33. The method according to any one of claims 1 to 23, wherein the compound or a pharmaceutically acceptable salt thereof binds to a molecular chaperone.

34. The method according to claim 33, wherein the molecular chaperone is a member of the Hsp10 family, Hsp40 family, Hsp60 family, Hsp70 family, Hsp90 family, or Hsp100 family.

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