Targeted degrader of abnormal tau based on pet tracers pbb3

Abstract

To provide improved techniques for combating diseases associated with abnormal tau.SOLUTION: Disclosed are bispecific compounds (degraders) that target tau protein for degradation. Pharmaceutical compositions containing the degraders and methods of using the compounds to treat neurodegenerative and neuropsychiatric diseases associated with abnormal tau are also disclosed.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62 / 991,359, filed March 18, 2020, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Tau protein is a microtubule-associated protein (MAP) abundantly expressed in neurons of the central nervous system (Okamura et al., Clin. Transl. Imaging 6:305-316 (2018)). Tau protein interacts with tubulin to stabilize microtubules and promote tubulin assembly into microtubules. Microtubule stabilization is regulated by isoforms or phosphorylation (Cleveland et al., J. Mol. Biol. 116:207-225 (1977)). Tauopathies are a group of neurodegenerative diseases pathologically defined by the presence of tau protein aggregates in the brain (Orr et al., Trends Pharmacol. Sci. 38:637-648 (2017)). Tau protein aggregates are composed of defective tau protein that no longer properly stabilizes microtubules, typically associated with hyperphosphorylation. Tau is involved in the pathogenesis of autism and related neurodevelopmental disorders, and tau reduction is a potential therapeutic strategy for treating these disorders (Tai et al., Neuron, pii:S0896-6273(20):30065-9(2020)). Currently, there are no FDA-approved small molecules or other therapeutic modalities that selectively target tau for the treatment of neurodegenerative, neuropsychiatric, neurological, and other disorders. Therefore, improved technologies are needed to overcome the challenge of combating diseases associated with abnormal tau. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Okamura et al.,Clin.Transl.Imaging 6:305-316(2018) [Non-patent document 2] Cleveland et al., J.Mol.Biol.116:207-225(1977) [Non-patent document 3] Orr et al.,Trends Pharmacol.Sci.38:637-648(2017) [Non-patent document 4] Tai et al.,Neuron,pii:S0896-6273(20):30065-9(2020) Summary of the Invention [Means for solving the problem]

[0004] A first aspect of the present invention is a compound of formula (I): [ka] wherein the targeting ligand represents a moiety that binds tau, the degron represents a moiety that binds an E3 ubiquitin ligase, or the degron is an autophagy recruitment tag (i.e., a tag that targets or targets a substrate for selective autophagy), and the linker represents a moiety that covalently links the degron and the targeting ligand, or a pharmaceutically acceptable salt or stereoisomer thereof; [ka] is the formula TL-1 or TL-2: [ka] represents wherein each X1, X2, X3, and R1 is as defined herein, or a pharmaceutically acceptable salt or stereoisomer thereof.

[0005] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bispecific compound of Formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0006] In another aspect of the invention, methods of making bispecific compounds are provided.

[0007] A further aspect of the present invention relates to a method of treating a disease or disorder characterized by or mediated by abnormal tau protein activity, comprising administering to a subject in need thereof a therapeutically effective amount of a bispecific compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0008] While not intending to be bound by any particular theory of operation, the bispecific compounds of Formula (I) (also referred to herein as PROTACs or degraders) are believed to promote the degradation of tau protein via the cellular ubiquitin / proteasome system, which functions to conventionally identify and remove damaged proteins. After the destruction of tau molecules, the degraders are released and remain active. Thus, by engaging with and utilizing the body's own natural protein disposal system, the bispecific compounds of the present invention may represent a potential improvement over conventional small molecule tau inhibitors. Therefore, the effective intracellular concentration of the degraders may be significantly lower than that of small molecule tau inhibitors. The bispecific compounds of the present invention may be more potent tau protein inhibitors than known inhibitors.

[0009] Therefore, the bispecific compounds of the present invention may provide at least one additional advantage, including improved pharmacodynamics. Using targeted degradation technology, E3-ligase adaptor proteins are recruited to tau protein aggregates via the bispecific compounds, leading to ubiquitination and clearance by the proteasome. In summary, the bispecific compounds of the present invention may represent an advance in the field. [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 is an immunoblot showing degradation of P-tau S396 in tau-A152T neurons after 24 hours of treatment with bispecific compounds 2 and 4 of the invention and a negative control at the concentrations (μM) indicated. [Figure 2] FIG. 2 is a series of graphs showing dose effects in tau-A152T neurons for bispecific compounds 2 and 4 and a negative control. [Figure 3] FIG. 3 is an immunoblot showing degradation of P-tau S396 in tau-P301L neurons after 24 hours of treatment with bispecific compounds 2 and 4 and a negative control at the indicated concentrations (μM). [Figure 4] FIG. 4 is a series of graphs showing dose effects in tau-P301L neurons for bispecific compounds 2 and 4 and a negative control. DETAILED DESCRIPTION OF THE INVENTION

[0011] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of this specification belongs. As used in this specification and the appended claims, unless specified to the contrary, the following terms have the meanings set forth to facilitate understanding of the invention.

[0012] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "composition" includes mixtures of two or more such compositions, reference to an "inhibitor" includes mixtures of two or more such inhibitors, and so forth.

[0013] Unless otherwise specified, the term "about" means within 10% (eg, within 5%, 2%, or 1%) of the particular value modified by the term "about."

[0014] The transitional term "comprising" is synonymous with "including," "containing," or "characterized by" and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes elements, steps, or ingredients not specified in the claim. The transitional phrase "consisting essentially of" limits the claim to certain materials or steps, "and which do not materially affect the basic and novel characteristics" of the claimed invention.

[0015] To the extent that the following terms are used herein with respect to and to further describe the compounds of the present invention, the following definitions apply.

[0016] As used herein, the term "alkyl" refers to a saturated straight-chain or branched-chain monovalent hydrocarbon radical. In one embodiment, an alkyl radical is C-C 18 It is the base. In other embodiments, the alkyl radical is C0-C 6、 C0-C 5、 C-0-C 3、 C1-C 12、 C1-C 8、 C1-C 6、 C1-C 5、It is a C1-C4 or C1-C3 group (C0 alkyl refers to a bond). Examples of alkyl groups include methyl, ethyl, 1-propyl, 2-propyl, i-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl. In some embodiments, the alkyl group is a C1-C3 alkyl group. In some embodiments, the alkyl group is a C1-C2 alkyl group or a methyl group.

[0017] As used herein, the term "alkylene" refers to a straight or branched divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing no unsaturation, having 1 to 12 carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, etc., that connects the rest of the molecule to a radical group. The alkylene chain can be attached to the rest of the molecule through a single bond or to the radical group through a single bond. In some embodiments, an alkylene group contains 1 to 8 carbon atoms (C1-C8 alkylene). In other embodiments, an alkylene group contains 1 to 5 carbon atoms (C1-C5 alkylene). In other embodiments, an alkylene group contains 1 to 4 carbon atoms (C1-C4 alkylene). In other embodiments, an alkylene group contains 1 to 3 carbon atoms (C1-C3 alkylene). In other embodiments, an alkylene group contains 1 to 2 carbon atoms (C1-C2 alkylene). In other embodiments, an alkylene group contains one carbon atom (C1 alkylene).

[0018] As used herein, the term "alkenyl" refers to a straight- or branched-chain monovalent hydrocarbon radical having at least one carbon-carbon double bond. Alkenyl includes radicals having "cis" and "trans" orientations, or alternatively, "E" and "Z" orientations. In one example, an alkenyl radical is C2-C 18 In other embodiments, the alkenyl radical is a C-C 12 , C2-C 10 , C2-C8, C2-C6 or C2-C3 groups. Examples include ethenyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl and hexa-1,3-dienyl.

[0019] As used herein, the term "alkynyl" refers to a linear or branched monovalent hydrocarbon radical having at least one carbon-carbon triple bond. In one example, an alkynyl radical is C2-C 18 In another example, the alkynyl radical is a C-C 12 , C2-C 10 , C2-C8, C2-C6 or C2-C3. Examples include ethynylprop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl and but-3-ynyl.

[0020] The term "alkoxyl" or "alkoxy" as used herein refers to an alkyl group having an oxygen radical attached to the alkyl group defined above, where the oxygen radical is the point of attachment. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy, and the like. An "ether" is two hydrocarbyl groups covalently linked by an oxygen. Thus, the alkyl substituent that makes the alkyl an ether is an alkoxyl or similar to an alkoxyl, and can be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-alkynyl.

[0021] As used herein, the term "halogen" (or "halo" or "halide") refers to fluorine, chlorine, bromine, or iodine.

[0022] As used herein, the term "cyclic group," used alone or as part of a larger moiety, broadly refers to any group containing saturated, partially saturated, or aromatic ring systems, such as carbocyclic groups (cycloalkyl, cycloalkenyl), heterocyclic groups (heterocycloalkyl, heterocycloalkenyl), aryl groups, and heteroaryl groups. A cyclic group can have one or more (e.g., fused) ring systems. Thus, for example, a cyclic group can contain one or more carbocyclic, heterocyclic, aryl, or heteroaryl groups.

[0023] As used herein, the term "carbocycle" (also "carbocyclyl"), used alone or as part of a larger moiety, refers to a group, alone or as part of a larger moiety (e.g., an alkyl carbocyclic group), containing saturated, partially unsaturated, or aromatic ring systems having from 3 to 20 carbon atoms. The term carbocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged, and spirocyclic ring systems, and combinations thereof. In one embodiment, a carbocyclyl contains from 3 to 15 carbon atoms (C3-C 15 In one embodiment, the carbocyclyl contains 3 to 12 carbon atoms (C3-C 12In another embodiment, the carbocyclyl is C3-C 8、 C3-C 10 or C5-C 10 In another embodiment, the carbocyclyl, as a monocycle, includes C3-C8, C3-C6, or C5-C6. In some embodiments, the carbocyclyl, as a bicycle, includes C7-C 12 In another embodiment, the carbocyclyl, as a spiro system, includes C5-C 12 Includes. Representative examples of monocyclic carbocyclyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, perdeuteriocyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, phenyl, and cyclododecyl; bicyclic carbocyclyls having 7 to 12 ring atoms include [4,3], [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, naphthalene, and bicyclo[3.2.2]nonane. Representative examples of spirocarbocyclyl include spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane, and spiro[4.5]decane. The term carbocyclyl includes aryl ring systems as defined herein. The term carbocycyl also includes cycloalkyl rings (e.g., saturated or partially unsaturated monocyclic, bicyclic, or spirocarbocyclic rings). The term carbocyclic group also includes carbocyclic rings fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., aryl or heterocyclic rings), where the radical or point of attachment is on the carbocyclic ring.

[0024] Thus, the term carbocycle as used herein means R c is an alkylene chain of formula --R cThe term carbocyclyl, as used herein, also encompasses carbocyclylalkyl groups, which refer to the group R c is an alkylene chain of the formula --O--R c Also included are carbocyclylalkoxy groups, which refer to groups attached through the oxygen atom of a -carbocyclyl.

[0025] As used herein, the term "aryl," used alone or as part of a larger moiety (e.g., "aralkyl" where the terminal carbon atom on an alkyl group is the point of attachment, e.g., a benzyl group), "aralkoxy" where an oxygen atom is the point of attachment, or "aroxyalkyl" where the point of attachment is on an aryl group), refers to a group containing a monocyclic, bicyclic, or tricyclic carbocyclic ring system, including fused rings, in which at least one ring in the system is aromatic. In some embodiments, an aralkoxy group is a benzoxy group. The term "aryl" may be used interchangeably with the term "aryl ring." In one embodiment, aryl includes groups having 6 to 18 carbon atoms. In another embodiment, aryl includes groups having 6 to 10 carbon atoms. Examples of aryl groups include phenyl, naphthyl, anthracyl, biphenyl, phenanthrenyl, naphthacenyl, 1,2,3,4-tetrahydronaphthalenyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, naphthyridinyl, and the like, which may be substituted or independently substituted with one or more substituents described herein. A particular aryl is phenyl. In some embodiments, an aryl group comprises an aryl ring fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocyclic or heterocyclic rings), where the radical or point of attachment is on the aryl ring.

[0026] Thus, the term aryl refers to R c is an alkylene chain such as methylene or ethylene; c-aryl groups, including aralkyl groups (e.g., benzyl). In some embodiments, the aralkyl group is an optionally substituted benzyl group. The term aryl, as used herein, refers to an R c is an alkylene such as methylene or ethylene; c Also included are aralkoxy groups, which refer to groups bonded through the oxygen atom of an -aryl.

[0027] As used herein, the term "heterocyclyl," used alone or as part of a larger moiety, refers to a "carbocyclyl" and includes saturated, partially unsaturated, or aromatic ring systems in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by a heteroatom (e.g., O, N, N(O), S, S(O), or S(O)). The term heterocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged, and spirocyclic ring systems, and combinations thereof. In some embodiments, heterocyclyl refers to a 3- to 15-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a 3- to 12-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a saturated ring system, such as a 3- to 12-membered saturated heterocyclyl ring system. In some embodiments, heterocyclyl refers to a heteroaryl ring system, such as a 5- to 14-membered heteroaryl ring system. The term heterocyclyl also includes C3-C8 heterocycloalkyl, which are saturated or partially unsaturated monocyclic, bicyclic, or spirocyclic ring systems containing 3 to 8 carbons and one or more (1, 2, 3, or 4) heteroatoms.

[0028] In some embodiments, heterocyclyl groups contain 3 to 12 ring atoms, including monocyclic, bicyclic, tricyclic, and spirocyclic systems, where the ring atoms are carbon and 1 to 5 ring atoms are heteroatoms such as nitrogen, sulfur, or oxygen. In some embodiments, heterocyclyls contain 3 to 7-membered monocyclic rings having one or more heteroatoms selected from nitrogen, sulfur, or oxygen. In some embodiments, heterocyclyls contain 4 to 6-membered monocyclic rings having one or more heteroatoms selected from nitrogen, sulfur, or oxygen. In some embodiments, heterocyclyls contain 3-membered monocyclic rings. In some embodiments, heterocyclyls contain 4-membered monocyclic rings. In some embodiments, heterocyclyls contain 5 to 6-membered monocyclic rings. In some embodiments, heterocyclyl groups contain 0 to 3 double bonds. In any of the foregoing embodiments, heterocyclyls contain 1, 2, 3, or 4 heteroatoms. Any nitrogen or sulfur heteroatom may optionally be oxidized (e.g., NO, SO, SO), and any nitrogen heteroatom may optionally be quaternized (e.g., [NR]). + Cl - , [NR4] + OH -Representative examples of heterocyclyl include oxiranyl, aziridinyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydropyranyl, dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, Thiazinanyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinonyl , oxazolidinonyl, imidazolidinonyl, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazole[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, thiophenyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl, 1-pyrrolyl nyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiapyranyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinonyl, pyrimidinedionyl, pyrimidine-2,4-dionyl, piperazinonyl, piperazinedionyl, pyrazolidinylimidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.2.1]octanyl, 8-azabicyclo[3.2.1]octanyl, 2-azabicyclo[2.2.2]octanyl, 8-azabicyclo[2.2.2]octanyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl, 1-azaspiro[4.5]decane-2-only, azaspiro[5.5]undecanyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocyclyls containing a sulfur atom or an oxygen atom and 1 to 3 nitrogen atoms include thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl, such as oxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocyclyls containing 2 to 4 nitrogen atoms include imidazolyls such as imidazol-2-yl; triazolyls such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and tetrazolyls such as 1H-tetrazol-5-yl. Representative examples of benzo-fused 5-membered heterocyclyls are benzoxazol-2-yl, benzthiazol-2-yl, and benzimidazol-2-yl. Examples of 6-membered heterocyclyls contain 1 to 3 nitrogen atoms and optionally sulfur or oxygen atoms, for example, pyridyl, such as pyrid-2-yl, pyrid-3-yl, and pyrid-4-yl; pyrimidyl, such as pyrimid-2-yl and pyrimid-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, especially pyridazin-3-yl, and pyrazinyl. Pyridine N-oxide and pyridazine N-oxide and pyridyl, pyrimid-2-yl, pyrimid-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl groups are further examples of heterocyclyl groups. In some embodiments, heterocyclic groups include heterocyclic rings fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocyclic or heterocyclic rings), where the radical or point of attachment is on the heterocyclic ring, and in some embodiments, the point of attachment is a heteroatom contained in the heterocyclic ring.

[0029] Thus, the term heterocycle, as used herein, includes N-heterocyclyl groups, which refer to heterocyclyl groups containing at least one nitrogen atom, where the heterocyclyl group is attached to the rest of the molecule through a nitrogen atom in the heterocyclyl group. Representative examples of N-heterocyclyl groups include 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl. The term heterocycle, as used herein, also includes C-heterocyclyl groups, which refer to heterocyclyl groups containing at least one heteroatom, where the heterocyclyl group is attached to the rest of the molecule through a carbon atom in the heterocyclyl group. Representative examples of C-heterocyclyl radicals include 2-morpholinyl, 2-, 3-, or 4-piperidinyl, 2-piperazinyl, and 2- or 3-pyrrolidinyl. The term heterocycle refers to a heterocyclic ring, as disclosed above, c is an alkylene chain of formula --R c -heterocyclyl groups, which refer to the group heterocyclyl, are also included. The term heterocycle as used herein refers to R c is an alkylene chain of the formula --O--R c Also included are heterocyclylalkoxy groups, which refer to radicals attached through the oxygen atom of a -heterocyclyl.

[0030] As used herein, the term "heteroaryl," used alone or as part of a larger moiety (e.g., "heteroarylalkyl" (also "heteroaralkyl") or "heteroarylalkoxy" (also "heteroaralkoxy"), refers to a monocyclic, bicyclic, or tricyclic ring system having 5 to 14 ring atoms, in which at least one ring is aromatic and contains at least one heteroatom. In one embodiment, heteroaryl includes 5- to 6-membered monocyclic aromatic groups in which one or more ring atoms is nitrogen, sulfur, or oxygen. Representative examples of heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, imidazopyridine, and the like.

[0023] The term "heteroaryl" includes diphenyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolo[1,5-b]pyridazinyl, purinyl, deazapurinyl, benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, indolyl, 1,3-thiazol-2-yl, 1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl, 1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl, 1,2,3-triazol-5-yl, and pyrid-2-yl N-oxide. The term "heteroaryl" also includes groups in which the heteroaryl is fused to one or more cyclic (e.g., carbocyclyl or heterocyclyl) rings, and the radical or point of attachment is on the heteroaryl ring.Non-limiting examples include indolyl, indolizinyl, isoindolyl, benzothienyl, benzothiophenyl, methylenedioxyphenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzodioxazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or tricyclic. In some embodiments, a heteroaryl group comprises a heteroaryl ring fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocyclic or heterocyclic rings), where the radical or point of attachment is on the heteroaryl ring, and in some embodiments, the point of attachment is a heteroatom contained in the heterocyclic ring.

[0031] Thus, the term heteroaryl, as used herein, encompasses N-heteroaryl groups, which refer to heteroaryl groups, as defined above, containing at least one nitrogen, where the point of attachment of the heteroaryl group to the rest of the molecule is through a nitrogen atom in the heteroaryl group. The term heteroaryl, as used herein, also encompasses C-heteroaryl groups, which refer to heteroaryl groups, as defined above, where the point of attachment of the heteroaryl group to the rest of the molecule is through a carbon atom in the heteroaryl group. The term heteroaryl, as disclosed above, encompasses R c is an alkylene chain as defined above; c The term heteroaryl, as used herein, also encompasses heteroarylalkyl groups, which refer to groups of R -heteroaryl. c is an alkylene chain as defined above, c Also included are heteroaralkoxy (or heteroarylalkoxy) groups, which refer to groups attached through the oxygen atom of a -heteroaryl.

[0032] Any of the groups described herein can be substituted or unsubstituted. As used herein, the term "substituted" refers broadly to all permissible substituents, with the implicit proviso that such substitution is consistent with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, and the like. Representative substituents include halogens, hydroxyl groups, and any other organic group containing any number of carbon atoms, e.g., 1 to 14 carbon atoms, and may include one or more (e.g., 1, 2, 3, or 4) heteroatoms, such as oxygen, sulfur, and nitrogen, grouped in the form of a straight-chain, branched-chain, or cyclic structure.

[0033] Unless otherwise disclosed for any particular group, representative examples of substituents include alkyl, substituted alkyl (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), substituted alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), haloa, alkyl (e.g., CF3), alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), cyclic (e.g., C3-C 12 , C5-C6), substituted cyclic (e.g., C3-C 12 , C5-C6), carbocyclic (e.g., C3-C 12 , C5-C6), substituted carbocyclic (e.g., C3-C 12 , C5-C6), heterocyclic (e.g., C3-C 12 , C5-C6), substituted heterocyclic (e.g., C3-C 12, C5-C6), aryl (e.g., benzyl and phenyl), substituted aryl (e.g., substituted benzyl or phenyl), heteroaryl (e.g., pyridyl or pyrimidyl), substituted heteroaryl (e.g., substituted pyridyl or pyrimidyl), aralkyl (e.g., benzyl), substituted aralkyl (e.g., substituted benzyl), halo, hydroxyl, aryloxy (e.g., C6-C 12 , C6), substituted aryloxy (e.g., C6-C 12 , C6), alkylthio (e.g., C1-C6), substituted alkylthio (e.g., C1-C6), arylthio (e.g., C6-C 12 , C6), substituted arylthio (e.g., C6-C 12 , C6), cyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, thio, substituted thio, sulfinyl, substituted sulfinyl, sulfonyl, substituted sulfinamide, substituted sulfinamide, sulfonamide, substituted sulfonamide, urea, substituted urea, carbamate, substituted carbamate, amino acid, and peptide groups.

[0034] The term "binding," when referring to the interaction between a targeting ligand and a target protein, in this case tau protein and mutant or misfolded forms thereof, refers to an intermolecular interaction that is substantially specific, in that binding of the targeting ligand to other proteinaceous entities present within the cell may not be functionally significant. The bispecific compounds of the present invention bind and recruit tau protein for selective degradation.

[0035] The term "binding" with respect to the interaction between a degron and an E3 ubiquitin ligase typically refers to an intermolecular interaction that may or may not exhibit an affinity level that is equal to or exceeds the affinity between a targeting ligand and a target protein, but the affinity is nonetheless sufficient to achieve recruitment of the ligase to the target for degradation and selective degradation of the target protein.

[0036] Generally, the bispecific compounds of the present invention have the formula (I): [ka] wherein the targeting ligand represents a moiety that binds tau, the degron represents a moiety that binds an E3 ubiquitin ligase, or the degron is an autophagy recruitment tag (i.e., a tag that targets a substrate for selective autophagy), and the linker represents a moiety that covalently links the degron and the targeting ligand, or a pharmaceutically acceptable salt or stereoisomer thereof; [ka] is represented by the formula TL-1 or TL-2, [ka] During the ceremony, each X1 is independently C, CH, or N; X2 is NH, S, or O; X3 is CH or N; each R1 is independently hydrogen or C1-C3 alkyl; When X2 is NH, X3 is CH; When X3 is N, X2 is S or O.

[0037] Thus, in some embodiments, the bispecific compounds of the present invention have formula I-1: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0038] Thus, in another embodiment, the bispecific compounds of the invention have formula I-2: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0039] In some embodiments, the formula of TL-1 is TL-1a to TL-1o: [ka] is.

[0040] In some embodiments, TL-1 is [ka] is.

[0041] In some embodiments, the formula of TL-2 is TL-2a through TL-2o: [ka] is.

[0042] In some embodiments, R1 is hydrogen.

[0043] In some embodiments, R1 is methyl.

[0044] Linker The linker ("L") provides a covalent bond between the targeting ligand and the degron. The structure of the linker may not be important as long as it does not substantially interfere with the activity of the tau targeting ligand or the degron. In some embodiments, the linker comprises an alkylene chain (e.g., having 1 to 20 alkylene units). In other embodiments, the linker can comprise an alkylene chain or a divalent alkylene chain, any of which is -O-, -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N( R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -O S(O)2-, -S(O)2O-, -N(R')S(O)2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, C3-C 12 It may be interrupted and / or terminated (at either or both termini) by at least one of carbocyclene, 3- to 12-membered heterocyclene, 5- to 12-membered heteroarylene, or any combination thereof, where R' is H or C1-C6 alkyl, and the interrupting group and one or both terminus groups may be the same or different.

[0045] In some embodiments, the alkylene chain has 1 to 18 alkylene units. In some embodiments, the alkylene chain has 1 to 12 alkylene units. In some embodiments, the alkylene chain has 1 to 10 alkylene units. In some embodiments, the alkylene chain has 1 to 8 alkylene units. In some embodiments, the alkylene chain has 1 to 6 alkylene units. In some embodiments, the alkylene chain has 1 to 4 alkylene units. In some embodiments, the alkylene chain has 1 to 2 alkylene units. In some embodiments, the alkylene chain is interrupted by and / or terminates (at one or both ends) in at least one of -N(R')-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)O-, -OC(O)N(R')-, -S(O)-, -N(R')S(O)-, -S(O)N(R')-, or combinations thereof. In some embodiments, the alkylene chain is interrupted by and / or terminates (at one or both ends) in -N(R')-. In some embodiments, the alkylene chain is interrupted by and / or terminates (at one or both ends) in -C(O)-. In some embodiments, the alkylene chain is interrupted by -C(O)O- and / or terminated (at one or both ends) by -C(O)O-. In some embodiments, the alkylene chain is interrupted by -C(O)N(R')- and / or terminated (at one or both ends) by -C(O)N(R')-. In some embodiments, the alkylene chain is interrupted by -N(R')S(O)- and / or terminated (at one or both ends) by -N(R')S(O)-.

[0046] In some embodiments, the linker has 1 to 10 alkylene units, [ka] The term "alkylene chain" includes an alkylene chain interrupted by or terminated at a group.

[0047] "Carbocyclene" refers to an optionally substituted divalent carbocycle radical.

[0048] "Heterocyclene" refers to an optionally substituted divalent heterocyclyl radical.

[0049] "Heteroarylene" refers to a divalent heteroaryl radical that may be optionally substituted.

[0050] Representative examples of alkylene linkers that may be suitable for use in the present invention include: [ka] wherein n is an integer of 1 to 12 ("of" means containing), for example, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, 9-10 and 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, examples of which are: [ka] are listed; Alkylene chains terminated with various functional groups (as described above), such as: [ka] [ka] are listed; Alkylene chains (as above) interrupted by various functional groups, such as: [ka] are listed; An alkylene chain interrupted or terminated by a heterocyclene group, e.g. [ka] In the formula, m and n are independently integers of 0 to 10, examples of which include: [ka] are listed; Alkylene chains interrupted by amide groups, heterocyclene groups and / or aryl groups, examples of which include: [ka] are listed; Heterocyclene and aryl groups as well as alkylene chains interrupted by heteroatoms, examples of which include: [ka] and Alkylene chains interrupted by heteroatoms such as N, O or B, e.g. [ka] wherein each n is independently an integer from 1 to 10, for example, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8, 8 to 10, 8 to 9, 9 to 10, and 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; and R is H or C1 to C4 alkyl, examples of which include: [ka] There is.

[0051] In some embodiments, the linker is -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2- , -S(O)2O-, -N(R')S(O)2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, C 3-12 It may comprise a polyethylene glycol chain that may be terminated (at one or both ends) by at least one of a carbocyclene, a 3- to 12-membered heterocyclene, a 5- to 12-membered heteroarylene, or any combination thereof, where R' is H or C1-C6 alkyl, and one or both end groups may be the same or different.

[0052] In some embodiments, the polyethylene glycol chain has 1 to 10 -(CH2CH2-O)- units. In some embodiments, the polyethylene glycol chain has 1 to 5 -(CH2CH2-O)- units. In some embodiments, the polyethylene glycol chain has 1 to 2 -(CH2CH2-O)- units. In some embodiments, the polyethylene glycol is interrupted by and / or terminated (at one or both ends) in at least one of -N(R')-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)O-, -OC(O)N(R')-, -S(O)2-, -N(R')S(O)2-, -S(O)2N(R')-, or combinations thereof. In some embodiments, the polyethylene glycol chain is interrupted by -N(R')- and / or terminated (at one or both ends) with -N(R')-. In some embodiments, the polyethylene glycol chain is interrupted by and / or terminated (at one or both ends) with -C(O)-. In some embodiments, the polyethylene glycol chain is interrupted by and / or terminated (at one or both ends) with -C(O)O-. In some embodiments, the polyethylene glycol chain is interrupted by and / or terminated (at one or both ends) with -C(O)N(R')-. In some embodiments, the polyethylene glycol chain is interrupted by and / or terminated (at one or both ends) with -N(R')S(O)-.

[0053] In some embodiments, the linker has 2 to 8 PEG units. [ka] It comprises a polyethylene glycol chain terminating in

[0054] Examples of linkers comprising polyethylene glycol chains include: [ka] In the formula, n is an integer of 1 to 10, and examples include the following. [ka]

[0055] In some embodiments, the polyethylene glycol linker may terminate in a functional group, examples of which are as follows: [ka]

[0056] In some embodiments, the bispecific compounds of Formula (I) comprise a linker represented by any one of the following structures: [ka]

[0057] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0058] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0059] Degron The ubiquitin-proteasome pathway (UPP) is a critical cellular pathway that regulates key regulatory proteins and degrades misfolded or abnormal proteins. The UPP is central to multiple cellular processes. The covalent attachment of ubiquitin to specific protein substrates is achieved through the action of E3 ubiquitin ligases. These ligases comprise over 500 different proteins and are divided into multiple classes defined by the structural elements of their E3 functional activity.

[0060] In some embodiments, the degron binds the E3 ubiquitin ligase, cereblon (CRBN), and has the following structure: [ka] [ka] is represented by one of wherein Y is NH or O.

[0061] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0062] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0063] Still other degrons that bind to cereblon and may be suitable for use in the present invention are disclosed in U.S. Patent Application Publication No. 2018 / 0015085 (e.g., indolinones such as isoindolinones and isoindoline-1,3-diones encompassed by formulas IA and IA' therein, and bridged cycloalkyl compounds encompassed by formulas IB and IB' therein).

[0064] In some embodiments, the E3 ubiquitin ligase bound by the degron is the von Hippel-Lindau (VHL) tumor suppressor. See Iwai et al., Proc. Nat'l. Acad. Sci. USA 96:12436-41 (1999).

[0065] Representative examples of degrons that bind VHL include: [ka] wherein Z is a cyclic group; and [ka] or a stereoisomer thereof.

[0066] In certain embodiments, Z is [ka] is.

[0067] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0068] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0069] Still other degrons that bind VHL and may be suitable for use in the present invention are disclosed in U.S. Patent Application Publication No. 2017 / 0121321 A1.

[0070] In some embodiments, the E3 ubiquitin ligase bound by the degron is an inhibitor of apoptosis protein (IAP). Representative examples of degrons that bind IAPs and may be suitable for use in the present invention have the following structure: [ka] or a stereoisomer thereof.

[0071] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0072] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0073] Still other compounds that bind IAPs and may be suitable for use as degrons in the present invention are disclosed in International Patent Application Publication Nos. WO2008128171, WO2008016893, WO2014060768, WO2014060767, and WO2015092420.

[0074] In some embodiments, the E3 ubiquitin ligase bound by the degron is mouse double minute 2 (MDM2). A representative example of a degron that binds MDM2 and may be suitable for use in the present invention has the following structure: [ka] or a stereoisomer thereof.

[0075] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0076] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0077] Still other compounds that bind MDM2 and may be suitable for use as degrons in the present invention are disclosed in U.S. Patent No. 9,993,472 B2. MDM2 is known in the art to function as a ubiquitin E3 ligase.

[0078] Autophagy is a homeostatic process important for balancing energy resources during critical periods of development and in response to nutritional stress. Autophagy also plays a central role in removing misfolded or aggregated proteins and clearing damaged organelles (Glick et al., J. Pathol., 221(1):3-12(2010)). Autophagy-mediated clearance serves as a cellular waste disposal system.

[0079] In some embodiments, the degron is an autophagy recruitment tag (i.e., a tag that ends up or targets a substrate for selective autophagy). Exemplary autophagy recruitment tags that may be suitable for use in the present invention have the following structure: [ka] or a stereoisomer thereof.

[0080] Thus, in some embodiments, the bispecific compounds of the invention have the following structure: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0081] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0082] Still other compounds that can function as autophagy recruitment tags and may be suitable for use as degrons in the present invention are disclosed in U.S. Patent Application Publication No. 2019 / 0290778.

[0083] Thus, in some embodiments, the bispecific compounds of the present invention are represented by any of the structures produced by combining the structures of the degrons described herein, including structures TL1-TL2, L1-L10, and D1-D5, or pharmaceutically acceptable salts or stereoisomers thereof.

[0084] In some embodiments, the bispecific compound of the present invention has the following structure: [ka] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0085] The bispecific compounds of the present invention may be in the form of a free acid or free base, or a pharmaceutically acceptable salt. As used herein, the term "pharmaceutically acceptable" in the context of a salt refers to a relatively non-toxic salt of a compound that does not abolish the biological activity or properties of the compound. That is, a compound in salt form can be administered to a subject without causing undesirable biological effects (such as dizziness or stomach upset) or interacting in a deleterious manner with any of the other components of the composition in which it is contained. The term "pharmaceutically acceptable salt" refers to the product obtained by reacting a compound of the present invention with an appropriate acid or base. Examples of pharmaceutically acceptable salts of the bispecific compounds of the present invention include those derived from appropriate inorganic bases, such as Li, Na, K, Ca, Mg, Fe, Cu, Al, Zn, and Mn salts. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, 4-methylbenzenesulfonate, or p-toluenesulfonate. Certain bispecific compounds of the present invention can form pharmaceutically acceptable salts with various organic bases, such as lysine, arginine, guanidine, diethanolamine, or metformin. Suitable base salts include aluminum, calcium, lithium, magnesium, potassium, sodium, or zinc salts.

[0086] The bispecific compounds of the present invention may have at least one chiral center and therefore, as used herein, may be in the form of stereoisomers, which encompass all isomers of individual compounds that differ only in the orientation of their atoms in space. The term stereoisomer includes enantiomers (enantiomers containing the (R-) or (S-) configuration of a compound), mixtures of enantiomers of a compound (physical mixtures of enantiomers, and racemates or racemic mixtures), geometric (cis / trans or E / Z, R / S) isomers of a compound, and isomers of a compound with multiple chiral centers that are not mirror images of one another (diastereoisomers). The chiral centers of a compound may undergo epimerization in vivo. Therefore, for these compounds, administration of the (R-) form of the compound is considered equivalent to administration of the (S-) form of the compound. Thus, the bispecific compounds of the present invention may be prepared and used in the form of a single isomer, substantially free of other isomers, or in the form of mixtures of various isomers, such as racemic mixtures of stereoisomers.

[0087] In some embodiments, the bispecific compounds are isotopic derivatives because they have at least one desired isotopic substitution of an atom at a level above the natural abundance of the isotope, i.e., enriched. In one embodiment, the compound contains deuterium or multiple deuterium atoms. Deuterium, i.e., 2 Substitution with heavier isotopes, such as H, may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be advantageous in some circumstances.

[0088] Thus, the term bispecific compound of Formula (I) encompasses the free base form of the bispecific compound, as well as isotopic derivatives, N-oxides, crystalline forms (also known as polymorphs), active metabolites of bispecific compounds having the same type of activity, prodrugs of bispecific compounds, tautomers, and unsolvated forms as well as solvated (e.g., hydrated) forms with pharmaceutically acceptable solvents such as water, ethanol, and the like.

[0089] Synthesis method In another embodiment, the present invention relates to a method for preparing a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof. Generally, the compounds of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof, can be prepared by any process known to be applicable to the preparation of chemically-related compounds. The compounds of the present invention will be better understood in connection with the synthetic schemes described in the various examples, which illustrate non-limiting methods by which the compounds of the present invention may be prepared.

[0090] Pharmaceutical Composition Another aspect of the present invention relates to a pharmaceutical composition containing a therapeutically effective amount of a bispecific compound of Formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier," as known in the art, refers to a pharmaceutically acceptable material, composition, or vehicle suitable for administering a compound of the present invention to a mammal. Suitable carriers can include, for example, liquids (both aqueous and non-aqueous, and combinations thereof), solids, encapsulating materials, gases, and combinations thereof (e.g., semi-solids), and gases, which function to carry or transport a compound from one organ or part of the body to another. A carrier is "acceptable" in the sense of being physiologically inert, compatible with the other ingredients of the formulation, and not harmful to the subject or patient. Depending on the type of formulation, the composition may contain one or more pharmaceutically acceptable excipients.

[0091] In general, the bispecific compounds of Formula I can be formulated into a given type of composition according to conventional pharmaceutical practice, such as conventional mixing, dissolving, granulating, dragee-making, wet-milling, emulsifying, encapsulating, entrapping, and compression processes (see, e.g., Remington: The Science and Practice of Pharmacy (20th ed.), ed. A.R. Gennaro, Lippincott Williams & Wilkins, 2000, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J.C. Boylan, 1988-1999, Marcel Dekker, New York). The type of formulation depends on the mode of administration, which may include enteral (e.g., oral, buccal, sublingual, and rectal), parenteral (e.g., subcutaneous (sc), intravenous (iv), intramuscular (im)), and intrasternal injection, or infusion techniques, intraocular, intraarterial, intramedullary, intrathecal, intracerebroventricular, transdermal, intradermal, intravaginal, intraperitoneal, mucosal, nasal, intratracheal instillation, bronchial instillation, and inhalation), and topical (e.g., transdermal). Generally, the most appropriate administration route will depend on various factors, including, for example, the nature of the agent (e.g., its stability in the gastrointestinal environment) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration). For example, parenteral (e.g., intravenous) administration may also be advantageous in that the compound can be administered relatively quickly, such as in single-dose treatments and / or acute conditions.

[0092] In some embodiments, the bispecific compounds are formulated for oral or intravenous administration (eg, systemic intravenous injection).

[0093] Thus, the bispecific compounds of the present invention can be formulated into solid compositions (e.g., powders, tablets, dispersible granules, capsules, cachets, and suppositories), liquid compositions (e.g., solutions in which the compound is dissolved, suspensions in which solid particles of the compound are dispersed, emulsions, and solutions containing liposomes, micelles, or nanoparticles, syrups, and elixirs); semisolid compositions (e.g., gels, suspensions, and creams); and gases (e.g., propellants for aerosol compositions). The compounds can also be formulated for rapid, intermediate, or sustained release.

[0094] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the bispecific compound may be combined with a carrier such as sodium citrate or dicalcium phosphate, and additional carriers or excipients, such as a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) cross-linked polymers (e.g., cross-linked polyvinylpyrrolidone (crospovidone), cross-linked They are mixed with disintegrating agents such as sodium carboxymethylcellulose (croscarmellose sodium), sodium starch glycolate, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retardants such as paraffin; f) absorption accelerators such as quaternary ammonium compounds; g) wetting agents such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite; i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents. Similar types of solid compositions can also be used as fillers for soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings. They may further contain opacifying agents.

[0095] In some embodiments, the bispecific compounds of the present invention can be formulated into hard or soft gelatin capsules. Exemplary excipients that can be used include pregelatinized starch, magnesium stearate, mannitol, sodium stearyl fumarate, anhydrous lactose, microcrystalline cellulose, and croscarmellose sodium. The gelatin shell can contain gelatin, titanium dioxide, iron oxide, and colorants.

[0096] Liquid dosage forms for oral administration include solutions, suspensions, emulsions, microemulsions, syrups and elixirs.In addition to the bispecific compound, liquid dosage forms can contain aqueous or non-aqueous carriers commonly used in the art (depending on the solubility of the compound), such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.Oral compositions can also contain excipients, such as wetting agents, suspending agents, coloring agents, sweeteners, flavoring agents and fragrances.

[0097] Injectable preparations may include sterile aqueous or oleaginous suspensions, which may be formulated according to standard techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, USP, and isotonic sodium chloride solution. Additionally, sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any mild fixed oil, including synthetic mono- or diglycerides, may be used. Additionally, fatty acids, such as oleic acid, are used in the preparation of injectables. Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. The effect of a compound can be prolonged by delaying its absorption, which can be achieved by using poorly water-soluble liquid suspensions or crystalline or amorphous materials. Sustained absorption of a compound from parenterally administered formulations can also be achieved by suspending the compound in an oil vehicle.

[0098] In certain embodiments, the bispecific compounds of Formula (I) can be administered locally rather than systemically, for example, by injecting the conjugate directly into an organ, often in a depot or sustained-release formulation. In certain embodiments, long-acting formulations are administered by infusion (e.g., subcutaneously or intramuscularly) or intramuscular injection. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide, poly(orthoesters), and poly(anhydrides). The compound release rate can be controlled by varying the ratio of compound to polymer and the nature of the particular polymer used. Depot injectable formulations are also prepared by encapsulating the compound in liposomes or microemulsions that are compatible with body tissues. Furthermore, in other embodiments, the compound is delivered in a targeted drug delivery system, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to and selectively taken up by the organ.

[0099] The bispecific compounds can be formulated for buccal or sublingual administration and examples include tablets, lozenges and gels.

[0100] The bispecific compounds can be formulated for administration by inhalation. Various forms suitable for administration by inhalation include aerosols, mists, or powders. The pharmaceutical composition can be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas). In some embodiments, the dosage unit of the pressurized aerosol can be determined by providing a valve to deliver a metered amount. In some embodiments, capsules and cartridges containing, for example, gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0101] The bispecific compounds of Formula (I) can be formulated for topical administration, which as used herein refers to intradermal administration by application of the formulation to the epidermis. These types of compositions are typically in the form of ointments, pastes, creams, lotions, gels, solutions and sprays.

[0102] Representative examples of carriers useful for formulating bispecific compounds for topical application include solvents (e.g., alcohols, polyalcohols, water), creams, lotions, ointments, oils, plasters, liposomes, powders, emulsions, microemulsions, and buffer solutions (e.g., hypotonic or buffered saline). For example, creams can be formulated using saturated or unsaturated fatty acids, such as stearic acid, palmitic acid, oleic acid, palmitoleic acid, cetyl, or oleyl alcohol. Creams can also contain nonionic surfactants, such as polyoxy-40-stearate.

[0103] In some embodiments, topical formulations can also contain excipients, such as penetration enhancers.These agents can transport pharmacologically active compounds through the stratum corneum, preferably to the epidermis or dermis with little or no systemic absorption.A wide variety of compounds have been evaluated for their effectiveness in improving the penetration rate of drugs through the skin.For example, see Percutaneous Penetration Enhancers, Maibach HI and Smith HE (eds.), CRC Press, Inc., Boca Raton, Fla. (1995), which summarizes the use and testing of various skin penetration enhancers, and Buyuktimkin et al., Chemical Means of Transdermal Drug Permeation Enhancement in Transdermal and Topical Drug Delivery Systems, Gosh TK, Pfister WR, Yum SI (Eds.), Interpharm Press Inc., Buffalo Grove, Ill. (1997). Representative examples of penetration enhancers include triglycerides (e.g., soybean oil), aloe compositions (e.g., aloe vera gel), ethyl alcohol, isopropyl alcohol, octriphenyl polyethylene glycol, oleic acid, polyethylene glycol 400, propylene glycol, N-decylmethyl sulfoxide, fatty acid esters (e.g., isopropyl myristate, methyl laurate, glycerol monooleate and propylene glycol monooleate), and N-methylpyrrolidone.

[0104] Representative examples of other excipients that may be included in topical and other types of formulations (to the extent they are compatible) include preservatives, antioxidants, moisturizers, emollients, buffers, solubilizers, skin protectants, and surfactants. Suitable preservatives include alcohols, quaternary amines, organic acids, parabens, and phenols. Suitable antioxidants include ascorbic acid and its esters, sodium bisulfite, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, and chelating agents such as EDTA and citric acid. Suitable moisturizers include glycerin, sorbitol, polyethylene glycol, urea, and propylene glycol. Suitable buffers include citrate buffer, hydrochloric acid buffer, and lactic acid buffer. Suitable solubilizers include quaternary ammonium chloride, cyclodextrin, benzyl benzoate, lecithin, and polysorbates. Suitable skin protectants include vitamin E oil, allatoin, dimethicone, glycerin, petrolatum, and zinc oxide.

[0105] Transdermal formulations typically use transdermal delivery devices and transdermal delivery patches, where the compound is formulated in a lipophilic emulsion or buffered aqueous solution and dissolved and / or dispersed in a polymer or adhesive. Patches can be constructed for continuous, pulsatile, or on-demand delivery of pharmaceuticals. Transdermal delivery of compounds can be achieved by iontophoretic patches. Transdermal patches can provide controlled delivery of compounds, and the absorption rate can be slowed by using rate-controlling membranes or by trapping the compound within a polymer matrix or gel. Absorption enhancers can be used to increase absorption, and examples include pharmaceutically acceptable absorbable solvents that aid passage through the skin.

[0106] Ophthalmic preparations include eye drops.

[0107] Preparations for rectal administration include enemas, rectal gels, rectal foams, rectal aerosols, and retention enemas, which may contain conventional suppository bases such as cocoa butter or other glycerides, and synthetic polymers such as polyvinylpyrrolidone, PEG, etc. Compositions for rectal or vaginal administration can also be formulated as suppositories, which can be prepared by mixing the compound with suitable non-irritating carriers and excipients, such as cocoa butter, a mixture of fatty acid glycerides, polyethylene glycol, suppository wax, and combinations thereof.All of these are solid at ambient temperature but liquid at body temperature, so they melt in the rectum or vaginal cavity and release the compound.

[0108] Dosage As used herein, the term "therapeutically effective amount" refers to an amount of a bispecific compound of Formula (I) of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof, that is effective to produce a desired therapeutic response in a patient suffering from a disease or disorder mediated by abnormal tau protein activity. Thus, the term "therapeutically effective amount" includes an amount of a bispecific compound or a pharmaceutically acceptable salt or stereoisomer thereof, when administered, that is sufficient to induce positive alterations in the disease or disorder being treated, or to prevent the onset or progression of the disease or disorder, or to alleviate to some extent one or more of the symptoms of the disease or disorder being treated in a subject, or simply to kill or inhibit the growth of diseased cells, or to reduce the amount of abnormal tau protein in diseased cells.

[0109] The total daily dosage of the bispecific compound and its use can be determined according to standard medical practice, for example, by the attending physician using sound medical judgment. The specific therapeutically effective dose for any particular subject will depend on various factors, including the disease or disorder being treated and its severity (e.g., its current condition); the activity of the bispecific compound used; the specific composition used; the subject's age, weight, general health, sex, and diet; the administration time, administration route, and excretion rate of the bispecific compound used; the duration of treatment; drugs used in combination with or simultaneously with the specific compound used; and similar factors known in the medical field (see, for example, Hardman et al., eds., Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 10th Edition, McGraw-Hill Press, 155-173, 2001).

[0110] Bispecific compounds of Formula (I) and their pharmaceutically acceptable salts and stereoisomers may be effective over a wide dosage range. In some embodiments, the total daily dosage (e.g., for an adult) may range from about 0.001 to about 1600 mg, 0.01 to about 1000 mg, 0.01 to about 500 mg, about 0.01 to about 100 mg, about 0.5 to about 100 mg, 1 to about 100 to about 400 mg / day, about 1 to about 50 mg / day, about 5 to about 40 mg / day, and in other embodiments, about 10 to about 30 mg / day. Individual doses may be formulated to contain the desired dosage depending on the number of times the compound is administered per day. For example, capsules may be formulated containing about 1 to about 200 mg of compound (e.g., 1, 2, 2.5, 3, 4, 5, 10, 15, 20, 25, 50, 100, 150, and 200 mg). In some embodiments, the compound may be administered at a dose ranging from about 0.01 mg to about 200 mg / kg body weight per day. In some embodiments, doses of 0.1 to 100, e.g., 1 to 30 mg / kg per day may be effective, with one or more doses per day. By way of example, suitable doses for oral administration may range from 1 to 30 mg / kg body weight per day, and suitable doses for intravenous administration may range from 1 to 10 mg / kg body weight per day.

[0111] How to use The present invention relates to treating diseases or disorders characterized by or mediated by abnormal tau protein activity (e.g., high levels of tau, or otherwise functional abnormalities or dysfunctions, e.g., dysregulation of tau levels) (collectively referred to as "diseases or disorders mediated by abnormal tau activity"). A "disease" is generally considered to be a state of health in a subject in which the subject is unable to maintain homeostasis and in which the subject's health will continue to deteriorate if the disease is not ameliorated. In contrast, a "disorder" in a subject is a state in which the subject is able to maintain homeostasis, but in which the subject's health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decline in the subject's health.

[0112] The term "subject" (or "patient") as used herein includes all members of the animal kingdom susceptible to or suffering from the indicated disease or disorder. In some embodiments, the subject is a mammal, such as a human or non-human mammal. The method is also applicable to companion animals such as dogs and cats, as well as livestock such as cows, horses, sheep, goats, pigs, and other farm animals, and wild animals. A subject "in need" of treatment may be "suffering from or suspected of having" a particular disease or disorder, may have been positively diagnosed, or may otherwise exhibit a sufficient number of risk factors, or a sufficient number or combination of signs or symptoms, such that a medical professional can diagnose or suspect the subject has the disease or disorder. Thus, subjects suffering from a particular disease or disorder and subjects suspected of having the disease are not necessarily two distinct populations.

[0113] In some embodiments, the disease or disorder is a neurodegenerative disease or disorder.

[0114] Exemplary types of neurodegenerative diseases or disorders that may be suitable for treatment with the bispecific compounds of the invention include Parkinson's disease, prion diseases, Huntington's disease, Alzheimer's disease, multiple system atrophy, Pick's disease, progressive supranuclear palsy (PSP), frontotemporal dementia (FTD), corticobasal degeneration (CBD), chronic traumatic encephalopathy, argyrophilic grain disease, tangle-dominant dementia, and primary age-related tauopathy (PART).

[0115] In some embodiments, the neurodegenerative disease is Alzheimer's disease.

[0116] In some embodiments, the disease or disorder is a neuropsychiatric disease or disorder.

[0117] Exemplary types of neuropsychiatric diseases or disorders that may be suitable for treatment with the bispecific compounds of the invention include autism, schizophrenia, bipolar disorder, attention deficit disorder, cognitive impairment, paralysis, and depression.

[0118] In some embodiments, the neuropsychiatric disorder is autism.

[0119] In some embodiments, the disease or disorder is a neurological disease or disorder.

[0120] Exemplary types of neurological diseases or disorders that may be suitable for treatment with the bispecific compounds of the invention include infantile tauopathies (e.g., tuberous sclerosis hemimegalencephaly, focal cortical dysplasia type 2b, ganglioglioma, or Niemann-Pick disease).

[0121] In some embodiments, the disorder is epilepsy or a seizure disorder.

[0122] In some embodiments, the disorder is a retinal disorder. In some embodiments, the retinal disorder is glaucoma.

[0123] The methods of the invention can involve administering a bispecific compound of the invention or a pharmaceutical composition thereof to a patient in a single dose or multiple doses (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 10, 15, 20 or more doses). For example, the dosing frequency can range from once daily to approximately once every 8 weeks. In some embodiments, the dosing frequency varies from about once daily for 1, 2, 3, 4, 5, or 6 weeks, and in other embodiments involves at least one 28-day cycle including daily dosing for 3 weeks (21 days) and a 7-day "rest" period. In other embodiments, the bispecific compound can be administered twice daily (BID) over a period of 2.5 days (5 doses total), or once daily (QD) over a period of 2 days (2 doses total). In other embodiments, the bispecific compound can be administered once daily (QD) for 5 days.

[0124] Combination therapy The bispecific compounds of Formula (I) and their pharmaceutically acceptable salts and stereoisomers can be used in combination with or simultaneously with at least one other active agent in the treatment of diseases and disorders. In this context, the terms "in combination" and "concurrently" mean that the agents are administered simultaneously, including substantially simultaneous administration, in the same or separate dosage forms, by the same or separate administration modes, or sequentially, e.g., as part of the same treatment regimen or by a continuous treatment regimen. Thus, when given sequentially, the first of the two compounds may still be detectable at effective concentrations at the treatment site at the time the second compound begins to be administered. The order and time intervals can be determined so that they can act together (e.g., to synergistically provide a greater benefit than if they were administered otherwise). For example, the therapeutic agents can be administered simultaneously or at different times in any order. However, if not administered simultaneously, they can be administered sufficiently close in time to provide the desired therapeutic effect, which may be in the form of a synergistic effect. Thus, these terms are not limited to administering the active agents exactly at the same time.

[0125] In some embodiments, a treatment regimen may include administering a bispecific compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer in combination with one or more additional therapeutic agents known for use in treating a disease or disorder (e.g., a neurodegenerative disease). The dosage of the additional therapeutic agent may be the same as or lower than the known or recommended dose. See Hardman et al., eds., Goodman & Gilman's The Pharmacological Basis of Basis of Therapeutics, 10th Edition, McGraw-Hill Press, 2001.

[0126] In some embodiments, the bifunctional compound of the present invention and the additional therapeutic agent are administered at intervals of less than 5 minutes, less than 30 minutes, less than 1 hour, about 1 hour, about 1 to about 2 hours, about 2 to about 3 hours, about 3 to about 4 hours, about 4 to about 5 hours, about 5 to about 6 hours, about 6 to about 7 hours, about 7 to about 8 hours, about 8 to about 9 hours, about 9 to about 1 hour, or at intervals of less than 5 minutes, less than 30 minutes, less than 1 hour, about 1 hour, about 1 to about 2 hours, about 2 to about 3 hours, about 3 to about 4 hours, about 4 to about 5 hours, about 5 to about 6 hours, about 6 to about 7 hours, about 7 to about 8 hours, about 8 to about 9 hours, or at intervals of less than 9 hours. The therapeutic agents may be administered at intervals of 0 hours, at intervals of about 10 to about 11 hours, at intervals of about 11 to about 12 hours, at intervals of about 12 to 18 hours, at intervals of 18 to 24 hours, at intervals of 24 to 36 hours, at intervals of 36 to 48 hours, at intervals of 48 to 52 hours, at intervals of 52 to 60 hours, at intervals of 60 to 72 hours, at intervals of 72 to 84 hours, at intervals of 84 to 96 hours, or at intervals of 96 to 120 hours. Two or more therapeutic agents may be administered during a single patient visit.

[0127] In some embodiments, bispecific compounds of the invention may be used in combination with one or more of levodopa, sinemet, safinamide, ropinirole, pramipexole, rotigotine amantadine, Artane, Cogentin, Eldepryl, Zelapel, and Azilect (e.g., for Parkinson's disease). In some embodiments, bispecific compounds of the invention may be used in combination with one or more of Aricept, Exelon, Razadyne, Namenda, and Namzalic (e.g., for Alzheimer's disease). In some embodiments, bispecific compounds of the invention may be used in combination with one or more of Xenazine, Haldol, chlorpromazine, Risperdal, Seroquel, Keppra, Klonopin, Celexa, Prozac, Epitor, and Depacon (e.g., for Huntington's disease). In some embodiments, bispecific compounds of the invention may be used in combination with one or more of Zoloft, Luvox, Zyprexa, and Seroquel (e.g., for Alzheimer's disease). Representative examples of other active agents known to treat neurodegenerative diseases and disorders and that may be used in conjunction with the bispecific compounds of the invention include dopaminergic therapeutics (e.g., carbidopa-levodopa, pramipexole (Mirapex), ropinirole (Requip), and rotigotine (Neupro, given as a patch)). Apomorphine and monoamine oxidase B (MAO-B) inhibitors (e.g., selegiline (Eldepryl, Zelapar), rasagiline (Azilect), and safinamide (Xadago)) for PD and movement disorders, cholinesterase inhibitors (e.g., benztropine (Cogentin) or trihexyphenidyl) for cognitive impairment, antipsychotics for behavioral and psychological symptoms of dementia, and agents such as riluzole to delay the onset of diseases such as ALS, cerebellar ataxia, and Huntington's disease, nonsteroidal anti-inflammatory drugs for Alzheimer's disease, and caffeine A2A receptor antagonists and CERE-120 (adeno-associated virus serotype 2-neurturin) for neuroprotection in Parkinson's disease.

[0128] Medicine Kit The compositions of the present invention can be assembled into kits or pharmaceutical systems. The kits or pharmaceutical systems according to this aspect of the present invention include a carrier or package, such as a box, carton, or tube, in which one or more containers, such as vials, tubes, ampoules, or bottles, containing the compounds or pharmaceutical compositions of the present invention are tightly enclosed. The kits or pharmaceutical systems of the present invention can also include printed instructions for using the bispecific compounds and compositions.

[0129] These and other aspects of the present invention will be further understood in light of the following examples, which are intended to illustrate particular embodiments of the invention but are not intended to limit its scope, which is defined by the claims. [Example]

[0130] general 1 H and 13 C NMR spectra were recorded on a Bruker AV-III-400 or 500 MHz NMR spectrometer. Chemical shifts are reported as δ values ​​in ppm downfield from TMS as the internal standard. 1 H NMR data are reported as follows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, b=broad, m=multiplet, quint=quintet), coupling constant (Hz), integral. 13 C chemical shifts are reported as δ values ​​in ppm downfield from TMS as the internal standard. Low-resolution mass spectra were obtained on a Waters Acquity™ UltraPerformance LC® by injecting samples into a steady flow of 1 mM ammonium acetate in 20% water-acetonitrile at a rate of 0.2 mL / min using electrospray ionization and an SQ detector. Compound purity was determined by Shimadzu Prominence-HPLC with an ELSD PDA multi and analytical HPLC performed on a Hypersil BDS C-18 column (250 × 4.6 mm, 5 μm) using mobile phase (A) acetonitrile and mobile phase (B) 5 mM ammonium acetate in water at a flow rate of 1.0 mL / min with a gradient of B / A (0 min, 80%), (25 min, 30%), (30 min, 10%), (31 min, 80%), and (36 min, 80%). Analytical thin-layer chromatography was performed on 250 μM silica gel F 254 Preparative thin-layer chromatography was performed on 1000 μM silica gel F 254 was carried out on the plate. Flash column chromatography was performed using 230-400 mesh silica gel.

[0131] Example 1: Synthesis of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-ol [ka] (ai) Acrolein diethyl acetal, Pd(OAc)2, Bu4NAc, K2CO3, KCl, DMF, 110 °C, 33%; (a-ii) Diethyl chlorophosphate, n-BuLi, iPr2NH, THF, -78 °C, 77%; (b) NaH, THF, 0 °C, 53%; (c) Fe powder, HCl, EtOH, 70 °C, 89%; (d) para-formaldehyde, NaBH4, NaOMe, MeOH, reflux, 80%; (e) BBr3, DCM, -10 °C, 77%. *E,E; Z,Z; and / or E,Z isomers. The E,E isomer has been reported to photoisomerize upon exposure to fluorescent light (Hashimoto et al., J. Nucl. Med., 55(9):1532-1538 (2014)). Thus, the intermediate is obtained as a mixture of isomers, with E,E being probably the major isomer known for the 1,4-diaryl-1,3-butaidene series.

[0132] (E)-3-(6-nitropyridin-3-yl)acrylaldehyde A suspension of 5-bromo-2-nitropyridine (2.01 g, 10.0 mmol), acrolein diethyl acetal (4.01 g, 30.0 mmol), K2CO3 (2.03 g, 15.0 mmol), tert-butyl acetoacetate (TBAA) (6.01 g, 20.0 mmol), KCl (740 mg, 10 mmol), palladium(II) acetate (224 mg, 1.0 mmol), and dimethylformamide (DMF) (25 mL) was stirred at 110 °C for 22 h under a N2 atmosphere. The reaction mixture was cooled, diluted with 2 N HCl solution (30 mL), and stirred for 30 min. The solution was filtered through a pad of Celite® and washed with ethyl acetate (EtOAc). The combined filtrate was extracted with EtOAc (2 × 40 mL). The combined organic layers were washed with saturated NaHCO3 solution (2 x 20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to give 1.7 g of a brown solid. The crude product was purified by silica gel column chromatography (0-2% MeOH in CHCl3) to give the title compound as a pale yellow solid (0.67 g, 32%). 1 H NMR(400 MHz,CDCl3)δ 9.83(d,J=7.6 Hz,1H),8.81(d,J=2.4 Hz,1H),8.36(d,J=8.4 Hz,1H),8.22(dd,J=2.0,8.4 Hz,1H),7.58(d,J=16.0 Hz,1H),6.90(dd,J=7.2,16.0,Hz,1H).

[0133] Diethyl ((6-methoxybenzo[d]thiazol-2-yl)methyl)phosphonate To a stirred solution of n-BuLi (2.5 M in hexane, 25 mL, 62.5 mmol) in anhydrous THF (25 mL) was added diisopropylamine (6.55 g, 65.5 mmol) under a N atmosphere at −78° C. The mixture was stirred at this temperature for 50 minutes. A solution of 6-methoxy-2-methylbenzothiazole (4.7 g, 26.1 mmol) in anhydrous THF (40 mL) was added dropwise to the mixture, and the resulting reddish solution was stirred at −78° C. for 30 minutes. A solution of diethyl chlorophosphate (5.16 g, 30.0 mmol) in anhydrous THF (20 mL) was then added dropwise, and the reaction mixture was stirred at −78° C. for 10 minutes. The reaction mixture was then warmed to room temperature (rt) and stirred at room temperature for 1 hour. The reaction was quenched with 1N aqueous NH4Cl (100 mL) and extracted with CHCl3. The combined organic layers were washed with 2% aqueous Na2CO3, brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by column chromatography (0 to 100% EtOAc in hexanes) to afford the title compound as a red oil (7.0 g, 77%). 1 H NMR(CDCl3)δ 7.88(d,J=8.8 Hz,1H),7.30(d,J=2.4 Hz,1H),7.07(dd,J=2.4,8.8 Hz,1H),4.15(quint,J=7.2 Hz,4H),3.87(s,3H),3.69(d,J=21.6 Hz,2H),1.31(t,J=7.2 Hz,6H).

[0134] 6-Methoxy-2-((1E,3E)-4-(6-nitropyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazole To a stirred solution of diethyl ((6-methoxybenzo[d]thiazol-2-yl)methyl)phosphonate (1.93 g, 6.1 mmol) in anhydrous THF (40 mL) was added NaH (60% dispersion in mineral oil, 460 mg, 11.5 mmol) at 0° C. under an argon atmosphere. The mixture was stirred for 30 minutes, after which (E)-3-(6-nitropyridin-3-yl)acrylaldehyde (1.01 g, 5.6 mmol) was added portionwise over 5 minutes. The reaction mixture was allowed to warm to room temperature while stirring overnight. The suspension was concentrated, resuspended in MeOH, and filtered. The filter cake was washed with MeOH and dried under vacuum to give the title compound as an orange solid (1.05 g, 53%). 1 H NMR(400 MHz,DMSO-d6)δ 8.82(d,J=2.0 Hz,1H),8.41-8.31(m,2H),7.88(d,J=8.8 Hz,1H),7.69(d,J=2.4 Hz,1H),7.64-7.54(m,1H),7.46-7.36(m,1H),7.18(d,J=15.2 Hz,2H),7.12(dd,J=2.8,8.8 Hz,1H),3.85(s,3H).

[0135] 5-((1E,3E)-4-(6-methoxybenzo[d]thiazol-2-yl)buta-1,3-dienyl)pyridin-2-amine To a stirred solution of 6-methoxy-2-((1E,3E)-4-(6-nitropyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazole (610 mg, 1.88 mmol) in EtOH (50 mL) was added iron powder (500 mg) and 1.0 M aqueous HCl (3 mL) and stirred at 70 °C for 4 h. The reaction mixture was filtered through a pad of Celite®, and the Celite® pad was washed with 10% MeOH in CHCl3 (50 mL). The combined organic layers were concentrated and purified by silica gel column chromatography (0–15% MeOH in CHCl3 with 0.5% NH4OH as an additive) to give the title compound as a yellow solid (500 mg, 89%). 1H NMR(400 MHz,DMSO-d6)δ 8.08(d,J=2.4 Hz,1H),7.84(d,J=9.2 Hz,1H),7.66(dd,J=2.4,8.4 Hz,1H),7.29(d,J=2.4 Hz,1H),7.20(dd,J=10.0,15.2 Hz,1H),7.05(dd,J=2.8,9.2 Hz,1H),6.85(d,J=9.6 Hz,1H),6.82-6.68(m,2H),6.52(d,J=8.4 Hz,1H),4.88(bs,2H),3.88(s,3H).

[0136] 5-((1E,3E)-4-(6-methoxybenzo[d]thiazol-2-yl)buta-1,3-dienyl)-N-methylpyridin-2-amine To a stirred solution of 5-((1E,3E)-4-(6-methoxybenzo[d]thiazol-2-yl)buta-1,3-dienyl)pyridin-2-amine (470 mg, 1.52 mmol) and paraformaldehyde (180 mg, 6.0 mmol) in anhydrous MeOH (25 mL) was added NaOMe (810 mg, 15.0 mmol). The reaction mixture was heated to reflux for 2 h, then cooled to 0 °C, and NaBH (240 mg, 6.0 mmol) was added. After 20 min, the mixture was heated to reflux for 2 h. The reaction was quenched with cold water (25 mL) and extracted with CHCl (2 × 50 mL). The organic layers were combined, dried over anhydrous NaSO, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (0-3% MeOH in CHCl3) to give the title compound as a yellow solid (370 mg, 80%). 1 H NMR(500 MHz,DMSO-d6)δ 8.10(d,J=2.0 Hz,1H),7.80(d,J=9.0 Hz,1H),7.70(dd,J=2.5,9.0 Hz,1H),7.62(d,J=2.5 Hz,1H),7.28(dd,J=10.0,15.5 Hz,1H),7.07(dd,J=3.0,9.0 Hz,1H),6.98-6.84(m,4H),6.49(d,J=9.0 Hz,1H),3.83(s,3H),2.81(d,J=5.5 Hz,3H).

[0137] 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-ol To a cooled (−10 °C) suspension of 5-((1E,3E)-4-(6-methoxybenzo[d]thiazol-2-yl)buta-1,3-dienyl)-N-methylpyridin-2-amine (1.05 g, 3.1 mmol) in dry CHCl (10 mL) was added a 1 M solution of BBr in CHCl (20 mL, 20 mmol) and stirred at −10 °C for 1 h, then allowed to warm to room temperature over 18 h. The reaction mixture was cooled in an ice bath, quenched with cold water (10 mL), and stirred for 20 min. The resulting heterogeneous mixture was basified with saturated NaHCO solution, and the precipitate was filtered. The filter cake was washed with water and dried under reduced pressure to give the title compound as a yellow solid (760 mg, 77%). 1 H NMR(400 MHz,DMSO-d6)δ 9.83(s,1H),8.09(d,J=2.0 Hz,1H),7.73-7.67(m,2H),7.32(d,J=2.4 Hz,1H),7.22(dd,J=10.0,15.2 Hz,1H),7.01-6.81(m,5H),6.50(d,J=8.8 Hz,1H),2.81(dJ=4.8 Hz,3H).MS(ESI + ) m / z 310.1 (M+H) + .

[0138] Example 2: Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetic acid [ka] (a) 3-Hydroxyphthalic anhydride, Et3N, toluene, reflux, 92%; (b) tert-butylbromoacetate, KI, K2CO3, DMF, 60 °C, 75%; (c) HCl in dioxane, room temperature, quantitative.

[0139] 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione To a mixture of 3-hydroxyphthalic anhydride (420 mg, 2.5 mmol) and 3-aminopiperidine-2,6-dione hydrochloride (420 mg, 2.5 mmol) was added dry toluene (20 mL) and triethylamine (0.4 mL, 2.8 mmol). The resulting solution was refluxed for 18 h. The reaction mixture was concentrated and purified by silica gel column chromatography (0-10% MeOH in CHCl3) to give the title compound as a yellowish solid (630 mg, 92%). 1 H NMR(400 MHz,DMSO-d6)δ 11.19(s,1H),11.10(s,1H),7.66(dd,J=7.2,8.4 Hz,1H),7.33(d,J=6.8 Hz,1H),7.26(d,J=8.4 Hz, 1H), 5.08 (dd, J=5.2, 12.8 Hz, 1H), 2.95-2.84 (m, 1H), 2.64-2.52 (m, 2H), 2.07-1.98 (m, 1H).

[0140] tert-Butyl 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetate To a mixture of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (170 mg, 0.62 mmol), tert-butyl bromoacetate (150 mg, 0.77 mmol), and K2CO3 (110 mg, 0.79 mmol), dry DMF (2 mL) and KI (10 mg, 0.06 mmol) were added. The mixture was stirred at 60 °C for 18 h. The reaction mixture was concentrated, diluted with EtOAc (50 mL), and washed with water (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4, concentrated, and purified by silica gel column chromatography (0-3% MeOH in CHCl3) to give the title compound as a white solid (180 mg, 75%). 1H NMR(400 MHz,DMSO-d6)δ 11.12(s,1H),7.83-7.77(m,1H),7.49(d,J=6.8 Hz,1H),7.39(d,J=8.4 Hz,1H),5.14-5.08(dd,J=5.2,12.8 Hz, 1H), 4.97 (s, 2H), 2.96-2.84 (m, 1H), 2.64-2.52 (m, 2H), 2.08-1.99 (m, 1H), 1.43 (s, 9H).

[0141] 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetic acid tert-Butyl 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetate (220 mg, 0.57 mmol) was dissolved in 4 M HCl in dioxane (3 mL, 12 mmol) and stirred at room temperature for 3 h. The reaction mixture was concentrated to dryness and co-evaporated with methanol (2 × 5 mL) to give the title compound as a white solid (190 mg, quantitative). 1 H NMR(500 MHz,DMSO-d6)δ 13.24(bs,1H),11.10(s,1H),7.79(dd,J=7.0,9.0 Hz,1H),7.48(d,J=7.0 Hz,1H),7.40(d,J=9.0 Hz,1H),5.11(dd,J=5.5,12.0 Hz,1H),4.99(s,2H),2.94-2.84(m,1H),2.64-2.52(m,2H),2.07-2.01(m,1H).

[0142] Example 3: Boc protection and mesylation [ka] (i)Boc2O,Et3N,CH2Cl2;(ii)MsCl,Et3N,CH2Cl2

[0143] 2-(2-(2-tert-butoxycarbonylaminoethoxy)ethoxy)ethyl methanesulfonate To a cooled solution of 6-aminohexanol (300 mg, 2.0 mmol) in CHCl (2.5 mL) was added Boc anhydride (450 mg, 2.0 mmol) in CHCl (2.5 mL) and stirred for 18 h. The reaction mixture was concentrated and purified by silica gel column chromatography (20 to 100% EtOAc in hexanes) to give 2-(2-(2-tert-butoxy-carbonylaminoethoxy)ethoxy)ethanol as a yellow oil (430 mg, 86%).

[0144] To a cooled solution of 2-(2-(2-tert-butoxycarbonylaminoethoxy)ethoxy)ethanol (300 mg, 1.2 mmol) in CHCl (5 mL) was added triethylamine (0.4 mL, 2.8 mmol), methanesulfonyl chloride (0.15 mL, 1.9 mmol), and a catalytic amount of 4-dimethylaminopyridine (10 mg). The reaction mixture was stirred for 2 h, then diluted with CHCl (50 mL) and washed with saturated NaHCO solution (10 mL) and saturated brine solution (10 mL). The organic phase was concentrated and purified by silica gel column chromatography (0–80% EtOAc in hexanes) to give the title compound as a yellow oil (400 mg, quantitative). 1 H NMR(400MHz,CDCl3)δ 4.94(bs,1H),4.42-4.37(m,2H),3.79-3.75(m,2H),3.69-3.65(m,2H),3.64-3.59(m,2H),3.53(t,J=5.2 Hz,2H),3.35-3.28(m,2H),3.07(s,3H),1.44(s,9H).

[0145] 8-(tert-butoxycarbonylamino)octyl methanesulfonate To a cooled solution of 8-aminooctanol (210 mg, 1.5 mmol) in CHCl (5 mL) was added Boc anhydride (360 mg, 1.65 mmol) in CHCl (2.5 mL) and stirred for 3 h. The reaction mixture was concentrated and purified by silica gel column chromatography (0–60% EtOAc in hexanes) to give 8-(tert-butoxycarbonyl-amino)octan-1-ol (265 mg, 72%) as a white solid.

[0146] To a cooled solution of 8-(tert-butoxycarbonylamino)octan-1-ol (250 mg, 1.02 mmol) in CHCl (2.5 mL) was added triethylamine (0.4 mL, 2.8 mmol), methanesulfonyl chloride (175 mg, 1.53 mmol), and a catalytic amount of 4-dimethylaminopyridine (20 mg). The reaction mixture was stirred for 2 h, then diluted with CHCl (50 mL) and washed with saturated NaHCO solution (10 mL) and saturated brine solution (10 mL). The organic phase was concentrated and purified by silica gel column chromatography (0–50% EtOAc in hexanes) to give the title compound as a clear oil (280 mg, 86%). 1 H NMR(500 MHz,CDCl3)δ 4.50(bs,1H),4.22(t,J=6.5 Hz,2H),3.14-3.07(m,2H),3.00(s,3H),1.78-1.71(m,2H),1.51-1.36(m,13H),1.36-1.28(m,6H).

[0147] 6-(tert-butoxycarbonylamino)hexyl methanesulfonate To a cooled solution of 6-aminohexanol (350 mg, 3.0 mmol) in CHCl (2.5 mL) was added Boc anhydride (650 mg, 3.0 mmol) in CHCl (2.5 mL) and stirred for 18 h. The reaction mixture was concentrated and purified by silica gel column chromatography (0-80% EtOAc in hexanes) to give 6-(tert-butoxycarbonyl-amino)hexan-1-ol as a white solid (640 mg, 100%).

[0148] To a cooled solution of 6-(tert-butoxycarbonylamino)hexan-1-ol (420 mg, 2.0 mmol) in CHCl (5 mL) was added triethylamine (0.8 mL, 5.6 mmol), methanesulfonyl chloride (0.25 mL, 3.2 mmol), and a catalytic amount of 4-dimethylaminopyridine (20 mg). The reaction mixture was stirred for 2 h, then diluted with CHCl (50 mL) and washed with saturated NaHCO solution (10 mL) and saturated brine solution (10 mL). The organic phase was concentrated and purified by silica gel column chromatography (0–50% EtOAc in hexanes) to give the title compound as a white solid (490 mg, 88%). 1 H NMR(400 MHz,CDCl3)δ 4.52(bs,1H),4.22(t,J=6.4 Hz,2H),3.16-3.08(m,2H),3.00(s,3H),1.80-1.71(m,2H),1.54-1.46(m,2H),1.44(s,9H),1.43-1.31(m,4H).

[0149] Example 4 Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)hexyl)acetamide (2) [ka] X = (CH2)6; (a) K2CO 3、 DMF, 100 °C, 63%; (b) 2 M HCl in methanol, room temperature, quantitative; (c) BOP / pyBOP reagent, Et3N, DMF, 100%. *E,E; Z,Z; or E,Z isomer. E,E is probably the major known isomer for the 1,4-diaryl-1,3-butaidene series.

[0150] tert-Butyl 6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)hexylcarbamate To a solution of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-ol (110 mg, 0.35 mmol) in DMF (2.5 mL) was added K2CO3 (100 mg, 0.71 mmol) and 6-(tert-butoxycarbonylamino)hexyl methanesulfonate (133 mg, 0.45 mmol), and the reaction mixture was stirred at 70 °C for 6 h. The reaction mixture was concentrated and then diluted with CHCl3 (20 mL) and water (2 mL). The organic layer was washed with saturated brine solution (10 mL), dried over Na2SO4, concentrated, and purified by silica gel column chromatography (0-40% acetone in hexanes) to give the title compound as a yellow solid (115 mg, 63%). MS (ESI) + ) m / z 509.3(M+H) + . [ka]

[0151] 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)hexyl)acetamide To a cooled solution of tert-butyl 6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)hexyl-carbamate (100 mg, 200 μmol) in MeOH (0.5 mL) was added 2 M HCl in methanol (2.5 mL, 5.0 mmol) and stirred at room temperature for 3 hours. The reaction mixture was concentrated to dryness and coevaporated with methanol (2 × 5 mL) to give a yellow solid residue. To the residue were added 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetic acid (60 mg, 180 μmol), DMF (2.0 mL), triethylamine (100 mg, 1.0 mmol), and pyBOP reagent (100 mg, 192 μmol). The solution was stirred at room temperature for 6 hours, then concentrated to dryness and triturated with water followed by hexane. The resulting residue was diluted with CHCl (50 mL) and washed with water (2 × 10 mL). The organic layer was dried over NaSO, concentrated, and purified by silica gel column chromatography (0-10% MeOH in CHCl) to give the title compound as a yellow solid (130 mg, 100%).

[0152] Example 5 Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)octyl)acetamide (3) [ka] X = (CH2)8; (a) K2CO 3、 DMF, 100 °C, 51%; (b) 2 M HCl in methanol, room temperature, quantitative; (c) BOP / pyBOP reagent, Et3N, DMF, 80%. *E,E; Z,Z; or E,Z isomer. E,E is probably the major known isomer for the 1,4-diaryl-1,3-butaidene series.

[0153] tert-Butyl 6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)octylcarbamate To a solution of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-ol (95 mg, 0.30 mmol) in DMF (2.5 mL) were added CsCO (180 mg, 0.55 mmol) and 8-(tert-butoxycarbonylamino)octyl methanesulfonate (130 mg, 0.40 mmol), and the reaction mixture was stirred at 70 °C for 6 h. The reaction mixture was concentrated and then diluted with CHCl (20 mL) and water (2 mL). The organic layer was washed with saturated brine solution (10 mL), dried over NaSO, concentrated, and purified by silica gel column chromatography (0–40% acetone in hexanes) to give the title compound as a yellow solid (86 mg, 51%). (400 MHz,DMSO-d6)δ 8.09(d,J=2.0 Hz,1H),7.78(d,J=9.2 Hz,1H),7.70(dd,J=2.0,8.98 Hz,1H),7.60(d,J=2.4 Hz,1H),7.27(dd,J=9.6,15.2 Hz,1H),7.05(dd,J=2.4,8.8 Hz,1H),6.98-6.82(m,4H),6.76(t,J=5.2 Hz,1H),6.49(d,J=8.8 Hz,1H),4.02(t,J=6.4 Hz,2H),2.92-2.86(m,2H),2.80(d,J=4.8 Hz, 3H), 1.77-1.69(m, 2H), 1.48-1.19(m, 19H). [ka]

[0154] 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)octyl)acetamide To a cooled solution of tert-butyl 6-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)octylcarbamate (80 mg, 150 μmol) in MeOH (1.0 mL) was added 2 M HCl in methanol (2.0 mL, 4.0 mmol) and stirred at room temperature for 3 hours. The reaction mixture was concentrated to dryness and coevaporated with methanol (2 × 5 mL) to give a yellow solid residue. To the residue were added 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetic acid (50 mg, 155 μmol), DMF (2.0 mL), triethylamine (140 mg, 1.4 mmol), and pyBOP reagent (88 mg, 170 μmol). The solution was stirred at room temperature for 6 hours, then concentrated to dryness and triturated successively with water and hexane. The resulting residue was diluted with CHCl (50 mL) and washed with water (2 × 10 mL). The organic layer was dried over NaSO, concentrated, and purified by silica gel column chromatography (0–10% MeOH in CHCl) to afford the title compound (90 mg, 80%) as a yellow solid.

[0155] Example 6 Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(2-(2-(2-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)ethoxy)ethoxy)ethyl)acetamide (4) [ka] X=(CH2)2O(CH2)2O(CH2)2;(a)K2CO 3、 DMF, 100 °C, 58%; (b) 2 M HCl in methanol, room temperature, quantitative; (c) BOP / pyBOP reagent, Et3N, DMF, 12%. *E,E; Z,Z; or E,Z isomer. E,E is probably the major known isomer in the 1,4-diaryl-1,3-butaidene series.

[0156] tert-Butyl-2-(2-(2-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)ethoxy)ethoxy)ethylcarbamate To a solution of 2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-ol (25 mg, 0.08 mmol) in DMF (2.0 mL) was added K2CO3 (28 mg, 0.20 mmol) and 2-(2-(2-tert-butoxycarbonylaminoethoxy)ethoxy)ethyl methanesulfonate (31 mg, 0.09 mmol), and the reaction mixture was stirred at 100 °C for 3 h and then cooled to room temperature over 18 h. The reaction mixture was concentrated and then diluted with CHCl3 (20 mL) and water (2 mL). The organic layer was washed with saturated brine solution (10 mL), dried over Na2SO4, concentrated, and purified by preparative TLC (8% MeOH in CHCl3) to give the title compound as a yellow solid (25 mg, 58%). MS(ESI + ) m / z 541.5(M+H) + . [ka]

[0157] 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)-N-(2-(2-(2-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)ethoxy)ethoxy)ethyl)acetamide To a cooled solution of tert-butyl-2-(2-(2-(2-((1E,3E)-4-(6-(methylamino)pyridin-3-yl)buta-1,3-dienyl)benzo[d]thiazol-6-yloxy)ethoxy)ethoxy)ethylcarbamate (25 mg, 46 μmol) in MeOH (1.0 mL) was added 2 M HCl in methanol (2.0 mL, 8.0 mmol) and stirred at room temperature for 3 h. The reaction mixture was concentrated to dryness and co-evaporated with methanol (2 × 5 mL) to give a yellow solid residue. To the residue was added pyBOP reagent (27 mg, 61 μmol), a solution of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yloxy)acetic acid (16 mg, 50 μmol) in DMF (1.0 mL), and triethylamine (20 mg, 200 μmol). The solution was stirred at room temperature for 18 hours and then concentrated to dryness. The crude product was purified by preparative TLC (10% MeOH in CHCl) to give the title compound as a yellow solid (35 mg), which was further purified by preparative HPLC (74:18:8% heptane:IPA:MeCN, Viridis® 2-ethylpyridine column) to give the title compound as a yellow solid (4.2 mg, 12%). MS (ESI + ) m / z 755.1(M+H) + .

[0158] Example 7: Effect of administration on tau in A152T and P301L neurons

[0159] The assay was performed according to the protocol described in Silva et al., eLife, 8:e45457 (2019). Each of the bispecific compounds, along with a control parent scaffold, was evaluated for its ability to degrade tau protein in cultured human differentiated frontotemporal dementia (FTD) neurons.

[0160] For each condition, one well of differentiated neurons (6-well plate) was washed, collected in PBS, pelleted, and lysed in radioimmunoprecipitation assay (RIPA) buffer (Boston BioProducts) containing 2% SDS (Sigma), protease inhibitors (Roche cOmplete™ Mini tablets), and phosphatase inhibitors (Sigma). The cells were then sonicated for 5 minutes in a water sonicator (Bransonic® Ultrasonic Baths, Thomas Scientific) and centrifuged at 20,000 g for 15 minutes. The supernatant was transferred to a new tube, and total protein concentration was quantified using the Pierce™ BCA Protein Assay Kit (Thermo Scientific™). SDS-PAGE gels were loaded with 10 μg of total protein per well (pre-boiled samples). Western blots were performed using the Novex NuPAGE™ SDS-PAGE gel system (Invitrogen™). All samples were separated in 7% Tris-acetate gels containing Tris-acetate running buffer (Invitrogen™). Blots were probed with antibodies against total tau (TAU5) and phosphorylated tau (Ser396), along with β-actin as a loading control. This was followed by detection with the corresponding HRP-conjugated secondary antibody (Cell Signaling Technology™) and SuperSignal™ West Pico Chemiluminescent Substrate (Thermo Scientific™). Membranes were exposed to autoradiography film (LabScientific), and the film was scanned using an Epson® Perfection V800 Photo Scanner. Protein band intensity (pixel mean intensity) was quantified using the Adobe Photoshop® CS5 histogram function.

[0161] Vehicle alone (DMSO) had no effect on tau. However, variable degradation activity was observed among specific compounds, with overall similar relative levels of degradation activity between tau-A152T and tau-P301L neurons. Bispecific compounds were added at the indicated doses (either 1 μM or 10 μM). Lenalidomide (1 μM or 10 μM) was also evaluated as a control, as it binds to CRBN but does not affect tau levels, as confirmed by its lack of involvement with tau. Bispecific compounds 2 and 4 significantly degraded hyperphosphorylated tau and / or total tau in human tau-A152T neurons (6-week differentiated) and tau-P301L neurons (6-week differentiated) after 24 hours of treatment with the bispecific compounds (Figures 1-4).

[0162] All patent and non-patent publications are indicative of the level of skill of those skilled in the art to which this invention pertains. All these publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

[0163] Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention. It is therefore to be understood that many modifications can be made to the exemplary embodiments and that other arrangements can be devised without departing from the spirit and scope of the present invention as defined by the appended claims.

Claims

1. Formula (I): 【Chemistry 1】 wherein the targeting ligand represents a moiety that binds tau and the degron represents a moiety that binds an E3 ubiquitin ligase, or the degron is an autophagy-recruitment tag and the linker represents a moiety that covalently links the degron and the targeting ligand, or a pharmaceutically acceptable salt or stereoisomer thereof; 【Chemistry 2】 is represented by formula TL-1 or TL-2: 【Transformation 3】 [In the formula, each X １ are independently C, CH, or N; X ２ is NH, S or O, X ３ is CH or N, Each R １ are independently hydrogen or C １ -C ３ is alkyl, X ２ When is NH, X ３ is CH, Furthermore, X ３ If N, then X ２ is S or O.

2. TL-1 has the structures TL-1a to TL-1o: 【Chemistry 4】 【Transformation 5】 2. The bispecific compound of claim 1, represented by any one of:

3. TL-1 【Transformation 6】 3. The bispecific compound of claim 2, wherein:

4. TL-2 has the structures TL-2a to TL-2o 【Transformation 7】 2. The bispecific compound of claim 1, represented by any one of:

5. R １ The bispecific compound of any one of claims 1 to 4, wherein is hydrogen.

6. R １ The bispecific compound of any one of claims 1 to 4, wherein is methyl.

7. The linker comprises an alkylene chain or a divalent alkylene chain, any of which is —O—, —S—, —N(R′)—, —C≡C—, —C(O)—, —C(O)O—, —OC(O)—, —OC(O)O—, —C(NOR′)—, —C(O)N(R′)—, —C(O)N(R′)C(O)—, —C(O)N(R′ )C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C (NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O) ２ -, -OS(O)-, -S(O)O-, -S(O)-, -OS(O) ２ -, -S(O) ２ O-, -N(R')S(O) ２ -, -S(O) ２ N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O) ２ N(R')-, -N(R')S(O)N(R')-, C ３ -C １２ may be interrupted and / or terminated at either or both termini by at least one of a carbocyclene, a 3- to 12-membered heterocyclene, a 5- to 12-membered heteroarylene, or any combination thereof; where R' is H or C １ -C ６ 7. The bispecific compound of any one of claims 1 to 6, wherein the interrupting group and one or both terminal groups are alkyl and may be the same or different.

8. the linker comprises an alkylene chain having 1 to 10 alkylene units; 【Transformation 8】 8. The bispecific compound of claim 7, wherein said bispecific compound is interrupted and / or terminated by

9. The linker is selected from the group consisting of -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O) ２ -, -OS(O)-, -S(O)O-, -S(O)-, -OS(O) ２ -, -S(O) ２ O-, -N(R')S(O) ２ -, -S(O) ２ N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O) ２ N(R')-, -N(R')S(O)N(R')-, C ３－１２ and a polyethylene glycol chain which may terminate in at least one of a carbocyclene, a 3- to 12-membered heterocyclene, a 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or C. １ ~C ６ 7. The bispecific compound of any one of claims 1 to 6, wherein one or both terminal groups are alkyl and may be the same or different.

10. the linker has 2 to 8 PEG units; 【Chemistry 9】 10. The bispecific compound of claim 9, comprising a polyethylene glycol chain terminating in

11. The following structure: 【Chemistry 10】 2. The bispecific compound of claim 1, represented by any one of:

12. The bispecific compound of any one of claims 1 to 11, wherein the degron binds cereblon.

13. The degron has the structure (D1-a to D1-i): 【Chemistry 11】 where:

13. The bispecific compound of claim 12, wherein Y is NH or O.

14. The bispecific compound of any one of claims 1 to 11, wherein the degron binds von Hippel-Landau (VHL).

15. The degron has the structure D2-a to D2-d: 【Chemistry 12】 【Chemistry 13】 wherein Z is a cyclic group; or 【Chemistry 14】 13. The bispecific compound of claim 12, represented by:

16. The bispecific compound of any one of claims 1 to 11, wherein the degron binds an inhibitor of apoptosis protein (IAP).

17. The degron has the structure D3-a to D3-d: 【Chemistry 15】 17. The bispecific compound of claim 16, represented by:

18. 12. The bispecific compound of any one of claims 1 to 11, wherein the degron binds mouse double minute 2 (MDM2).

19. The degron has the structure D4-a to D4-b: 【Chemistry 16】 19. The bispecific compound of claim 18, represented by:

20. The bispecific compound of any one of claims 1 to 11, wherein the degron is an autophagy-recruitment tag.

21. The degron has the structure D5-a to D5-b: 【Chemistry 17】 21. The bispecific compound of claim 20, represented by: 【Request Item 22】 【Chemistry 18】 【Chemistry 19】 2. The bispecific compound of claim 1, which is:

23. 23. A pharmaceutical composition comprising a therapeutically effective amount of a bispecific compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

24. 24. The pharmaceutical composition of claim 23, wherein the pharmaceutically acceptable carrier is a solid.

25. 24. The pharmaceutical composition of claim 23, wherein the pharmaceutically acceptable carrier is a liquid.

26. 23. A method of treating a disease or disorder characterized or mediated by abnormal activity of tau protein, comprising administering to a subject in need thereof a therapeutically effective amount of a bispecific compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt or stereoisomer thereof.

27. 27. The method of claim 26, wherein the disease or disorder is a neurodegenerative disease or disorder.

28. 28. The method of claim 27, wherein the neurodegenerative disease is Parkinson's disease, prion disease, Huntington's disease, Alzheimer's disease, multiple system atrophy, Pick's disease, progressive supranuclear palsy (PSP), frontotemporal dementia (FTD), corticobasal degeneration (CBD), chronic traumatic encephalopathy, argyrophilic grain disease, tangle-dominant dementia, or primary age-related tauopathy (PART).

29. 27. The method of claim 26, wherein the disease or disorder is a neuropsychiatric disorder.

30. 30. The method of claim 29, wherein the neuropsychiatric disease or disorder is autism, schizophrenia, bipolar disorder, attention deficit disorder, cognitive deficit disorder, paralysis, or depression.

31. 27. The method of claim 26, wherein the disease or disorder is a neurological disease or disorder.

32. 32. The method of claim 31, wherein the neurological disease or disorder is an infantile tauopathy.

33. 27. The method of claim 26, wherein the disorder is epilepsy or seizures.

34. 27. The method of claim 26, wherein the disease or disorder is a retinal disorder.

35. 35. The method of claim 34, wherein the retinal disease or disorder is glaucoma.

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